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Summer 2014 NIDDK Medical Student Research Symposium Student Life Center Vanderbilt University Nashville, TN August 6 and 7, 2014 Funded by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Table of Contents Description of the Program ......................................................................................................... 2 Agenda .................................................................................................................................. 3 - 4 Students Participating in the Program ................................................................................... 5 - 9 Medical Schools of Student Participants .................................................................................. 10 Visiting Professors for Research Symposium .................................................................... 11 - 13 Vanderbilt Faculty Participating In Moderating the Poster Sessions and Lunches ……………14 NIDDK-Funded Diabetes Centers ............................................................................................ 15 Supporting Training Grants for Diabetes Program .................................................................... 16 Advisory Panel for Program ...................................................................................................... 17 Abstracts from Students ......................................................................................................... 18+ 1 Description of the Programs Supporting The NIDDK Medical Student Research Symposium The National Institutes of Health (NIH) through the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) supports programs to encourage physicians to enter careers involving research. These programs offer first- and second-year medical students the opportunity to conduct mentored research. The research opportunities range from basic laboratory studies to clinical studies in humans. Students also participate in a core curriculum of seminars on research approaches as well as a variety of clinical and research topics in diabetes and kidney disease. These programs help students gain an improved understanding of career opportunities in biomedical research. At the conclusion of the summer, each student presents his/her work at the NIDDK Medical Student Research Symposium, which provides a venue for the students to meet, share their research, and to learn about career pathways and opportunities in NIDDK-related professions. The Vanderbilt Diabetes Center and the NIDDK-funded, Vanderbilt Diabetes Research and Training Center are hosting the sixth annual NIDDK Medical Student Research Symposium on August 6 and 7, 2014 in Nashville, Tennessee. For the summer of 2014, the 126 students from 69 medical schools who are attending the NIDDK Medical Student Research Symposium were supported by the following programs: • NIDDK Medical Student Research Program in Diabetes in which students conducted research at one of 15 NIDDK-funded Diabetes Research Centers • Vanderbilt Student Research and Training Program in which students conducted research in diabetes, obesity, renal disease, or GI disease at Vanderbilt University • Diabetic Complications Consortium in which students conducted research on complications of diabetes at a NIDDK-funded Diabetes Research Center • Vanderbilt Center for Kidney Disease in which students conducted research in renal disease at Vanderbilt University • Summer Program in Obesity, Diabetes and Nutrition Related Research Training in which students conducted research at the University of Maryland 2 Symposium Agenda for NIDDK Medical Student Research Program Symposium Student Life Center, Vanderbilt University August 6 – 7, 2014 Tuesday, August 5 7:00 pm Opening reception in Centennial Ballroom, Holiday Inn Vanderbilt Wednesday, August 6 8 – 8:45 am Poster set up in the Student Life Center 9:00 Welcome and introduction Alvin C. Powers, MD Joe C. Davis Chair in Biomedical Science Professor of Medicine and Molecular Physiology & Biophysics Chief, Division of Diabetes, Endocrinology & Metabolism Director, Vanderbilt Diabetes Center Vanderbilt University Medical Center 9:15 – 10:00 Presentation – Ballroom C Richard G. Kibbey, MD, PhD Associate Professor of Medicine, Division of Endocrinology and of Cellular & Molecular Physiology Yale School of Medicine 10:00 – 11:30 Poster session 1 (moderated) 11:45 – 1:00 Lunch with visiting professors in small groups 1:00 – 1:45 Presentation – Ballroom C Andrea L. Cherrington, MD Associate Professor, Department of Medicine, Division of Preventative Medicine University of Alabama at Birmingham 1:45 – 3:15 1:45 – 2:30 Poster Session A (open) Odd numbered posters present Even numbered posters visit Even numbered posters present Odd numbered posters visit 2:30 – 3:15 3:15 Reassemble for Program Information – Ballroom C 3:30 - 5:00 Poster Session 2 (moderated) 5:00 Reception 3 Thursday, August 7 9:00 am Welcome – Ballroom C Mark de Caestecker, MB, BS, PhD Associate Professor of Surgery, Medicine, Division of Nephrology and Cell & Developmental Biology Vanderbilt University Medical Center 9:05 – 10:15 Poster Session B (open) Even numbered posters present Odd number posters visit 10:15 – 11:00 Approaches to Residency Selection and Application John A. McPherson, MD, FACC, FACP Associate Professor of Medicine Vice-Chair for Education, Department of Medicine Director, Internal Medicine Residency Program Vanderbilt University Medical Center and Rebecca Swan, MD Director, Pediatric Residency Program Associate Professor of Pediatrics, Division of General Pediatrics Assistant Dean of Graduate Medical Education Vanderbilt University Medical Center 11:00 – 12:30 Poster Session 3 (moderated) 12:30 – 1:15 Lunch with visiting professors in small groups 1:15 – 2:15 Poster Session C (open) Odd numbered posters present Even number posters visit 2:30 – 3:15 Presentation – Ballroom C Art Castle, PhD Director, Metabolomics and Informatics Program, Division of Diabetes, Endocrinology & Metabolic Diseases National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health and Alvin C. Powers, MD Joe C. Davis Chair in Biomedical Science Professor of Medicine and Molecular Physiology & Biophysics Chief, Division of Diabetes, Endocrinology & Metabolism Director, Vanderbilt Diabetes Center Vanderbilt University Medical Center 3:15 Concluding remarks 6:15 6:30 9:00 Meet the bus in front of the Holiday Inn Vanderbilt for transport to restaurant Dinner in downtown Nashville Approximate end to dinner; students on own for return to hotel Friday, August 8 Students depart on own 4 Participants 2014 Abburi, Nandini, S Altshuler, Courtney Amezquita, Joel Asfaw, Elizabeth, K Beardsley, Ryan, D Beltre, Melba A Bian, Rachel R Binns, Thomas (Chad) Bloom, Benjamin, A Brooksbank, Jeremy Buendia, Matthew A Butler, Erin Cai, Steven Card, Casey, E Choudhury, Natasha Chun, Hyun, B Clancey-Rivera, Marie G. Cole, Heather Contreras, Ignacio J Couzens, Erica, C Cross, Benjamin R Das, Anjali David, Sthuthi Deshpande, Sagar, S Diao, Xavier Downes, Laura Estrada, Kathleen, L Abst # 10 69 86 38 81 101 122 14 68 22 66 2 44 40 25 80 91 9 70 39 52 59 20 125 1 35 6 5 University of Kentucky Michigan State University University of Toledo University of Southern California University of California - Los Angeles University of California - Los Angeles University of Illinois - Champaign-Urbana New York Medical College University of Michigan University of Arkansas University of Maryland University of Tennessee University of Tennessee Eastern Virginia Medical School Rosalind Franklin University National University of Ireland - Galway Meharry Medical College Stony Brook Medicine Ponce School of Medicine and Health Sciences University of Tennessee Columbia University Loma Linda University University of Nebraska Rutgers NJ Medical School - Newark University of Rochester University of Michigan University of Maryland Medical School Vanderbilt U Michigan U Michigan Vanderbilt Yale Washington U U Chicago BADERC U Michigan Vanderbilt JHopkins/U Maryland Vanderbilt Vanderbilt U Maryland Vanderbilt Yale Vanderbilt UCSD Yale Vanderbilt Columbia U Washington UCSD Vanderbilt Vanderbilt U Michigan U Maryland Center for Summer Email address [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] 2014 Participants in the NIDDK Medical Student Research Program in Diabetes and Obesity Participants 2014 Etiel, Kelsey Foggia, Megan, J Fowler, Zachary, G Fox, Kacie, R Freeman, Casey Galletta, Thomas Gandhi, Shriya Ghosh, Sujata Gonsalves, Zenobia, T González Bouza, Beatriz Guzman, Herodes, G Han, Amy Yuh Jung Harrison, Bradley Hoover, Elizabeth, A Ikizler, Halil, O Jain, Tarun Jin, Hope, L Jones, Terrence T Jose, Charles Kelly, Emily Kim, Janice, S Kim, Jessica Kissel, Suzanne Kuhlmann, Paige, K Landreth, Hannah Leiva, Orly Lepard, Morgan Abst # 124 19 43 30 34 119 112 31 110 5 108 46 73 97 113 76 55 37 85 67 83 26 114 17 53 16 111 6 Loyola University - Chicago University of Nevada University of North Dakota University of Hawai'i University of Kansas Northeast Ohio Medical University University of Illinois - Chicago Texas A&M Health Science Center Rosalind Franklin University University of Puerto Rico University of North Carolina Case Western Reserve University Mercer University University of Iowa University of Vermont Indiana University New York Medical College University of Tennessee University of Nevada Florida Atlantic University Washington University University of Nevada Indiana University University of Missouri - Columbia University of Oklahoma Boston University University of Tennessee Medical School Vanderbilt UCLA Columbia Joslin Vanderbilt Vanderbilt U Chicago Vanderbilt U Washington Washington U UCSF U Pennsylvania Vanderbilt Columbia U Washington Vanderbilt Joslin UAB Vanderbilt Vanderbilt Yale Vanderbilt Vanderbilt Washington U Vanderbilt Joslin UAB Center for Summer Email address [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] 2014 Participants in the NIDDK Medical Student Research Program in Diabetes and Obesity 36 50 103 28 107 18 64 45 3 47 102 12 116 24 63 100 4 62 11 87 94 71 48 27 56 74 84 Abst # 7 Leung, Sophie, L Li, Jennifer Li, Muyi Liao, Si (Sherry) Lindauer, Katherine L. Linder, Alexandra, N Liu, Mingchun Love, Heather R. Ludi, Nicolas Lyons, James Magee, Ronald, R Marcaccio, Christina, L Marks, Bianca Martinez, Jordan A Matar, Regina, N Mauch, Scott, C Mazza, Dayna Mehregan, Christian, A Milligan, Ian, L. Mizera, Megan Morones, Nohemy Mottla, Jay, L Moutos, Christopher, P. Mulero Morales, Maris Nevarez, Nicole M. Nguyen, Kerrie, K Nguyen, Ngoc, B Participants 2014 Dartmouth University Rosalind Franklin University Medical University of South Carolina New York Medical College Michigan State University Tulane University University Alabama - Birmingham Tufts University University of Tennessee University Alabama - Birmingham Brown Uniersity University of Pennsylvania Meharry Medical College Jefferson Medical College Michigan State University Michigan State University University of Maryland Wayne State University Saint Louis University University of Central Florida Columbia University Georgetown University University of Arkansas Ponce School of Medicine and Health Sciences University of Michigan Tulane University University of Washington Medical School BADERC UCLA Yale Vanderbilt U Chicago Columbia UAB Joslin Vanderbilt UAB JHopkins/U Maryland U Pennsylvania Vanderbilt U Pennsylvania Columbia U Michigan U Maryland U Michigan Washington U Vanderbilt UCSF BADERC JHopkins/U Maryland Vanderbilt U Michigan UCSD U Washington Center for Summer Email address [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] 2014 Participants in the NIDDK Medical Student Research Program in Diabetes and Obesity Participants 2014 Nigam, Tina Oelsner, Kathryn Oelsner, William Olecki, Elizabeth Order, Kaitlyn, E Ortiz Baez, Vivian Owens, Sarah Pahouja, Gaurav Panosian, Jennifer, A Parris, Donique A Patel, Feenalie, N Penchev, Radostin Popli, Tanav Prasad, Pooja Pujare, Deepti, U Rabbani, Zaheen Rai, Dilreet Rai, Puja Ramirez, Lourdes, G Ran, Nina, A Reed, Thomas Rockas, Mary, Demetra Roos, Vanessa Scheidemantle, Brooke Sharma, Ellora, V. Sherwood, Pamela, R Shin, Won Seok Abst # 7 95 99 72 58 123 42 29 93 77 90 65 75 78 79 92 117 96 109 54 13 98 33 88 15 126 32 8 Michigan State University Medical University of South Carolina Medical University of South Carolina Northeast Ohio Medical University Georgetown University San Juan Bautista School of Medicine University Alabama - Birmingham Northeast Ohio Medical University Tufts University University of Maryland University of Tennessee George Washington University Indiana University University of Rochester New York Medical College University of Louisville University Missouri - Kansas City University of Connecticut University of Central Florida University of Pennsylvania George Washington University University of Michigan Medical University of South Carolina Thomas Jefferson Medical College Northeast Ohio Medical University Tufts University Eastern Virginia Medical School Medical School U Michigan Vanderbilt Vanderbilt Vanderbilt BADERC Vanderbilt UAB JHopkins/U Maryland Joslin JHopkins/U Maryland BADERC Vanderbilt Vanderbilt UCSF UCSF U Washington U Washington Coumbia Joslin U Pennsylvania Vanderbilt U Michigan JHopkins/U Maryland U Chicago JHopkins/U Maryland BADERC Vanderbilt Center for Summer Email address [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] 2014 Participants in the NIDDK Medical Student Research Program in Diabetes and Obesity 49 21 51 106 23 61 105 89 115 118 57 60 8 104 121 82 120 41 Abst # 9 Silva, Alejandro, J Tanaka, Mari Tarun, Samiksha Thakur, Sneha Theprungsirikul, Poy Vangala, Divya Venn, April, M.R. Verlage, Kenneth Wang, Eileen Weintraub, Michael Welch, Andrew, A Wesson, Emily Whitlock, Scott M Xie, Juliana, L Xu, Cassie Young, Verona Zhong, Le Zhu, Linjue Participants 2014 Rutgers NJ Medical School - Newark Georgetown University Saint Louis University New York Medical College Dartmouth University Marshall University Dartmouth University Texas Tech University Rush Medical College Rutgers Robert Wood Johnson - Piscataway Midwestern University - Glendale University Alabama - Birmingham Eastern Virginia Medical School Pennsylvania State University Indiana University Stony Brook University University of Miami Medical College of Georgia Medical School BADERC Vanderbilt Washington U Washington U Joslin Joslin Vanderbilt Vanderbilt Vanderbilt Vanderbilt U Washington UAB JHopkins/U Maryland Joslin Vanderbilt Columbia Vanderbilt Vanderbilt Center for Summer Email address [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] [email protected] 2014 Participants in the NIDDK Medical Student Research Program in Diabetes and Obesity Medical Schools of Students Participating in the 2014 NIDDK Medical Student Research Symposium Boston University Brown University Case Western Reserve University Columbia University Dartmouth University Eastern Virginia Medical School Florida Atlantic University George Washington University Georgetown University Indiana University Jefferson Medical College Loma Linda University Loyola University - Chicago Marshall University Medical College of Georgia Medical University of South Carolina Meharry Medical College Mercer University Michigan State University Midwestern University - Glendale National University of Ireland - Galway New York Medical College Northeast Ohio Medical University Pennsylvania State University Ponce School of Medicine Rosalind Franklin University Rush Medical College Rutgers NJ Medical School - Newark Rutgers Robt Wood Johnson - Piscataway Saint Louis University San Juan Bautista School of Medicine Stony Brook Medicine Texas A&M Health Science Center Texas Tech University Thomas Jefferson Medical College Tufts University Tulane University University Alabama - Birmingham University Missouri - Kansas City University of Arkansas University of California - Los Angeles University of Central Florida University of Connecticut University of Hawai'i University of Illinois - Champaign-Urbana University of Illinois - Chicago University of Iowa University of Kansas University of Kentucky University of Louisville University of Maryland University of Miami University of Michigan University of Missouri - Columbia University of Nebraska University of Nevada University of North Carolina University of North Dakota University of Oklahoma University of Pennsylvania University of Puerto Rico University of Rochester University of Southern California University of Tennessee University of Toledo University of Vermont University of Washington Washington University Wayne State University 10 Visiting Professors/Faculty for the NIDDK Medical Student Research Symposium Art L. Castle, PhD Program Director Metabolomics and Informatics National Institute of Diabetes & Digestive & Kidney Diseases National Institutes of Health 6707 Democracy Blvd., Rm 791, MSC 5460 Bethesda, MD 20892-5460 [email protected] Andrea L. Cherrington, MD Associate Professor Division of Preventative Medicine Department of Medicine FOT 725 University of Alabama at Birmingham Birmingham, AL 35294 [email protected] Richard G. Kibbey, MD PO Box 208056 Yale School of Medicine 333 Cedar Street New Haven, CT 06520-8056 Richard.kibbey@yale. Alvin C. Powers, MD Joe C. Davis Chair in Biomedical Science Professor of Medicine, Molecular Physiology & Biophysics Director, Vanderbilt Diabetes Research and Training Center 802 Light Hall Vanderbilt University Medical Center 2215 Garland Avenue Nashville, TN 37232-0202 [email protected] Mark de Caestecker, MB, BS, PhD Associate Professor of Medicine, Division of Nephrology, and of Cell & Developmental Biology and Surgery S-3223 Medical Center North Vanderbilt University Medical Center Nashville, TN 37232-23372 [email protected] 11 Biographies of NIDDK Visiting Professors and Faculty Art L. Castle, PhD is the Director of the Metabolomics and Informatics Program, Division of Diabetes, Endocrinology and Metabolic Diseases at the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. Arthur L. Castle has a PhD in Physiology from University of Texas, Austin. He did his postdoctoral research training study glucose transporters at NIDDK/NIH before working as a Manager in Bioinformatics for Gene Logic Inc. His research experience includes diabetes, metabolism, genomics, bioinformatics, and computer simulation/modeling. Dr. Castle joined the Division of Diabetes, Endocrinology and Metabolic Diseases, NIDDK, NIH, DHHS in 2004, where he is current the program director for fellowships (“F”) awards, institutional training grants, metabolomics and informatics. Andrea L. Cherrington, MD, MPH, is an Associate Professor in the Division of Preventive Medicine at the University of Alabama at Birmingham where she is a clinician trained in health services research and community-based participatory research methods. After completing Internal Medicine residency training in 2003, she attended the University of North Carolina-Chapel Hill as a fellow in the Robert Wood Johnson Clinical Scholar’s Program (CSP) and obtained an MPH from the UNC School of Public Health. Following the CSP, she obtained funding for a third year of fellowship through an NRSA T32 mechanism. In 2006, Dr. Cherrington returned to Alabama as an Assistant Professor of Medicine at UAB. She has received funding for her research through pilot grants from the UAB Minority Health and Research Center, the UAB Clinical Nutrition Research Center, the UAB Diabetes Research and Training Centers, and the American Diabetes Association. Her research focuses on developing, implementing and evaluating community-based interventions to promote weight loss and improve the outcomes of disadvantaged populations at high risk for developing chronic disease, specifically diabetes. Additionally, she obtained two career development awards, one through the Robert Wood Johnson Foundation and the other an internal K12 in Comparative Effectiveness Research. Both projects focus on use of the Community Health Worker model for the reduction of diabetes related health disparities. Dr. Cherrington co-directs the Intervention and Translational Core within the UAB Diabetes Research Center. She is currently involved in several projects that seek to advance methods of community-based intervention delivery by combining the Community Health Advisor model with multi-component, technology-assisted support systems comprising health professionals. In keeping with her research interests, she has a continuity clinic two half days per week within Cooper Green Health System, the county’s safety net system. Mark de Caestecker, MB, BS, PhD, received his undergraduate degree from Cambridge University and medical degree (M.B., B.S.) from The Middlesex Hospital (now University College) in London, England. After training in internal medicine at the Middlesex and Charring Cross Hospitals in London, Dr. de Caestecker trained in Nephrology, obtained his PhD in Medicine at the University of Manchester, and underwent post-doctoral training on a Wellcome Trust Clinical Scientist Fellowship at the NIH under the mentorship of Dr. Anita Roberts. He is currently an Associate Professor in the Division of Nephrology at Vanderbilt, and conducts NIH funded research on regenerative therapies in acute and chronic kidney injury, and on hereditary vascular diseases. He 12 currently serves on the Respiratory and Integrative Biology and Translational Research study section. Dr. de Caestecker is Director of the Vanderbilt-HHMI Certificate Program in Molecular Medicine, and Director of the Vanderbilt Center for Kidney Disease education program. He has directed the summer research-training program in kidney disease for the last four years. Richard G. Kibbey, MD, PhD is an Associate Professor in the Departments of Internal Medicine Division of Endocrinology and Cellular & Molecular Physiology at Yale University School of Medicine. He attended Trinity University in San Antonio where he obtained a degree in Music and a degree in Biochemistry. Next he completed his M.D. Ph.D. training at UT Southwestern in Dallas in the laboratories of Drs. Lila Gierasch and Richard Anderson where he studied NMR structural biology. From there he short-tracked through internal medicine residency and endocrinology at Yale to later do a postdoctoral fellowship in the laboratory of Dr. Gerald Shulman. During his fellowship he obtained support from the Clinical and Scientific Training Award, American Diabetes Association Junior Faculty Awards, and a NIH K08. Since then he has obtained additional funds from ADA research grants, NIH R03 and R01 awards, and several industry-sponsored awards. At Yale he runs an independent research lab studying the metabolism of diabetes with particular interest in human islet biology. Here he has identified a novel mitochondrial energy sensing mechanism that regulates glucose homeostasis and has developed state-of-the-art techniques for measuring intracellular metabolic fluxes. He is currently working closely with the pharmaceutical industry to translate these findings into new therapies. As a practicing Endocrinologist he also sees patients in a continuity clinic ½ day per week and attends on the Endocrine Consult Service as well as General Internal Medicine 1-1.5 months a year. In addition he teaches medical physiology to first year medical students. Alvin C. Powers, MD, the Joe C. Davis Chair in Biologic Science and Professor of Medicine, Molecular Physiology and Biophysics at Vanderbilt University, is the Director of the Vanderbilt Diabetes Center, the Chief of the Vanderbilt Division of Diabetes, Endocrinology, and Metabolism, and the Director of the Vanderbilt Diabetes Research and Training Center, a NIH-funded center that facilities the diabetes-related research of more than 120 Vanderbilt scientists. He conducts diabetes-related research that focuses on pancreatic islet biology, development, function, and transplantation. His research is or has been supported by the NIH, the VA Research Service, the Juvenile Diabetes Research Foundation (JDRF), and the American Diabetes Association (ADA). Dr. Powers is the Director of the Vanderbilt Medical Student Research Training Program in Diabetes, Endocrinology, and Metabolism and the Coordinator for the NIDDK Medical Student Research Program in Diabetes. These two programs enable more than 100 medical students to conduct diabetes-related research each summer at a NIDDK-supported Diabetes Research Center. Dr. Powers, a physician at Vanderbilt University Medical Center and the VA Tennessee Valley Healthcare System, is listed by Castle Connolly Medical Ltd as one of “America’s Top Doctors.” Dr. Powers received his undergraduate degree from the University of Virginia and his medical degree from the University of Tennessee Center for the Health Sciences. After training in internal medicine at Duke University Medical Center, Dr. Powers trained in Endocrinology and Diabetes at the Joslin Diabetes Center, the Massachusetts General Hospital, and Harvard Medical Schooll. 13 Vanderbilt Faculty Participating In Moderating the Poster Sessions and Lunches Nathan Bingham, MD, PhD Milam A. Brantley, MD, PhD Jessica Devin, MD Raymond C. Harris, MD J. Matthew Luther, MD James M. May, MD Owen P. McGuinness, PhD Daniel J. Moore, MD Kevin D. Niswender, MD, PhD Richard M. Peek, MD William (Bill) E. Russell, MD Ashley H. Shoemaker, MD John M. Stafford, MD, PhD James W. (Tom) Thomas, MD Melissa F. Wellons, MD Keith T. Wilson, MD Lan Wu, MD 14 NIDDK-Sponsored Diabetes Research Centers Boston Area Diabetes Center DK057521 Columbia University DK063608 Johns Hopkins University and University of Maryland DK079637 Joslin Diabetes Center DK036836 University of Alabama at Birmingham DK079626 University of California at San Diego and University of California at Los Angeles DK063491 University of California at San Francisco DK063720 University of Chicago DK020595 University of Michigan DK020572 University of Pennsylvania DK019525 University of Washington DK017047 Vanderbilt University DK020593 Washington University DK020579 Yale University DK045735 15 Training Grants Supporting 2014 NIDDK Medical Student Research Program in Diabetes Grant Number T32 DK007770-13 T32 DK007328-35 T32 DK007751-18 T32 DK007260-38 T32 DK007028-40 T32 DK062710-11 T32 DK007494-30 T32 DK007418-34 T32 DK007011-40 T32 DK007245-38 T32 DK007314-34 T32 DK007247-37 T32 DK007061-40 T35 DK007383-35 T32 DK007120-40 PI of Training Grant Melmed, Shlomo Gottesman, Max Radovick, Sally Blackwell, Thomas Keith Avruch, Joseph Allison, David Witztum, Joseph L German, Michael Refetoff, Samuel Auchus, Richard Birnbaum, Morris Schwartz, Michael May, James Powers, Alvin Semenkovich, Clay Institution Cedars Sinai Medical Center Columbia University Johns Hopkins University Joslin Diabetes Center Massachusetts General Hospital University of Alabama Birmingham University of California San Diego University of California San Francisco University of Chicago University of Michigan University of Pennsylvania University of Washington Vanderbilt University Vanderbilt University Washington University T32 DK007058-40 Wysolmerski, John Yale University Some students were supported by the Diabetic Complications Consortium U24 DK076169-03 McIndoe, Richard Georgia Regents University Some students were supported by the Summer Program in Obesity, Diabetes and Nutrition Related Research Training (SPORT) T35 DK095737-02 Steinle, Nanette University of Maryland Some students were supported by the Vanderbilt Center for Kidney Disease The NIDDK Medical Student Research Program and Symposium were coordinated and hosted by the Vanderbilt Diabetes Research and Training Center P60 DK020593-37 Powers, Alvin Vanderbilt University 16 NIDDK Medical Student Program Advisory Panel – 2014 Art L. Castle, PhD Program Director Metabolomics and Informatics National Institute of Diabetes & Digestive & Kidney Diseases National Institutes of Health 6707 Democracy Blvd., Rm 791, MSC 5460 Bethesda, MD 20892-5460 [email protected] James F. Hyde, PhD Senior Advisor, Career Development & Diabetes Centers Programs Division of Diabetes, Endocrinology and Metabolic Diseases National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health 6707 Democracy Blvd., Room 789 Bethesda, MD 20892-5460 [email protected] Steven E. Kahn, MB, ChB Professor of Medicine Division of Metabolism, Endocrinology & Nutrition Director, Diabetes Research Center University of Washington VA Puget Sound Health Care System 1660 S. Columbia Way (S-151) Seattle, WA 98108 [email protected] Alvin C. Powers, MD Joe C. Davis Chair in Biomedical Science Professor of Medicine, Molecular Physiology and Biophysics Director, Vanderbilt Diabetes Research and Training Center 802 Light Hall Vanderbilt University Medical Center 2215 Garland Avenue Nashville, TN 37232-0202 [email protected] Michael R. Rickels, MD, M.S. Assistant Professor of Medicine Division of Endocrinology, Diabetes & Metabolism University of Pennsylvania School of Medicine 763 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104-6149 [email protected] Louis H. Philipson, MD, PhD Director, The University of Chicago Kovler Diabetes Center Professor of Medicine University of Chicago Pritzker School of Medicine Chicago, IL [email protected] Sally Radovick, M.D. Lawson Wilkins Professor of Pediatrics Director, Division of Pediatric Endocrinology The Johns Hopkins Hospital Room 3125 200 N. Wolfe Street Baltimore, MD 21287-2520 [email protected] 17 Abstracts from the Posters of the Students Participating in the 2014 NIDDK Medical Student Research Program (For abstract number see Table of Student Participants: pages 5 - 9) 18 Association between total cholesterol and at-risk psychosis symptoms among helpseeking adolescents. Xavier Diao1, Caroline Demro2, Elizabeth Thompson2, Emily Kline2, Kristin Bussell1, Jason Schiffman2*, Gloria Reeves1 1 University of Maryland School of Medicine, 2University of Maryland, Baltimore County *Drs. Schiffman and Reeves served as Co-Mentors on this project. Schizophrenia is a potentially devastating mental illness, characterized by perceptual disturbances (e.g. hallucinations), delusional thoughts, and/or disorganized behavior. The primary pharmacologic treatment, antipsychotic medication, can also cause physical health problems through metabolic side effects (e.g. weight gain, new-onset diabetes). However, there is evidence that metabolic abnormalities may be present in individuals with first-episode psychosis prior to antipsychotic medication exposure, including hyperglycemia. It is unclear how early these metabolic abnormalities can be detected over the course of mental illness, and also if there is any association between specific psychotic symptoms and metabolic parameters. In this study, we assess metabolic syndrome criteria in a sample of 51 antipsychotic-naïve adolescents (15.45 ± 2.64 yrs old) receiving mental health services. Youth were screened for “at-risk” psychosis symptoms (e.g. suspiciousness without overt paranoia) using the Structured Interview for Psychosis Risk Syndromes (SIPS), and metabolic parameters (total cholesterol, HDL-C, LDL-C, glucose, insulin, triglycerides, weight, height, waist circumference) were measured after an overnight fast. We examined the relation between at-risk psychosis symptoms and fasting metabolic parameters. We found that disorganized symptoms were significantly correlated with total cholesterol (r = –0.39, p = .005) and LDL-C (r = –0.29, p = .040). The correlation with total cholesterol remained significant after controlling for waist circumference (r = –0.35, p = .015), and when participants with current stimulant use were excluded from analyses (r = –0.41, p = .015). There was no significant association between atrisk psychosis symptoms and fasting glucose. The results of this pilot study, if replicated, indicate a possible association between total cholesterol and disorganized symptoms that can be detected in adolescence. Longitudinal studies are needed to assess the causal relation between this metabolic parameter and psychiatric symptom over a course of illness. Abstract 1 Mobile Health Messaging as a Tool for Behavior Change in Diabetes Management Erin Butler, BS (Eastern Virginia Medical School), Charlene C. Quinn, RN, PhD, University of Maryland School of Medicine, Department of Epidemiology & Public Health Mobile health (mHealth) is rapidly expanding in healthcare allowing for patient care outside the physician’s office to help prevent and manage chronic medical conditions. The Mobile Diabetes Intervention Study (MDIS) evaluated a mobile communication system for patients with Type 2 diabetes. The primary study outcome was 1.9% mean change in HbA1c (mean difference from control 1.2% (P<0.001) over a one year treatment period. Patients used a secure web portal to record blood glucose, medication, and carbohydrate intake, and exchange messages with diabetes educators. This secondary data analysis used a qualitative approach to describe patient engagement, defined by content and quantity of messages, to determine if patient engagement is related to change in HbA1c. This study is a mixed methods-qualitative, quantitative analysis of the patient messages (n=3188) of three stepped treatment groups (n=107); coach-only (CO), coach PCP portal (CPP), and coach PCP portal with decision support (CPDS). Seven code themes (total 38 codes) were defined based on the American Association of Diabetes Educators Self Care Behaviors for diabetes management, plus a code for technical issues. Each message was coded based on content, and a quantitative analysis of the results was performed. Modeling to determine the relationship of patient engagement and HbA1c was performed. The patient engagement pattern followed the initial results of the study, with CPDS group being the most engaged, followed by CO and CPP, respectively. Patients messaged more about monitoring (mean 8.37 messages per patient) and healthy eating (mean 6.23 messages per patient) overall. Technical problems comprised a large proportion of messages (mean 6.36 messages per patient). Accounting for non-substantive messages, patients across the treatment groups sent 2.81 healthy eating substantive messages per patient and 7.10 monitoring substantive messages. While further analysis of patient engagement prediction of change in HbA1c is pending, these qualitative results indicate patients used mhealth to engage with providers on self-care behaviors. This information can help further develop mobile patient engagement tools. Abstract 2 Examining Ethnic Disparities in Insulin Administration among Patients with Type 2 Diabetes Nicolas Ludi (University of Tennessee Health Science Center) and Russell Rothman MD MPP, Vanderbilt University, Nashville, TN Latinos have a higher prevalence of T2DM and are disproportionately burdened with diabetes complications. Insulin is an important treatment to optimize glycemic control in patients with Type 2 Diabetes (T2DM), particularly for those with poor diabetes control. To date, little research has examined the prevalence of insulin administration among Latinos with diabetes versus non-Latinos. We believe that Latinos are less likely to be on insulin even after adjustment for factors such as A1C, duration of diabetes, and insurance status. To test our hypothesize we performed a cross sectional analysis of baseline data from a cluster randomized trial examining the role of a literacy sensitive intervention to improve diabetes care in patients seen at 10 separate Tennessee county health departments for the Vanderbilt PRIDE study. The study partnered with the Tennessee Department of Health and included a total patient population size of 400. Our analysis focused on the 98 patients who were self-identified as Latino’s and compared them to the remaining 302 non-Latino individuals. The baseline data collected included: patient characteristics, ethnicity from patient self-report, Subjective Literacy Skills (SLS-3) test scores, Diabetic Numeracy Test (DNT-5) scores, diabetes medications, BMI, and Hemoglobin A1C (from medical records.) Our results showed that almost 2/3 of T2DM patients in this vulnerable population were on insulin therapy. However, Latino patients were significantly (62% vs. 51%) less likely to be on insulin therapy. Lower likelihood of insulin usage among Latinos remained even after adjustment for potential confounders. Additional research is needed to examine potential etiologies for disparities in insulin therapy. Future interventions could aim to reduce these disparities to optimize insulin therapy for Latino patients with diabetes. The identification of insulin disparities among Latino’s is important because it is a factor that is potentially modifiable. We hope that health care professionals can examine the data we collected and use it to focus their care and improve the outlook of Latino patients. Abstract 3 Prenatal drug exposure, stress reactivity, and adolescent drug experimentation, problem behaviors and weight status. Dayna Mazza (University of Maryland School of Medicine), Stacy Buckingham-Howes Ph.D. and Maureen Black Ph.D., University of Maryland School of Medicine Toxic stress, such as prenatal drug exposure (PDE), can alter the Hypothalamic-PituitaryAdrenal (HPA) axis, resulting in dysregulated stress reactivity, with long-term problems in child health, development, and growth (Shonkoff et. al., 2012, Kirschbaum and Hellhammer 1989). Although findings are controversial, some adolescents with PDE have a higher likelihood of engaging in problem behaviors, including drug experimentation, and have a higher weight status than non-exposed (NE) adolescents (Frank et. al., 2011, Lambert et. al., 2013, Shankaran et. al., 2010). Little is known about the mechanisms, including the role of stress reactivity, underlying these associations (Chaplin et. al., 2010, Lester et. al., 2010, Buckingham-Howes et. al., in press). We hypothesize that among the NE group, typical stress reactivity is associated with few problem behaviors and low BMI while no such associations are expected in the PDE group. The study was conducted among adolescents mean age = 14.17 years (SD = 1.17; range = 11.9316.64) who varied in PDE exposure (PDE, n=76; NE, n=61) and had been followed since delivery (PDE group), age 5 (NE group), or age 14 (additional NE group). The sample was 50% male and 99% African American. Measured height, weight, cortisol (stress reactivity), caregiver report of risk behaviors, and youth report and urine samples for drug use were collected. Overall, 45.3% of the sample was overweight/obese (BMI for age > 85th percentile) with no differences in PDE status. The NE group was significantly more likely to react to a mild stressor than the PDE group (26% vs. 12%; x2=4.49, p=.03), and the PDE group was more likely than the NE group to experiment with tobacco/alcohol (25% vs. 10%; x2=4.76, p<.05). Within each group (PDE vs. NE), we used linear or logistic regression to examine the association between stress reactivity and problem behaviors and weight. The adolescents in the NE group with typical stress reactivity engaged in fewer problem behaviors (any drug use: aOR=0.49, CI: 0.25-0.94, p<.05; aggression: b=-4.99, p<.05) than adolescents with atypical stress reactivity. There was no identifiable pattern within the PDE group. There was no association between stress reactivity and weight status in the NE group (B=-.156, p=>.05); however, there was marginal significance within the PDE group, showing an association between less stress reactivity and higher BMI zscore (B=-.871, p=.08). These results suggest that there may be a disruption of the HPA axis in adolescents who are PDE, which blunts stress reactivity and interferes with expected associations between stress reactivity and adolescent problem behaviors and weight status. Future research is needed to further understand how PDE relates to the HPA axis and the role that the stress reactivity system plays on behavioral and biological outcomes in youth exposed to toxic stress. Abstract 4 Effects of Vitamin D Supplementation on the Quality of Life in Patients with Type 2 Diabetes Mellitus. Beatriz González Bouza (University of Puerto Rico School of Medicine)., Amy E. Riek, M.D., Robin R. Bruchas, MSW, Carlos Bernal-Mizrachi, M.D. Washington University School of Medicine in St. Louis, MO. Evidence suggests that vitamin D may influence both insulin secretion and sensitivity, and those patients with type 2 diabetes mellitus (T2DM) that have a good glycemic control have reported a better quality of life. The hypothesis was that vitamin D supplementation over 4 months will help improve the quality of life of patients with T2DM and who have low levels of vitamin D. A double blind, placebo controlled trial was conducted and patients participating were randomized to receive either 4000 IU of vitamin D or a placebo tablet. Both groups received 1.2g of calcium carbonate daily. Data collecting visits were preform at baseline, 2 months and 4 months. Participants had their fasting glucose and A1C’s measured and completed the Daily Quality of Life Survey. Results showed that there were no significant differences in the answers between the groups and that only the questions regarding the time to determine the blood sugars and the frequency of pain caused by the treatment for diabetes showed a tendency to improve after the treatment in group A. More patients are required to clarify the tendencies in the differences observed between the groups. Abstract 5 Effect of Sestrin2 on signaling pathways of liver cancer cells. Kathleen Estrada (Michigan State University), Hwan-Woo Park, Jun Hee Lee, University of Michigan Sestrin2 protein has been shown to prevent the development of type 2 diabetes and non-alcohol fatty liver disease in mice placed on a high fat diet (HFD). During HFD-induced obesity, Sestrin2 was shown to suppress ER stress signaling and promote AKT signaling, both of which contributes to the maintenance of metabolic homeostasis in liver. It has been recently shown that HFD can promote liver carcinogenesis in a chemical (diethylnitrosamine, DEN)-induced liver cancer model. Thus, we investigated how Sestrin2 affects liver carcinogenesis and ER stress/AKT signaling pathways. We compared different ER stress markers and other signaling markers in the liver tumors and non-tumor liver tissues of wild type mice and Sestrin2 knockout mice that were fed HFD. We found that the expression of p-AKT was reduced in knockout mice compared to wild type mice. However, ER stress markers p-eIF2α and BIP were elevated in the knockout mice. These results confirm known relationships between Sestrin2, ER stress and AKT signaling pathways in liver cancer cells. More studies will have to be done to fully understand Sestrin2’s role in liver carcinogenesis. Abstract 6 Pro-inflammatory pathways in the human colon: Relationships between toll-like receptor 4 (TLR4) and prostaglandin formation. Tina Nigam (Michigan State University College of Human Medicine), Elkhansa Sidahmed, Jianwei Ren, D. Kim Turgeon, Mack T. Ruffin, Dean E. Brenner, and Zora Djuric University of Michigan The Mediterranean diet has been shown to reduce the risk of cardiovascular disease and diabetes, and this diet could have beneficial effects on preventing colon cancer as well. A proinflammatory state in the colon, with a key role in prostaglandin (PG) E2 formation, is known to increase colon cancer risk. A Mediterranean diet, conversely, is thought to be anti-inflammatory. This study evaluated the effects of a Mediterranean diet on genes involved in the proinflammatory response in the human colon. The hypothesis of this study was that exposure to intestinal bacteria, as evidenced by induction of toll-like receptor 4 (TLR4), would be decreased by a Mediterranean diet. Activation of TLR4 should increase PGE2, expression of enzymes involved in PGE2 synthesis, and expression of the MUC2 gene, which is involved in formation of mucin that protects the colon epithelial lining. To test this hypothesis, we used samples from a clinical trial that randomized 120 high risk individuals into two diet groups (a Mediterranean diet or a Healthy Eating diet) for 6 months. Colon biopsies were obtained by flexible sigmoidoscopy without prior preparation of the bowels. Quantitative Real Time PCR using the standard curve method was utilized to quantify gene expression. Expression of TLR4 and MUC2 was fairly high in colon biopsies; however, neither gene showed significant changes in expression after dietary intervention. At baseline, TLR4 expression was significantly and positively correlated with MUC2 expression and PGE2 concentration. Expression levels of both MUC2 and TLR4 were correlated with expression of several genes in the PGE2 pathway: PTGS1, PTGES1, PTERG2, and PTGER4. The especially strong correlation of TLR4 gene expression with enzymes in the PG pathway was surprising and indicates the possible importance of exposure to intestinal bacteria in stimulating pro-inflammatory pathways in the colon. Future work should better characterize the factors that induce TLR4 and the mechanisms by which TLR4 activation could induce the eicosanoid pathway in the human colon. Abstract 7 Metabolic conditions extensively modulate the mRNA expression and circulating levels of inflammatory cytokines. Scott M. Whitlock (Eastern Virginia Medical School), Pia S. Petersen, X. Lei, M.M. Seldin, S. Rodriguez, M.S. Byerly, A. Wolfe and G.W. Wong, Johns Hopkins School of Medicine Cytokines play diverse and critical roles in innate and acquired immunity, and a few have been shown to act in the central nervous system and peripheral tissues to modulate energy metabolism. The extent of crosstalk between the immune and endocrine systems is not well understood and has not been systematically examined. We hypothesized that circulating levels of cytokines are dynamically regulated in response to physiologically relevant, short-term (fasting and refeeding) or long-term (chronic high-fat feeding), metabolic perturbations. We used a multiplex approach to profile changes in seventy-one circulating mouse cytokines in response to acute (fasting and refeeding) and chronic (high-fat feeding) alterations in wholebody metabolism, and then used trizol RNA extraction and real-time PCR to analyze the alterations in cytokine mRNA expression. Strikingly, refeeding significantly decreased serum levels of IL-22, IL-1α, sIL-2Rα, sVEGFR2, and sVEGFR3, but markedly increased G-CSF, IL-16, IL-1β, CXCL1, sIL-1RI, lipocalin-2, pentraxin-3, TIMP-1, and SAP relative to the fasted state; these changes were accompanied, in part, by alterations in cytokine mRNA expression. In the context of chronic high-fat feeding, circulating levels of CXCL1, SAA3, TIMP-1, and Alpha-2macroglobulin (A2M) were increased, whereas IL-12p40, CCL4, sCD30, sRAGE, CCL12, CCL20, IL-16, IL-22, and haptoglobin were decreased relative to mice fed a control low-fat diet; these changes accompanied alterations in cytokine mRNA expression. Our results demonstrate that short- and long-term changes in whole-body metabolism extensively alter circulating cytokine levels and gene expression. This study highlights the potential functional crosstalk between the immune and endocrine systems, and provides a foundation to functionally characterize the potential physiological role of cytokines in the context of metabolic perturbations. Abstract 8 Chronic Kidney Disease: Is Fibrin-Mediated Inflammation a Cause of Renal Capillary Rarefaction? Heather Cole BS1,2 Emily Smith2, Leslie Gewin MD3, Jonathan Schoenecker MD, PhD2 University of Tennessee Health Science Center, Memphis, TN 1, Vanderbilt University Department of Orthopedics 2 and Department of Medicine3, Nashville, TN Chronic kidney disease (CKD) affects one out of every seven adults in the United States that ultimately involves peritubular capillary rarefaction (vascular regression) followed by interstitial fibrosis and tubular atrophy. Despite numerous investigations into the mechanisms of renal fibrosis, peritubular capillary rarefaction remains incompletely understood. Previous work has established that persistent fibrin deposition within tissue results in increased local and systemic inflammation. Because fibrinogen has been shown to be elevated in patients with chronic renal failure and has been linked as a mitogen for tubulointerstial fibroblasts, we hypothesized that inordinate fibrin within renal tissue potentiates chronic kidney disease through peritubular capillary rarefaction. Using plasminogen-deficient mice as a model for defective fibrinolysis, we performed angiographies to assess renal vascularity in relation to fibrin deposition (immunofluorescence) and inflammatory response (IL-6 ELISA and F4/80 macrophage immunohistochemistry). Wild-type mice (C57/Bl6), plasminogen-deficient mice (Plg-/-) 1, and mice deficient in plasminogen and the αMβ2 binding site of fibrinogen (Plg-/Fbgγ390-396A) _ENREF_102 were sacrificed at 20 weeks of age and analyzed. Our findings demonstrate that impaired fibrinolysis leads to a pro-inflammatory environment with elevated systemic IL-6 and fibrinogen levels, fibrin deposition in renal tissue, and significant increase in F4/80 macrophages compared to WT. Interestingly, these findings are ameliorated in the Plg-/Fbgγ390-396 mice. Additionally, renal angiographies demonstrate a significant decrease in renal vascularity in the Plg-/- mice that is normalized in the Plg-/- Fbgγ390-396 (15 weeks). As inflammation has been identified as a contributing factor to capillary rarefaction and has been associated with mortality in patients with chronic kidney disease, our findings suggest that fibrinmediated inflammation may be an inciting factor for chronic kidney disease. Abstract 9 Effects of hyperglycemia and hyperlipidemia on mitochondrial dynamics in sensory neurons Nandini S. Abburi, B.S. (University of Toledo College of Medicine), Samuel W. Jackson, B.S., Gideon E. Levinson, B.S., Tatsuhito Himeno, Ph.D., Lucy M. Hinder, Ph.D., John M. Hayes, B.S., Eva L. Feldman, M.D., Ph.D., and Stephen I. Lentz, Ph.D., University of Michigan – Ann Arbor Diabetic peripheral neuropathy (DPN) is the most common complication of diabetes and is characterized by a length-dependent loss of sensory nerve function. The mechanism by which diabetes promotes the development and progression of neuropathy remains unknown. Mitochondria are the primary source for cellular energy, and mitochondrial dysfunction has been associated with a number of neurodegenerative diseases, including chemotherapy- and diabetes-induced peripheral neuropathies. Studies from our laboratory and others have identified hyperglycemia and diabetes-induced changes in mitochondrial dynamics, but recent studies have also implicated hyperlipidemia as a potential disruptor of normal mitochondria function. This current study investigates the effects of both hyperglycemia and hyperlipidemia on mitochondrial trafficking in sensory neurons as a mechanism for sensory loss associated with DPN. Live cell fluorescence microscopy was used to study the movement of individual mitochondria within sensory neurites of primary cultured mouse dorsal root ganglion neurons. Mitochondria were labeled with Mito-GFP (BacMam 2.0 encoding mitochondrial-targeted green fluorescent protein; Life Technologies) and imaged every 2.5 sec for 2.5 min with a 40x oil (1.3 N.A) objective on a Nikon A1 confocal microscope. Kymographs (MetaMorph, Molecular Devices) were used to quantify the percent motility, and the proportion and velocity of anterograde and retrograde trafficking were for the following treatment groups: control, 0.5% fatty-acid-free BSA solution control (BSA), 50 mM glucose (G), 0.5 and 2 mM palmitate (P), 25 mM glucose + 0.25 or 1 mM palmitate (G+P). All conditions and time points showed a similar density of mitochondria within sensory neurites, with approximately 2-3 mitochondria per 10 microns of neurite. After 12 hours of treatment, P and G+P significantly lowered the percentage of motile mitochondria compared to control, while BSA and G had no significant influence on mitochondrial motility. After 12 and 24 hours of treatment, the P and G+P conditions showed primarily anterograde movement, while retrograde motion dropped off entirely by 24 hours. The results demonstrated that hyperlipidemia deleteriously reduced mitochondrial trafficking, suggesting that dyslipidemia may be more influential in the development and progression of DPN. Abstract 10 Using Genotypes to Compare Clinical Presentation of Symptoms in Patients with Wolfram Syndrome Ian Milligan (Saint Louis University School of Medicine), Dr. Fumihiko Urano, MD, PhD, Washington University in St. Louis Wolfram syndrome (WS), also known as DIDMOAD, is a rare (~1/700,000), primarily autosomal recessive disease characterized by the adolescent onset of diabetes mellitus and optic atrophy, as well as the development of deafness, diabetes insipidus, and eventual neurodegeneration in the cerebellum and brainstem. Mutations in the WFS1 gene have been implicated as the cause for WS since the protein product, wolframin, has been associated with relieving endoplasmic reticulum stress, thereby reducing cell apoptosis. Wolframin accomplishes this by regulating cellular calcium levels and assisting with the proper folding of proteins such as proinsulin, a precursor to insulin. The progression of disease in WS is highly variable, and has been hypothesized to be associated with the type of mutations present in a patient’s WFS1 gene. Blood samples taken from patients and their family members were sequenced to look for mutations in the WFS1 gene and subsequently characterized. Genetic information was correlated with a patient database constructed using REDCap software, which had questionnaires regarding the onset and progression of symptoms. Mutations were found throughout the WFS1 gene, with missense mutations being the most prevalent (39%, n=58), and dominant mutations also being present (3.06%). Having two missense mutations was the most common genotype, and was found to have delayed onset of diabetes mellitus (10.17 ± 5.87 years, n=6) and insipidus (18.00 ± 1.00 years, n=2) as compared to other genotypes. Missense mutations were also found to be more prevalent (75%, n=2) in the slowly progressing patient group, those patients diagnosed with diabetes mellitus after the age of 20. Dominant mutations had an increased prevalence (50%, n=3) in the group of patients whose first diagnosed DIDMOAD symptom was hearing loss. These patients also developed hearing loss earlier (1.33 ± 1.04 years) when compared to all other patients (12.55 ± 9.06 years, n=28). Overall, missense mutations appear to be associated with a milder phenotype, whereas dominant mutations appear to be associated with deafness at a younger age. However, a larger patient population is needed to more accurately depict disease progression and detect differences between genotype and disease presentations. Abstract 11 Application of SOMAscanTM Technology for Biomarker Discovery in Peripheral Arterial Disease among Type 2 Diabetics Christina Marcaccio (Perelman School of Medicine, University of Pennsylvania), Benjamin Krieger, Kevin Trindade, Stephen R Master, Scott M Damrauer, Emile R Mohler, and Daniel J Rader, University of Pennsylvania Peripheral arterial disease (PAD) is a common manifestation of atherosclerosis and is a significant cardiovascular complication of both type 1 and type 2 diabetes. Importantly, diabetes is associated with increased risk for symptomatic PAD and higher morbidity and mortality rates. Currently, there are few preventive and treatment therapies for PAD among diabetics because the etiology and progression of PAD are largely unknown. Further, whether diabetic PAD is fundamentally different that non-diabetic PAD remains poorly understood. Previous efforts to gain a mechanistic understanding of diabetic PAD have used mass spectrometry-based proteomic methods to assess circulating proteins in disease samples; however, this traditional strategy is limited by issues of sensitivity, specificity, reproducibility, throughput, and cost. In this study, we use a novel multiplexed proteomics technology for the discovery of circulating biomarkers in diabetic PAD. Proteins are measured using a Slow Off-rate Modified Aptamer (SOMAmer)—based capture assay called “SOMAscan” (SomaLogic, Inc). This approach uses chemically modified nucleotides to transform each individual protein concentration into a corresponding SOMAmer concentration, which is then quantified using relative fluorescence on microarrays. The SOMAscan assay analyzes levels of 1129 circulating human proteins representing a diverse set of molecular functions and disease processes across a wide range of concentrations. We hypothesized that use of SOMAscan to assess the plasma proteome in type 2 diabetic subjects with PAD will identify novel protein biomarkers significantly altered in this disease. To test this hypothesis, we identified from the extensive prospectively-consented PennMedicine Biobank four patient subgroups: (1) T2 diabetic subjects with documented PAD; (2) matched non-diabetic subjects with documented PAD; (3) matched T2 diabetic subjects without PAD; (4) matched non-diabetic subjects without PAD. We use SOMAscan to analyze plasma proteins in all four groups. Our primary aim is to identify plasma proteins that exhibit significant differences in concentrations between group (1) and each of the control groups (2-4). Novel proteins that differ in expression between groups will be further evaluated through ELISA assays to confirm and accurately quantify the difference in protein concentrations between T2 PAD subjects and matched controls. Future studies will involve mechanistic assessment of newly identified proteins to categorize molecular pathways that may be altered in diabetic PAD. We hope that the identification of novel circulating biomarkers in PAD may advance understanding of disease pathophysiology, identify novel therapeutic targets, and improve disease management. Abstract 12 Medication Management of Dippers Vs. Non-Dippers in the Systolic Blood Pressure INTervention Trial (SPRINT) Thomas Reed, George Washington University School of Medicine, Jamie Dwyer, M.D. Vanderbilt University School of Medicine Blood pressure (BP) is higher during the daytime (wake periods), and lower during the nighttime (sleep periods). Recent studies have suggested that nocturnal systolic hypertension may be more useful for prognosis than conventional BP measurements. We hypothesized that nondippers require more classes of medications to reach their target BP than dippers and would be on higher dosage of medication at month 27 than dippers. We also hypothesized that nondippers will be at target BP for fewer study visits. Data was gathered on a host of variables such as demographics, BP readings, medications, dosage, and a 24-hour ambulatory BP reading. Using SPSS, data was analyzed using independent sample T-tests. Non-dippers spent on average 5.1 study visits at target BP, whereas dippers spent on average 5.6 study visits at target BP. Non-dippers were on an average of 3.0 classes of anti-hypertensive medicines per visit, whereas dippers were on an average of 2.6 classes of anti-hypertensive medicines per visit. Non-dippers had fewer visits at goal, more medications per visit, and a higher dosage of medication at month 27. However, there was no significant difference in classes of medications, time spent at target BP, or dosage of medication at month 27 between dippers and non-dippers. Abstract 13 Renal interstitial cell plasticity underlies the hypoxic erythropoietin response. Chad Binns (University of Arkansas for Medical Sciences), Hanako Kobayashi, Volker Haase, Vanderbilt University Medical Center One of the kidney’s responses to systemic hypoxia is to increase the production of erythropoietin (EPO), a glycoprotein hormone responsible for erythropoiesis. Recombinant EPO therapy has been utilized for patients suffering from anemia for the past two decades. Despite its effectiveness, recent studies have raised questions concerning the safety of recombinant EPO. Studying the kidney’s response to hypoxia may provide insight into how endogenous EPO production may be boosted, resulting in the development of safer alternatives to recombinant EPO therapy. Endogenous EPO production is tightly regulated through the hypoxia-inducible factor (HIF)/HIF prolyl-4-hydroxylase pathway. Under conditions of normoxia, constitutively expressed HIFα is hydroxylated by prolyl-4-hydroxylase domain (PHD) proteins and subsequently tagged for proteasomal degradation via von Hippel-Lindau (VHL) E3 ubiquitin ligase. Hypoxia impedes HIF hydroxylation and consequent degradation, thus allowing for its nuclear translocation and the transcription of hypoxia-regulated genes such as EPO. The kidney controls EPO production by modulating the number of renal EPO-producing cells (REPCs). REPCs are peritubular interstitial fibroblasts found in the renal cortex and outer medulla. Fate-tracing studies have identified forkhead box D1 (FOXD1) expressing cells as renal interstitial cell progenitors, a cell population containing all REPCs. Neural/glial antigen 2 (NG2) expression, a characteristic of renal pericytes, likely defines a subpopulation of FOXD1derived cells and, thus, REPCs. We tested the hypothesis that the REPC population is more heterogeneous than currently characterized by comparing EPO expression in FOXD1 and NG2 cre-reporter mouse models. Crosses with floxed PHD2 lines were also utilized in order to assess PHD isoform contributions to EPO expression in relation to these two REPC populations. Studying EPO expression in these mouse models under normoxic and hypoxic conditions allowed the comparison of REPC subpopulations. Our initial findings, generated using a novel RNA in situ hybridization technique, indicate heterogeneity within the REPC population such that knockout of PHD2, the primary PHD isoform responsible for HIF-2α stabilization, is not sufficient to stimulate EPO production in all REPCs – i.e. another hypoxia-dependent stimulus is required to induce EPO transcription in some REPCs. Oxygen sensing mechanisms must, therefore, differ between REPC subpopulations. These findings are reflected in FOXD1-lineage and potentially in NG2-lineage REPC populations. While previous studies have compared PHD isoform sensitivity in wholekidney homogenizations, how oxygen sensing mechanisms differ has yet to be shown in a spatial context. This study comparing FOXD1-lineage and NG2-lineage REPC subpopulations will provide desired spatial resolution to the REPC population and, by extension, proof-ofprinciple that the REPC population is more heterogeneous than currently characterized. Abstract 14 Sleep Disturbances and Hyperglycemia in Adults with and without Type II Diabetes Ellora Sharma (Northeast Ohio Medical University), Emily A. Knapp, Jonathan C. Jun, MD, Hsin-Chieh Yeh, PhD, Johns Hopkins University School of Medicine Sleep disruptions and disorders are highly prevalent in today’s society and have been linked to diabetes and the metabolic syndrome. Sleeping disturbances, deprivation, and sleep-disordered breathing have been shown to decrease insulin sensitivity by activating the sympathetic nervous system, increasing nonREM sleep duration, and increasing the expression of the enzymes of fatty acid synthesis. The purpose of this study was to characterize diabetic status and sleep issue status in the National Health and Nutrition Examination Survey participants and analyze the 2011-2012 data set to determine if a significant relationship exists between the two statuses. Self-reported diabetic condition and clinically obtained HbA1c levels were used to categorize individuals into 6 diabetic categories ranging from no diabetes to poorly controlled diabetes. Patients were categorized into either a sleep disturbance group, a sleep disorder group, or a no issue group based on their responses to questions in NHANES’ sleep disorders questionnaire. STATA 13 software was used to run logistic regression tests to evaluate the association between diabetic status and sleep issue status. Results showed that having a sleep disturbance was significantly associated with nearly all diabetic statuses and odds ratios for having a sleep disturbance increased with severity of diabetic status. Patients with poorly controlled diabetes were 1.77 times more likely than non-diabetic patients to have a sleep disturbance and 2.60 times more likely to have a sleep disorder. Sleep disturbances were reported in 43.2% of people with poorly controlled diabetes and sleep disorders were reported in 25.7% of people in this category. Having poorly controlled diabetes presented the greatest likelihood of having sleep issues. Patients’ sleeping habits should be an area of additional concern when physicians are treating diabetes and metabolic syndrome components. Abstract 15 T Cell Based Detection of Autoreactivity In Type 1 Diabetes Orly Leiva (Boston University School of Medicine), Russell Eason, Yulan Ai, Jason Gaglia, Joslin Diabetes Center Type 1 diabetes (T1DM) is caused by the autoimmune destruction of the insulin-producing pancreatic beta cells. Currently, assessment of the risk of developing T1DM is done through the detection of autoantibodies against beta cell antigens (βCAs) in the blood; however this approach does not provide a direct insight into the intensity of the autoimmune response against these cells at a given moment. Our lab is currently developing a T cell based assay to better reflect disease activity at the point of testing. In order to produce sufficient quantities of relevant βCAs , we are utilizing the BL21 Star strain of E. coli bacteria. We are currently working on scaling up the synthesis of three proteins: chloramphenicol acetyltransferase (CAT), human chorionic gonadotropin (hCG), and pre-pro-insulin (PPI). CAT will be used as a control in the Tcell assay, while hCG will act as a model protein for testing purposes and PPI is a βCA. We transfected the bacteria with a plasmid containing: a T7 promoter, the sequence for our desired protein (CAT, hCG, PPI), a lumio-tag for inducible fluorescence, a His-tag for purification and finally a ampicillin (AMP) resistance gene for selection. The lumio-tag consists of the peptide sequence, Cys-Cys-Pro-Gly-Cys-Cys, which binds to a biarsenical detection reagent allowing visualization of the protein. Cells were transfected with the appropriate plasmid and grown overnight in 50 mL of LB media with 1% AMP selection. The culture was then transferred to 1 L of 2YT media with 1% AMP and was grown for approximately 2-3 hours to an OD 600 of 0.5-0.8. PPI cultures were also supplemented with 1% glucose. To induce expression of the desired protein, 0.5 mM (1 mM for PPI) isopropyl β-D-1-thiogalactopyranoside (IPTG) was used. Protein expression resulted in inclusion body aggregates, which were then purified before purifying the protein itself. The inclusion bodies were solubilized in 6 M guanidine-HCl solution and ran through a column containing a Cobalt chelate resin. The presence of a His-tag on the denatured proteins specifically allows for binding to the resin and the removal irrelevant proteins. Proteins were eluted with a series of elution buffers with decreasing pH. Proteins were then tested for endotoxin contamination. Protein production via this method will prove to be valuable in order to acquire the protein needed for the new T cell assay the lab is developing to better test for autoreactivity for T1DM. Abstract 16 Short-term DPP4 inhibition improves glucose tolerance, reduces an inflammatory marker and liver enzyme levels in HIV+ adults with glucose intolerance. Paige Kuhlmann (University of Missouri School of Medicine), Heidi Struthers, Erin Laciny, Michael Royal, Dominic Reeds, Kevin Yarasheski, Washington University in St. Louis Introduction & Background. People infected with HIV are at higher risk for developing metabolic complications such as insulin resistance, diabetes, and cardiovascular disease than the general population. Chronic low-grade inflammation and treatment with highly active antiretroviral therapy (HAART) contribute to the metabolic complications. Dipeptidyl peptidase-4 inhibitors (DPP4i) are glucose-lowering medications with potential anti-inflammatory and pleiotropic actions that may benefit HIV+ people, but these beneficial actions have not been tested in HIV+ adults with glucose intolerance. Hypothesis: In HIV+ adults with glucose intolerance, DPP4i will improve indexes of glycemic control and ß-cell function, and reduce the concentration of circulating inflammatory markers typically elevated in HIV+ men and women (sCD14, CXCL10). Methods. We conducted a randomized, placebo-controlled, double-blind 8week trial of the DPP4 inhibitor sitagliptin (100mg/d) in HIV+ adults with glucose intolerance. We measured oral glucose tolerance at baseline and week 8, and plasma markers of inflammation (CXCL10), immune activation (sCD14), lipids/lipoproteins, and liver enzyme levels at baseline, week 4 and 8 in placebo (n=12) and sitagliptin (n=16) recipients. Results. In comparison to baseline, sitagliptin reduced plasma glucose concentrations during the oGTT (P< 0.03). Oral glucose insulin sensitivity index (OGIS) improved in the sitagliptin group after 8 wk (P=0.029), and there was a trend for this improvement to be greater than the change in the placebo group (P=0.072). Other glucose-insulin indexes (HOMA-IR, HOMA-ß, IGI) were unchanged after treatment in both groups. Plasma CXCL10 concentrations declined more in the sitagliptin group than in placebo (p<0.007). Plasma sCD14 concentrations did not change significantly after treatment in either group. Liver enzyme levels (SGOT, SGPT) were reduced more in the sitagliptin group than in the placebo group after 8 weeks (P<0.005). No within or between group differences were observed for fasting serum triglycerides, total-, LDL-, or HDL-cholesterol levels. Conclusion. In HIV+ men and women, sitagliptin (100 mg/d for 8 wk) improved glucose tolerance, apparently by enhancing insulin sensitivity. Sitagliptin appeared to induce other effects that might benefit HIV+ individuals with cardiometabolic complications; it reduced liver enzyme levels, suggesting a beneficial effect on liver health, and it reduced a circulating marker of inflammation. Abstract 17 Use of Magnetic Resonance Spectroscopy to Measure Intrahepatic Lipid and Percentage Liver Fat in Nonobese Adolescents with Polycystic Ovary Syndrome Alexandra N. Linder (Tulane University School of Medicine), Dr. Sharon E. Oberfield, Tamara Cameo, Donald J. McMahon, Dr. Aviva B. Sopher. Division of Pediatric Endocrinology, Columbia University Medical Center, New York, NY. Polycystic ovary syndrome (PCOS) is a common endocrine disorder that affects adolescents and women of reproductive age. Insulin resistance is thought to be an underlying cause of PCOS. A combination of genetic predisposition, insulin resistance and elevated androgens seen in this population may lead to development of non-alcoholic fatty liver disease (NAFLD), which can be considered the hepatic manifestation of the metabolic syndrome. Magnetic resonance spectroscopy (MRS) is a non-invasive tool for evaluating the presence of NAFLD by quantifying intrahepatic lipid measurement (IHL) and highly correlates with the gold standard, liver biopsy. There have been several studies using MRS in obese adults with PCOS for evaluation of NAFLD but this method has not yet been used to evaluate non-obese adolescents with PCOS for NAFLD. We hypothesize that non-obese adolescents with PCOS have significantly higher percentage liver fat compared to age-, BMIz-, and ethnicity comparable- controls, and that percentage liver fat in PCOS is correlated with metabolic abnormalities including insulin resistance and dyslipidemia. We studied 34 non-obese adolescent girls between ages 13 and 21 (16 PCOS and 18 controls), with: physical exam with Ferriman-Gallwey hirsutism score, fasting blood tests for reproductive and adrenal hormones and lipids, two hour oral glucose tolerance test, whole body DXA for percentage body fat, and MRS for IHL. Percent liver fat was calculated from IHL data obtained by MRS. As expected, the PCOS group had significantly higher Ferriman-Gallwey scores, and in a menarcheal age adjusted analysis had higher T, free T, DHEAS, androstenedione, and FAI, and lower estradiol and SHBG than controls. Percentage liver fat was not significantly higher in PCOS compared to controls, nor was incidence of NAFLD. There was a statistical trend in the PCOS group for correlation between percentage liver fat and triglycerides (r=0.48, P=0.057) and percent body fat (r=0.49, P=0.057). There was a negative statistical trend in the control group for correlation between percentage liver fat and LDL (r=0.41, P=0.09), HOMA (r=-0.38, P=0.12), and percent body fat (r=-0.39, P=0.11). Although this small study did not find any significant differences in liver fat, the metabolic abnormalities, including insulin resistance and elevated triglycerides, found in adolescents with PCOS suggest that this population may be at risk of later developing other components of the metabolic syndrome, including NAFLD. These findings suggest that non-obese adolescents with PCOS should be monitored closely and that interventions with insulin sensitizing drugs such as metformin may be considered to prevent metabolic abnormalities and future cardiovascular disease and type 2 diabetes mellitus. Abstract 18 Polymorphisms in three genes may contribute to the expression of hypogonadotropic hypogonadism. Megan Foggia1, Cuiqi Zhou2, Ahmed Khattab3, Maria New3, Shlomo Melmed2 1 University of Nevada School of Medicine, Reno, NV 2Cedars-Sinai Medical Center, Los Angeles, CA, 3Mount Sinai Hospital, New York, NY Isolated hypgonadotropic hypogonadism (IHH) is a condition which occurs in the absence of gonadotropin-releasing hormone (GnRH) secretion from the hypothalamus, or a failure of the gonadotropins to act at the level of the testes and ovaries. Many cases of IHH have been linked to genetic polymorphisms in genes such as the GnRH receptor gene, KAL1, KISS1, and PROK2. In addition, there are a number of genes associated with the hypothalamic-pituitary axis which have not been extensively studied, but may play a role in the expression of IHH. This study focuses on a male patient who was diagnosed with IHH of unknown cause. Whole exon sequencing of the patient’s genomic DNA revealed more than 600 single-nucleotide polymorphisms (SNPs) in more than 70 different genes. Nine of these SNPs were selected for further validation based on their location in coding regions of genes associated with the hypothalamic-pituitary axis and the regulation of GnRH. The SNPs were confirmed via polymerase chain reaction, and were further validated via sequencing. After validation, the SNPs were evaluated, revealing that four out of the nine polymorphisms were uncommon amongst populations in the NCBI database. The four uncommon SNPs occurred in the following genes: TAF7L, MAMLD1, and NR0B1. These SNPs in TAF7L, MAMLD, and/or NR0B1 may be linked, directly or indirectly, to the expression of IHH in this patient. Further studies on the mutant proteins resulting from variations in TAF7L, MAMLD, and NR0B1 are needed to evaluate whether or not polymorphisms in these genes are associated with IHH. Abstract 19 Nutrition, Inflammation, and Insulin Resistance in End Stage Renal Disease Sthuthi David (University of Rochester School of Medicine and Dentistry), Serpil M. Deger MD, Gerald B. Denny MD, Feng Sha PHD, Charles D. Ellis PHD, Alp Ikizler MD, Vanderbilt University Medical Center. Patients receiving chronic hemodialysis (CHD) exhibit a metabolic derangement characterized by inappropriate loss of skeletal muscle mass, termed protein energy wasting (PEW). CHD patients who exhibit PEW have higher mortality and hospitalization rates. The mechanism of PEW is complex but may be related to insulin resistance and chronic inflammation. We hypothesize that there is a direct correlation between glucose and amino acid metabolism, related to diminished insulin action seen in the maintenance hemodialysis population. To test this hypothesis, we utilized the hyperinsulinemic euglycemic and euaminoacedemic dual clamp. So far, we have done this study on seven hemodialysis and five control patients. Within two weeks of taking part in the metabolic clamp study, subjects underwent a dual-energy x-ray absorptiometry (DEXA). Patients were also asked to eat based on current dietary recommendations during the two weeks before the study. At the beginning of the dual clamp study, baseline blood samples were drawn and patients received a bolus and continuous infusion of labeled leucine, phenylalanine, and glucose. After the equilibration period of two hours, subjects received an insulin infusion of 60 mU/m2/min and baseline glucose levels were maintained by infusions of dextrose. In the second study, a prime constant infusion of 10% balanced amino acid solution was begun to increase plasma leucine levels. Insulin sensitivity was assessed by the glucose disposal rate (GDR) derived from HEGC. Median GDR level was 4.9 for hemodialysis patients and 4.4 mg/kg/min for control patients (p=.905). Amino acid infusion induced a significant rise in GDR in hemodialysis patients. GDR was increased in controls but did not reach statistical significance. Leucine disposal rate (LDR) was similar in both groups and averaged .68 ±.23 and .44 ± .47 mg/kg/min in hemodialysis and control subjects, respectively (p=.268). There was a strong positive correlation between LDR and GDR (r=.883, p=.002). In conclusion, the results of this study indicate that the severity of insulin resistance to carbohydrate metabolism is associated with a similar resistance to anabolic actions insulin has on protein metabolism in CHD patients. This implies that insulin resistance may represent a potential target for prevention or treatment of PEW in CHD patients. Abstract 20 The Use of Electronic Communication Media and Evaluation of Living Kidney Donors Mari Tanaka (Georgetown University School of Medicine), Anthony Dreher, Powell Newbern, Irene Feurer PhD, and Heidi Schaefer MD, Vanderbilt University Medical Center A proposed reason why it has been difficult to increase the number of living kidney donors is that many potential donors lead busy lives and find the pre-donation process too lengthy and burdensome. Most transplant centers initially screen donors using phone or written surveys, which limit the time during which self-referring donors can initiate the process. With this in mind, Vanderbilt University Medical Center implemented a web-based application in 2011 where potential donors could privately initiate the process and proceed through the initial screen anytime from the comfort of their home and/or office resulting in an increase in the number of self-referrals. To test the hypothesis whether or not patients who interface through electronic tools with the transplant center have expedited living donor evaluations, surgeries, and increased conversion rate we conducted a single center retrospective study using patients who interfaced with the Vanderbilt transplant center between January 2011 – March 2014. Data were analyzed using t-tests, nonparametric tests of proportions, Kaplan-Meier survival, and multivariable Cox proportional hazards regression methods. Our results indicate that patients who contacted via the web are more likely to have a graduate degree and to live more than 400 miles away from the transplant center. There were no significant associations between contact methods and time to evaluation, decision regarding candidacy, or donation surgery between groups. Lastly, those who contacted via the phone were more likely to be turned down for donation while those who contacted via the web were more likely to decide not to donate. These findings suggest that the web-based application increases the overall number of living donor surgeries performed but has no effect on the times to evaluation, eligibility decision, or surgery. Future efforts should be aimed at those patients who decide not to donate and reasons why to determine if modifiable factors are present that could be addressed by the living donor program in hopes of further increasing the number of eligible living donors. Abstract 21 Myeloid lineage-specific overexpression of P110α induces vascular lesions in mice Jeremy Brooksbank, Vladimir R. Babaev, PhD, Sergey V. Ryzhov, PhD, Lei Ding, Youmin Zhang, and MacRae F. Linton, MD Phosphoinositide 3-kinases (PI3Ks) signaling is downstream of multiple cell-surface receptors that regulate cell proliferation, survival, and death. Activation of receptor tyrosine kinases and growth factors induces PI3Ks activity with formation of heterodimers of class Ia lipid kinases, composed of the P110 (catalytic) and p85 (regulatory) subunits. These heterodimers propagate secondary messenger signaling by phosphorylating phosphatidylinositols (PIP2 to PIP3) and initiating Akt signaling. Due to the association between P110α and pathologic angiogenesis, it is believed to be a promising drug target. In order to examine the role of PI3Ks in macrophage functions and atherogenesis, we recently generated mice with conditional myeloid lineage-specific overexpression of P110α (MS-P110a+) induced by the LysM Cre promoter. These mice exhibit a striking phenotype with massive venous enlargement in the extremities. Here we demonstrate that these MS-P110a+ mice develop venous enlargement in the tail, feet, and ears progressively over time with frequent hemorrhage of the varicose veins of the tail at 4-5 months of age. The endothelial cell lining of the dilated veins is preserved as demonstrated by staining with antibodies (Abs) to vonWillebrand factor and pan-endothelial cell antigen. Venous dilation and remodeling with increased vascular permeability promoted extravasation of erythrocytes into the interstitial space. With the phagocytosis of RBCs by tissue macrophages, Fe and hemosiderin were deposited around the lateral veins in the tail. This iron accumulation and enlargement of peripheral veins is consistent with chronic venous diseases in humans such as varicose veins. In addition to peripheral enlargement of veins, MS-P110α mice exhibit marked splenomegaly with effacement of normal architecture, increased granulocytes, and decreased numbers of Bcells. Macrophages isolated from MS-P110α+ mice constitutively express increased activity of Akt with a compensatory increase in the levels of phosphatase and tensin homolog (PTEN), which is an inhibitor of PIP3 and Akt and downstream signaling pathways. RT-PCR of gene expression in MS-P110α+ macrophages shows decreased levels of pro-inflammatory cytokines like TNF-α and IL-12 with elevated anti-inflammatory cytokines like IL-10, indicating that these macrophages are of the M2 phenotype. In addition, we found increased levels of matrixmetalloproteinase-2 (MMP-2) and inducible-nitric oxide synthase (iNOS) in macrophages, which may be critical for the pathogenesis of these vascular lesions. Finally, we used bone marrow transplantation (BMT) to examine the hypothesis that the myeloid-specific overexpression of P110α is sufficient to recapitulate the vascular phenotype. Examination of these mice at 4 and 8 weeks post-BMT revealed the presence of similar vascular lesions in both tail and spleen to the original MS-P110a+ mice. These results strongly support the hypothesis that myeloid specific overexpression of P110a is sufficient to induce the venous malformations and highlight a previously unappreciated role of macrophages in vascular remodeling. Thus, we have developed a new murine mouse model for chronic venous disease that should allow the identification of novel targets for pharmacologic intervention to slow or prevent the development of varicose veins. Abstract 22 Identification of BBMP-3 as a betatrophin binding membrane protein Poy Theprungsirikul (Geisel School of Medicine at Dartmouth), Qian Huang, Ph.D., and Peng Yi, Ph.D., Joslin Diabetes Center-Affiliate of Harvard Medical School Replenishing the insulin-producing β-cell mass is considered to be a potential cure for diabetes. One possible avenue to replenish β-cell mass is via replication of pre-existing β-cells in the pancreas. Betatrophin, a secreted protein in liver and fat that belongs to the angiopoietin-like protein family, has been found to cause an increase in pancreatic β-cell replication and improve glucose tolerance in mouse models. However, the mechanism of action of betatrophin is not yet fully understood. The goal of this study was to identify betatrophin binding protein(s)/receptor(s) in hope to understand the mechanism behind how betatrophin induces pancreatic β-cell proliferation and β-cell mass expansion. To identify betatrophin binding protein(s), we prepared cDNA expression libraries from mouse liver and fat, and screened the libraries with an alkaline phosphatase-mouse betatrophin (AP-mBT) fusion protein. We identified Betatrophin Binding Membrane Protein (BBMP-3) in the mouse liver. The identified BBMP-3 is a major subunit of a single transmembrane glycoprotein receptor that is expressed primarily in liver. It plays a role in serum glycoprotein homeostasis by mediating the endocytosis and lysosomal degradation of glycoproteins with exposed terminal galactose or N-acetylgalactosamine residues. We speculate that BBMP-3 may play a role in regulating serum betatrophin level. The identification of BBMP-3 as a betatrophin binding membrane protein may have important implications for the development of betatrophin receptor modulators for treating both type I and type II diabetes. Abstract 23 The Prevalence of Copper Deficiency before and after Bariatric Weight Loss Surgery Jordan Martinez (Jefferson Medical College), John LaBarra, Joe Boullata, & Charlene Compher, The University of Pennsylvania Bariatric surgery is one of the only current, long term treatment options used to increase the quality of life within the obese population. This surgical option allows patients to attain considerable weight loss and also helps improve several comorbidities associated with obesity such as diabetes, high blood pressure and cholesterol, and arthritis. Even though it is very clear that there are many benefits associated with bariatric surgery, these operations also have quite a few potential complications. Particularly, patients who have received bariatric surgery have shown to be at risk for a variety of important nutritional deficiencies. Serum copper deficiency is one of the nutritional deficiencies that has been reported in the literature within the postoperative obese population. The purpose of this clinical study is to understand how serum copper levels change within patients who undergo bariatric surgery. We want to know the prevalence of being copper deficient both before and after surgery. By studying new patients and reviewing the current literature, the revision of current recommendations and treatment options can be prepared and any uncertainties in the relevant literature can be identified and serve as targets for future research pertaining to bariatric patients and their serum copper levels. The ultimate goal for this clinical study is to be able to help these bariatric patients attain the highest quality of life by ensuring they receive the necessary copper supplementation that is needed to prevent deficiency and any associated symptoms. Abstract 24 CRISPR-Cas9 mediated MMP10 deficiency may impact macrophage polarization in THP-1 cells. Natasha Choudhury (Meharry Medical College, School of Medicine), Barbara Fingleton, Vanderbilt University MMP10 (matrix metalloproteinase 10 or stromelysin-2) deficiency has been implicated in depressed resolution of colonic inflammation and increased risk of colitis-associated cancer development. In a mouse model of colitis, MMP10 expression was shown to be predominant in macrophages. The phenotype of MMP10-null mice in this model was deficient in macrophages over time, with a skewing toward the M1 (classically activated) macrophage profile. This may suggest that macrophages may be required for wound healing in models of colonic damage; and M2 (alternatively activated) macrophages may be protective against colitis. To test the hypothesis that MMP10 deficiency in undifferentiated monocytes results in a skewing toward the M1 profile and significant decrease in development toward the M2 profile during polarization; we utilized the CRISPR-Cas9 system, an RNA-guided targeted genome editing technology, to induce an MMP10-null line of THP-1 monocytic cells and characterize their polarization. First, we used a standard monocyte/macrophage cell line, Raw 264.7 murine cells, to serve as a control for analysis of MMP10 expression and polarization ability. Next, we analyzed THP-1 monocytic cells for base levels of MMP10 expression and polarization ability. Transfection of THP-1 cells was achieved by addition of plasmid DNA-lipid complexes (MMP10-Cas9-GFP plasmid; Lipofectamine 2000 Reagent) to cells; GFP expression was used to assess transfection efficiency. Finally, transfected cells were serially diluted in order to establish clonal cells suitable for analysis of MMP10 expression and polarization ability. MMP10 expression was shown to be present in both Raw 264.7 and THP-1 cells; polarization ability in Raw 264.7 cells yielded predictable results. The CRISPR-Cas9 system proved to be simple, rapid, and efficient; and yielded high transfection efficiency (~85-90%) in THP-1 cells. However, analysis of MMP10 expression and polarization ability of transfected THP-1 cells depends on further expansion of clonal cells. Therefore, the relationship between MMP10 and macrophage polarization in models of colitis has yet to be clarified further. Abstract 25 KrasG12D mutation in pancreatic duct cells results in the development of pancreatic ductal adenocarcinoma precursor lesions Jessica N. Kim (University of Nevada School of Medicine), Fong C. Pan, R. Daniel Beauchamp, Christopher V. Wright, Anna L. Means, Vanderbilt University Medical Center 90% of human pancreatic carcinomas have a Kras mutation; murine models of pancreatic ductal adenocarcinoma (PDAC) have demonstrated that Kras mutation and subsequent inflammation result in the development of pancreatic cancer precursors (PanINs). While PanINs and PDAC have ductal characteristics, acinar cells can convert to duct cells following an inflammatory event, making it unclear whether acinar or duct cells give rise to pancreatic cancer. We hypothesize that Kras mutation in duct cells results in the development of PanIN lesions in the setting of clinically relevant inflammation. Mice with Kras mutation in either duct or acinar cells underwent partial duct obstruction via partial ligation of the main pancreatic duct, mimicking a type of damage observed in humans, and analyzed at different time points. When Kras mutation was introduced into duct cells, ducts gave rise to dysplastic lesions but when Kras mutation was introduced into acinar cells, the tissue resembled wild-type pancreas with most KrasG12D cells lost by 4 months post-ligation. We found that Kras mutation supplies a selective advantage when introduced to duct cells in the setting of inflammation as early as 2 weeks after injury: Kras mutant cells were enriched in damaged parts of the pancreas compared to undamaged parts. To understand this selective advantage, double IF studies with PHH3 (a marker of proliferation), histone γH2AX (a marker of double strand DNA breaks), and cleaved caspase-3 showed that a survival, rather than proliferative, advantage is conferred to duct cells with KrasG12D. Finally, we found that Kras mutation in duct but not acinar cells maintained a fibrotic microenvironment similar to that found in human PanINs, confirming the ductal origin of these precancerous lesions. In summary, KrasG12D in duct cells provides these cells a survival advantage, resulting in the development of PanIN lesions and the maintenance of a fibrotic tumor microenvironment. These observations have implications for the pathway in which Kras mutant cells survive, potentially revealing possible drug targets and diagnostic biomarkers of PDAC. Abstract 26 Women with the Comorbid Condition of Obesity and Depression Have Improved Body Composition, Eating Behaviors, Inflammation and Insulin Resistance Upon Consuming a Balanced High Fat Diet María E. Mulero Morales, Ponce School of Medicine & Health Sciences; Heidi J. Silver, Ph.D., R.D., Vanderbilt University School of Medicine Background: Obesity and depression are comorbid conditions; obese adults are 55% more likely than normal weight adults to become depressed and depressed adults are 58% more likely to become obese. This bidirectional relationship is influenced by sex, age, dietary behaviors, body size, body image, inflammation, and insulin resistance. Moreover, both conditions are associated with increased risk for type 2 diabetes and cardiovascular disease. Hypothesis: We hypothesized that consuming a balanced high fat diet, previously shown to decrease weight, body fat, and retain lean body mass in obese premenopausal women, would improve eating behaviors, inflammation, oxidative stress, insulin resistance, and thereby, promote decreased prevalence of depression and improved quality of life. Methods: 144 premenopausal women aged 21-50 with Class I/II obesity (BMI 30-39.9 kg/m2) were enrolled in a 16-week higher fat diet intervention where type of fat was balanced as 1/3 saturated, 1/3 monounsaturated, 1/3 polyunsaturated. History of depression was self-reported and verified by review of medical record diagnosis and/or anti-depressive medication prescription. Dietary intake was assessed by computerized 24-hour recall interviews at random time-points using NDSR software. Body composition was assessed by DEXA. Eating behavior was assessed using the Three Factor Eating Questionnaire (TFEQ) and the Eating Attitudes Test (EAT-26). Inflammatory markers (hsCRP, PAI-1 antigen, pro-inflammatory cytokines), fasting glucose and fasting insulin were assayed at the Vanderbilt Pathology Lab. Urinary F2-Isoprostane metabolite was assayed at the Roberts Lab. Insulin resistance was estimated by calculating HOMAIR score. Quality of life was assessed using the Impact of Weight on Quality of Life-Lite (IWQOL-Lite) questionnaire. Results: At baseline, 63 (46.1%) of subjects had a history of depression. At week 16, women with depression had significant improvements in weight, BMI, VAT, TFEQ scores, EAT-26 oral control, pro-inflammatory cytokines (IFNγ, IL-1α, IL-6), F2-Isops, glucose, insulin, HOMAIR score, and quality of life (all ps < 0.05). In addition, women with depression had significantly greater improvements compared to non-depressed women in fasting glucose (p = 0.04), IL-6 (p = 0.06), TFEQ disinhibition (p = 0.05) and TFEQ total score (p = 0.007). Overall, prevalence of depression decreased from baseline to week 16 by 6.1% (p = 0.001). Conclusions: Effective lifestyle interventions for obese adults with depression are much needed. In this cohort of obese women, both body weight and body composition improved substantially upon consuming a higher fat diet where the type of fat was balanced. In addition, depressed women had significant improvements in eating behaviors, inflammatory markers, insulin resistance, and quality of life. Moreover, the balanced high fat diet appeared to have a significant impact on the prevalence of depression. Future work with dietary intervention is needed using a standardized method for determining a meaningful change in depressive symptoms and remission rate. Abstract 27 Quantification of urinary exosomal NPT2a co-transporter expression may serve as a marker for phosphate reabsorption Si Liao (New York Medical College), Taopheeq Ayodele Mustapha, James Matthew Luther, Vanderbilt University Enrichment of proteins isolated from urinary exosomes may offer insight into the pathophysiological state of renal tubular cells and the potential for non-invasive detection, classification, and monitoring of renal disease. We have previously identified proteins, including NPT2a (type IIa sodium-dependent phosphate co-transporter), TRPV5 (epithelial calcium channel), and klotho in urinary exosomes via Multidimensional Protein Identification Technology (MudPIT). These proteins play an essential role in calcium and phosphate handling. Phosphate handling is impaired in chronic kidney disease (CKD) where increased serum phosphate levels is associated with increased cardiovascular mortality and decreased in kidney transplant patients who experience phosphate wasting. Renal phosphate reabsorption is primarily a function of hormonal regulation of NPT2a expression on the apical membrane of proximal tubule cells, and has been shown to increase with insulin. Therefore, we hypothesized that insulin increases renal phosphate reabsorption and expression of NPT2a in urinary exosomes. To validate exosomal NPT2a as a marker for phosphate reabsorption, differential ultracentrifugation was performed to isolate exosomes from human urine samples collected before and after high-dose hyperinsulinemic clamps (120 mU/m2/min), followed by an in-solution trypsin digest in preparation for multiple reaction monitoring (MRM) coupled with stable isotope dilution mass spectrometry. The tubular reabsorption of phosphate (TRP) was calculated from plasma and urine concentrations of phosphate and creatinine before and after insulin infusion in seven subjects. Mean plasma phosphate concentration decreased from 1.62±0.057 mM preinsulin infusion to 1.39±0.050 mM post-insulin infusion (P<0.001). Mean TRP increased from 89.86±6.40% pre-insulin infusion to 91.61±6.70% post-insulin infusion (p=0.48). Trypsin digests of urinary exosomes have been prepared and are awaiting analysis for NPT2a peptide fragments. NPT2a expression in urinary exosomes may allow assessment of novel treatment of renal disease, including phosphate retention in CKD and phosphate wasting in kidney transplant patients. Abstract 28 The Effects of Low Dose Metformin on Hepatic Glucose Production Gaurav Pahouja (Northeast Ohio Medical University), Sara M. McMillin, Fredric E. Wondisford, Johns Hopkins University School of Medicine One of the leading chronic illnesses in the United States is type 2 diabetes, afflicting nearly 1 in every 10 people. Metformin is considered the first line therapy and standard of care for type 2 diabetics. However, the exact mechanism by which metformin lowers blood glucose values is still under debate. To date, the majority of rodent studies have utilized high doses of metformin given in a manner (i.e. intravenous injection) that does not accurately represent the normal oral delivery of metformin in patients. Therefore, we set out to investigate the effects of low dose metformin given orally to better reflect the patient model and normal metabolism of the drug. Mice that contained a floxed version of the gene for the PKA regulatory subunit R1a were treated with adenovirus coding for CRE recombinase to knockout the R1a regulatory subunit in the hepatocytes and achieve constitutive activity of PKA. Subsequently, a group of the Ad-CRE treated mice were given a low dose, 12.5 mg/kg/day, of metformin in the drinking water. When compared to mice given Ad-CRE alone, the mice that received Ad-CRE and metformin showed significantly lower blood glucose levels. In addition, a pyruvate challenge test also showed a significantly lower rise in blood glucose values in the mice treated with metformin as compared to the mice receiving only the virus. Finally, the activation of AMPK, tracked by the presence of Phospho-AMPKα (Thr 172), increased in the hepatocytes of the mice treated with metformin but did not increase in the skeletal muscle. These results indicate that metformin was active in the liver, even at such a low dose. A larger scale study is currently underway to confirm these novel findings. Abstract 29 Effects of Maternal Obesity during Pregnancy on Insulin Signaling in Human Umbilical Vein Endothelial Cells Kacie Fox, John A. Burns School of Medicine- University of Hawaii, Elvira Isganaitis, Joslin Diabetes Center There is now a substantial body of evidence linking maternal obesity during pregnancy and gestational diabetes to metabolic dysfunction in offspring. It is likely that in addition to genetics and the postnatal environment, the intrauterine environment programs metabolic activity in offspring tissues, resulting in increased risk for obesity and type 2 diabetes. However, the mechanisms are still not well understood. We hypothesized that maternal obesity alters insulin signaling in fetal cells. To test this hypothesis, we obtained umbilical cords from a cohort of 35 women across the BMI spectrum undergoing scheduled C-sections at Brigham and Women’s Hospital. Human Umbilical Vein Endothelial Cells (HUVEC) were isolated and grouped according to maternal weight status: lean – BMI < 25, overweight – BMI 25-30, and obese – BMI > 30. Primary HUVEC were stimulated with insulin (1 and 10 nmol) before extracting protein, RNA & DNA. We examined the expression and phosphorylation of IR, IRS1, AKT, ERK, and FoxO1 proteins by Western blotting. Our preliminary analyses indicate that these proteins respond to insulin in a dose responsive fashion and that protein expression of phosphorylated IRS-1 is increased in fetal cells exposed to maternal obesity. Examination of transcriptional responses to insulin is underway. Our data will help advance the understanding of mechanisms by which maternal weight status during pregnancy influences risk of metabolic dysfunction in offspring. Abstract 30 Patients’ Satisfaction with a Medication Adherence Support Website: Diabetes MAP Sujata Ghosh, MPH (Texas A&M Health Science Center), Magaela C. Bethune, MPA, Chandra Y. Osborn, PhD, MPH Vanderbilt University Among adults with type 2 diabetes mellitus (T2DM), nonadherence to medications is common, and is associated with worse glycemic control, hospitalizations, and pre-mature death. Effective medication adherence promotion interventions have not been feasible and sustainable in busy clinic settings. Web-delivered interventions are a viable alternative. However, their effectiveness depends on patient use, which is determined by many factors, including user satisfaction. We conducted a mixed methods study to understand patients’ satisfaction with a medication adherence promotion intervention website called Diabetes MAP. We recruited adult patients with T2DM who were prescribed diabetes medications (N=32) from an academic medical center for a three-part usability evaluation. Participants completed an initial survey (N=32), were instructed to independently use Diabetes MAP for two weeks (n=29), complete a follow-up survey, and participate in a focus group session (n=27). Positive comments from focus group transcripts were extracted, coded, and analyzed using thematic analysis. Survey data were analyzed descriptively, and linked to transcripts to characterize qualitative data. Most participants reported being satisfied with using Diabetes MAP in the follow-up survey (69%) and in the focus group session (66%). Focus group participants (35%) expressed the most satisfaction with viewing videos that addressed user-specific barriers to medication adherence. Younger participants (45.3 ± 9.7 vs. 54.2 ± 22) were more satisfied with using dose and refill reminder text messages than older participants, whereas older participants (56.0 ± 8 vs. 49.9 ± 24) were more satisfied with reading about their medications (e.g., drug interactions) than younger participants. Patients were satisfied with Diabetes MAP overall, and satisfaction with the site’s features varied by age and duration of diabetes. These findings are limited by our mixed methods approach, specific population, and single website. Satisfaction with a web-delivered diabetes intervention is audience dependent, and user age and diabetes duration may be important predictors of website satisfaction among adults with diabetes. Abstract 31 Helicobacter pylori strains from high and low risk cancer regions of Colombia do not differentially induce nitric oxide production from RAW 264.7 cells Won Seok Shin (Eastern Virginia Medical School), Dana M. Hardbower, Keith T. Wilson, Vanderbilt University Medical Center Helicobacter pylori is a Gram negative, microaerophilic bacterium that infects an estimated 50% of the global population. H. pylori infection is known to be the principal risk factor for gastric cancer, the third leading cause of cancer-related deaths. H. pylori is able to evade the host immune system and persist over the lifetime of the host. It indirectly attenuates nitric oxide (NO) production, by the enzyme inducible nitric oxide synthase (iNOS), such that effective clearance of pathogen is thwarted. Epidemiological studies have shown that two distinct geographical regions in Colombia exhibit vastly different gastric cancer prevalences. Individuals from the high risk region of Colombia are typically infected with more highly virulent strains of H. pylori than individuals in the low risk regions. We chose one prototypical strain from the low and high risk regions, termed 5009 and 5056 respectively, for our studies. Our lab has previously demonstrated that 5056, the high risk strain, induces more DNA damage in infected cells than 5009, the low risk strain. Since H. pylori-mediated gastric carcinogenesis requires both DNA damage and chronic inflammation, we hypothesize that 5009 and 5056 also differ in their ability to induce NO production. To test this hypothesis, we infected RAW 264.7 cells, a murine, macrophage cell line with 5009, 5056 and PMSS1, a laboratory reference strain, for 6 or 24 h. Experiments were performed with and without the use of Transwell filter supports in order to assess the affects of both direct contact between the bacterium and the cell and the effect of soluble H. pylori-produced factors. We assessed iNOS mRNA expression using quantitative real-time PCR after 6 h, and measured NO production using a Griess Assay after 24 h. When the RAW 264.7 cells and the H. pylori strains were in direct contact, no significant differences in iNOS mRNA expression and NO production were observed. Infection with 5009 resulted in a 702-fold increase in iNOS expression, while 5056 led to similar levels of iNOS expression with a 725-fold increase (n=1) in iNOS expression. 5009 induced 11.41 ± 3.043 mM NO2-, while 5056 induced 16.35 ± 1.891 mM NO2- (n = 3). Likewise, both strains induced similar levels of iNOS expression and NO production when separated from the cells by Transwell filter supports. Infection with 5009 induced 388.1 ± 151.1-fold increase in iNOS expression (n = 4). Similarly, infection with 5056 induced 355.4 ± 175.7-fold increase in iNOS expression (n = 4). RAW 264.7 cells infected with 5009 and 5056 produced 8.040 ± 5.791 mM NO2- and 9.654 ± 6.109 mM NO2(n = 3), respectively. Again, no significant differences were observed in iNOS mRNA expression and NO production after infection with 5009 and 5056. These data indicate that the high versus low risk status of different H. pylori strains does not affect NO production by macrophages. However, there are many other immunological functions that macrophages utilize to combat H. pylori infection. Other parameters, such as phagocytosis, anti-microbial peptide production and cytokine/chemokine expression should be assessed in future experiments. Strains from high and low risk regions of Colombia may yet differentially regulate one or more of the aforementioned macrophage functions. Abstract 32 The Role of Hepatic Estrogen Receptor in the Regulation of Glucose Metabolism Vanessa Roos (Medical University of South Carolina), Andrew Wolfe, Johns Hopkins University School of Medicine Reproductive function has long been known to be sensitive to changes in metabolic status with states of negative energy balance associated with decreased reproductive function. It is becoming increasingly clear that states of excess energy can also contribute to reproductive dysfunction. Less understood is how reproductive function can influence metabolism. We proposed that gonadal steroid hormones may directly regulate metabolic function. Since the liver plays an essential role in regulating energy metabolism and expresses the estrogen receptor, we hypothesized that estrogen action via the ERα (ESR1) in the liver regulates glucose metabolism. To test this hypothesis, floxed Esr1 (flEsr1) mice, developed in our laboratory, were injected via tail vein with a CRE expressing adeno-associated virus (AAV8.TBG.PI.-Cre.rBG) to produce a hepatic Esr1 knock out mouse (LEsr1KO mouse). Control mice were injected with a GFP expressing adeno-associate virus (AAV8.TBG.PI.eGFP.WPRE-.bGH). To assess the efficacy of the injection, fluorescent microscopy was used to observe the expression of GFP (green fluorescent protein) in the GFP virus injected mice livers and in control CRE injected mice livers. The results demonstrated wide spread expression of GFP with highest expression near the portal vein. Successful knockout of Esr1 receptor in LEsr1KO mice was assessed using a TaqMan quantitative real-time PCR assay. GAPDH expression was also measured as a reference gene. Glucose tolerance tests were performed on LEsr1KO and control mice using a standard glucometer and levels of insulin 30 minutes post-glucose injection (GSIS) were measured using a Luminex Insulin assay. Glucose tolerance proved to be significantly improved in male CRE injected mice but not female CRE injected mice compared to GFP injected controls. GSIS levels were increased in both male and female CRE injected mice compared to controls, although not significantly. These data suggest that estrogen plays an important role in liver metabolic function and that changes in glucose tolerance during the reproductive cycle may be due to estrogen action in the liver. Abstract 33 Does a High Fat Diet Induce an Inflammatory Response and Accelerated Amyloid Pathology in APP/PS1 Mice? Casey L Freeman1, Fiona E Harrison2, Jennifer M Walker2; 1 University of Kansas School of Medicine 2 Division of Diabetes, Endocrinology & Metabolism, Vanderbilt University Medical Center Obesity has been linked to many chronic diseases; however, the molecular mechanisms of obesity and many of its metabolic consequences are complex and not well understood. Our research investigates the effects of a high fat diet on expression of a number of genes relating to inflammatory response and amyloid neuropathology in a mouse model of Alzheimer’s disease. Our hypothesis is that obesity drives inflammatory response and oxidative stress in the brain, and that these changes will be magnified in the brains of APP/PS1 mice. Wild-type and APP/PS1 littermates were maintained on a high fat diet (HFD: 60% fat from lard) or a low fat diet (LFD: 10% fat from lard) starting at 8 weeks of age. At 8 months of age, a glucose tolerance test was performed and the mice were sacrificed. Specific brain areas (cortex, hippocampus) were excised and used for qPCR for measurements of a number of genes including inflammatory markers, insulin resistance, and Alzheimer’s neuropathology, as well as determination of lipid peroxidation and amyloid beta. HFD led to obesity in wild-type and APP/PS1 mice, but HFD APP/PS1 mice showed significantly impaired glucose clearance as compared to the two LFD groups and the HFD wild-type group. Gene expression for leptin, TNFα, and INSR (insulin receptor) differed between hippocampus and cortex and indicated that both HFD and APP/PS1 genotype modified expression, although not all trends reached significance. More work is needed to confirm trends observed in these data, but this preliminary study does support our hypothesis that APP/PS1 mice may be more affected by HFD, and obesity can drive changes in hippocampal gene expression. The main limitation at this point is in terms of group sizes. In the future, more mice will be added to each group and a diet reversal will be performed to assess the effects on leptin, TNFα, INSR, and several other oxidative stress, inflammatory, insulin resistance, and Alzheimer’s neuropathological markers. Abstract 34 TcBuster transposase aggregation in kidney cells. Laura M. Downes(University of Kentucky College of Medicine) Lauren E. Woodard, and Matthew H. Wilson, Vanderbilt University. Transposons, or “jumping genes”, are transposable elements used for gene therapy. We are investigating TcBuster, a transposon/transposase system in the hAT superfamily whose transposase tends to aggregate in a rodlet shape in kidney cell nuclei. We set up both cell-free and HEK-293 cell assays to probe how the rodlets form over time. Through observation of timelapsed images, we found that aggregation appears to occur additively at both rodlet ends, that the rodlets are very mobile within the cell nucleus, and that the rodlets also tend to clump with other rodlets within the nucleus to create larger structures. From our cell free assay we conclude that in the presence of DNA transposon, the transposase tends to aggregate with the DNA into structures similar to the octameric structure found to form by the hAT transposase Hermes. The polymerized TcBuster transposase appears to be seeded to the surface by the octameric DNA transposase. Our findings suggest that rodlet formation is a visible sign of overproduction inhibition, which is when the transposase fails to transpose when present at a high concentration. Most transposases are subject to overproduction inhibition so it is important to obtain a better understanding of this phenomenon for gene transfer success. Abstract 35 In vitro interferon regulatory factor 3 (IRF3) regulation of thermogenesis. Sophie Leung (Geisel School of Medicine at Dartmouth), Manju Kumari, Evan Rosen, Beth Israel Deaconess Medical Center Regulators of thermogenesis are sought after therapeutic targets for the treatment of type 2 diabetes and obesity due to their potential to increase energy expenditure and decrease adiposity. In preliminary studies in IRF3 knock out mice, we observed increases in energy expenditure, brown adipose tissue, cold tolerance, and thermogenic gene expression. Further, in wild type mice we observed hypophosphorylation of IRF3 (which is inhibitory) at S135 after cold exposure. These together suggest that IRF3 is a negative regulator of thermogenesis. To further characterize IRF3’s regulatory role, we created two IRF3 mutants containing a S171A or S135A substitution thereby eliminating an activational phosphorylation site. In future studies we will overexpress these mutants separately in murine adipocyte and pre-adipocyte cell lines and stromal vascular cells to determine the effect on Ucp1, Pgc1α, and Prdm16 transcription upon cold exposure. In light of our preliminary data, we expect that the mutations will have a disinhibitory effect leading to upregulation of thermogenic gene expression. Abstract 36 Mitochondrial DNA haplogroup is associated with cellular oxidant production and mitochondrial DNA damage. Terrence T. Jones (University of Tennessee Health Science Center), Jamelle A. Brown, Tanja Dudenbostel, David A. Calhoun and Scott W. Ballinger, University Alabama at Birmingham Diabetes affects 29.1 million people or 9.3% of the U.S. population. Interestingly, AfricanAmericans and Hispanics have an increased susceptibility to type-2 diabetes than Caucasian Americans; the mechanisms underlying this disparity are not known. One factor often overlooked is the contribution of mitochondrial genetics to disease susceptibility. Mitochondria are both sources and targets of oxidative stress, which has been shown to contribute to endothelial dysfunction and diabetes. Mitochondrial DNA (mtDNA) damage has also been associated with diabetes. We hypothesize that cells harboring different mitochondrial haplogroups will produce different amounts of oxidants under conditions of stress and will accumulate different levels of mitochondrial DNA damage. Overall these processes may result in differential susceptibility to type-2 diabetes. In this light, we hypothesized that cells with African mtDNA haplogroups (e.g. haplogroup “L”) will generate more oxidants and have more mtDNA damage than those with northern European origins. To test this, human umbilical vein endothelial cells (HUVECs) and lymphocytes from healthy adult controls were haplogrouped, and separated into African or Eurasian cohorts based on restriction fragment length polymorphism (RFLP) analysis of mtDNA PCR products. HUVECs were treated with TNFα, a proinflammatory cytokine that mediates mitochondrial oxidant production, and stained with dihydrorhodamine 123 to observe oxidant production. Basal mtDNA damage was also assessed in both HUVECs and healthy adult lymphocytes. Results show that HUVECs with African mtDNAs generated more oxidants when treated with TNFα compared to HUVECs harboring Eurasian mtDNAs; this oxidant generation also co-localized with mitochondria, suggesting mitochondrial origin. Finally, both HUVEC and lymphocytes having African haplogroup mtDNAs displayed increased mtDNA damage compared to those carrying Eurasian mtDNAs. Consequently, increased basal expression of oxidants and mtDNA damage associated with mtDNA haplogroup may explain the differential susceptibility to diabetes. Abstract 37 rpL13a SnoRNAs Mediate Lipotoxicity in Diabetic Cardiomyopathy Elizabeth K. Asfaw (Charles R. Drew University/David Geffen School of Medicine at UCLA), Chris Holley, Jiyeon Lee, and Jean Schaffer Washington University in St. Louis School of Medicine, St. Louis, MO Introduction: Diabetic patients have an increased risk of heart failure. Pathologically, failing hearts from diabetic patients are characterized by steatosis that is thought to contribute to cardiomyocyte cell death and dysfunction. To model this metabolic cardiomyopathy, the MHCACS (ACS+) mouse was generated by cardiomyocyte overexpression of long-chain acyl-CoA synthetase, which leads to heart failure through lipid-induced oxidative stress and inflammation. Small nucleolar RNAs (snoRNAs) from the rpL13a locus have been shown to be important mediators of lipotoxic and oxidative stress in cell culture models. Mice with loss of function of the rpL13a snoRNAs (sno-/-) have recently been generated to study the in vivo role of these non-coding RNAs, and this model has normal heart function. Methods: To determine if rpL13a snoRNAs mediate cellular damage and contribute to cardiomyopathy in the MHC-ACS cardiomyopathy model, ACS+ and sno-/- mice were crossed. Heart function, gene expression, and histology were assessed in the progeny (ACS+/sno-/-). Results: In ACS+ mice, left ventricular hypertrophy and heart failure are detected as early as 6 weeks of age. Echocardiographic analysis showed left ventricular mass was decreased in the ACS+/sno-/- mice compared to the ACS+ mice at both the 6 and 8-week time points. Fractional shortening in the ACS+/sno-/- mice was also increased compared to the ACS+ mice, suggesting a significant improvement in left ventricular systolic function. To examine mechanisms underlying improved cardiac function in the ACS+/sno-/- mice, gene expression was analyzed for markers of inflammation, heart failure and fibrosis. Cytokines and cellular markers of inflammation were induced in the ACS+ mice at 6 and 8 weeks. A similar pattern was observed ACS+/sno-/- mice, although IL-6 was decreased compared to the ACS+ mice. Compared to ACS+ mice, ANP levels were significantly reduced in both 6 and 8-week ACS+/sno-/- mice, consistent with a decrease in heart failure. Markers of fibrosis (Col1A1, Col3A, CTGF) were induced in ACS+ and ACS+/sno-/mice at similar levels at 6 and 8 weeks. Trichrome staining of formalin-fixed samples revealed no evidence of fibrosis at 6 weeks in ACS+ and ACS+/sno-/- mice (8 week studies pending). Enlarged cardiomyocytes and mononuclear infiltrates were observed and in both ACS+ and ACS+/sno-/- mice. Conclusion: Loss of function of rpL13a snoRNAs improves cardiomyopathy in the MHC-ACS mouse model of metabolic cardiomyopathy. Given similar markers of inflammation in ACS+ and ACS+/sno-/-, it appears that the rpL13a snoRNAs act downstream of lipid induced inflammation in this model. Abstract 38 HDL-mediated eNOS activation falls before the onset of T2DM in obese non-human primates Erica Couzens (Loma Linda University), Francis Kim, University of Washington One of the protective actions of HDL is to stimulate the production of NO in endothelial cells via eNOS phosphorylation and activation. In T2DM we found that there is decreased HDLmediated eNOS activity. Decreased eNOS has been proposed to increase risk for the development of T2DM. We hypothesized that HDL-mediated eNOS activation is impaired prior to the onset of T2DM during obesity. Evidence for this was seen in a group of non-human primates on a high fat diet for 24 months that became obese and subsequently developed insulin resistance and T2DM. We utilized blood samples at baseline, 4, 8, 12, 18, and 24 months, isolated the HDL, and completed a cell-based assay in which HDL was incubated with endothelial cells, and eNOS phosphorylation, which is a marker of eNOS activity, was measured by in-cell western. In high fat fed primates, eNOS activity fell by 31% (p=0.0025) after 4 months and remained decreased between 4-24 months. At the end of 24 months, obese primates that underwent roux-en-y gastric bypass showed a 16% increase (p=0.1765) in eNOS activity. These data indicate that HDL-mediated eNOS activation falls prior to the onset of T2DM and may be altered following bariatric surgery. Proteomic experiments are ongoing to determine the changes in HDL proteins and lipids that may cause decreased eNOS activity. Abstract 39 Influence of Glucose Setpoint and Pre-Meal Bolus on PID-Based Closed-Loop System Performance Casey Card (National University of Ireland, Galway), Jennifer L Sherr, Eda Cengiz, Camille I Michaud, Neha Patel, Miladys M Palau-Collazo, Lori Carria, Eileen Tichy, William V Tamborlane, Stuart A Weinzimer, Yale University Closed-loop (CL) artificial pancreas systems combine insulin pump and continuous glucose monitor (CGM) technologies to control blood glucose (BG) levels automatically without human intervention. Proportional-Integral-Derivative (PID)-based computer controller algorithms employed in the design of such systems determine the appropriate insulin doses in response to BG levels and BG rate of change. The optimal glucose setpoint and need for manual pre-meal insulin boluses to optimize such a system have not been established. We analyzed blood glucose profiles from five closed-loop protocols utilizing a PID controller with insulin feedback to explore these questions. Data for 41 subjects (23 female, age 19±4y, A1c 7.3±0.7%) were analyzed to assess the effect of glucose setpoint (100mg/dl, [G100] vs 120mg/dl [G120]) and use of 0.05-0.08 u/kg pre-meal insulin bolus (PMB+ or PMB-) on CL system performance. Mean blood glucose (BG); BG levels within (70-180mg/dL), below (<70mg/dL), or above (>180mg/dL) target range, prandial BG excursion and prandial BGAUC; and hypo- (BG<60mg/dL) and hyperglycemia (BG>250mg/dL) were calculated for each condition. Compared to 30 G120 subjects, 11 G100 subjects had a significantly lower average BG (132±33mg/dL vs 146±33mg/dL, p<0.01); spent significantly more time in target range (83.8±9.8%, vs 74.9±10.5%, p<0.02) and less time above target range (13.0±9.6% vs 23.7±11.4%, p<0.01), with no significant increase in time <70mg/dL (3.2±3.5% vs 1.4±2.4%, p=0.07) or <60 mg/dL (1.1% vs 0.3%, p=0.11). 15 PMB+ subjects had smaller prandial BG excursion and BGAUC than 26 PMB- subjects (94 vs 105mg/dL and 207.4 vs 254.2mg*hr/dL, respectively), but these differences only reached statistical significance post-lunch (64±8 vs 102±7mg/dL, p<0.01 and 112±20 vs 225±29mg/dL, p<0.01, respectively). Using a sensor glucose setpoint of 100mg/dL rather than 120mg/dL in a PID-based CL system results in a lower average BG and improved time in target range, without a significant increase in hypoglycemia. The use of a small pre-meal bolus did not equivocally demonstrate benefit in mitigating prandial glycemic excursions; larger boluses or other strategies will likely be necessary to minimize post-prandial glucose levels. Abstract 40 Interleukin 17A induces renal proximal tubule NHE3 expression in hypertension Linjue Zhu (Medical College of Georgia – Georgia Regents University), Allison Norlander, Meena Madhur, Dave Harrison, Vanderbilt University School of Medicine T cells are essential to the development of Angiotension II (Ang II) induced hypertension. Specifically, the TH17 class of CD4+ T Helper cells produces a unique cytokine Interleukin 17 (IL17) that when knocked out in mice, confers the mice a 20mmHg a reduction in blood pressure following Ang II treatment as compared to controls by day 21. We hypothesized that IL17 increases the activity of Sodium Hydrogen Exchanger 3 (NHE3) in the kidney proximal tubule through intermediary SGK1 signaling to increase Na+ and water reabsorption, therefore causing a rise in blood pressure. We used a quantitative Western method to quantify the expression of NHE3 in human kidney proximal tubule epithelium (HK2) cells following IL17 treatment. We found that following treatment with IL17A, an isoform of the IL17 cytokine family, NHE3 expression increased by as much as 2.5 times as compared to untreated controls. To investigate the role of serum glucocorticoid kinase 1 (SGK1) in the IL17-NHE3 signal transduction pathway, we treated cells with a SGK1 inhibitor as well as IL17A and subsequently examined the NHE3 expression. We found that at 1mM concentration of SGK1 inhibitor, NHE3 expression was decreased. Altogether, these results suggest that IL17A signals through SGK1 to induce NHE3 expression in the kidney proximal tubule epithelium. These results may shed light on a novel function of SGK1 and NHE3 in modulating blood pressure in addition to their traditional roles in regulating epithelium sodium channels and tubular pH, respectively. Abstract 41 Attitudes and Beliefs about Diabetes Medications and Illness Acceptance in the Rural Black Belt Sarah Owens (University of Alabama School of Medicine); Emily Wesson; Susan Andreae; Monika M. Safford, MD; Division of Preventive Medicine, UASOM The Black Belt Action Committee reported that diabetes mortality was 38.7/100,000 population, contrasted to the US rate of 24.6/100,000 in 2007. The rate is 50% higher among Alabama’s rural African American population compared to whites, and studies show that people living with diabetes in this population report almost two times higher medication non-adherence than the national average. The Chronic Illness Trajectory Model by Corbin and Strauss posits that a feeling of wellness is achieved when there is a balance between 3 central domains: Body, Biography, and Conception of Self (the BBC Chain). Diagnosis of chronic illness, such as diabetes, can profoundly destabilize this BBC Chain, making it difficult for patients to accept their illness and understand the role of medications in living healthy lifestyles long-term. To gain insights into community members’ lived experience of diabetes and barriers to acceptance of illness and medication adherence, we conducted focus groups in Alabama’s low income Black Belt area. Using modified Grounded Theory, a focus group topic guide was created, eligible participants were recruited through respondent driven sampling, and focus groups were moderated, transcribed, and analyzed using Nvivo 10 software and open coding. Common themes were revealed throughout these focus groups, which will help inform the development of a peer support program that aims to improve diabetes medication adherence and health outcomes using peer storytelling and diabetes education. Major themes included: 1) low knowledge and misperceptions of both diabetes as a disease entity and the efficacy of diabetes medication; 2) disease acceptance and resolve to take medications triggered by a physical exacerbation of the participants’ body symptoms and occurrence of diabetes complications; 3) improved medication adherence with the realization that pharmacologics were a vital link to maintaining a participants’ sense of biography and conception of self by preserving body functions that could not be maintained through self-management behaviors (e.g., diet and exercise) alone; and 4) the physical complications of diabetes manifestations in close family and friends often served as the emotional impetus to participants’ disease acceptance and motivation to take medications. Further focus groups will be conducted until no new themes emerge and saturation is reached. Education to address diabetes and medication misperceptions as well as focused storytelling addressing these themes will be integrated into the intervention arm of the study. Abstract 42 Human Islet Cells Replicate and Transdifferentiate in Response to Hyperglycemia in an Islet Transplant Mouse Model Zachary Fowler (University of North Dakota), Xiaojuan Chen, Columbia University Pancreatic islet cell replication and transdifferentiation are potential mechanisms through which beta cell mass that is lost in the diabetic state can be restored. The factors that stimulate these processes in human islets are not well understood. To test the hypothesis that human islet cells can replicate and transdifferentiate in response to glucose, isolated human islets from deceased donors were transplanted under the kidney capsule of immune-deficient mice. Streptozotocin was administered to induce hyperglycemia, and islet graft sections taken three and eight days post hyperglycemia-induction were examined by immunofluorescence. BrdU and Ki67 were used as markers of proliferation and were readily detectable in both alpha and beta cells exposed to hyperglycemic conditions, while the islets under normoglycemic conditions had minute positive staining for these markers. The highest levels of beta cell proliferation were seen after three days of hyperglycemia. Additionally, hyperglycemia induced a colocalization of insulin and glucagon, suggesting that glucose is a stimulus for lineage reprogramming in pancreatic endocrine cells. In conclusion, using immunohistochemistry, we observed the occurrence of both replication and transdifferentiation in human islets under hyperglycemia. Both of these phenomena have important implications for beta cell regeneration in patients with type I diabetes and some forms of type II diabetes, as well as improving islet mass in patients with type I diabetes undergoing treatment with islet transplantation. Abstract 43 Bile diversion to the distal intestine achieves weight loss and metabolic improvements similar to Roux-en-Y gastric bypass. Steven Cai (Chicago Medical School), Robb Flynn (Department of Surgery, Vanderbilt University), Naji Abumrad (Department of Surgery, Vanderbilt University) Bariatric surgery, specifically Roux-en-Y gastric bypass (RYGB), achieves durable weight loss and imparts both immediate and long-term metabolic improvements by mechanisms that are not entirely understood. We investigated in diet-induced obese (DIO) mice the specific role of increased circulating bile acids (BA) frequently observed after RYGB using a novel surgical procedure where gallbladder (GB) bile was diverted to the duodenum (GB-D, sham surgery), the jejunum (GB-J) or ileum (GB-IL) and compared metabolic outcomes to RYGB mice. Mice receiving GB-IL lost more body weight, preferentially through fat mass, than GB-D or GB-J mice and even surpassed weight loss in RYGB mice. GB-IL mice also sustained improvements in glycemic control and hepatic steatosis that was comparable to RYGB mice. Circulating bile acids, specifically tauro-β-muricholic acid (TβMCA), were increased in GB-IL but not RYGB mice. Increased TβMCA, a known FXR antagonist, was concurrent with decreased BA receptor (FXR) expression in the liver and increased TGR5 and GLP-1 expression in the ileum. These results suggest that altered enterohepatic circulation of BA contributes to metabolic improvements seen in bariatric procedures such as RYGB. The GB-IL procedure, a technically simpler procedure, may serve as a potential therapeutic in treating obesity. Abstract 44 Influence of Childbearing on Women with Type 1 Diabetes: The 50 Year Medalists Heather Love, MPH, Tufts University School of Medicine, Liane J. Tinsley, MPH, Jennifer Sun, MD MPH, Stephanie D’Eon, MS ATC, Stephanie Hastings, BA, George King, MD, Hillary Keenan, PhD, Joslin Diabetes Center, Boston MA Pregnancy is a documented independent risk factor for progression of pre-existing diabetic retinopathy (DR), nephropathy (DN) and cardiovascular disease (CVD). Throughout pregnancy, hormonal readjustments in maternal endocrine tissues alter glucose tolerance, and increased cardiovascular demands result in vascular changes and potential preeclampsia. In the early days of type 1 diabetes treatment, women were discouraged from getting pregnant and having children due to these potential complications. This analysis examines long term influence of childbearing on women with 50 or more years of type 1 diabetes (T1D) duration in female participants of the Joslin 50-Year Medalist Study, a cross-sectional study collecting clinical, selfreport and laboratory data (n=465). These women have an average age of 64±7.1 years, T1D duration 54.5±5.3 years, duration at time of pregnancy 13.8±7.6 years and year of first pregnancy of 1971 [1966, 1977]. A significance was found in A1c (7.4±1.1% v. 7.2±0.9%, p<0.01), waist to hip risk ratio (WHR) (66% v. 81% p<0.01), antihyperlipidemic medication use (58.9% v. 74.9% p=0.02) , HDL (69.8±20.6 mg/dL v. 73.2±18.8 mg/dL p=0.04), LDL (85.3±20.6 mg/dL v. 79.89±18.8 mg/dL p<0.01), c-reactive protein (2.7±4.8 mg/dL v. 1.1±2.4 mg/dL, p=0.03) and body mass index (24.5±4.7 kg/m2 v. 25.5±4.8 kg/m2, p=0.04) between non childbearing and childbearing groups. No significant difference was seen in the prevalence of DR (59% v. 41%, p=0.9), DN (15.3% v 10.8%, p=0.2) or CVD (34.0% v. 29.8% p=0.4) between groups. Interestingly, there was a difference in the prevalence of self-reported cardiovascular disease by births (1: 21.9%, 2: 26.2%, 3: 53.3%, 4: 50.0%, p=0.02).This was not explained by a difference in duration (p=0.12), HbA1c (p=0.7), DN (p=0.1), c-peptide (p=0.7), antihypertensive medication use (p=0.9), antihyperlipidemic medication use (p=0.08), HDL (p=0.2), WHR (p=0.8), GAD65 (p=0.7), or IA2 autoantibodies (p=0.4). Although vascular endpoints did not vary significantly between women who did and did not have children, the number of children borne appears to influence the rate of CVD in this group, without influence from traditional diabetes related CVD risk factors. Abstract 45 Metabolic phenotypes of obesity-prone and obesity-resistant mice Amy Y. Han (Case Western Reserve University School of Medicine), Charles Addo-Yobo, Xiaoyan Yin, Fred Anokye-Danso, Rexford Ahima, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania Worldwide obesity epidemic has resulted in an increased prevalence of metabolic disorders such as type 2 diabetes and hyperlipidemia. Both environmental factors and genetic predispositions play significant roles in the development of obesity. Similar to humans, certain mice strains are obesity prone while some remain obesity resistant when fed on similar high fat diets. We studied obesity prone C57Bl/6J mice and obesity resistant SWR/J mice in pre-obese states for differences in their energy homeostasis. Male C57BL/6J mice and SWR/J mice, (Jackson Laboratory, Bar Harbor, Maine), age 10 weeks, were housed in 12 hour light-dark cycles (lights on at 7 am) with free access to chow and water. After one week adjustment period, body composition was measured with 1H-MRS, and energy homeostasis was assessed with Comprehensive Laboratory Animal Monitoring System (CLAMS). Thermal imaging as well as changes in energy expenditure in response to β3-adrenergic agonist treatment were assessed. Lastly, glucose homeostasis was measured using glucose tolerance test. Data analysis showed that the C57Bl/6J mice weighed significantly more than the SWR/J mice despite similar food intake. Both strains demonstrated nocturnal increases in food intake, oxygen consumption, respiratory exchange ratio (RER), and locomotor activity. SWR/J mice demonstrated lower RER, but higher energy expenditure during the light phase indicating an increase in fat oxidation. In addition, SWR/J mice showed greater energy expenditure in response to β3-adrenergic agonist treatment and more glucose tolerant than C57Bl/6J mice. In conclusion, the obesity resistant genetic background of SWR/J mice may be associated with enhanced fat oxidation, thermogenesis, and glucose homeostasis Abstract 46 The Clinical and Biochemical Characterization of Pancreatic Diabetes James Lyons (UAB School of Medicine), Fernando Ovalle MD, UAB Pancreatic diabetes is a phenotype of diabetes mellitus that is caused by pancreatic destruction. Disorders that lead to this condition include, among others: chronic pancreatitis, cystic fibrosis and pancreatectomy. Presently, this disease has not been well characterized and we believe that by analyzing the data of patients with pancreatic diabetes we will discover distinct parameters that will aid in the diagnosis of the disorder and will help clinicians distinguish it from other types of diabetes, most importantly types 1 and 2 diabetes. The data of 1257 consecutive patients from their first visit to a tertiary care center diabetes specialty clinic was recorded and the date from 27 patients who were diagnosed with pancreatic diabetes was compared to that from a matched group of patients diagnosed with type 1 and type 2 diabetes. The type 1 and 2 patients used for comparison were matched based on gender, race, and duration of diabetes. The results show that patients with pancreatic diabetes have similar BMI, systolic blood pressure, diastolic blood pressure, and HDL levels to those with type 1 diabetes, while their triglyceride levels were similar to patients with type 2 diabetes. C-peptide levels were distinct in patients with pancreatic diabetes and islet cell autoantibodies were frequently present in patients with pancreatic diabetes probably representing an epiphenomenon. These novel findings could help clinicians avoid making a wrong diagnosis of autoimmune diabetes in these patients. Abstract 47 Insulin Use and Long-Term Weight Change in U.S. Adults with Type 2 Diabetes Mellitus: Analysis of NHANES 2009-2012 Christopher Moutos (University of Arkansas for Medical Sciences), Hsin-Chieh “Jessica” Yeh, and Nisa Maruthur, Johns Hopkins University School of Medicine Standard treatment for achieving glycemic control in type 2 diabetes mellitus is to begin with lifestyle modification. If an optimal glycemic target is not achieved, oral antidiabetic medications are used, followed by insulin therapy if necessary. After initiating insulin therapy, patients tend to gain weight, which exacerbates the already fading insulin sensitivity of these individuals. This can lead to a greater need for insulin and potentially more weight gain over time. However, few data are available on long-term weight change after insulin initiation. We used the National Health and Nutrition Examination Survey (NHANES) to explore how different stages of type 2 diabetes treatment relate to weight change. Using this nationally representative sample, we characterized insulin use in adults with type 2 diabetes and to determine the association between duration of insulin use and body mass index (BMI). In survey years 2009-2012, we identified 1286 individuals who were diagnosed with diabetes at age 30 or older with available BMI data. These individuals were divided into five groups based on the type of diabetes treatment they received: (1) no treatment (2) oral antidiabetic treatment only (3) insulin use ≤ 1 year (4) insulin use 1-5 years (5) insulin use > 5 years. The age of the individuals characterized showed an overall upward trend from a mean of 58.6 years in the non-treated group, to a peak of 65.9 years in those having been on insulin for > 5 years. Similarly, the lowest values for duration of disease and mean HbA1c levels were both seen in the non-treated group (6.2 years and HbA1c =6.7%), with the highest values in patients treated with insulin for > 5 years (19.2 years and HbA1c = 8.2%). The trend in gender followed that in patients not currently on treatment, the majority (57.4%) were female, while in patients who had been on insulin > 5 years, the majority (56.9%) were male. After adjusting for current age, gender, and race, compared to patients without medications, individuals on oral medication and insulin had a higher BMI (adjusted beta-coefficients (95% CIs) of BMI for oral meds only, insulin use ≤1 yr, insulin use 1-5 yrs, and insulin use >5 yrs: 2.4 (0.9-4.0), 2.7 (0.2-5.3), 4.7 (1.4-8.1)and 3.0 (1.15.0), respectively) . In all, these results indicate that weight gain after initiation of insulin therapy may not be a transient event. Instead, this increase in weight may continue even up to 5 years after beginning insulin. Abstract 48 The Impact of Peripheral Artery Disease on Skeletal Muscle Gene Expression Alejandro Silva1, Katelyn Hughes2, Alison Burkart2, 3, Jonathan Dreyfuss2, Mark Creager3,4, and Mary-Elizabeth Patti2.4 1 Rutgers NJMS, 2Joslin Diabetes Center, 3Brigham and Women’s Hospital, 4Harvard Medical School Peripheral artery disease (PAD) is a manifestation of atherosclerosis which typically affects arteries in the lower extremities. Arterial narrowing can lead to functional impairment with pain and reduced ability to walk, in parallel with altered skeletal muscle metabolism. In addition, PAD is associated with insulin resistance, limiting the uptake of glucose into skeletal muscle. To test the hypothesis that PAD alters gene expression, we analyzed gene expression in human skeletal muscle in patients with PAD (n=21) as compared with healthy controls (n=9) using GeneChip® PrimeView™ Human Gene Expression microarrays. We identified differentially expressed genes and analyzed their functional ontology, focusing on those potentially impacting metabolic function and vascularization. 3,431 probe sets were differentially expressed in PAD vs. control (pnom<0.05), with 1953 upregulated and 1478 downregulated. To determine if the expression of these genes was normalized by exercise, we conducted a similar analysis comparing sedentary PAD patients (n = 7) to PAD patients following an exercise program for 6 weeks (n = 3). 3,478 probe sets were differentially expressed in PADsedentary vs PADexercise (pnom<0.05), with 1,611 downregulated and 1,867 upregulated in exercising PAD patients. In both PAD vs control and PADsedentary vs PADexercise, we identified candidate genes for further investigation, notably PLA2G16, PKM2, AK3, SFRP4, and RNASE4. These data demonstrate gene expression patterns differ in patients with PAD, and that exercise may normalize expression of some of these genes. Abstract 49 Germline zebrafish AIP mutant may serve as a potent disease model for the study of pituitary adenoma Jennifer Li (Chicago Medical School-Rosalind Franklin University of Medicine and Science), Ning-Ai Liu and Shlomo Melmed, Cedars-Sinai Medical Center Pituitary Adenomas are monoclonal, benign tumors of the anterior pituitary gland. Germline heterozygous mutations in the Aryl hydrocarbon receptor interacting protein (AIP) gene are reportedly associated with pituitary adenomas, in particular somatotropinomas, prolactinomas, and/or mix of both tumor types. To elucidate the role of AIP in pituitary development and tumor formation, we investigated embryonic pituitary development in a germline zebrafish AIP (zAIP) mutant. A nonsense mutation within exon 4 of zAIP gene was generated previously using customized transcription activator-like effector nucleases. Heterozygous zAIP mutant zebrafish were then crossed, and the 3-day post fertilization (dpf) zebrafish embryos (consist of wild-type, heterozygous-, and homozygous-AIP mutants) were used to study for the differential expression of growth hormone (GH), prolactin (PRL), and pro-opiomelanocortin (POMC, a precursor for ACTH). Following in-situ hybridization, we examined the phenotype and identified the genotypes of these zebrafish embryos. At 3 days post-fertilization, heterozygous-zAIP-mutant embryos (GH expression intensity 185.63±8.82, n=24) show induced GH expression compared to wildtype (178.97±13.9, n=11) and homozygous-zAIP mutant (176.22±12.7, n=11) embryos. In contrast, we did not find significant differences in prolactin expression between wild-type (PRL expression intensity 195.09±8.43, n=8), heterozygous- (191.83±14.01, n=28), and homozygouszAIP mutant (194.78±8.28, n=14) embryos. Similarly we did not find significant differences in POMC expression between the wild-type (183.20±10.41, n=15), heterozygous (184.07±9.73, n=22), homozygous zAIP mutant (177.60±7.17, n=11) embryos. These results suggest that at an early stage in zebrafish embryo development (3 days post-fertilization), the non-sense mutation on exon 4 of zAIP may induce GH-producing cells in the pituitary gland to express increased levels of GH. The results also indicate that this particular mutation does not alter prolactin and ACTH producing cells. To validate this animal model as an accurate disease model for pituitary adenomas (specifically for somatotropinomas), we are continuing to study developmental stage expression of these hormones 5 and 7 days post-fertilization. Abstract 50 Metabolic Effects of Hydroxychloroquine in Patients with Type 2 Diabetes Mellitus Samiksha Tarun (Saint Louis University School of Medicine), C. Rachel Kilpatrick, MD, Stacy Hurst, BSN, CDE, Janet B. McGill, MD, and Clay Semenkovich, MD, Washington University in St. Louis, Division of Endocrinology, Metabolism and Lipid Research Hydroxychloroquine (HCQ) is a quinolone antimalarial drug with poorly defined antiinflammatory properties that is currently used to treat rheumatologic conditions such as rheumatoid arthritis and systemic lupus erythematosis. HCQ use has been associated with reduced incidence of type 2 diabetes mellitus (T2DM) in non-diabetic individuals with rheumatic conditions and prevents worsening of hyperglycemia in T2DM patients. HCQ has also been shown to lower low density lipoprotein (LDL). However, HCQ’s metabolic effects in T2DM patients are not clearly understood. The primary hypothesis of this study is: clinically relevant doses of HCQ given to patients with T2DM who are taking metformin with or without other oral antihyperglycemic drugs, will reduce insulin resistance and reduce hepatic glucose output. The secondary hypothesis, the focus of this interim analysis, is: HCQ use in these patients will reduce fasting plasma glucose and fasting lipids. This study is a randomized, double blind, placebo controlled clinical trial where subjects were given either 200 mg of HCQ twice daily or placebo for 4 weeks. Hyperinsulinemic-euglycemic clamp testing was done at baseline and after four weeks of treatment. Unpaired and paired two-tailed T tests with equal variance were used to determine significance of treatment. A total of 16 patients have thus far been randomized to placebo (n=9) and HCQ (n=7). Amongst the HCQ treatment group, reductions were observed in diastolic blood pressure (P=0.030), HbA1c (P=0.047), and LDL (P=0.039). Mean change between baseline and follow-up clamp between HCQ and placebo group indicate trends toward decreased fasting glucose, HbA1c, triglycerides, total cholesterol, and LDL. Thus, these interim results suggest a possible trend toward beneficial metabolic effects of HCQ for patients with T2DM. Future studies are warranted to further characterize metabolic effects of HCQ use in T2DM patients. Abstract 51 Repression of LSD1 in adult pancreatic β cells leads to hyperinsulinemia and coincident hypoglycemia Benjamin Cross (University of Nebraska Medical Center), Matthew Wortham, Maike Sander, University of California, San Diego Epigenetic influences on gene expression are determined by various histone modifications, and methylation of histone H3 at lysine 4 (H3K4) is often indicative of an active promoter site. LSD1 (lysine-specific demethylase 1) demethylates mono- and dimethylated H3K4 (H3K4me1/2), and thereby suppresses gene expression. LSD1 has been proven essential for metazoan development, but its pathophysiological function in diabetes has remained essentially uncharacterized. In this study, we used both a mouse model and adult human β islets to determine the role of LSD1 on β cell function. In the mouse, we inactivated LSD1 in an inducible and cell-specific manner via breeding Pdx1-CreER; LSD1fl/fl mice. After isolating β islets from the mouse pancreas, LSD1ΔAdultβ and control islets were subjected to glucose stimulated insulin secretion (GSIS) assays, and insulin secretion was measured by ELISA. Additionally, adult human β cell islets were subjected to these same assays, with LSD1 being inhibited pharmacologically by 2µM tranylcypromine (TCP). We found that in LSD1ΔAdultβ mice, basal insulin secretion was enhanced coincident with hypoglycemia. It was shown that these mice had normal glucose tolerance compared to control mice, and the hypoglycemia was not due to enhanced insulin sensitivity. Human islets showed the same increase in basal insulin secretion upon treatment with TCP. Interestingly however, human islets from a patient with type 2 diabetes mellitus (T2D) showed no response to TCP, indicating there may be epigenetic differences that are LSD1-independent in T2D β cells. These data suggest that LSD1 plays an important role in suppressing the triggering pathway for insulin secretion, both in mouse and human islets. There are obvious potential implications for diabetes, with further mechanistic studies needed to determine specific targets of LSD1 in the β cell. Abstract 52 Sleep Behaviors in Pseudohypoparathyroidism Hannah Landreth (University of Oklahoma College of Medicine), Dr. Ashley Shoemaker, MD, MSCI, Department of Pediatrics, Division of Endocrinology and Diabetes, Monroe Carell Jr. Children’s Hospital at Vanderbilt Background: Pseudohypoparathyroidism (PHP) is an extremely rare, heterogeneous group of disorders. Etiology derives from a mutation in the GNAS gene, which encodes for the Gs-alpha subunit of G protein coupled receptors, and leads to absence of downstream second messenger effects via cAMP. Imprinting of GNAS results in two subtypes— PHP-Ia (maternally inherited PHP with obesity and multihormone resistance) and pseudopseudohypoparathyroidism (PPHP—paternally inherited PHP)— both of which share features of the Albright’s Hereditary Osteodystrophy (AHO) phenotype including short stature, brachydactyly and subcutaneous ossification. We have noted that PHP patients often complain of sleep apnea/sleep disturbances, but this has not been evaluated systematically. Objective: To determine the prevalence of sleep apnea and sleep behaviors in patients with PHP. We hypothesize that patients with PHP have a higher prevalence of sleep apnea/sleep disturbances than the general population. Methods: Twenty patients with PHP were recruited from Vanderbilt clinics and online advertisements. A validated, self-reported sleep quality survey was administered via REDCap (Children’s Sleep Habits Questionnaire (CSHQ) for parents of patients 4-13 years old or the Pittsburg Sleep Quality Index (PSQI) for participants >13 years). Normative data from healthy 4.5 year old children was used as control data for the CSHQ. Polysomnography records were obtained if available. Results: Nine patients 4-12 years old (all PHP-1a, age 8.9 ± 2.7 years) and 11 patients >13 years old (age 29.6 ± 11.4 years, PHP1a n= 8, PPHP n= 3) completed the study. The CSHQ total score was 2.59 ± 0.47 in the PHP-1a group versus 2.04 ± 0.41 in controls (p= <.001 by t-test). The PSQI global score was 8.33 ± 4.30, greater than the PSQI standardized global score cutoff of 5 for disordered sleeping. The scores were highest in the “disrupted subjective sleep quality” and “daytime dysfunction” subcategories. Scores were not associated with age or diagnosis. Five of the pediatric patients had polysomnography records available. Four patients were diagnosed with sleep apnea (1 with obstructive sleep apnea (OSA), 1 with central sleep apnea (CSA) and 2 with both OSA and CSA). All pediatric patients had undergone tonsillectomy/ adenoidectomy without resolution of their sleep apnea. Conclusions: Patients with PHP may have a higher prevalence of sleep apnea and sleep disturbance than the general population. This finding may be a complication of obesity or directly attributable to PHP and further studies are needed. Abstract 53 Effect of Real-Time Continuous Glucose Monitoring on Glucose Counterregulation in Long Standing Type 1 Diabetes Nina Ran (Perelman School of Medicine at the University of Pennsylvania), Amy Peleckis, Cornelia Dalton-Bakes, Shannon O’Brien, Nora Rosenfeld, Huang-Lan Nguyen, Mark H. Schutta, Michael R. Rickels. Institute for Diabetes, Obesity, and Metabolism at the University of Pennsylvania Perelman School of Medicine. Patients with long-standing type 1 diabetes are at high risk for severe hypoglycemia due to defects in both glucose counterregulation mechanisms: intra-islet signaling and the sympathoadrenal system. This clinical trial investigates whether avoidance of hypoglycemia – as achieved using real-time continuous glucose monitoring (RT-CGM) – can restore glucose counterregulation. Specifically, the study included eight participants with longstanding type 1 diabetes and severe hypoglycemia unawareness who underwent paired hyperinsulinemic (1.0 mU/kg/min) euglycemic (plasma glucose targeted to 90 mg/dl for 240 minutes) and hypoglycemic (glucose targeted to 80 mg/dl at 60 minutes, 65 mg/dl at 120 minutes, 55 mg/dl at 180 minutes, and 45 mg/dl at 240 minutes) clamps at baseline and six months to assess endogenous glucose production in response to insulin-induced hypoglycemia. The control group consisted of eight aged-matched healthy participants, and baseline characteristics were comparable between the two cohorts (average age of 43-44 years, BMI of 25-26 kg/m2; four males in each group). At six months, participants on RT-CGM experienced statistically significant improvements in Clarke score (from 5.8 to 4.5; p<0.05) and Hypoglycemia Score (from 1,952 to 714; p<0.05). A glucagon response was not detected; however we observed a significant difference in glucagon secretion between the six-month eu- and hypoglycemic clamps (p<0.05) that only trended toward significance at baseline (p = 0.07). Participants exhibited a trend for increased endogenous glucose production during the final hour of hypoglycemia at six months (p = 0.08); furthermore, we observed a difference in glucose production between the six-month eu- and hypoglycemic clamps (p< 0.05) that was absent at baseline. Lastly, participants experienced an increase in free fatty acids in response to hypoglycemia at six months (p<0.05); however, no change was detected in pancreatic polypeptide. These results suggest a minor effect of RT-CGM on glucose counterregulation, which may be further elucidated in the 18-month data. Abstract 54 Genetics and the Brain: A Preliminary Investigation into GLUL Hope Jin (New York Medical College), Gail Musen, Alessandro Doria, Richie Huynh, Joslin Diabetes Center Many studies have shown both poorer cognitive functioning and structural brain differences in individuals with diabetes and increased cardiovascular disease risk, though the mechanisms underlying these conditions are unclear. Recently, a single-nucleotide polymorphism (rs10911021) was shown to be significantly associated with coronary heart disease (CHD) in patients with type 2 diabetes. This SNP was linked to decreased glutamine synthase expression and altered levels of γ-glutamyl cycle intermediates, suggesting a relationship to vascular health. To conduct a preliminary study into whether the C risk allele at rs10911021 was related to changes in brain function and structure in diabetic individuals, an existing dataset of brain function, structure, and cognition for type 2 diabetic subjects and controls was analyzed. Previously collected research subject samples were genotyped by the Joslin DERC Genetics Core. Cognitive test data and brain imaging data of subjects were then analyzed to look for differences between control and diabetic individuals of C/C genotype or C/T or T/T (protective allele) genotype. Among the dataset, there were 19 C/T and T/T control subjects, 22 C/C control subjects, 11 C/T diabetic subjects, and 14 C/C diabetic subjects. No significant differences were found for cognition. Significant decreased cortical thickness was seen in the middle frontal gyrus of control subjects with the C/C risk allele genotype. This region is known to involved in working memory and executive function, indicating a possible mechanistic link between the rs10911021 genetic variant and poorer cognition in individuals. Future studies will recruit a larger sample size of subjects homozygous for the C/C risk allele and homozygous for the T/T protective allele as well as utilize magnetic resonance spectroscopy to measure concentrations of glutamate and glutamine within the brain. Abstract 55 Is SUDOSCAN a sensitive tool for the assessment of small autonomic neuropathy in type 1 diabetes? Nicole Nevarez (University of Michigan Medical School), Lynn Ang MD, Mamta Jaiswal PhD, and Rodica Pop-Busui MD, PhD, University of Michigan Cardiovascular autonomic neuropathy (CAN) is a common, prevalent, and serious complication in both type 1 (T1DM) and type 2 (T2DM) diabetes and an independent predictor of mortality. Sudomotor dysfunction may also appear early in the course of diabetes. The control of cardiovascular and sudomotor function is largely regulated by the autonomic nervous system via the small postganglionic, unmyelinated sympathetic cholinergic fibers. Currently available methods to detect autonomic dysfunction although reliable and sensitive are too expensive, invasive, time-consuming, or require large sample sizes. The goal of this study was to evaluate whether the non-invasive sudomotor function testing using the SUDOSCAN (Impeto Medical; Paris, France) is a sensitive and specific test for the diagnosis of autonomic dysfunction in patients with diabetes. We tested this hypothesis in a cross-sectional study in subjects with T1DM and age-matched healthy control subjects. 38 subjects with T1DM as defined by the American Diabetes Association diagnostic criteria and 10 age-matched healthy control subjects were enrolled and analyzed. The electrochemical skin conductance (ESC) of the hands and feet was measured with the SUDOSCAN through reverse iontophoresis. CAN was assessed in these subjects by evaluating the left ventricle sympathetic innervation and measuring the retention index (RI) with positron emission tomography (PET) scans with the sympathetic analogue [11C] meta-hydroxyephedrine (HED) and by heart rate variability (HRV) testing. In initial analyses, mean RI was not correlated with either mean hands or mean feet ESC. Additionally, none of the measures of HRV significantly correlated with means hands or feet ESC with the exception of EI ratio, which only significantly correlated with mean hands ESC but not mean feet ESC. These preliminary data suggest SUDOSCAN may not be the ideal tool for assessing small autonomic neuropathy in type 1 diabetes. However, these findings may be partly due to the limited number of subjects and large population studies with subgroup analysis need to be done in order to show the effectiveness of SUDOSCAN as a method assessing autonomic neuropathy. Abstract 56 The Novel Angiotensin (1-7) Cleavage Product Angiotensin (1-4) Increases GlucoseStimulated Insulin Secretion and Cell Viability in Cultured Mouse Islets Andrew Welch (Midwestern University Glendale, AZ) Breanne Barrow, Sakeneh Zraika, VA Puget Sound Health Care System and University of Washington Background: Type 2 diabetes is characterized by islet β-cell dysfunction. Recent studies demonstrate a novel branch of the renin-angiotensin system in which angiotensin converting enzyme 2 (ACE2) cleaves angiotensin I to generate the Mas receptor (MasR) agonist angiotensin (1-7) (Ang(1-7)). Upregulation of this ACE2/Ang(1-7)/MasR axis has multiple beneficial effects on pancreatic islets including enhanced glucose-stimulated insulin secretion (GSIS) and increased islet cell viability. While studies have focused on Ang(1-7) as the potential mediator of these beneficial effects, our lab has shown that peptidases present in the islet can further cleave Ang(1-7) to angiotensin(1-4) (Ang(1-4)). It is unknown whether Ang(1-4) acts as an agonist or antagonist of the MasR. Aim: To determine whether Ang(1-4) increases GSIS, insulin content, and islet cell viability. Methods: Islets were isolated from 10-week old C57BL/6J mice and cultured in the presence or absence of 1 nM Ang(1-4). After 48-hour culture, insulin secretion in response to 2.8 mM (basal) and 20 mM (GSIS) glucose, as well as islet insulin content were measured by ELISA (n=4). Islet cell viability was determined using an XTT assay which measures mitochondrial dehydrogenase activity in metabolically active cells (n=3). Results: Islets exposed to Ang(1-4) tended to secrete more insulin in response to 20 mM glucose compared to islets not exposed to the peptide [123.8±47.4 vs 81.7±25.3 pM/5 islets/h; p=0.16], while basal insulin secretion remained unchanged [26.1±3.5 vs 24.5±4.5 pM/5 islets/h; p=0.23]. This resulted in a 4.9±1.4 fold increase above the basal insulin response for Ang(1-4) exposed islets versus 3.0±0.7 fold with no peptide [p=0.09]. Further, insulin content did not differ between islets exposed to Ang(1-4) versus no peptide [62.9±7.0 vs 58.4±3.9 nM insulin/5 islets; p=0.66]. However, cell viability was significantly increased by 24±9% in islets exposed to Ang(14) [p=0.03]. Conclusion: Ang(1-4) increases GSIS and islet cell viability, suggesting it could be utilized as a novel therapeutic agent to treat β-cell dysfunction in type 2 diabetes. Abstract 57 Effect of Adipose-specific Inhibition of BCKDH Activity on Hepatic BCAA Metabolizing Enzymes and Adipocyte Morphology Kaitlyn Order (Georgetown University School of Medicine) J. Lee1, A. Vijayakumar1, Y. Xu2, O. D. Peroni1, C. J. Lynch 2, M.A. Herman1 and B.B. Kahn11Beth Israel Deaconess Medical Center and Harvard School of Medicine, Boston, MA and 2 Penn State University College of Medicine, Hershey, PA. Elevated levels of circulating branched chain amino acids (BCAAs) (valine, leucine, isoleucine) have been associated with insulin resistance in humans. In rodents, adipose tissue contributes largely to whole body BCAA metabolism. The branched chain α-ketoacid dehydrogenase E1β (BCKDH E1β) is the enzymatic subunit that regulates the irreversible decarboxylation step in BCAA metabolism. Our aim was to determine whether deletion of BCKDH E1β in white adipose tissue (WAT) is sufficient to cause insulin resistance and impair hepatic BCAA disposal. We measured hepatic protein expression of BCAA catabolism enzymes: BCKDH complex (E2, E1α, E1β), BCKDH Kinase, Protein Phosphatase 2Cm (PP2CM), and mitochondrial branched-chain aminotransferase (BCATm) from control chow-fed mice, and high-fat high sucrose (HFHS)-fed control and adipose-specific BCKDH knock-out (Ad-BCKDH-/-) mice. Additionally, adipocyte morphology (number and size) from perigonadal and subcutaneous WAT was characterized from these mice. HFHS-fed mice developed obesity compared to control mice, but relative fat mass, circulating BCAA levels, and insulin resistance were not further worsened in Ad-BCKDH/- mice. Protein levels of hepatic BCAA catabolic enzymes were reduced in HFHS-fed mice, but was not further attenuated in Ad-BCKDH-/- mice. Adipocyte size and number were not altered by HFHS-feeding or genotype. BCKDH activity in WAT is important for whole body BCAA metabolism, but circulating BCAA levels are not higher in Ad-BCKDH-/- on HFHS compared to control HFHS. One explanation is that other tissues such as the liver are compensating in AdBCKDH-/- mice to regulate whole body BCAA levels. Since hepatic protein levels and phosphorylation of BCAA catabolizing enzymes are not affected in our knockout model, muscle will be a key target organ of focus for future experiments. Abstract 58 Elevated Glucose Alters Epoxyeicosatrienoic Acid-related Enzymes in Human Retinal Cells Anjali J. Das (Rutgers New Jersey Medical School), Megan E. Capozzi (Departments of Molecular Physiology & Biophysics), and John S. Penn (Departments of Molecular Physiology & Biophysics, and Ophthalmology and Visual Sciences) Vanderbilt University School of Medicine, Nashville, TN Diabetic retinopathy (DR) is a vision-threatening complication of both type 1 and type 2 diabetes. The mechanism by which diabetes causes these changes is largely undefined. However, experimental evidence suggests that inflammatory cytokines and protein mediators may be involved. Previous studies have shown that several retinal cell types, including Müller glia, endothelial cells, and pericytes produce and/or respond to inflammatory signals under diabetesrelevant conditions. One approach to attenuate the chronic inflammation caused by diabetes may be to increase levels of epoxyeicosatrienoic acids (EETs), an anti-inflammatory lipid produced naturally in the body as a result of arachidonic acid metabolism by various cytochrome P450 isoforms (CYPs). However, the benefits of EETs are limited because of their rapid conversion to DHETs by soluble epoxide hydrolase (sEH). Particularly in diabetes, the levels of these enzymes may be altered to accelerate the development of retinopathy. To test this hypothesis, expression of CYP isoforms CYP2C8, CYP2C9, and CYP2J2, along with sEH (gene name EPHX2) was assessed using qRT-PCR against the housekeeping gene β-actin under normal glucose conditions (5.5 mM), L-glucose or mannitol (25 mM) as an osmotic control, or elevated D-glucose (25 mM) in primary cultures of human retinal microvascular endothelial cells (hRMECs), retinal pericytes, and Müller cells. Elevated D-glucose significantly increased EPHX2 transcription but did not change the levels of CYPs in HRMECs. In Müller cells, elevated glucose reduced the expression of CYP genes. This suggests that EET levels are reduced, limiting their bioavailability, and subsequently promoting inflammatory signaling during diabetes. In pericytes, high glucose stimulus did not change the expression of CYPs and sEH was not expressed in quantifiable amounts. Overall, this suggests that increasing production of EETs upstream by increasing the expression of CYPs or by preventing their conversion into DHETs by decreasing expression/activity of sEH may hold therapeutic promise. Abstract 59 Qualities of Diabetes Patients with Medication Adherence in Rural Alabama Emily Wesson (University of Alabama School of Medicine); Sarah Owens; Lynn Andreae, MPH; Susan Andreae, MPH; Monika M. Safford, MD; Division of Preventive Medicine, UASOM Diabetes is one of the most prevalent diseases in the United States, affecting an estimated 8.3% of the U.S. population in 2010. In rural Alabama, diabetes is compounded by medication non-adherence, which was found to be a problem in 56% of the population in these areas. One model of chronic illness that may explain non-adherence is Corbin and Strauss’s illness trajectory framework, which posits that disease states disrupt the balance between three elements – body, biography, and conception of self – that, when aligned, creates a sense of well being. In disease states such as diabetes, where the individual experiences a body failure, the conception of self must be reconfigured. This process may be especially stressful and lead to denial of the disease, negatively impacting optimal self-management behaviors. To understand the influences that allowed individuals with diabetes to move past their body failure toward achieving a balance with their new biography and conception of self, we interviewed four individuals from rural south Alabama who reported being adherent to their medications. These interviews revealed several common qualities among adherent participants, including both internal and external qualities. Internal qualities noted among participants included a high level of self efficacy and an internal locus of control. They also believed that the complications of diabetes were severe and that they themselves were susceptible to these complications. Adherent participants had a high belief that the medication would help to prevent diabetes complications. With regard to external qualities, these patients had strong social support networks and had heard about others’ experiences with diabetes. These findings reveal attitudes and beliefs regarding diabetes and diabetes medications that may be helpful for future interventions. Further work with non-adherent patients is needed in order to compare the qualities of non-adherent participants to those of adherent patients found in this study. Abstract 60 Patient Perspectives on Complications in Type 1 Diabetes Divya Vangala, M.A. (Marshall University Joan C. Edwards School of Medicine WV), Tara MacNeil, H.Dip.; Fernanda Lain, M.D.; Katie Weinger, Ed.D.,R.N. Diabetes Center; Harvard Medical School, Boston, MA Huntington, Joslin Successful treatment of diabetes is aimed at delaying the onset and/or slowing the progression of severe complications while maximizing the patient’s quality of life. Dynamic patient-provider collaboration is necessary to optimize treatment; however, there is little evidence on which to build patient-centered approaches to the discussion of complications. The purpose of this study was to gain a better understanding of patient concerns, feelings, and preferences about complications across the duration of diabetes and how these influence selfmanagement. We used a qualitative survey and validated questionnaires to assess patient perceptions about complications, diabetes self-care, and diabetes related distress. Medical records were accessed to obtain the most recent A1c level. The sample included patients with type 1 diabetes (n=60, 63% female, 56.4±12.5 years old, 15.6±1.7 years of education, 32.6±12.1 years duration, and HbA1c 7.6± 1.1) Primary concerns about developing complications include the inability to do things that were able to be done prior to having complications (80%), increased dependence on others (70%) and premature death (53%). 73% had already been told by their health care provider that they have at least one complication, with eye disease as the most prevalent (84%). The majority of our sample (98%) was able to identify their emotional response to diagnosis with a complication with fear (40%), feeling it was a “wake up call” (37%) and sadness (30%) being common. The response to coping with complications varies; however, more than half (55%) feel capable of minimizing complications, maintaining a positive attitude, and trying to maintain their independence. We found no significant differences in self-reported self-care or self-reported distress when comparing participants with an A1c <7.5% to participants with an A1c≥7.5%. 60% would like to discuss complications with their provider shortly after diagnosis. Patient preference on how often this conversation should occur was distributed among once or twice a year (30%), more often (32%) and others would base it on the patient (35%). Our findings suggest that there are common concerns about complications among patients with type 1 diabetes; however, the responses to cope and recommendations to providers varied. This was a cross-sectional qualitative study that encompassed people who have long-term diabetes; future research should look at people across the various phases of diabetes from early diagnosis to when complications dominate. Studies on the provider’s perspective should also be done in order to develop methods to enhance patient-centered care. Abstract 61 Increased BMI May be Associated With Increased Risk of Proliferative Diabetic Retinopathy in Persons With Type 1 Diabetes Christian A. Mehregan (Wayne State University School of Medicine), Taylor S. Blachley, Thomas W. Gardner (University of Michigan Department of Ophthalmology and Visual Sciences) Introduction and Background: The prevalence of overweight and obesity has increased in non-diabetic and diabetic persons over the past 20 years but the relationship of excess BMI to retinopathy has not been examined in depth. Hypothesis: Individuals with type 1 diabetes who are also overweight or obese are more likely to develop diabetic retinopathy. Methods: We conducted a retrospective study of patients with type 1 diabetes at the University of Michigan Kellogg Eye Center. Dilated eye fundus examination results were used to determine diabetic retinopathy status. BMI, HbA1c levels, blood pressure, serum triglycerides, and demographics were collected. Results: Logistic regression analysis found association between duration of diabetes and presence of retinopathy (Odds Ratio (95%CI):1.129 (1.082-1.179); p < .0001). Logistic regression model assessing for presence of proliferative diabetic retinopathy (PDR) found an association between duration of diabetes and PDR: (1.139 (1.08-1.201); p < .0001) and relationships approaching significance between BMI and PDR: (1.08 (0.992-1.177); p =.076) and most recent HbA1c and PDR: (1.468 (0.96-2.244); p =.076). Chi-squared analysis found significant differences in the likelihood of developing PDR in individuals with elevated BMI (p=.033). The relative risk of developing PDR in overweight patients compared to normal weight patients is 1.33 and 2.42 in obese patients. Conclusion: These data suggest increasing BMI may be associated with a higher risk of developing PDR. These findings imply that overweight and obese persons with type 1 diabetes may require additional treatment considerations. Abstract 62 Hepatic Nuclear Hormone Receptor Profiling in Nonalcoholic Fatty Liver Disease Regina Matar (Michigan State University College of Human Medicine), Joel Lavine, Columbia University The worldwide increase in nonalcoholic fatty liver disease (NAFLD) is attributed to the rise in obesity, insulin resistance, dyslipidemia, and hypertension. Usually nuclear hormone receptors coordinate enzymatic cascades to regulate energy and lipid metabolism, but when this system is disrupted, the risk for hepatic steatosis and metabolic syndrome increases. Similar to mouse models of cachexia, we project that the human nuclear receptor profile will show variable expression levels that will be diagnostic of the disease state. To test this hypothesis, we utilized quantitative real-time reverse-transcription PCR (qpCR) to find an average of all 48 human nuclear hormone receptor expression levels in about 300 hepatic biopsies with over 200 unique individuals. Receptor hormone expression profiles were then compared amongst groups including men to women, boys to girls, adults to children, those with more advanced NAFLD to those with simple steatosis or normal liver, those children with type I NASH to those with type II, and those treated to those untreated. Normalized mRNA levels for PPAR-δ were significantly lower in patients who showed steatohepatitis (30.8 ± 1.84, N=19) in comparison to patients with no steatosis (32.2 ± 1.68, N=21), in patients with fibrosis (30.6 ± 1.89, N=13) compared to patients with an insignificant fibrosis status (32.0 ± 1.72, N=27), and in patients with a high amount of lobular inflammation (30.4 ± 1.75, N=13) compared to patients with little to no lobular inflammation (32.0 ± 1.72, N=27). RAR-β and FXR-α had normalized expression levels that were significantly reduced in patients with steatosis >33% (28.7 ± 1.36, N=18) and (21.2 ± 0.99, N=18) compared to patients with steatosis ≤33% (30.2 ± 2.14, N=22) and (22.1 ± 1.25, N=22), respectively. Normalized mRNA levels for VDR were lower in patients with fibrosis (31.4 ± 2.17, N=13) in comparison to patients with insignificant fibrosis (51.5 ± 42.0, N=27). Nuclear hormone receptor expression profiles were also reduced in patients who had severe lobular inflammation compared to patients with low to no lobular inflammation; this included (33.0 ± 1.42, N=13) compared to (44.7 ± 24.8, N=27) in PR, (30.4 ± 1.75, N=13) compared to (32.0 ± 1.72, N=27) in PPAR-γ, and (26.8 ± 1.71) compared to (29.8 ± 3.33) in PPAR-γ2. These findings suggest that disruption of certain nuclear hormone receptors expression levels is correlated with the pathological progression of NAFLD. These observations allow for diagnostic and/or prognostic signatures regarding forms of human fatty liver disease. Abstract 63 High prevalence of type 2 diabetes mellitus exists in resistant hypertensive patients with primary aldosteronism. Mingchun Liu (University of Alabama School of Medicine), Timothy Wang, Bin Zhang PhD, Peng Li PhD, Tanja Dudenbostel MD, Suzanne Oparil MD, University of Alabama at Birmingham (UAB) Resistant hypertension (RHTN) is uncontrolled blood pressure (BP>140/90 mmHg) on 3 different antihypertensive drug classes. A subset of RHTN patients may have primary aldosteronism (PA), defined as aldosterone-renin ratio (ARR)>30 or 24-h urinary aldosterone (UAldo)>12 mcg/24hr. Previous work has shown that aldosterone stimulates insulin resistance. This led us to hypothesize that PA patients have a higher prevalence of type 2 diabetes mellitus (T2DM). To test our hypothesis, we retrospectively analyzed the EMR of RHTN patients who had been worked-up for PA. Patients taking spironolactone, have end-stage renal disease (ESRD), or had incomplete urine collections were excluded. Those included in the analysis (n=2043) were categorized into two groups, PA (n=442) and non-PA (n=1601). Prevalence of T2DM was assessed for both groups if they meet one of the following criteria: 1) previous diagnosis or current anti-diabetic medication(s), 2) fasting glucose≥126 or, 3) HbA1C≥6.5. Statistical analysis revealed that PA patients have a significantly higher prevalence of T2DM than non-PA patients (22.1% vs. 15.4%, p=0.0001). Therefore, PA may be a predictor of T2DM risk in patients with RHTN. This clinical study provides a basis for further exploration into how these disease processes interact. Ultimately, it may help in the development of more effective therapeutic treatments for targeting these chronic conditions. Abstract 64 A role for Retinoic Acid Signaling in Macrophage Phenotypic Switching During Acute Kidney Injury Radostin Penchev, George Washington University School of Medicine, Takuto Chiba, Nataliya Skrypnyk MD, and Mark de Caestecker M.B., B.S., Ph.D, Vanderbilt University Medical Center Acute Kidney Injury (AKI) is characterized by sudden and rapid decline in GFR from baseline that is independently associated with increased risk of death and an in-hospital mortality rate of 40-50%. During the early stages of AKI pronounced hypoxia triggers the activation and infiltration of proinflammatory M1 macrophages and further tissue damage. With time, the adaptive immune system is mobilized and M2 macrophages promote tissue repair and downregulate the acute M1 response that is damaging to the kidney. Preliminary studies in our lab have shown that inhibition of Retinoic Acid (RA) Signaling with the antagonist BMS-493 exacerbates kidney injury and leads to downregulation of M2 and upregulation of M1 macrophage markers. We therefore hypothesized that RA signaling regulates macrophage phenotypic switching after AKI. To address this we first utilized an ischemia and reperfusion mouse model of acute kidney injury where we detected gradual increase in the level of the RA producing enzyme RALDH3 within the interstitium of the outer medulla at 6 hours, 12 hours and 24 hours after injury followed by a gradual decrease from day 3 to day 7 after injury whereas Raldh3 was virtually absent in uninjured control. Using a RARE-LacZ reporter mouse, a LacZ reporter line for RA signaling activation, we observed active RA signaling response after ischemia and reperfusion injury in proximal tubular epithelial cells and interstitial cells of an uncertain cell type that morphologically resembled infiltrating monocytes. We demonstrated that cultured renal fibroblasts are capable of producing RA when co-cultured with F9-RARE-LacZ reporter cells. Furthermore, we showed that there is a significant upregulation (3.2 +/- 1.2 fold increase, p=0.002) of the M2 marker Arginase-1 in cultured macrophage-like Raw264.7 cells incubated with conditioned media from renal proximal tubular cells (RPTC) treated with all trans retinoic acid (ATRA) compared to Raw264.7 cells treated with RPTC media and ATRA alone. Based on these findings we propose a new model in which infiltrating peritubular cells secrete RA early after injury which promotes secretion of an as yet unidentified factor by proximal tubular epithelial cells which induces macrophage switching to a pro-regenerative phenotype. Early activation of RA during AKI may therefore limit M1 macrophage differentiation and tissue damage. Abstract 65 Basal insulin treatment with detemir improves glycemic control but does not potentiate weight loss in obese individuals with type 2 diabetes mellitus on a hypocaloric diet Matthew A. Buendia (University of Tennessee Health Science Center – College of Medicine), BettyAnn Chodkowski, Hakmook Kang, Malcolm J. Avison, Heidi J. Silver, and Kevin D. Niswender, Vanderbilt University School of Medicine Lowering blood glucose level is a primary treatment strategy used to avoid long-term complications of type 2 diabetes mellitus (T2DM). As T2DM progresses, patients typically require insulin to maintain glycemic control; however, insulin therapy is often associated with weight gain, which is contrary to clinical goals in diabetes care. The goal of this study is to quantify improvements in glycemic control associated with low dose basal insulin detemir therapy and weight loss in obese insulin-naïve individuals with T2DM. This is a 26-week, interventional, randomized, controlled trial. Subjects were randomized to receive insulin detemir (IDet) treatment or no treatment (No IDet). During the first 4-week period, participants were asked to maintain a constant body weight. After the 4 weeks, both groups began the weight loss phase, which consisted of 20 weeks of low-calorie diet intervention (Diet) in addition to continuation of insulin if in the insulin group. Mean change in body weight from baseline did not differ between No IDet (-5.20 ± 0.88 kg) and IDet (-5.52 ± 0.85 kg) groups (p=0.4559) over the 24-weeks. Glycemic control assessed by fasting plasma glucose (FPG), hemoglobin A1c (HbA1c), and mixed meal tolerance testing (MMTT) improved in both groups, but occurred earlier in the IDet treatment group, as seen by significant differences between groups at Week 14. Interestingly, 4 weeks of IDet alone in the absence of weight loss significantly lowered FPG (-21.71 ± 7.43 mg/dL) compared to no IDet (1.08 ± 3.65 mg/dL; p=0.0018). Glycemic control (FPG, HbA1c, and MMTT) was significantly improved in the subjects receiving both 14 weeks of IDet + 10 week Diet compared to the No IDet + 10 week Diet group even though the groups did not differ in weight change from baseline. By the end of the study, both groups had significant improvements in glycemic control and weight loss from baseline. IDet treatment for 4 weeks, independent of weight loss, improved specifically FPG, and potentially accelerated FPG, HbA1c and MMTT improvements by Week 14 compared to Diet alone. At the end of the study, week 24, IDet+Diet was not superior to Diet alone in terms of mean changes in glycemic control and body weight. Our results suggest that low dose basal insulin detemir accelerates improvements in glycemic control compared to weight loss alone, thereby reducing the risk of long-term T2DM complications. These observations have implications when developing appropriate and timesensitive drug regimen and treatment plans for patients with T2DM and obesity. Abstract 66 The role of CSF-1 in macrophage proliferation and differentiation in acute kidney injury Emily F. Kelly,1 Bing Yao,2 Yinqui Wang,2 Ming-Zhi Zhang,2 Raymond C. Harris,2 1. Florida Atlantic University, Charles E. Schmidt College of Medicine, Boca Raton, FL, USA. 2. Division of Nephrology, Department of Medicine, Vanderbilt University, Nashville, TN, USA. Acute kidney injury (AKI) leads to injury of kidney epithelial cells, especially proximal tubule cells, and to the activation of monocytes and macrophages. Following AKI, M2 or wound healing macrophages are alternatively activated and signaled to proliferate by CSF-1 produced by the renal proximal tubule cells. Upon M2 activation paracrine growth factors are produced that target epithelial cells and fibroblasts allowing for cellular growth and cytoskeletal repair and regeneration. We hypothesize that CSF-1 is critical for the recovery form AKI by acting as a proliferation signal for M2, wound healing, and macrophage differentiation. Deletion of CSF-1 in the renal proximal tubule cells will delay recovery from AKI by decreasing M2 macrophage proliferation and, as a result, decrease tissue repair and regeneration. Diphtheria toxin receptor (DTR) mice, with human HB-EGF cDNA construct, selectively express the DTR receptor within the renal proximal tubular cells. DTR mice were crossed with CSF-1 floxed mice to generate CSF-1flox/flox DTR (wild type) mice. Additionally, knockout mice were created by crossing the DTR mice with γ-GT Cre mice generating γ-GT Cre; CSF-1flox/flox DTR (knockout) mice in which CSF1 in the renal proximal tubule epithelial cells was deleted. When injected with the diphtheria toxin these DTR mice (CSF-1flox/flox DTR mice and γ-GT Cre; CSF-1flox/flox DTR) developed AKI. Through blood urea nitrogen (BUN) levels, flow cytometry, and qRT-PCR, recovery from AKI and the degree of macrophage proliferation can be measured. CSF-1 knockout mice showed increased BUN levels which correlated to the delayed rate of recovery from AKI. Through flow cytometry it was shown that blocking CSF-1 caused diminished M2 (F4/80+, CD206+, CD11b+) cellular proliferation in CSF-1 KO mice (P < 0.01; n=4). qRT-PCR revealed that CSF-1 depleted mice (KO) displayed marked reduction in mRNA levels of M2 dependent growth factors (P < 0.1; n=4). Depletion of M2 macrophages resulted in delayed renal tissue repair and increased kidney fibrosis. Overall, CSF-1 plays an essential role in M2 polarization. When the CSF-1 signal is absent, M2 macrophage proliferation declines resulting in a decreased production of M2 growth factors and consequently delayed recovery from AKI. CSF-1, therefore, is critical for the activation of M2 macrophages and their role in repair and differentiation after AKI. Abstract 67 Can website information be used to identify guideline-concordant weight loss programs in the community? Benjamin Bloom (University of Maryland School of Medicine), Dr. Ambereen Mehta, Dr. Kimberly Gudzune, Johns Hopkins University, Welch Center for Epidemiology, Prevention and Clinical Research Background: Weight loss has been demonstrated to prevent diabetes mellitus. Guidelines recommend that primary care providers (PCP) refer patients with obesity to high intensity weight loss interventions; however, many PCPs may be unaware of locally available programs. Objective: Our objective was to determine the reliability of web-based information regarding guideline-concordant practices of community weight loss programs in Maryland. Methods: We conducted a systematic online search for community-based weight loss programs, and performed a content analysis to abstract weight management practices from their websites. We then randomly selected 50 of these programs for a telephone survey to obtain actual program components and practices. We compared the accuracy of web and telephone data among this vanguard group to determine reliability of web-based information using crosstabulations. Results: We identified 191 programs and recruited 41 for the telephone survey. Among this vanguard, 41% were physician delivered and 98% were in-person interventions. From the website, we could abstract program intensity (frequency of contacts) reliably among 37% of programs. Most misclassified programs were graded as a lower intensity as compared to phone report. Content analysis correctly identified programs containing dietary components (71%), exercise components (53%), and any behavioral component (51%). Both exercise and behavioral components were often not reported on the websites, despite use of these guidelinebased practices as reported by phone. We accurately identified 90% of programs that used FDA-approved anti-obesity medications; however, 29% of programs that dispensed nutraceuticals were misclassified. Typically, websites failed to mention the use of these nonguideline-supported supplements (43%), despite endorsement by program staff of their use. Significance: Evaluation of weight loss programs’ web-based content can reliably reflect some measures of guideline-concordant care and underestimates other aspects. This information could potentially be used to create a decision aid to facilitate PCPs’ assessment of local weight loss programs, thus enhancing referrals to guideline-concordant programs. Abstract 68 Development of CD8+ Tregs for in vivo Islet Protection. Courtney N. Altshuler (Keck School of Medicine – USC), Blair T. Stocks, Andrew F. Marshall, and Daniel J. Moore, Vanderbilt University Medical Center, Nashville, TN Islet transplantation for the cure of Type 1 diabetes is challenged by two fundamental immunologic barriers: recurrent autoimmunity against islet beta cells and alloimmunity against MHC mismatches and novel antigens within the graft. Both of these deleterious processes are driven by ongoing interactions between T and B lymphocytes. It is now known that suppression of direct, ongoing T cell–B cell interactions can be executed by CD8+ Tregs, which are highly potent regulators of autoimmunity. We investigated the hypothesis that CD8+ Tregs are required for the induction of transplantation tolerance to islets in T1D and that their insufficiency predisposes to autoimmunity and graft rejection. We tested this by developing an assay to measure alloantibody production in mice challenged with MHC-mismatched splenocytes, measured the responses of different cell types (CD8+, CD4+ TFH, and germinal center B cells) at 7-day intervals, and tested to see whether fully activated CD8+ Tregs could suppress this alloantibody response. Our results show that B6 mice immunized with MHC-mismatched C3H splenocytes develop a measurable alloantibody response (anti-C3H IgG1) by day 7, and peaks by day 14, remaining stable at day 21. Based on subset analysis, we detected an increase in naïve CD8+ T cells, a decrease in memory CD8+ T cells, and no significant change in CD8+ Treg levels at 14 and 21 days vs. 7 days. In vivo, activated CD8+ Tregs were able to delay the development of diabetes in a stringent, adoptive transfer model; we are extending these findings to islet transplantation. Overall, we propose that development and activation of CD8+ Tregs will be a critical new target to promote the success of islet transplantation in patients with T1D. Abstract 69 Identification of the cellular source of hepatic Notch signaling in obesity and nonalcoholic steatohepatitis. Ignacio J. Contreras (College of Physicians and Surgeons, Columbia University), Changyu Zhu, Utpal Pajvani, Columbia University Notch signaling is an essential pathway that determines cell fate during development. Recently, Notch has been shown to be highly activated in patients with non-alcoholic steatohepatitis (NASH). NASH is a condition where the liver undergoes lipid accumulation, inflammation and fibrosis. It can ultimately lead to cirrhosis or hepatocellular carcinoma.1 Its pathogenesis is not yet well understood. We hypothesized that hepatic Notch signaling to be a contributing factor to the pathogenesis of NASH. We tested this visually by using immunostaining in Nicastrinknockout mice and wild-type mice that were fed on methionine-and-choline-deficient diet (MCDD), which is a useful animal model of NASH. Nicastrin is a protein that makes up a part of the gamma-secretase, an enzyme that cleaves the Notch receptor within the plasma membrane. We generated liver-specific Nicastrin knockout mice by crossing Nicastrin-floxed mice with Albumin-Cre mice (Nicastrinfl/fl;Albumin-Cre genotype). We confirmed the efficacy of deletion of Nicastrin by using quantitative PCR (qPCR) and immunofluorescence. Compared with controls, liver-specific Nicastrin knockout mice showed a great reduction of Nicastrin mRNA and protein in the livers. We co-stained liver sections with Notch target protein Hey1 and cell-type specific markers that would allow us to visually identify the cell type(s) that display Notch activity within the liver, be it a hepatocyte, Kupffer cell, endothelial cell, or stellate cell. We found that Hey1 is expressed in hepatocytes in control mouse livers, but its level is decreased in hepatocytes in Nicastrin knockout livers. Interestingly, we noticed that other cell types, such as endothelial cells, might be a source of Hey1 activity. We did not have enough time to adequately test out all possibilities, given the fact that not all antibodies work for immunofluorescence, although we remain optimistic to find more conclusive data that point to the fact that Notch signaling is behind NASH. 1 Ertle, J. et al. Non-alcoholic fatty liver disease progresses to hepatocellular carcinoma in the absence of apparent cirrhosis. Int J Cancer 128, 2436-2443, doi:Doi 10.1002/Ijc.25797 (2011). Abstract 70 Relationship between skeletal muscle mitochondrial stereomorphometry and insulin resistance 1 Jay Mottla (Georgetown University School of Medicine), 2Caroline Suresh, 2,3,4Steven K. Grinspoon, MD, 2,3,4Hideo Makimura, MD, PhD 1 Georgetown University School of Medicine, Washington, D.C., 20007, 2Program in Nutrition Metabolism, Massachusetts General Hospital, 3Neuroendocrine Unit, Massachusetts General Hospital, 4Harvard Medical School, Boston, MA 02114 Currently 32% of Americans are obese with a body mass index (BMI) ≥30kg/m2. Insulin resistance may contribute to metabolic diseases associated with obesity. Recent studies have implicated mitochondrial dysfunction in mediating insulin resistance. We hypothesize insulin resistance is associated with a decrease in skeletal muscle mitochondrial number, therefore leading to decreased mitochondrial function. Thus, we investigated the relationship between characteristics of mitochondrial stereomorphometry and measures of insulin resistance. As an exploratory endpoint, we assessed the relationship between mitochondrial stereomorphometry and body composition. We recruited 20 abdominally obese men and women from the Boston metropolitan area. Skeletal muscle tissue was obtained via percutaneous muscle biopsy. A novel, semi-automated software program analyzed electron microscope images of the muscle tissue. The program generated stereomorphometry parameters including total number of pixels occupied by mitochondria, total density of mitochondria, number of mitochondria and average density of individual mitochondria. Insulin sensitivity of each subject was determined via the “gold-standard” hyperinsulinemic-euglycemic clamp as well as fasting insulin, glucose, HOMA and HbA1c. Subjects were 70% male, 44.9±1.58 years old, with BMI of 39.5±1.07 kg/m2, median HOMA index of 4.38 [3.31, 5.21], median glucose infusion rate (GIR) of 3.16 [2.26, 3.62] on hyperinsulinemic-euglycemic clamp, and median mitochondrial number of 56.9 [53.7, 61.5]. GIR was positively associated with increased total mitochondrial pixels (ρ=0.54, P=0.02), density (ρ=0.52, P=0.02), and number (ρ=0.52, P=0.02). However, neither fasting glucose, insulin, HOMA nor HbA1c were associated with descriptive parameters of mitochondria (all P>0.05). In addition, total body fat percentage was negatively correlated with mitochondrial pixels (ρ=-0.59, P=0.006), density (ρ=-0.57, P=0.009) and average density (ρ=-0.46, P=0.04). Total lean mass percentage was positively correlated with mitochondrial pixels (ρ=0.60, P=0.005), density (ρ=0.58, P=0.007) and average density (ρ=0.52, P=0.02). In conclusion, we demonstrated a significant positive association between the parameters of mitochondrial stereomorphometry and insulin resistance using hyperinsulinemic-euglycemic clamp but not with more crude measures of insulin sensitivity. The observed relationship is consistent with previous publications, but the determination of mitochondrial parameters via semi-automated software demonstrates objective and reproducible evidence for the association. Additionally, a previously unpublished relationship between mitochondrial parameters and body composition is reported. Abstract 71 Cell Culture Optimization for in vitro Chondrocyte Differentiation Elizabeth J. Olecki (Northeast Ohio Medical University), Matthew R. Karolak, and Florent Elefteriou, Vanderbilt Center for Bone Biology, Vanderbilt University Many diseases affecting growth and bone repair are the result of genetic mutations in chondrocytes. One way to study the etiology of these diseases is in vitro culture of differentiating chondrocytes. Unfortunately, because chondrocyte proliferation and differentiation is highly dependent on precise spacio-temporal interactions between various cell types, cytokines and matrix proteins, it is very difficult to differentiate chondrocytes in culture. Because chondrocytes are embedded in an ion and growth factor rich matrix, we hypothesized that chondrocytes will more reliably differentiate if the hypoxic and osmotic conditions of the growth plate are mimicked in vitro. To test this hypothesis, we harvested chondrocytes from P5 murine ribs and plated them in micropellets to mimic hypoxic conditions while also growing the cells in a standard media or standard media supplemented with 100 mOsm NaCl or 200 mOsm NaCl. Chondrocyte pellets were harvested at 0, 2, 5, 10, and 15 days following plating and were then stained for matrix production with Alcian blue and assessed for differentiation by quantitative PCR. We found that matrix production was increased in day 2-15 differentiated WT chondrocytes grown in +100 mOsm and +200 mOsm when compared to the standard media. Using Col2, Acan, and Col10 as gene expression markers of chondrocyte commitment and hypertrophic differentiation, we found that a +200 mOsm supplementation resulted in the most reliable differentiation and maintenance of chondrocyte commitment. We also used Nf1-/chondrocytes (taken from Nf1Col2-/- mice) to confirm that chondrocyte differentiation, as assessed by Col10 gene expression, to hypertrophy is impaired when Nf1 is deleted. As expected, expression of Col2 and Acan were not affected by Nf1 loss-of-function. Finally, we found that chondrocytes plated in micropellets immediately following harvest from P5 pups versus chondrocytes expanded for one week in culture prior to pelleting showed the best maintenance of chondrocyte commitment and differentiation to hypertrophy (Col2 and Col10 gene expression, respectively). In conclusion, mouse rib chondrocytes showed the best overall differentiation when grown in vitro with +200 mOsm NaCl conditions and not expanded in culture. Abstract 72 Medication Adherence: An Important Factor in Understanding Racial Disparities in Glycemic Control Bradley W. Harrison, BS (Mercer University School of Medicine); Lindsay S. Mayberry, MS, PhD; Chandra Y. Osborn, PhD, MPH; Vanderbilt University Medical Center Among adults with Type 2 diabetes mellitus (T2DM), non-White race has been consistently associated with less medication adherence and worse glycemic control. Less medication adherence has also been associated with worse glycemic control. Few studies have explored less medication adherence as a mechanism by which non-Whites have worse glycemic control. In an effort to build upon this literature, we tested the hypothesis. We collected data from 314 adults with T2DM who were prescribed oral anti-hyperglycemic medication(s) and/or insulin and presented for an appointment at a Federally Qualified Health Center in Nashville, TN. Racial (White vs. non-White) differences on sociodemographic and clinical characteristics were examined with Fisher’s exact and Mann Whitney U tests. We used indirect effect tests with biascorrected bootstrapping (5000 samples) to test for mediation. We conducted an unadjusted model and a model adjusted for apriori covariates (i.e., age, gender, education, income, insulin status, duration of diabetes). The average participant was 51.8 ± 11.7 years old. Nearly twothirds were female (65%), whereas non-Whites constituted 63%; the average years of education was 11.9 ± 2.9; 71.2% reported annual income below $15,000. In unadjusted analysis, nonWhite race was associated with less medication adherence (coefficient = -1.36, p<0.05) and worse glycemic control (coefficient = 0.92, p<0.001), less medication adherence was associated with worse glycemic control (coefficient = -0.09, p<0.001), and less medication adherence mediated the association between non-White race and worse glycemic control (unadjusted indirect effect = 0.128 [0.021,0.296]; adjusted indirect effect = 0.076 [0.001,0.223]). The adjusted model explained 19.9% of the variance in glycemic control (F = 8.079, p<0.001). Our results suggest medication nonadherence is one mechanism underlying racial disparities in glycemic control. Study limitations include sampling from a single clinic, under-representation of White male participants, and the use of self-reported measures. Interventions targeting medication nonadherence may also help reduce disparities in glycemic control among lowincome patients with T2DM. Abstract 73 Type 2 Diabetes may affect the potential effects of age and BMI on markers of skeletal muscle mitochondrial structure and function. Kerrie Nguyen (Tulane University School of Medicine), Theodore Ciaraldi, PhD, and Robert R. Henry, MD. VA San Diego Healthcare System and University of California, San Diego The skeletal muscle (SkM) of type 2 diabetes mellitus (DM2) patients is known to be structurally and functionally compromised, which is believed to contribute to insulin resistance and enhanced fatigability. The presence of DM2 may involve perturbations in SkM mitochondrial size, number, and efficiency, resulting in the SkM’s inability to properly oxidize glucose and fatty acids. We hypothesized that age and weight may affect mitochondrial structure and function, and that these effects are different in the presence of DM2. To test this we evaluated the expression of proteins involved in mitochondrial biogenesis (mitochondrial transcription factor A, Tfam), structure (porin and optic atrophy 1, OPA1) and activity (citrate synthase, CS) in SkM tissue obtained from non-diabetic (ND) and DM2 human subjects of varying ages (18-64 yrs) and BMIs (20.6-43.0 kg/m2). The two groups did not differ with regard to average age or BMI. In ND there was a tendency (p=0.054) for a negative association between age and Tfam protein content; all other markers were independent of age. However, in DM2 subjects, CS (p=0.0519) and OPA1 (p=0.0433) protein content were positively correlated with age, while there was no such association with Tfam. The only significant relationship between BMI and the mitochondrial markers examined was a positive one seen in ND subjects for the CS/porin ratio (p=0.0246), a surrogate for mitochondrial efficiency, but not in DM2 subjects (p=0.019 between groups). There were no significant differences between the ND and DM2 groups in the mitochondrial structure and function markers examined. In summary, there were modest effects of age and BMI on indicators of SkM mitochondrial structure and function. However, the nature of these effects differed in some instances between ND and DM2 subjects, providing partial support for our hypothesis of responses unique to DM2 subjects to age and BMI. Abstract 74 The role of ER and oxidative stress in overnutrition-induced β-cell neogenesis Tanav Popli (Indiana University School of Medicine, Indianapolis, IN), Wenbiao Chen, Lisette Maddison, Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN Pancreatic β-cells exhibit two mechanisms to compensate for chronically increased blood glucose levels: increased insulin biosynthesis and secretion and increases in total β-cell mass. Individuals with robust β-cell compensatory mechanisms are thought to be protected from developing Type 2 Diabetes, as opposed to individuals whose pancreatic islets have a limited capacity for mass increase. Previous work demonstrates that the increase in β-cell mass is due to the induction of β-cell, however, the specific intracellular events involved in this process are not yet well-characterized. Further work by Dr. Chen’s group points to a role for ER and oxidative stress in β-cell induction. To test the hypothesis that activation of ER and oxidative stress response pathways in pancreatic β-cells of zebrafish larvae plays a role in compensatory β-cell neogenesis, we created 3 different knockout lines of zebrafish, atf3-/-, hsf1-/-, and sirt1-/- to disrupt the ER and oxidative stress response in β-cells exposed to excess nutrition. Differences in β-cell compensatory capacity were assessed by manual quantification of islet β-cell number in 5% yolk fed and unfed control larvae 6 days post fertilization. No significant difference was found between any of the four atf3 larval groups (overfed atf3-/-, 36.1±8.7 cells, n=14, overfed atf3+/- or atf3+/+,36.6±9.2 cells, n =54, unfed atf3-/-, 35.5±6.7, n=18, unfed atf3+/- or atf3+/+, 38.2±8.0, n=47). Overfed hsf1-/- (40.6±5.4 cells, n=17) had significantly more β-cells than unfed hsf1-/- larvae (33.5±6.2 cells, n=7), though β-cell number in overfed hsf1 nonmutants (38.8±7.3 cells, n=27) was not significantly different than unfed hsf1 nonmutants (36.4±6.5 cells, n=61), and no difference was found between mutants and nonmutants in either treatment group. Similarly, the four sirt1 larval treatment groups exhibited no significant differences in β-cell number between genotypes or overfed and underfed sirt1 nonmutants (overfed sirt1+/- or sirt1+/+,41.8± cells, n =30, unfed sirt1+/- or sirt1+/+, 41.0±, n=17), but overfed sirt1-/- larvae (45.4±6.9 cells, n=11) had significantly more β-cells than unfed sirt1-/- larvae (39.9±8.0, n=20). This experiment suggests that knockouts of atf3, hsf1, and sirt1 do not affect β-cell neogenesis. However, the lack of previously documented effects between the control larvae implies more data is needed to definitively draw this conclusion. The possibility of a heterozygous effect on βcell neogenesis that may be confounding the data cannot be discounted, and is being explored. Abstract 75 Ascorbate recycling in human brain vascular pericytes Tarun Jain (Indiana University School of Medicine, Indianapolis, IN), William H. Parker, M. Elizabeth Meredith, Zhi-chao Qu, James M. May, Department of Medicine, Division of Diabetes, Endocrinology, and Metabolism, Vanderbilt University, Nashville, TN One of the major complications of diabetes mellitus is diabetic retinopathy; according to the Centers for Disease Control and Prevention (CDC), 30% of people with diabetes over the age of 40 are affected, with blindness occurring in 5% of those with diabetes. The condition is characterized by loss of vision due to microvascular damage in the retina, which results from oxidative stress caused by high blood glucose concentrations. A chronic increased superoxide concentration will lead to depletion of cellular antioxidants, including Vitamin C (ascorbate), which in turn leads to protein, nucleic acid, and lipid damage. The resulting apoptosis of pericytes is one of the earliest steps in microvascular damage, leading to loss of structural support and increased permeability of the endothelial barrier. Our studies have demonstrated that ascorbate supplementation reduces the permeability of the endothelial barrier, as well as prevents apoptosis in endothelial cells and pericytes. Ascorbate acts as a one-electron donor, and the cell must regenerate ascorbate in order to maintain part of its antioxidant activity. While this reduction or recycling of oxidized ascorbate has been studied in human umbilical vein endothelial cells (EA.hy926), the purpose of this study is to characterize the process in human brain vascular pericytes (HBVP). This would serve both as a measure of ascorbate recycling and total cellular antioxidant reserve. When pericytes were loaded with ascorbate, increasing concentrations of ascorbate were hypothesized to lead to increased antioxidant activity of pericytes. Futhermore, because pericytes should be able to recycle ascorbate, antioxidant activity of pericytes was hypothesized to continue over time, and total antioxidant activity was hypothesized to increase with relation to time. Recycling and antioxidant activity of ascorbate was measured by ascorbate-dependent ferricyanide reduction; cells cultured in 6-well plates were treated with varying concentrations of ascorbate, after which they were treated with 500 µM potassium ferricyanide. This assay is an indirect measure of ascorbate activity through the transfer of an electron from ascorbate to ferricyanide via a transmembrane protein. In both EA.hy926 and HBVP, an increase in the concentration of the initial ascorbate treatment lead to an increase in the concentration of ferrocyanide, the reduced form of ferricyanide. However, endogenous ascorbate was present in HBVP, and thus these cells were able to reduce a higher concentration of ferricyanide to ferrocyanide both initially and overall over time, regardless of initial ascorbate treatment. In order to account for this difference, both EA.hy926 and HBVP were loaded with 250 µM ascorbate, which both the literature and our studies have shown should saturate the cells. Interestingly, while the initial increase in ferrocyanide concentration was higher in HBVP cultures, the rate of increase slowed down over time relative to EA.hy926 cultures. Nevertheless, the increase in the concentration of ferrocyanide in both cell groups over time indicated that ascorbate activity had been maintained. These results suggest that pericytes act similarly to endothelial cells in regulation of intracellular ascorbate concentrations, although the rates of reduction or recycling may vary. Future experiments would focus on other aspects of ascorbate recycling, such as reduction of dehydroascorbate (DHA), as well as the behavior of other antioxidants within pericytes. Results from these studies can be used to elucidate the mechanisms behind ascorbate’s ameliorative effects on the maintenance of the blood-retinal barrier under oxidative stress, both in vitro and in vivo. Abstract 76 Replication of association of a loss-of-function mutation in HSL with metabolic traits in the Amish Donique Parris (University of Maryland School of Medicine), Coleen Damcott PhD, Alan Shuldiner MD, University of Maryland School of Medicine Lipolysis is a metabolic pathway that regulates energy homeostasis through degradation of intracellular triacyglycerol (TAG) and release of fatty acids for use as energy substrates or lipid mediators in cellular processes. Both excess (e.g. obesity) and insufficient (e.g. lipodystrophy) white adipose tissue (WAT) stores, the primary energy depot, are associated with insulin resistance and increased risk for type 2 diabetes. Thus, maintenance of normal WAT function is critical in preserving whole body insulin sensitivity and energy homeostasis. Hormone-sensitive lipase (HSL) is a key lipolytic enzyme with high affinity for diacylglycerol, but which also hydrolyzes TAG, cholesterol esters, and retinol esters. Recently, a 19-base pair deletion in the LIPE gene, which encodes HSL, was identified in the Amish. The mutation was associated with absence of the HSL protein, dyslipidemia, hepatic steatosis, systemic insulin resistance, and type 2 diabetes. The objective of this study was to identify additional carriers of the HSL mutation in a second Amish cohort and determine its effects on metabolic traits, such as fasting total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol. We genotyped 1,076 DNA samples from Amish participants in the Wellness Study and identified 33 heterozygote carriers (ID) and 1 homozygote carrier (DD). Association analyses were then performed to determine which metabolic traits were significantly associated with the HSL mutation while taking relatedness of the participants into account. Of the metabolic traits analyzed, triglycerides and HDLs were the only two that had a statistically significant association with the HSL mutation. The presence of the D allele was associated with higher serum triglycerides (p=0.009) and lower HDL cholesterol levels (p=0.03). These data provide replicating evidence of the reported effects of the HSL mutation on metabolic traits and further highlight the importance of HSL in maintaining systemic lipid homeostasis. Abstract 77 A peptide barcoding strategy to study insulin secretion from mixed cell populations. Pooja Prasad (University of Rochester), Thomas Hennings, Gregory Ku, University of California San Francisco While a fraction of the numerous genes expressed in beta cells have been shown to be involved in insulin secretion, we hypothesize that many more unidentified genes regulate insulin secretion. A better understanding of insulin secretion has valuable applications for conversion of an embryonic stem cell to an insulin-secreting cell and for drug targeting. Pooled RNA interference and Cas9/CRISPR screens would allow the identification of novel genes involved in insulin secretion. Towards that aim, our study’s goal was to develop and test a peptide barcoding strategy to measure the amount of insulin being secreted from a mixed population of cells with unique genes knocked down. We hypothesized that adding engineered peptide sequences detectable by mass spectroscopy to C-peptide would allow us to monitor insulin secretion. To test this system, we produced lentivirus containing a unique 20 amino acid tag and a FLAG epitope tag appended to C-peptide and infected mouse pancreatic beta cells. Infection rates were measured using droplet PCR from genomic DNA. We then performed a western blot on cell lysates using an anti-FLAG antibody to detect the tagged C-peptide. Our western blot showed a FLAG reactive band whose size is consistent with proinsulin, suggesting that the presence of these tags may inhibit insulin maturation. To test whether the band of unexpected size is due to incomplete processing, we are constructing a plasmid containing only the tagged C-peptide, rather than tagged C-peptide within the context of preproinsulin. Before sending cells to mass spectroscopy for detection of peptide tags and utilizing this method for studying insulin secretion, we must ensure that the peptide sequences serving as a barcode are not affecting secretion of insulin. Abstract 78 Using Human Embryonic Stem Cells to Derive Pancreatic Beta Cells In Vitro Deepti Pujare (New York Medical College), Audrey V. Parent, Holger A. Russ, Matthias Hebrok, University of California- San Francisco A novel approach to treating diabetes is to transplant hESC-derived beta cells into patients. Current methods of treating diabetes include pancreatic transplantation or infusion of pancreatic islets into the bloodstream. However, these methods have various limitations such as the lack of sufficient donors. Our hypothesis is that we can create functional pancreatic beta cells in vitro from human embryonic stem cells (hESCs). The hESC line MEL1/INS-GFP was differentiated in vitro using two different protocols (protocol 1 and 2). Immunofluorescence and flow cytometry were used to characterize and compare the cells generated using both protocols at various stages of differentiation. Both methods generated different proportions of pancreatic progenitors as well as cells that express markers characteristic of beta cells (C-peptide, PDX1, NKX6.1, NKX2.2) and other pancreatic cell types (Glucagon, Somatostatin). Importantly, cells differentiated using protocol 2 had more cells with characteristics of mature beta cells (expressed important transcription factors such as NKX6.1 and PDX1 and did not express other hormones). The proliferation of pancreatic progenitors and beta-like cells was also assessed using Ki67. Our study suggests that hESCs can be induced to differentiate into more mature beta-like cells in vitro using an improved differentiation protocol. Abstract 79 ApoE Receptor 2 (LRP8) Cleavage Product Dependent Transcriptional Programs of Learning and Memory Hyun Chun (Stony Brook Medicine), Patricia Montilla Pérez, Francesca Telese, Michael Geoff Rosenfeld Department of Medicine, UCSD, La Jolla, CA Neuroimaging studies on diabetes type 2 patients show increased cortical and hippocampal atrophy correlated with cognitive dysfunction, independent of hypertension. Interestingly, preliminary studies show that the ApoE/reelin receptor (LRP8) is a new substrate of gammasecretase activity that releases an intracellular domain (ICD) upon binding of ligands. LRP8 knockout mice show deficits in hippocampal-dependent associative learning which prompts the investigation behind the molecular mechanisms of how LRP8-ICD affects cognitive abilities in the brain. To test the hypothesis that the intracellular domain of LRP8 plays a direct role in the regulation of transcriptional programs related to learning and memory formation, we combine global genomic approaches, such as RNA-seq and ChIP-seq, to behavioral paradigms using LRP8 KO mice as an in vivo model. Global genomic approaches are particularly useful in investigating transcriptional programs in an unbiased way. By RNA-seq, LRP8 KO mice have decreased expression of learning and memory related genes using gene ontology categorization. To further understand how the differential expression and cognitive dysfunction phenotype is mediated by LRP8 biochemically, subcellular protein isolation and Western blots were used to localize the gamma secretase cleavage product of LRP8 to the nucleus. Supporting our hypothesis, LRP8-ICD shows transcriptional activity when tested in a Gal4 luciferase assay. To investigate the genome-wide binding pattern of LRP8-ICD, a BLRP/BirA expression system in transgenic mice was combined with strepdavidin-ChIP-seq for LRP8-ICD. Though ChIP-seq results are pending, these data suggest that the gamma secretase cleaved intracellular domain of ApoE receptor influences transcriptional programs of learning and memory through an epigenomic regulation. These observations also have clinical implications for better understanding cognition in the context of disease such as diabetes and dyslipidemia while providing an opportunity for new therapeutics and biomarkers. Abstract 80 Bariatric surgery may increase expression of IR-beta and Akt in subcutaneous adipocytes. Ryan Beardsley (University of Illinois College of Medicine), Matt Piron, Milad Abusag, Matthew Brady, University of Chicago Bariatric surgery has been shown to improve insulin sensitivity in obese, insulin resistant patients as soon as 2 weeks after surgery, before any significant long term weight loss has occurred. This shows us that a significant amount of the anti-diabetic effects of bariatric surgery occurs in a mechanism independent of that induced solely by a decrease in adiposity, but the intracellular changes and molecular mechanisms that produce this benefit are not yet clear. We hypothesized that bariatric surgery increased expression of the insulin receptor beta subunit (IRbeta) and Akt (also known as PKB, or Protein Kinase B), two vital proteins in the insulin signaling pathway, supposing that such a change could explain at least part of the significant changes in insulin sensitivity that occur. Subcutaneous adipose tissue biopsies were taken approximately two weeks before and two weeks after bariatric surgery in 8 patients (all of whom had either a Roux-en-Y gastric bypass or duodenal switch procedure) and subjected to insulin signaling assays (being stimulated for 10 min by increasing concentrations of insulin) before being Western blotted for IR-beta, total Akt (tAkt), and S473 phosphorylated Akt (pAkt, a strong indicator of activation of the insulin signaling pathway), with beta-actin being blotted as a loading control. Densitometry was performed on the resultant images collected from the antibody visualization, and ratios of pAkt:tAkt, pAkt:beta-actin, tAkt:beta-actin, and IR-beta:beta-actin were calculated. The mean of each ratio was higher post-op than pre-op, but none were statistically significantly higher when taken as a whole. The mean post-op tAkt:beta-actin (1.82 ± 0.76) was higher than the pre-op tAkt:beta-actin (0.97 ± 0.13), and mean post-op IRbeta:beta-actin (2.08 ± 0.98) was higher than the pre-op IR-beta:beta-actin (1.17 ± 0.15), but due to the high variability from one patient to the next the post-op increases were not significant (p>0.05). The 0.5nM insulin stimulation post-op samples, though, had a significantly higher pAkt:tAkt than the 0.5nM insulin stimulation pre-op samples (p=0.032). For all other insulin stimulation concentrations the pAkt:tAkt and pAkt:beta-actin ratios were also at least greater post-op than pre-op, although no others were significantly higher. The variation between these patients prevents any solid conclusions from being drawn from this data, but it does suggest to some degree that expression of IR-beta and Akt in subcutaneous adipocytes generally increases after bariatric surgery. The apparent post-op trends towards increased expression of IR-beta and Akt, along with more dramatic increases in pAkt:tAkt, could reasonably be expected to be more pronounced in patients who are considered fully diabetic before bariatric surgery. Abstract 81 An Overview of the Recruitment Strategies for GRADE (Glycemia Reduction Approaches in Diabetes: A Comparative Effectiveness Study) Verona Young (Stony Brook University School of Medicine); Patricia Kringas RN, MA, CDE; Camille Hausheer BS; Diabetes And Endocrinology Research Center (DERC) Columbia University Medical Center Over the past century, type 2 diabetes has become an epidemic that is currently affecting up to 1 in 3 Americans. According to recent estimates, the incidence of type 2 diabetes increased to 1.9 million while the prevalence currently exceeds 24.5 million. When deciding which medication is most optimal at lowering and maintaining good glycemic control, practitioners often encounter a major challenge due to the surplus of glucose-lowering medication available on the market. The Glycemia Reduction Approaches in Diabetes: A Comparative Effectiveness (GRADE) Study plans to address this issue by comparing the longevity outcomes of four commonly used drugs through a randomized, open label clinical trial. Patients will be given the maximum tolerated dose of metformin then randomized to receive one of the following medications: the sulfonylurea glimepiride, the DPP-4 inhibitor sitagliptin, the GLP-1 agonist liraglutide, and the basal insulin glargine. Recruitment for clinical trials demands a multi-faceted approach. The Naomi Berrie Diabetes Center (NBDC) has embarked on multiple approaches including pre-screening the diabetes and nutrition classes, advocating that subjects become study ambassadors, fostering referrals from primary care physicians, building connections with neighborhood clinics and hosting health fairs and other community events. In-person recruitment, partnering with the American Diabetes Association, and publicizing in the media (i.e. creating newspaper ads and a NBDC Facebook newsletter) are amongst some of the other recruitment strategies implemented. This summer, we have focused on increasing daily pre-screening calls by extracting participant contact information from the center database as well as on-site recruitment by hosting a weekly Diabetes Alert Day in the main hospital. These strategies resulted in an increase in the overall recruitment numbers for the site, which we predict will continue to drive participant enrollment during the months to come. Abstract 82 Continuous glucose monitoring, hunger and food intake in non-obese, obese and diabetic free-living individuals. Janice S. Kim (Washington University School of Medicine), Janice J. Hwang M.D., Renata Belfort-De Aguiar M.D. Ph.D., Robert Sherwin M.D., Yale School of Medicine Background: Obesity and diabetes are serious problems in the U.S. that can be ameliorated through diet and exercise. Previous research has found that both rapid declines as well as absolute low levels of blood glucose are associated with increased hunger and food consumption; however, no study thus far has looked into the glucose profiles of free-living nonobese (N-OB), obese (OB) and diabetic (DM) individuals in relation to hunger and food intake. Hypothesis: We hypothesized that larger glucose declines, as opposed to absolute low levels of glucose, will be associated with increased hunger, increased calorie consumption and subsequent consumption of more calorie-dense foods. Methods: Eight subjects ((6 F/ 2 M), age 31.4±5.5, BMI 27.6±5.8, 2 N-OB/ 4 OB/ 2 T1DM patients) wore a continuous glucose monitor (Dexcom CGM) for five days while keeping a food diary of hunger ratings (from a scale of 1-10, 1 being none and 10 being very hungry) prior to meals and amounts of food consumed. Results: Preliminary findings suggest that mean subjective hunger ratings were 5.9±2.1 and that prior to each meal, mean glucose peaks were 132±46 mg/dL and mean glucose nadirs were 85±19 mg/dL. Hunger ratings were positively associated with glucose peaks (ρ=0.25, P=0.04) but not associated with glucose nadirs (P=NS). Glucose declines correlated with subsequent consumption of fat (ρ=0.34, P=0.005) and protein (ρ=0.32, P=0.009), but not carbohydrates (P=NS). Conclusion: In conclusion, increasing subjective hunger levels are more related to pre-prandial glucose peaks as opposed to nadirs and furthermore, declines in glucose are associated with the macronutrient content of subsequent meals. Our findings suggest that glycemic excursions following a meal may play a role in hunger and food choices for the subsequent meal. Abstract 83 Effect of reduced cardiac triglyceride turnover on insulin signaling and glucose homeostasis. Ngoc Bao Nguyen (University of Washington), Nathan Roe, PhD, Rong Tian, MD, PhD, Mitochondria and Metabolism Center, University of Washington Increased myocardial triglyceride (TG) accumulation is observed in patients with diabetes and obesity. However, recent evidence implicates TG turnover rate, rather than total TG levels, as a critical factor in cardiac metabolism and function. Preliminary findings in our lab demonstrate that a reduction in TG turnover increases fatty acid oxidation while decreasing glucose oxidation, although the mechanism governing this switch remains unclear. Altered fatty acid and TG metabolism has been implicated in insulin resistance; we therefore hypothesized that reducing TG turnover in the heart would reduce insulin sensitivity and lead to reduced glucose oxidation. To test our hypothesis, inducible cardiac specific diacylglycerol:acyltransferase 1 (DGAT1) deficient mice (iKO), a model of reduced TG turnover, were fasted for 16 hours and exposed to either acute insulin stimulation (10 mIU/g body weight, i.p., tissues harvested 20 minutes postinjection) or chronic high fat diet feeding (5 weeks on diet containing 60% kcal fat). The insulin signaling cascade in the heart was then evaluated via western blot. Compared to baseline measurements, insulin-stimulation lowered blood glucose to similar levels in both control and iKO animals. No difference in insulin-stimulated Akt or GSK-3β phosphorylation was observed in iKO compared to controls. Additionally, Akt and GSK-3β phosphorylation was unchanged in response to high fat feeding in both control and iKO animals, suggesting a minimal effect of DGAT1 on insulin signaling. Lactate levels in the heart were significantly increased with insulin stimulation in control animals, however this increase was not observed in iKO, suggesting a decrease in insulin stimulation of glycolysis (n = 3-6/group, p < 0.05). Similarly, glycogen content in control animals increased 2.6-fold with insulin over saline treatment, but no increase was seen in iKO (n = 3-6/group, p < 0.01). Collectively, our data suggests that reduced TG turnover does not significantly alter insulin signaling, but may affect insulin-stimulated glucose uptake and/or metabolism, providing a different mechanism through which TG turnover regulates glucose homeostasis. Abstract 84 The role of α2A adrenergic receptors via Gβγ-SNARE inhibition in exocytotic glucosestimulated insulin secretion. Charles Jose (University of Nevada School of Medicine), Zack Zurawski, Heidi Hamm, Vanderbilt University Gi/o-coupled GPCRs function through Gβγ subunits to inhibit exocytosis. This mechanism works via: 1) inhibition of calcium entry by binding to the voltage-dependent calcium channel and 2) Gβγ binding directly to the SNARE complex. α2A adrenergic receptors (α2AAR) utilize Gβγ-SNARE inhibition to reduce glucose-stimulated insulin secretion (GSIS) in pancreatic beta cells. Inhibited glucose-stimulated insulin secretion is an early sign of type 2 diabetes (T2D), a condition affecting millions throughout the world. We hypothesize that α2A adrenergic receptor signaling is overactive in T2D, and that this over activity is mediated via the interaction of Gβγ with SNAREs. To test our hypothesis that over-active inhibition of insulin secretion by the sympathetic system exacerbates T2D through decreased GSIS, we performed static culture assays using isolated pancreatic islet cells from lean and diet-induced obese (DIO) mice. Islets were incubated in 5.6mM and 16.7mM glucose static cultures with combinations of epinephrine (1uM); VU0451223, a Gβγ-SNARE inhibitor (50uM); and tolbutamide (1uM). GSIS was measured via the AlphaLISA insulin assay. In lean mice, islets exhibited reduced GSIS when incubated with epinephrine. Islets incubated in VU0451223 enhanced tonic GSIS but did not reverse the antagonistic effects of epinephrine on insulin secretion. Tolbutamide and VU0451223 did not enhance GSIS in an additive manner. In DIO mice, islets exhibit nondifferential GSIS under varying glucose concentrations. The DIO mice used in this experiment may have already experienced signs of beta cell failure, a condition that occurs in late-stage T2D. The impaired function of VU0451223 in inhibiting Gβγ-SNARE interactions may be attributed to either reduced potency or structural defects that prevent the small molecule inhibitor from fully penetrating the pancreatic islet cell. These data suggest that further screening and optimization of Gβγ-SNARE inhibitors is necessary to achieve the desired effect of maintaining tonic GSIS while simultaneously reversing the antagonistic effects of α2A adrenergic receptors on exocytosis. Suppressing α2AAR -mediated Gβγ-SNARE inhibition can enhance insulin secretion while maintaining the critical physiological role of insulin secretion by glucose and other modulators. Understanding the pathophysiological relevance of Gβγ-SNARE inhibition in exocytotic GSIS can identify novel therapeutic targets to improve T2D management. Abstract 85 Ventral Tegmental Area µ-Opioid Receptors are Important in Feeding Behavior in Obese Rats. Joel Amezquita (Charles Drew University/UCLA), Candace M. Reno, and Robert Sherwin, Yale University Mu opioid receptors (µORs) are widely expressed in the brain and are involved in regulation of feeding and food reward. A canonical two neuron model of reward posits that brain µ-opioid receptor activation produces reward by disinhibiting midbrain ventral tegmental area (VTA) dopamine neurons through inhibition of local GABAergic interneurons leading to increased dopamine neurotransmission and feelings of reward. Diet-induced obese (DIO) rats have increased expression of µORs in the VTA, as well as absent dopamine and GABA responses to re-feeding. It is hypothesized that knockdown of µOR expression in the VTA will decrease food intake, preference for highly palatable food, and weight gain by restoring the dopamine and GABA responses. To test this hypothesis, DIO adult, male, Sprague-Dawley rats underwent stereotaxic surgery to induce µOR knockdown in the VTA through use of an adeno-associated siRNA virus (AAV). Control groups received a random scrambled RNA AAV. Rats were placed into one of six groups: 1) normal chow diet with control virus (NC-ScrAAV; n=3), 2) normal chow diet with µOR knockdown (NC-µORKD; n=4), 3) high fat diet with control virus (HFD-ScrAAV; n=2), 4) high fat with µOR knockdown (HFD-µORKD; n=4), 5) a choice of both diets with control virus (Choice-ScrAAV; n=4), and 6) choice of both diets with µOR knockdown (Choice-µORKD; n=4). The normal chow diet was 70% carbohydrates, 10% fat and 3.85 kcal/g. The HFD was 35% carbohydrates, 45% fat and 4.73 kcal/g. Food intake and body weight were measured twice weekly for approximately two weeks before and after surgery. Daily body weight change decreased in all groups from basal to two weeks after virus injection: NC-ScrAAV (3.29 ± 0.3 vs 1.04 ± 0.9%), NC-µORKD (3.12 ± 0.2 vs 1.82 ± 0.2 %), HFD-ScrAAV (3.92 ± 0.2 vs 1.90 ± 0.1%), HFD-µORKD (4.61 ± 0.4 vs 2.17 ± 0.2%), Choice-ScrAAV (3.68 ± 0.6 vs 1.39 ± 0.3%), and Choice-µORKD (3.53 ± 0.3 vs 1.13 ± 0.37%). There were no differences in body weight change among the groups. Daily calorie intake two weeks after virus injection decreased compared to basal in the NC-µORKD group (82 ± 5 vs 60 ± 9 kcal/day; p < 0.04) and trended to be reduced in the HFD-µORKD (96 ± 5 vs 81 ± 7 kcal/day; p < 0.11) and Choice-µORKD groups (95 ± 8 vs 79 ± 7 kcal/day; p < 0.12). There were no differences in food preference for normal chow or HFD between the two choice groups. These results indicate that µOR knockdown in the VTA has no effect on body weight during the short time given in the protocol and that overall food intake is decreased in the presence of normal chow, HFD, or choice of both. Surprisingly, µOR knockdown had no effect on preference for HFD. This data suggests that µORs play an important role in satiety and reduction of food intake and may have implications for obesity treatment and prevention. Abstract 86 Assessing Correlates and Inter-rater Reliability of the Surprise Question in Non-dialysis Dependent CKD. Megan Mizera (University of Central Florida College of Medicine), T. Alp Ikizler, Khaled AbdelKader, Vanderbilt University Older adults with severe chronic kidney disease (CKD) frequently receive care at the end-of-life that conflicts with their values and preferences. Prognostic uncertainty is a critical barrier that prevents providers from engaging patients in timely prognostic and advance care planning discussions. The surprise question (SQ), “Would you be surprised if this patient died within 12 months?” has been advocated as a practical tool to identify patients at high risk for short term mortality; however, it’s validity and reliability in non-dialysis dependent CKD has not been explored. We hypothesized the SQ would associate with cardiovascular disease (CVD), congestive heart failure, comorbidity burden, and low albumin levels and have a moderate to high inter-rater reliability between attendings and fellows. We conducted a cross-sectional analysis of a prospective cohort study at Vanderbilt’s outpatient nephrology clinic. We asked enrolled nephrology providers the SQ immediately following an eligible patient visit. Responses were recorded using a simple dichotomous ‘Yes’/’No’ and a 5-point Likert Scale with lower scores indicating less surprise. Patient characteristics were abstracted from the electronic health record. SQ responses associated with multiple clinical markers in univariable analysis (n=76) including age (ß=-0.46, standard error [SE]=0.18, p=0.01), CHF with decreased LVEF (ß=-1.94, SE=0.54, p=0.001), hospitalization in the prior year (ß=-0.81, SE=0.31, p=0.01), Charlson Comorbidity Index (ß=-0.15, SE=0.06, p=0.01), hemoglobin (per 1g/dL increase) (ß=0.43, SE=0.11, p<0.001), and albumin (per 1g/dL increase) (ß=0.97, SE=0.44, p=0.03). The dichotomous SQ demonstrated moderate inter-rater reliability (kappa = 0.42, SE=0.17, p=0.008, n=29). These results suggest that the SQ response associates with known predictors of mortality in non-dialysis dependent CKD. In addition, the SQ appears to have moderate interrater reliability among nephrology providers at different stages in training/practice. Abstract 87 Elementary Students' Knowledge, Preferences, and Skills related to Food, Nutrition, and Cooking in Chicago Brooke Scheidemantle (Sidney Kimmel Medical College at Thomas Jefferson University), Alexandra De Sorbo-Quinn (Pilot Light), Michael Quinn PhD, Althera Steenes, Deborah Burnet MD (University of Chicago) Background: Limited data exists regarding cooking education programs to teach healthy nutrition to schoolchildren. “Pilot Light” is an existing program which works with Chicago Public School leaders to build food and nutrition education into regular classroom lessons like math, science, and English language arts, with cooking demonstrations by Chicago’s leading chefs. Lessons align with the Common Core State Standards and Chicago Public School Wellness Standards for nutrition. We used focus groups to understand baseline knowledge, preferences, and skills related to food, nutrition and living a healthy lifestyle for third through fifth graders in Chicago. These insights will be used to develop and expand the Pilot Light curriculum for implementation in additional schools. Methods: We developed partnerships with two summer camps in order to assess children’s knowledge, preferences and skills. We conducted focus groups with children attending a YMCA summer program on Chicago’s North Side and a Chicago Park District summer camp on Chicago’s South Side. Research staff conducted focus groups using a semistructured guide developed to elicit children’s knowledge of “healthy” foods, experience and interest in cooking, and influences on their eating behaviors. Focus groups were audio taped, professionally transcribed, and analyzed for themes. Results: 22 third- through fifth-grade children enrolled and participated in the study. Four focus groups (two at each site) were conducted with 4-7 participants per group. Participants comprised 11 girls and 11 boys, including 10 African Americans, 6 Latinos, and 6 Caucasians. Preliminary themes are as follows: children are knowledgeable and interested with food and nutrition, many children are engaged in cooking at home, and parents/media/role models influence the child’s food choices. Formal analysis is currently underway. Conclusions: This study provides insight into the perspectives and experience regarding food and nutrition among school-aged children in summer camps in Chicago. The beliefs, preferences and practices of these children can become potential leverage points in curriculum development and implementation for teaching nutrition and cooking skills to school children. Abstract 88 Calcium-activated potassium channels modulate pancreatic a-cell Ca2+ influx and glucagon secretion Kenneth R. Verlage1, Prasanna K. Dadi2, and David Jacobson2 1. Texas Tech University Health Sciences Center School of Medicine 2. Department of Molecular Physiology and Biophysics, Vanderbilt University Introduction: Ca2+ influx into pancreatic a-cells through voltage-dependent Ca2+ channels (VDCCs) is required for glucagon secretion and regulatory mechanisms of this process become defective during the pathogenesis of diabetes mellitus. Potassium channels are the primary regulators of membrane potential, and thus, regulate the activity of VDCCs. However, the roles of Ca2+ activated potassium channels (KCa2+) during a-cell Ca2+ entry and glucagon secretion have not been determined. Understanding how KCa2+ channels regulate glucagon secretion has the potential to illuminate therapeutic targets that could be utilized to reduce hyperglucagonemia and hyperglycemia in diabetic patients. Hypothesis: KCa2+ channels hyperpolarize the a-cell membrane potential which limits Ca2+ entry and glucagon secretion. Methods: Ca2+ imaging was performed on mouse and human pancreatic islets and single cells in low (1mM) and high (11mM) glucose conditions with and without inhibitors of small conductance (SK), intermediate conductance (IK) or large conductance (BK) KCa2+ channels (apamin, TRAM34 and iberiotoxin respectively). Glucagon secretion was assayed under similar conditions. Lastly, immunofluorescent imaging was used to detect IK expression in mouse and human pancreatic sections. Results: In mouse single a-cells, inhibition of SK and BK channels enhanced a-cell Ca2+ influx under low glucose conditions, increasing Ca2+ levels by 36.18% (p < .01) and 42.03% (p < .001) respectively. SK and BK channels also limited low glucose induced Ca2+ influx into a-cells within clusters of islet-cells and when inhibited increased Ca2+ influx (by 23.72% (p < .05) with SK channel inhibition and 27.31% (p < .05) with BK channel inhibition). IK channels are also expressed in pancreatic islets and we find that mouse and human pancreatic sections show the highest expression of IK channels in glucagon positive a-cells. Interestingly, inhibition of IK channels caused significant increases in a-cell Ca2+ influx under both low and high glucose conditions. IK channel inhibition caused a 41.76% (p < .001) increase for mouse single a-cells in low glucose and a 28.76% (p < .05) increase in high glucose. Human single a-cells mirrored this behavior, showing a 31.66% (p < .05) increase in Ca2+ influx in low glucose with IK channel inhibition; while showing a 33.28% (p < .05) increase in Ca2+ influx in high glucose with IK channel inhibition. Summary: SK, BK and IK channels limit Ca2+ influx during conditions that stimulate a-cell glucagon secretion. a-cell IK channel activity is unique in that it also helps to maintain glucose induced inhibition of Ca2+ influx. Thus, KCa2+ modulation of Ca2+ influx will influence glucose regulated glucagon secretion. Abstract 89 AdipoRon’s differential roles in autophagy and inflammation in human aortic endothelial cells exposed to excess nutrients Feenalie Patel (University of Tennessee Health Science Center), Dr. Karen Weikel, Dr. Neil Ruderman, Boston University Medical Center The risk for cardiovascular diseases such as atherosclerosis is nearly tripled in patients with type 2 diabetes compared to healthy individuals. The high concentrations of circulating glucose and fatty acids that are observed in these patients have been shown to increase oxidative stress and inflammation, and impair autophagy in vascular endothelial cells, thus contributing to atherogenesis. Adiponectin is a circulating cytokine secreted by adipose tissue that has been shown to attenuate nutrient-induced increases in oxidative stress and inflammation, as well as restore autophagy. In a mouse model of diet-induced diabetes, stimulation of adiponectin signaling using an adiponectin receptor agonist, AdipoRon, activated AMP-activated protein kinase (AMPK) and reduced oxidative stress and inflammation in the liver, muscle and adipose. These mice were also more insulin-sensitive and lived longer than controltreated mice. To determine whether AdipoRon can also protect human endothelial cells from the glucose- and fatty acid-mediated stresses associated with type 2 diabetes, we treated primary human aortic endothelial cells (HAECs) with AdipoRon, and evaluated its effects on autophagy and inflammation. AdipoRon increased expression of phosphorylated AMPK and acetyl CoA carboxylase (ACC) in HAECs, confirming its role as an AMPK activator. AdipoRon also activated autophagy in HAECs incubated in control conditions (5 mM glucose, 0 mM palmitate), but not in HAECs incubated in media containing excess nutrients (25 mM glucose + 0.4 mM palmitate). Interestingly, reducing glucose concentrations in the media of these nutrient-laden HAECs restored induction of autophagy by AdipoRon, suggesting that nutrient-induced changes in adiponectin receptor signaling may be reversible. Furthermore, AdipoRon reduced adhesion of THP-1 monocytes to HAECs incubated in control and hyperglycemic conditions. These data suggest that under control conditions, AdipoRon can activate AMPK, induce autophagy and reduce inflammation in HAECs. While AdipoRon’s actions on autophagy were not observed in the presence of excess nutrients, its anti-inflammatory actions were still observed under hyperglycemic conditions. In addition to its previously described actions in the liver, muscle and adipose, our data suggest that AdipoRon may also have a potential therapeutic role in the reduction of endothelial cell inflammation and atherogenesis among type 2 diabetes patients. Abstract 90 Assessing the Correlation between the Levels of Immunoglobulin and the Gut Microbiota Marie G. Clancey-Rivera (Ponce School of Medicine), Changyun Hu, Li Wen, Department of Internal Medicine, Division of Endocrinology, Yale University School of Medicine, New Haven, CT Obesity is a worldwide pandemic health issue with increasing prevalence in children and adults, which is the result of the interplay between environmental factors and genetic susceptibility. Previous studies imply that different Immunoglobulin isotypes may play different roles in dietinduced-obesity (DIO) and a higher ratio of Firmicutes/Bacteroidetes is also correlated with obesity. Activation induced deaminase (AID) plays an essential role in generating different immunoglobulin isotypes. To test the hypothesis that the presence of immunoglobulin affects gut bacteria, we profiled the immunoglobulin in sera and feces samples from 1) AID-/- C57B/6 mice that were naturally delivered and reared by AID+/- C57B/6 mom and 2) AID-/- C57B/6 mice that were naturally delivered by AID-/- C57B/6 mom but fostered by C57B/6 mom. Simultaneously, the ratio of Firmicutes/Bacteroidetes in feces samples was determined by qPCR. The result indicated that AID-/- mice could acquire immunoglobulins from mom through milk. Interestingly, the half life of IgG from sera of AID-/- babies from AID+/- mom (~3 week) is longer than that of B6 mom fostered AID-/- mice (~1 week). IgM was the main immunoglobulin detected in feces of AID-/- mice. In correlation to the immunoglobulin profile in feces, the ratio of Firmicutes/Bacteroidetes in the feces of B6 mom fostered AID-/- mice was significantly increased over time in comparison with AID+/- mom’s AID-/- babies. In conclusion, immunoglobulin plays a role in high fat diet induced obesity through the modulation of gut bacteria. Abstract 91 The Extracellular Matrix Component Laminin α4 Modulates Insulin Secretion Zaheen Rabbani (University of Louisville School of Medicine), Michael J. Peters, Joshua R. Willard, Rebecca L. Hull, VA Puget Sound Health Care System and the University of Washington, Seattle, WA Islet endothelial cells synthesize laminin, a component of the extracellular matrix (ECM). Laminin is a trimeric glycoprotein comprised of α, β, and γ subunits. Laminins have been implicated in facilitating insulin release, and signal through integrin receptors expressed by β cells. We have observed decreased expression of laminin-411 (α4, β1, γ1) in islets from diabetic C57BL/6.db/db mice. In addition, our data show a 45% decrease in expression of laminin α4, but no change in β1 and γ1, in islet endothelial cells cultured in high glucose. Signaling of the trimeric laminin glycoprotein requires binding of its α subunit to cell-surface integrins. Thus, we hypothesize that decreased laminin α4 expression by islet endothelial cells may contribute to decreased glucose-stimulated insulin secretion, as seen in type 2 diabetes.To test our hypothesis, immortalized mouse islet endothelial MS-1 cells were transfected with siRNA specific for laminin α4 (lamα4-i; 0.1 - 20 nM), scrambled oligonucleotide control (scr; 10 nM) or untreated control (n=3 for all conditions). We collected RNA and pre-conditioned media (CM) from our MS-1 cell cultures. mRNA levels of laminin-411 subunits (α4, β1 and γ1) were measured using quantitative PCR, normalized to the housekeeping gene cyclophillin and expressed relative to untreated control. C57BL/6 mouse islets were exposed to CM-lamα4-i or CM-scr for 48 hours, after which basal and glucose-stimulated insulin secretion were measured (n=1). A dose-dependent knockdown of laminin α4 was observed in lamα4-i-treated MS-1 cells, with 10 nM lamα4-i resulting in a 65% decrease of laminin α4 expression relative to scr (0.2±0.1 vs. 0.7±0.1; n=3, p=0.09). In contrast, laminin β1 mRNA was not changed (0.9±0.2 vs. 0.7±0.2) while laminin γ1 mRNA was somewhat decreased (0.7±0.1 vs. 1.1±0.1). Exposure of islets to CM-lamα4-i resulted in an approximately 33% reduction in glucose-stimulated insulin secretion relative to CM-scr.Based on these findings, we conclude that laminin α4 may be necessary for normal glucose-stimulated insulin secretion. Thus, decreased laminin α4 in the islet could contribute to impaired insulin secretion in type 2 diabetes. Furthermore, increasing islet laminin α4 in type 2 diabetes could be a therapeutic approach to improve β-cell function. Abstract 92 Changes in measures of physical function in obese type 2 diabetes patients after Rouxen-Y gastric bypass surgery vs. intensive diabetes and weight management: results of the SLIMM-T2D study Jennifer Panosian (Tufts University School of Medicine), Su-Ann Ding, M.D., Donald C. Simonson, M.D., M.P.H., Sc.D., Florencia Halperin, M.D., Marlene Wewalka, M.D., Kathleen Foster, R.N., Katherine Kelly, B.A., Ann Goebel-Fabbri, Ph.D., Osama Hamdy, M.D., Ph.D., Kerri Clancy, R.N., David Lautz, M.D., Ashley Vernon, M.D., Allison B. Goldfine, M.D., Joslin Diabetes Center Physical activity is an integral part of diabetes and weight management, however changes in physical functioning post-bariatric surgery compared with lifestyle education and group training is not well understood. This sub-analysis of the SLIMM-T2D trial seeks to compare measures of physical function in obese patients with type 2 diabetes who receive either Roux-en-Y gastric bypass (RYGB) or intensive medical diabetes and weight management (IMWM). We randomized 38 obese T2D (15 male / 23 female; BMI 36.3±3.4 kg/m²; age 52±6 yrs; HbA1c 8.5±1.3%) to RYGB (n=19) or IMWM (n=19). Changes in self-reported physical activity, health status (SF-36), impact of weight on quality of life (IWQOL), and 6-minute walk test metrics were recorded. After 2 years, RYGB had substantially greater weight loss and lowering of HbA1c, blood pressure, and lipids than IMWM. Self-reported exercise increased early in the IMWM group but was not sustained, while both cohorts improved to a similar magnitude at 2 years. Changes in SF-36 physical health did not differ between RYGB and IMWM at 2 years, however both were similarly improved from baseline at this time point. IWQOL total and its major component sub-score of physical function improved more in RYGB after 2 years, and this change in IWQOL directly related to greater weight loss. 6 minute walk test distance and oxygen saturation increased similarly in both groups at 2 years, while RYGB had greater reduction in pulse 1 minute after physical exertion. Pooled data between the groups revealed significant correlation between change in weight and heart rate recovery at 12 and 18 months postintervention. Both RYGB and IMWM produce improvements in self-reported and quantitative measures of physical function and perception of physical health at 2 years. Weight lossassociated improvements in quality of life measures and cardiovascular fitness should be considered when assessing medical and surgical options for weight loss and glycemic control. Abstract 93 Identifying new targets for β-cell regeneration using MEN1 as a model Nohemy Morones (Columbia University), Chester E. Chamberlain, PhD, University of California, San Francisco Patients with Type 1 and Type 2 diabetes have decreased beta cell mass, and ultimately need some form of beta cell replacement. One strategy to meet this need is to identify proteins that can be targeted to drive the proliferation of insulin-producing beta cells. Multiple Endocrine Neoplasia Type 1 (MEN1) is a rare autosomal dominant cancer syndrome characterized by beta cell tumors and other endocrine tumors derived from the pancreas, pituitary gland and parathyroid glands. Most MEN1 patients inherit a loss of function mutation in the MEN1 gene that encodes for the protein menin, which suppresses beta-cell proliferation. However, about 10% of MEN1 patients lack mutations in MEN1 and the causative gene remains unknown. We therefore hypothesized that these patients inherit a mutation in a gene other than MEN1 and that this gene also functions to suppress the proliferation beta cells and related MEN1 endocrine cell-types. To test this hypothesis, we performed next-generation exome-sequencing on genomic DNA isolated from four patients who have extreme MEN1 (endocrine tumors in all three MEN1-sensitive tissues: the pancreas, parathyroid and pituitary) but lack mutations in the MEN1 gene, and analyzed the data to determine whether these patients shared a common gene nullifying insertion/deletion mutation (indel). Of the 42,000 mutations identified, we found less than 20 frameshifting indels to be common in all four patients. We are continuing to analyze these genes and are developing a method based on CrispR technology to test the function of these genes in beta cell proliferation using human islets. Abstract 94 The Epigenetics of Childhood Obesity Kathryn Tully Oelsner (Medical University of South Carolina), Yan Guo, Holli Dilks, Shari Barkin MD, Vanderbilt University Medical Center Epigenetics is emerging as a lens to view physiologic changes related to obesity. Recent research identifies potential methylated candidate genes linked to specific biologically plausible mechanisms associated with increased adiposity. This proof of principle study compared DNA methylation in 92 saliva samples between Latino preschool children of normal weight mothers (BMI <25 and WC <90 cm) vs children of obese mothers (BMI >30 and WC >100 cm) using a candidate gene approach. After methodic selection, candidate genes were categorized by biological mechanism and labelled by the impact of the promoter methylation on the downstream mechanism to better quantify the impact of methylation on each potential obesogenic pathway. These samples were run using the Illumina HumanMethylation Beadchip 450K and analyzed using linear regression model, differential methylation, and pathway analysis. We hypothesized that Latino children of normal weight mothers versus those of obese weight mothers would show differential methylation patterns at candidate gene sites, reflective of their phenotypic adipocyte storage, hormone regulation, chemical balance, and other potential regulators of weight gain. 119 CpG probes and 4 genes (MTL5, XKR9, FAM7A2; FAM7A1, MAP6D1) had a significant effect change after stratification, using maternal BMI as the predictor and differential methylation as the outcome (padj value <.05). Other genes linked to an obesogenic trajectory (SEC14L2, TRIM61, NUDT4, APOL4, NUAK1) were differentially methylated in children born of normal weight mothers versus those born of obese weight mothers in quartile differential methylation analysis (p<0.025). Utilizing an obesogenic scoring protocol to assess the impact of methylation upon the functionality of the downstream product and overall physiologic mechanism, differential DNA methylation in genes involved with lipid and cholesterol metabolism was identified as the most impactful upon Latino preschool obesogenic trajectory. Utilizing saliva, a practical human tissue to obtain in naturalistic settings and in pediatric populations, was confirmed to be a viable medium for epigenetic testing, although not identical to results yielded from placenta. Comprehensive epigenetic analysis taking functionality into account has not yet been utilized in previous literature, so this scoring mechanism can be used as a model in future studies to yield relevant, comprehensive information from epigenetic analysis. Abstract 95 Differential Effects of Gastric Bypass versus Adjustable Gastric Banding on Body Composition Changes in Obese Subjects Puja Rai1 (University of Connecticut School of Medicine), Blandine Laferrère MD2, Roxanne Dutia PhD2 New York Obesity Research Center, Columbia University Medical Center, New York, NY. Introduction: Laparoscopic Roux-en-Y gastric bypass (GBP) and adjustable gastric banding (AGB) are 2 commonly performed bariatric surgeries to treat obesity and induce significant longterm weight loss. Evidence suggests that GBP leads to more favorable weight loss and metabolic outcomes over AGB. The aim of this study was to compare the effect of GBP and AGB on body composition changes in subjects with obesity and type 2 diabetes, using a novel 3-dimensional photonic scanner (3DPS) technology. Hypotheses: GBP results in greater weight loss and fat loss than AGB, as well as greater reduction in waist circumference and torso volume, after adjusting for differences in weight loss. Reduction of fat mass is correlated with improved insulin sensitivity. Methods: Subjects were studied before and 12 months after GBP and/or after 20% weight loss following AGB. Body composition data was collected using the 3DPS and insulin sensitivity was assessed using HOMA-IR values. Results: At baseline, GBP patients (n=18) were similar in age, weight and BMI compared to AGB patients (n=9). GBP induced significantly greater weight loss compared to AGB (31.3±6.1% vs. 18.4±10.3%, P<0.01). After adjusting for differences in weight loss, GBP resulted in a greater reduction in fat mass (40.4±13.5% vs. 32.0±13.8%, P=0.04) compared to AGB. Although GBP lead to a greater decrease than AGB in waist girth (24.0±6.9% vs. 14.9±7.2%, P=0.66) and torso volume (36.2±8.2% vs. 23.0±11.2%, P=0.98), these changes were not statistically significant. The association between percent loss in fat mass and percent decrease in HOMA-IR was stronger in AGB compared to GBP, although not significant (R2=0.355, P=0.12 vs. R2=0.041, P=0.47). Conclusions: GBP has more favorable effects on weight loss and results in greater fat mass reduction compared to AGB. However, once adjusting for weight loss differences, GBP did not cause a significantly greater reduction in other body composition parameters. The improvement in insulin sensitivity may not only be driven by loss in fat mass after GBP. 3DPS is a novel tool which has the potential to play a vital role in future studies to monitor and evaluate regional body composition changes in individuals with severe obesity that undergo bariatric surgery. Abstract 96 Increase in plasma FGF19 levels is dependent on weight loss method and independent of percentage weight loss. Elizabeth Hoover (University of Iowa Carver College of Medicine), Gerardo Febres, Irene M. Conwell, Donald J. McMahon, Clifton Jackness, Wahida Karmally, Leaque Ahmed, Marc Bessler, and Judith Korner, Columbia University Medical Center Fibroblast growth factor 19 (FGF19) is a hormone secreted in the ileum postprandially in response to the binding of bile acids to the Farnesoid X receptor. FGF19 treatment in high-fat diet or ob/ob mice reduces body weight and improves insulin sensitivity. Decreased levels of FGF19 have been noted in humans with obesity or type 2 diabetes mellitus (T2DM) with an increase in FGF19 in the fasted and postprandial states following Roux-en-Y gastric bypass (RYGB) surgery. It remains unknown whether this increase, which has been proposed as a mechanism for improvement in glycemia after RYGB, is secondary to weight loss or to the altered nutrient transit specific to the bypass procedure. The objective of this study was to evaluate the effect of different methods of weight loss on levels of FGF19 in obese individuals with and without T2DM after equivalent percentage of weight loss (7.9 ± 0.3%). Weight reduction was achieved by RYGB (n=24), vertical sleeve gastrectomy (SG, n=10), or calorie restriction (CR, n=28); each cohort included individuals with and without T2DM. Within the calorie restriction group, subjects either underwent laparoscopic adjustable gastric banding (LAGB, n=7) or followed a low-calorie-diet (LCD, n=21). Levels of FGF19 were quantified in the fasted state before and after weight loss. FGF19 increased significantly in subjects who underwent RYGB (97 ± 9 to 142 ± 9 pg/mL; P=0.0016) and SG (100 ± 14 to 179 ± 14 pg/mL; P=0.0005) but not in subjects who lost weight via CR (102 ± 9 to 115 ± 9 pg/mL; P=0.31). Across all interventions, baseline FGF19 levels did not differ based on T2DM status (P=0.99), but after weight loss subjects with T2DM had a blunted increase in FGF19 levels compared to subjects without T2DM, although this was not statistically significant. These data suggest that the increase in plasma FGF19 levels following weight reduction is independent of percentage weight loss and unique to anatomical alteration. This may further implicate FGF19 in the improvement in glycemic control commonly seen after RYGB and SG. Abstract 97 Integration and Utilization of Peer Leaders for Long-Term Diabetes Self-Management Support: Are Healthy Behaviors Associated with Improved A1C 1 Mary Rockas, BS and 1Gretchen A. Piatt, PhD, MPH 1 University of Michigan Medical School Evidence demonstrates that diabetes self-management education (DSME) is effective at decreasing diabetes-related complications, including micro and macrovascular disease, in part due to behavior changes that help lower A1C. However, certified diabetes educators are scarce and the effects of DSME are not sustainable in the long-term without ongoing self-management support programs (DSMS). Implementing Peer Leader (PL) led DSMS represents one possibility for helping people with diabetes to manage their A1C levels and make behavior changes. Therefore, we determined whether a PL DSMS model was as effective in achieving and maintaining improvements in glycemic control and behavior change following a DSME and DSMS intervention compared to a traditional DSME support model in 6 rural primary care practices in Pennsylvania. Additionally, we examined whether making a behavior change was associated with improved glycemic control over time. Practices and eligible participants (n=221; mean age: 63.0 years, 63.8% female, 96.8% Caucasian, 28.5% at or below poverty level, 32.5% using insulin, A1c ≥ 7%: 54.2%) were randomized to the intervention (DSME+PL SMS; n=119) or usual care (UC) group (DSME+traditional DSME support with no PL; n=102). Each intervention practice had one PL to support participants. Behavior change was defined as change in eating differently, eating less, eating less fat, eating less sugar, eating more vegetables, drinking less pop/juice, and increasing physical activity. Data were collected at baseline, 6 weeks, and 6 and 12 months. Marked decreases in A1c were observed in both groups following DSME (intervention: -0.24%, p<0.0001; UC: -0.31%, p=0.01); however, as SMS continued over time, intervention participants’ average A1c levels continued to significantly decrease at 6 months (-0.32%, p=0.04) and at 12 months (-0.37%, p=0.02) in comparison to UC which had increases in average A1c during the support period (6 months: +0.08%, p=0.09; 12 months: +0.02%, p=0.13). Greater than 80% of participants in both groups who made a behavior change at baseline maintained the behavior change at each follow-up time. When the association between behavior change and A1C was examined at each time point, healthy behaviors were associated with lower mean A1C. At baseline, average A1c in the intervention group was 8.1% for participants who ate less fat compared to 7.3% for those who did not eat less fat (p=0.04). Similar results were observed for eating less sugar (p=<.001) and eating more vegetables (p=0.04). In the UC group, significant associations were observed between A1c and physical activity (baseline: p=0.03, 6 weeks: p<0.001, 12 months: p=0.04), but not eating behaviors. These data demonstrate that PL DSMS is as effective as traditional DSME support in helping participants to maintain glycemic control and increase healthy behaviors in the longterm. In addition, lower mean A1c was associated with improvements in healthy eating behavior in the intervention group and physical activity in the UC group. These differences in associations may reflect provider site bias toward encouraging patients to change eating rather than physical activity and vice-versa. Future work will focus on examining factors, such as psychosocial outcomes, that may mediate the relationship between behavior change and glycemic control. Abstract 98 The Acute Phase Response in Total Joint Arthroplasty: is it predictable, consistent, and respective? William K. Oelsner Jr. (Medical University of South Carolina), Thomas J. An, Michael A. Benvenuti, Richard A. Jacobson, Gregory Polkowski MD, Jonathan G. Schoenecker MD PhD, Vanderbilt University Total joint arthroplasty is a major orthopedic surgery that has a high risk of postoperative thromboembolic events as a result of the acute phase response (APR). In response to tissue injury, fibroblasts, endothelial cells, keratinocytes, and monocytes release Interleukin-6 into circulation which induces the APR in the liver to release pro-coagulant factors and inflammatory metabolites including fibrinogen and C-reactive protein (CRP).1 D-dimer, a degradation product of plasmin and fibrin (a product of fibrinogen and thrombin), is a key biomarker for ongoing clot formation and fibrinolysis.2 Although the APR is essential for preventing bleeding and infection, a prolonged APR and coagulation imbalance can lead to life-threatening hemorrhage, wound hematoma, poor wound healing, deep venous thrombosis, pulmonary embolus, and disseminated intravascular coagulation if the APR is not resolved.3 Our hypothesis is that the APR response to a specific surgery is predictable, consistent, and respective for total joint arthroplasty. Lab values were retrospectively studied in 76 patients undergoing total knee arthroplasty (TKA) and 43 patients undergoing total hip arthroplasty (THA) by a single surgeon. Samples where measured pre-op, post-op day 1 to 4, 2 weeks post-op, and 6 weeks post-op. CRP had a 38.6 fold increase and peak on post-op day 2.796 +/- 3.213 for TKAs and a 20.2 fold increase and peak on post-op day 1.840 +/- 0.5360. Fibrinogen had a 1.6 fold increase and peaked on post-op day 6.135 +/- 8.711 for TKAs and a 1.5 fold increase and peak on post-op day 7.583 +/- 7.937 for THAs. D-Dimer had a 6.3 fold increase and peak on post-op day 5.675 +/- 3.031 for TKAs and a 6.0 fold increase and peak on 13.81 +/- 5.075 for THAs. CRP and Fibrinogen levels were resolved by week 2 and week 6 respectively, while D-Dimer was unresolved by week 6. Furthermore, resolution, as measured by the time from peak value to half-maximum value, was not statistically different between the TKA and THA cohorts for CRP and Fibrinogen. There was statistical difference between D-Dimer resolutions for the cohorts. Increased pre-operative CRP levels correlated with increased max Fibrinogen levels for TKAs, (p=0.0183, Spearman r = 0.3063) and for THAs (p= 0.0087, Spearman r = 0.4252). Increased pre-operative fibrinogen correlated with increased max CRP for TKAs (p=0.0013, Spearman r=0.3769). Pre-operative blood glucose levels correlated with max D-dimer depression (p= 0.0226, Spearman R = -0.2940) for TKAs but not for THAs. In conclusion, total joint arthroplasty demonstrated a consistent APR for both TKAs and THAs. The acute phase reactants had specific, unique temporal peaks and intensities which were not significantly different between cohorts. This data suggests that total joint arthroplasty induced APR is consistent and predictable for a non-Gaussian distribution of patients with few comorbidities and adverse outcomes. For this reason, future investigations into the APR for Gaussian distributions are merited: if the APR is predictable and consistent for patients with good outcomes as well as those with adverse outcomes, then it would allow clinicians to predict and treat before and during post-operative, APR complications. Abstract 99 Screening for Diabetes in Routine Clinical Practice Scott Mauch (Michigan State University College of Human Medicine), Laura N. McEwen PhD, William H. Herman MD, University of Michigan, Ann Arbor, MI In 1997, the American Diabetes Association recommended that diabetes screening be carried out at 3-year intervals for all non-diabetic individuals ≥ 45 years of age, with the preferred screening test being fasting plasma glucose. A 2004 study conducted by Ealovega et al. described screening and follow-up in the population of non-diabetic patients ≥ 45 years of age who utilized an academic health care system. Despite high screening rates and targeting of patients at high-risk for diabetes, follow-up of patients with abnormal screening results was low. The authors concluded that failure to follow-up on abnormal glucose levels was a barrier to diagnosis and treatment. In 2009, with the addition of HbA1c as the recommended screening test for diabetes, we hypothesized that screening, follow-up, and diagnosis of diabetes in routine clinical practice would improve. To test this hypothesis, we studied screening in non-diabetic patients ≥ 45 years of age enrolled in Blue Care Network (BCN), a large commercial HMO, who were assigned to University of Michigan Health System primary care providers. Screening was defined as the first test of glycemia performed between January 1, 2010 and December 31, 2013. Of 12,797 patients studied, 9,966 (78%) were screened. Laboratory glucose testing accounted for 86% (n = 8,522) of all screening tests. Women were more likely than men to be screened, and patients ≥ 65 years of age were more likely to be screened than those of lower ages. Of 9,966 patients screened, 39% (n = 3,843) had abnormal results. These data suggest that the addition of HbA1c as the recommended screening test has increased both the rate of screening in the study population and the use of HbA1c as a screening method. Chart reviews of patient medical records and BCN insurance claims to determine diabetes risk factors, rates of follow-up, and rates of diagnosis of diabetes are currently ongoing. Abstract 100 Genetic Variants Associated with Testosterone to Androstenedione Ratio in Polycystic Ovary Syndrome Melba Beltre1 (New York Medical College), Andrew Bjonnes2, Cindy Pau1, Richa Saxena2, and Corrine Welt1 The Reproductive Endocrine Unit1, Department of Anesthesia and Center for Human Genetic Research2, Massachusetts General Hospital, Boston, MA The phenotypic expression of polycystic ovary syndrome (PCOS) varies among women of different racial and ethnic backgrounds, particularly the levels of testosterone and androstenedione. Although the cause of PCOS is not clearly defined, there is evidence for a genetic basis. 17β-hydroxysteroid dehydrogenase type 5 (AKR1C3) and type 6 (SDR9C6) are involved in the conversion of androstenedione to testosterone in the adrenal gland and ovary. We tested the hypothesis that genetic variants in AKR1C3 and SDR9C6 are associated with testosterone to androstenedione (T:A) levels. Women recruited were 1) with PCOS, according to the NIH criteria (hyperandrogenism and irregular menses; n=294), and 2) Controls with regular menstrual cycles and no signs of hyperandrogenism (n=252). All women were of mixed European ethnicity in Boston, with normal TSH and prolactin, and no evidence of ovarian insufficiency or nonclassic congenital adrenal hyperplasia. Serum testosterone and androstenedione levels were measured using radioimmunoassay, and DNA was isolated for genetic analysis. Genotyping was performed using the Omni Express Plus Exome Bead Chip (Illumina). Serum T:A ratios were calculated and PLINK software was used to analyze associations between T:A and genetic variants in the candidate genes, as well as associations between T:A and genome-wide variants. Bonferroni corrections were applied to the p-values which were adjusted to avoid false positive associations from multiple testing. The candidate gene analysis did not show a significant correlation between variants in AKR1C3 (p>6x10-5) and SDR9C6 (p>2x10-4) and T:A ratios. The genome-wide analysis showed one SNP with a significant association (p<4x10-9). The SNP, rs9323945, is a missense variant in the ATG2B gene, resulting in a change from asparagine to aspartic acid. The amino acid is highly conserved, although the change is predicted to be benign. ATG2B has functions in the regulation of the metabolic profile and the association may indicate a relationship with androgenic or other phenotypic features of PCOS. The study will need to be replicated in an additional population. The data suggest that more genetic studies are needed to further assess the correlation between androstenedione and testosterone levels and SNP associations in PCOS. Abstract 101 Gender and income level influence dietary habits of African-Americans. Ronald R. Magee Jr. (The Warren Alpert Medical School of Brown University), Dr. Jessica Yeh, PhD; Dr. Pete Miller, PhD, MD; Johns Hopkins School of Medicine The relationship between diet and diabetes has fueled a number of recent studies, especially with regard to the interplay between the two and other societal factors within minority and disadvantaged communities. In this study, we examine the diets of an inner-city AfricanAmerican cohort to test the following three hypotheses: 1) adults with diabetes have better adherence to an advantageous diet than those without diabetes, 2) diabetic women have better adherence to a nutritious diet than men, and 3) higher socioeconomic status is positively correlated with adherence to advantageous dietary guidelines. Adherence to a DASH-OMNI diet and general nutritional guidelines prescribed by the United States Department of Agriculture (USDA) and United States Department of Health and Health Services (USDHHS) was measured from data acquired through a self-administered Block Fruit/Vegetable/Fiber screener and Block Dietary Fat Screener. Statistical analyses were arranged by diabetes status, gender, and yearly household income. 123 African Americans with controlled hypertension were included in the study, including 89 people with diabetes and 34 people without diabetes. Among individuals without diabetes, we found that women and people of lower socioeconomic status displayed statistically significantly greater adherence to USDA guidelines than men and people of higher socioeconomic status, respectively. Non-diabetics of higher socioeconomic status, on average, ate daily 10.92 more grams of fat (P=.04), 5 more grams of saturated fat (P=.01), and 47.39 more milligrams of cholesterol (P=.01) than their lower income counterparts. Non-diabetic men, on average, daily ate 5.13 more grams of saturated fat (P=.0099) and 69.05 more milligrams of cholesterol (P=.0001) than their female counterparts. The results of this study suggest that gender and income are associated with adherence to a health-maintaining diet in African Americans. The results of this study could have implications for designing a clinical diet therapy and nutritional diabetes prevention for inner city minority populations. Abstract 102 Impaired TUG protein proteolysis in insulin resistant adipose tissue. Muyi Li (Medical University of South Carolina), Estifanos Habtemichael, Don Li, Max Petersen, Gerald Shulman, Jonathan Bogan, Yale University. Insulin stimulates glucose uptake in adipose and muscle by triggering proteolytic cleavage of TUG proteins. In unstimulated cells, TUG sequesters GLUT4 glucose transporters intracellularly. Cleavage releases GLUT4 for translocation to the plasma membrane to increase glucose uptake. During the pathogenesis of type 2 diabetes, GLUT4 translocation is impaired, resulting in insulin resistance. Here, we tested the hypothesis that impaired TUG proteolysis contributes to impaired GLUT4 translocation in adipose tissue of insulin resistant rats. To induce insulin resistance, animals were fed a high-fat+sucrose diet for three days. Control and insulin-resistant rats were fasted, treated with or without a 20-minute infusion of insulin and glucose, and then sacrificed. Adipose tissue was harvested and protein lysates were immunoblotted to detect TUG. In control rats, insulin caused a decrease in the abundance of intact TUG protein to 43±7% of the abundance observed in fasting animals (p<0.05, n=4 in each group). In insulin resistant rats, this decrease was not observed, and TUG abundance was unaffected by insulin stimulation. These results indicate that TUG proteolysis was impaired in white adipose tissue of insulin resistant rats, compared to controls. Because TUG cleavage is independent of canonical insulin signaling through phosphatidyl-3-kinase to Akt, the data imply that diet-induced insulin resistance results in part from impairment of other pathways. Thus, the data reveal a novel mechanism for diet-induced insulin resistance, which may contribute to the development of type 2 diabetes. Abstract 103 Identifying novel antigenic targets and role of antigen modification for BDC-2.5 T cells in type I diabetes Juliana Xie (Penn State College of Medicine), Martin Thelin, and Tom Serwold. Research conducted at Joslin Diabetes Center, Boston. BDC-2.5 T cells are a T cell clone developed from nonobese diabetic (NOD) mouse that reliably induces autoimmune pancreatitis and have been used extensively in type I diabetes research. However, the identity of the autoantigen that these BDC-2.5 T cells specifically react to has remained elusive. More than 20 years later, in 2010, a candidate autoantigen called chromogranin A, had finally emerged. Interestingly, follow-up studies have shown that posttranslationally modified version of chromogranin A transfers autoantigenic. This is achieved by an enzyme called transglutaminase. These two important observations drive the two hypotheses in this study: 1) Transglutaminase modifies chromogranin A, making it autoantigenic to BDC-2.5 T cells; 2) Besides chromogranin A, there are other autoantigenic targets of BDC2.5 T cells. To test hypothesis 1, we aimed to co-express chromogranin A and transglutaminase in both the fibroblast and the NIT cell (pancreatic beta cells from NOD mice) systems. These cells are then co-cultured with BDC-2.5 splenocytes, and then assayed for the level of IL-2 cytokine production via ELISPOT (as a measure of T cell stimulation). To test hypothesis 2, we aimed to express seven novel pancreatic genes (individually) in both fibroblasts and NIT cells, then test for their ability to stimulate T cells by co-culturing with BDC-2.5 T cells, then IL-2 ELISPOT assay. During experimentation, three of the seven candidate genes were cloned and expressed in fibroblasts and NIT cells with success. These were chromogranin A, chromogranin B, and secretogranin 3, and they are components of secretory granules in pancreatic beta cells. Results (qualitative; quantitative to come): chromogranin A+-NIT cells induced higher number of spots in the ELISPOT assay than control NIT cells, while no significant difference was observed in chromogranin A+-fibroblasts versus control fibroblasts. There was no observable difference in the number of spots in chromogranin B+ and secretogranin 3+ cells when compared with their respective controls. Therefore, we conclude that chromogranin A is likely the epitope for BDC2.5 T cells, and that an unknown mechanism of post-translational modification specific to NITcells is at work to render the peptide more antigenic to BDC-2.5 T cells. Abstract 104 Association of Physical Activity Between Children and Their Mothers April Venn, Maciej Buchowski, PhD Dartmouth Geisel School of Medicine & Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine Vanderbilt University Medical Center Obesity is a rapidly growing epidemic in the US population affecting both adults and children that is thought to be caused by genetic and environmental factors. Our study explored some environmental factors by looking at the energy expenditure, physical activity, BMI and percent body fat of 21 children and their mothers to see if there was an association among energy expenditure, physical activity and BMI. Based on maternal influences and control over a child’s activities we hypothesized that a child’s physical activity level and time spent in sedentary behaviors and performing physical activity of various intensities would be similar to their mother’s. The participants had Actigraph accelerometers placed on their waists. The Actigraph recorded their physical activity (PA) in the free-living environment for a week. The PA was categorized as sedentary, low, moderate or vigorous intensity. Children and their mothers showed a moderate correlation in their total energy expenditure during low intensity PA. Weak correlation existed between total energy expenditure of children and their mothers in other activity levels. Mothers spent 81.6 % of their time sedentary, while children spent 75.3% of their time sedentary. Mothers spent 18.4 % of their time in low, moderate or vigorous physical activity. Children spent 24.7% of the time in low, moderate or vigorous physical activity. Overall the data showed a weak to moderate correlation between children and their mothers for energy expenditure, time spent performing various activities and total physical activities. Weak correlations were seen between BMI, body fat and total physical activity. The small sample size did not allow us to definitively assess the strength of the effects of these associations. Larger studies are needed to make stronger conclusions about the association among BMI, body fat, physical activity and energy expenditure between children and their mothers. Abstract 105 Volumetric brain changes detected over time in association with hypo- and hyperglycemia in youth with type 1 diabetes mellitus (T1DM) Sneha Thakur (New York Medical College), Heather Lugar, Tasha Doty, Amal Al-Lozi, Jonathan Koller, Neil White, and Tamara Hershey, Washington University in St. Louis Severe hypoglycemia, severe hyperglycemia, and chronic hyperglycemia in children and adults with type 1 diabetes mellitus (T1DM) have been associated with cross-sectional structural brain differences in magnetic resonance imaging (MRI), but it is unclear how and when these differences emerge. We performed a 2 year longitudinal structural MRI study on youth with T1DM and their non-diabetic siblings, examining their rates of change in subcortical and whole brain volumes. Previous analyses of these data used a voxel-wise approach that required multiple comparison corrections for all voxels. We sought to confirm and possibly reveal new findings by using Freesurfer’s established longitudinal processing stream which applies a different registration algorithm to anatomically define and measure volume in brain regions in individuals. Our analyses revealed that non-diabetic controls showed a greater increase in volume over time than T1DM youth in the brainstem (p=.045) and nucleus accumbens (p=.045). Within T1DM youth, we found significant differences in rates of volumetric change between T1DM youth exposed to hypoglycemic episodes (hypo) and chronic hyperglycemia (hyper) between time points (p’s<.05). Specifically, T1DM youth with any hypos between time points were associated with a smaller rate of increase in both cerebellar white (p=.016) and total cortical white matter volume (p=.031) over time than those with no hypos. Additionally, T1DM youth with chronically high hyperglycemia (mean A1c>9.0%) between time points showed a greater rate of decrease in total cortical gray matter volume than T1DM youth with mid (mean A1c=7.5-8.9%) or low (mean A1c<7.5%) hyper exposure (p=.001 and p=.003), or controls (p=.014). The high hyper exposure group also showed smaller rates of increase in corpus callosum volume than those with mid hyper exposure (p=.003) and controls (p<.001), while the low hypo exposure group showed a smaller rate of increase in corpus callosum volume than did controls (p=.041). Our findings complement previous VBM analyses by showing that exposure to glycemic extremes in T1DM youth was associated with suppressed white matter growth over as little as 2 years, and hyper exposure had the added effect of greater gray matter loss in the cortex but not in the subcortex. Further, we found novel volumetric changes over time in subcortical regions of T1DM youth compared to their non-diabetic controls. Our results may suggest that poor glucose control in the developing brains of T1DM youth is associated with decreased myelin growth (white matter) and increased neuronal pruning (gray matter). Abstract 106 Finding a Cause for Neonatal Diabetes Katherine Lindauer (Michigan State University College of Human Medicine), Graeme I. Bell, David Carmody, Siri Atma W. Greeley, University of Chicago Kovler Center for Diabetes Research When diabetes is diagnosed in patients younger than one year of age a genetic, rather than autoimmune, cause is often attributed to their high blood sugars. While there are currently several known causative genes, there are still many patients whose causative genetic variants have yet to be identified, and new gene-based etiologies must be sought. Based on an early diabetes diagnosis and the presence of other autoimmune symptoms, we hypothesized that six patients in the Monogenic Diabetes Registry were good candidates for testing of a causative variant in STAT3, a newly suspected causative gene that has been previously implicated in various forms of cancer. We also hypothesized more generally that patients in our registry were more likely to have an identifiable mutation if they received their diabetes diagnosis when they were less than six months of age. We used the University of Chicago’s Monogenic Diabetes Registry to a) select patients as STAT3 testing candidates and b) track genetic causes for all patients diagnosed at less than one year of age. DNA samples from the patients who were candidates for STAT3 testing were sent to the University of Exeter for sequencing. This testing is ongoing, but its results will determine whether these patients are at higher risk for having other autoimmune conditions in the future. If the results come back positive, that will also indicate that alterations in the sequence of STAT3 are causative of diabetes for these patients. In addition to the results concerning STAT3, we found that among all patients in the registry, individuals were more likely to have a known causative variant if they were diagnosed with diabetes when they were less than six months old; and that among these, mutations in the potassium channel gene KCNJ11 were most common. Abstract 107 Examining the role of β-oxidation in fatty acid-induced IRE1α signaling and NLRP3 inflammasome activation Herodes Guzman (University of North Carolina School of Medicine), Megan Robblee and Suneil Koliwad, University of California, San Francisco In obesity and type 2 diabetes, adipose tissue macrophages are exposed to excess fatty acids which have been shown to induce inflammatory activation and ER stress. Activation of the ER stress sensor IRE1α in macrophages treated with saturated fatty acids (SFAs) has been linked to the activation of the NLRP3 inflammasome, leading to the secretion of the cytokine IL1β. Recent studies link IRE1α activation to changes in ER membrane composition, suggesting that altering fatty acid flux to phospholipids may modulate SFA-induced IRE1α activation. We hypothesized that inhibition of fatty acid β-oxidation with etomoxir would potentiate SFA-induced IRE1α signaling and subsequent NLRP3 inflammasome activation. The efficacy of etomoxir was confirmed with the Seahorse Biosciences XF Analyzer, as the oxygen consumption rate of fatty acid-treated bone marrow derived macrophages was decreased in the presence of etomoxir. Etomoxir was found to heighten IRE1α activation in response to the SFA stearate, as measured using a luciferase reporter. This suggests that inhibiting beta-oxidation may divert SFAs to metabolic pathways that lead to IRE1α activation. Contrary to the potentiation of the SFA-induced IRE1α, etomoxir did not affect stearate-induced IL-1β secretion but unexpectedly reduced the secretion of IL-1β in response to another SFA, palmitate. These data suggest that blocking β-oxidation may affect NLRP3 inflammasome activation in a fatty acid-specific and IRE1α-independent manner. Abstract 108 Characterization of iPS cells for Wound Healing in Diabetic Patients Lourdes Ramirez (University of Central Florida), Mogher Khamaisi, M.D., Ph.D.; George L. King, M.D. Joslin Diabetes Center, Harvard Medical School, Boston, MA Background: Impaired wound healing is the most frequent cause of hospitalization in diabetes patients, often characterized by the development of diabetic foot ulcers, which precede 84% of diabetes-related lower extremity amputations. Many complex pathophysiological mechanisms involved are still unknown. The development of human inducible pluripotent stem cells (hiPSc) by reprogramming of somatic cells to a self-renewing and pluripotent state, has led to the potential for new treatment modalities in impaired wound healing and regenerative medicine. The aim of this study is to characterize hiPSc derived from Type 1 diabetic patients as compared to age-matched healthy controls in parameters significant in wound healing, namely, cell adherence, proliferation and VEGF production. The hypothesis of the study is that hiPSc derived from diabetic patients have reduced adhesion, proliferation, and VEGF expression in comparison to healthy controls. Methods: Three cell lines were used in this study: one hiPSc line derived from a 78 year-old healthy male control and two hiPSc lines (89 year-old male with 85-year duration of diabetes, and 66 year-old male with a 51-year duration of diabetes). To measure adhesion and proliferation, hiPSc were harvested after incubation for 1 h (adhesion), or every 48 h followed for up to 8 days after seeding (proliferation). VEGF mRNA levels were obtained by qRT-PCR after hiPSc stimulation by insulin (100 nM) and hypoxia (5% O2) for 24 h. mRNA results were corrected in comparison to human b-Actin house-keeping gene. Results: Cell adhesion is reduced in diabetic hiPSc to 56.2% and 43.6% from that seen in the healthy control hiPSc. VEGF mRNA expression is impaired in diabetic hiPSc (3.1 and 3.03-fold increase from basal conditions) following insulin stimulation in comparison to healthy controls (16-fold increase). VEGF mRNA levels in diabetic hiPSc increased 2.15 and 2.36-fold from basal conditions following hypoxia stimulation, but further work with the control is required for conclusions. Preliminary data suggest reduced proliferation rate in diabetic hiPSc compared to healthy controls, however, an increased sample size is necessary to measure significant differences. Summary: This study suggests that diabetic hiPSc show impaired phenotypes related to wound healing when compared to healthy hiPSc controls. Furthermore, diabetic hiPSc retain some of the impaired characteristics observed in the fibroblasts from which they were derived. Conclusion: These findings contribute to the long-term end-goal of developing therapies where transplantation of hiPSc, with defects corrected, into diabetic wounds may be used as a novel cell-based strategy for wound healing in diabetic patients Abstract 109 Initial evidence that hypothalamic injury is associated with obesity and insulin resistance. Zenobia Gonsalves (Chicago Medical School at Rosalind Franklin University of Medicine and Science), Susan J Melhorn, Mary K Askren, Vidhi Tyagi, Kenneth R Maravilla, Ellen A Schur; University of Washington In both rodent models and humans, obesity is associated with evidence of gliosis in the mediobasal hypothalamus (MBH), a region critical for energy balance, regulation of body mass and glucose homeostasis. Gliosis in the central nervous system (CNS) occurs in response to neuronal injury and is characterized by the recruitment, activation and proliferation of astrocytes and microglia. It may be detected using quantitative magnetic resonance imaging (MRI) techniques and identified as an increased T2 relaxation time or an elevated T2 signal ratio. Additionally, hypothalamic activity in response to an oral glucose load, measured using functional MRI (fMRI), has been shown to be altered in obesity and type 2 diabetes. However, it is unknown whether gliosis plays a role in this change. Therefore, two studies were implemented to test the hypotheses that 1) radiologic evidence of MBH gliosis in humans will be associated with obesity, insulin resistance, decreased physical activity, and eating behaviors that promote excess food consumption, and that 2) hypothalamic activity in response to glucose ingestion will be increased in those showing radiologic evidence of MBH gliosis. In a nested case-control design, 21 healthy, normal-weight (BMI 18.5-24.9) or obese (BMI>30) adults (7 males, 14 females) aged 18-50 years (mean 33.5 + 12.1) had a fasting blood draw, completed behavioral questionnaires and underwent an MRI in Study 1. Select participants from Study 1 (those with a T2 relaxation time less than 90.5 ms or greater than 95.5 ms) then completed Study 2, a pilot study to test the second hypothesis noted above. After an overnight fast, these subjects (n=2; 1 male, 1 female; 1 T2 relaxation time<90.5, 1 T2 relaxation time>95.5) had a fasting blood draw, answered behavioral questionnaires and completed an oral glucose tolerance test (OGTT) while undergoing a functional MRI (fMRI). From the 21 Study 1 participants, a group of cases with radiologic evidence of gliosis was identified as the tertile (n=7) with the longest T2 relaxation time in the left MBH and was compared to a group of controls identified as the tertile (n=7) with the lowest T2 relaxation time in the left MBH. Compared to controls, cases with radiologic evidence of left MBH gliosis tended to have higher BMIs (33.1+10.2 in cases vs. 23.7+7.3 in controls, p=0.07), a higher proportion of obese subjects (57% in cases vs. 14% in controls, p=0.09) and lower total physical activity (2008+2621 MET-min/week in cases vs. 3305+1713 MET-min/week in controls, p=0.30). Additionally, cases with radiologic evidence of left MBH gliosis tended to have higher fasting glucose levels (93.9+11.3 mg/dl in cases vs. 88.7+7.1 mg/dl in controls; p=0.33), higher fasting insulin levels (10.5+7.5 uU/ml in cases vs. 5.3+3.3 uU/ml in controls; p=0.13) and higher HOMA-IR values (2.5+2.0 in cases vs. 1.1+0.7 in controls, p=0.12). These data lend initial evidence that hypothalamic damage may play a role in obesity and insulin resistance and suggest that studies showing hypothalamic gliosis in rodent models of diet-induced obesity are relevant to the scientific understanding of obesity and insulin resistance in humans. Our current fMRI findings indicate a noticeable difference in hypothalamic activity in response to an oral glucose load between the two pilot study subjects. Participants will continue to be enrolled in both studies to better understand the potential link between hypothalamic gliosis, obesity, insulin resistance and the hypothalamic response to glucose ingestion. Abstract 110 A systematic literature review: Diabetes self-management interventions for patients with type 2 diabetes living in rural areas Morgan G. Lepard, BS (University of Tennessee Health Science Center); Alessandra Joseph (Brown University); April Agne, MPH; Andrea Cherrington, MD MPH, University of Alabama at Birmingham Background Patients living in rural areas have a higher prevalence of type 2 diabetes mellitus (T2DM) than their urban counterparts.1 Rural-dwelling patients face unique challenges and obstacles in accessing diabetes education programs and day-to-day diabetes management. Objective In order to develop an effective diabetes self-management program for rural Alabamians, we systematically reviewed the evidence for diabetes education and self-management interventions designed for rural-dwelling patients with T2DM. Methods A systematic electronic search was conducted on PubMed using the keywords: rural population, diabetes education, diabetes “self-management” education, diabetes “self-management”, and diabetes “self-management” support. Eligible studies had to describe interventions, have measured outcomes, occur within the last 10 years, and focus on a rural population. Studies without a control/comparison group, study designs with N<50, or studies that did not target adult patients with T2DM were excluded. Results The search yielded 1,512 abstracts. Twenty-seven studies were reviewed in their entirety and 9 studies met inclusion criteria and data abstracted. Over half were randomized controlled trials (n=5). Intervention strategies varied: group or individual DSME programs (n=5), telehealth interventions (n=3), and a web-based intervention (n=1). Outcomes were grouped into 3 main categories: biologic, behavioral, and diabetes-related knowledge. Five studies reported statistically significant outcomes (3 DSME, 1 telehealth, 1 web-based). Of the 7 studies examining changes in HbA1c, 2 demonstrated significant improvements at 1 year (1 DSME program and 1 telehealth program). A separate study testing a DSME intervention resulted in significantly decreased serum glucose at 6 months. Two DSME interventions resulted in statistically significant weight loss and reductions in BMI. Finally, a study comparing telehealth with face-to-face sessions found that these intervention strategies produced similar improvements in metabolic control after one year. Conclusions We identified a limited number of studies with rigorous study designs evaluating interventions for rural-dwelling patients with T2DM. Effective interventions involved more patient contact hours and a focus on DSM-related behaviors and weight management. Telehealth interventions may produce outcomes comparable with face-to-face DSME interventions. Additional studies are needed to identify the most efficient and effective modalities for delivery of DSME in rural areas. 1. O’Brien T, Denham SA. Diabetes care and education in rural regions. The Diabetes Educator. 2008; 34(2): 334-347. Abstract 111 Effect of High-Fat Diet on PKA Subunit Expression in Mice Shriya Gandhi (University of Illinois College of Medicine – Chicago), Ronald N. Cohen, Barton Wicksteed, University of Chicago The cyclic AMP (cAMP) signaling pathway has been shown to regulate multiple intracellular processes, including growth, metabolism, differentiation, proliferation, and gene transcription. One of the main downstream effectors of this pathway, cAMP-dependent protein kinase (PKA), plays a critical role in the mechanisms that regulate beta-adrenergic receptor-stimulated lipolysis in adipocytes. cAMP-mediated activation of PKA leads to phosphorylation of perilipin and hormone sensitive lipase (HSL), two proteins that are important for maintaining the balance between lipogenesis and lipolysis within adipocytes, and likely results in many other effects as well. The PKA holoenzyme is composed of two regulatory and two catalytic subunits. The two classes of regulatory subunits, RI and RII, have different affinities for cAMP and show differences in their intracellular localization. Studies with transgenic mice have demonstrated that alterations in PKA subunit expression can change lipid deposition in different fat depots. The aim of this study was to determine whether the expression of PKA subunits and PKA activity is altered in response to high-fat feeding. Adipose tissue from four fat depots―subcutaneous (SQ), mesenteric, epididymal, and brown adipose (BAT)―was harvested from 25 week-old male mice, which had been fed either a standard chow diet or a high-fat diet (60% fat) since weaning at 4 weeks of age. Protein and RNA was isolated from each fat depot and characterized respectively by immunoblotting and quantitative real-time polymerase chain reaction (qRT-PCR). SQ and BAT depots exhibited little change in protein expression of either PKA regulatory or catalytic subunits between chow and high-fat diet. However, in the epididymal depot, our data demonstrated an increase in PKA RIα and a decrease in PKA RIIβ protein expression, when mice were exposed to high-fat feeding. Ongoing studies will evaluate PKA subunit expression in mesenteric tissue, a second visceral fat depot. Our data suggest that in the epididymal depot, high-fat feeding could induce PKA signaling to become more reliant on the RIα subunit, which has a higher affinity for cAMP. This may cause PKA to release its catalytic subunits at lower cAMP levels and result in increased phosphorylation of downstream targets, possibly leading to increased rates of basal lipolysis. Such findings may demonstrate a novel mechanism by which high-fat feeding leads to increased free fatty acid circulation in obese individuals, a condition that has been shown to result in increased insulin resistance and lipotoxicity in other metabolic tissues. Abstract 112 Dietary acid load is associated with serum bicarbonate but not insulin resistance in chronic kidney disease. H. Omer Ikizler (University of Vermont College of Medicine), Leila Zelnick, Kristina Utzschneider, Steven Kahn, Jonathan Himmelfarb, Ian de Boer, University of Washington School of Medicine Chronic metabolic acidosis is a common complication of chronic kidney disease (CKD) that may contribute to the progression of CKD and its numerous comorbidities, including increased insulin resistance. Dietary acid load may be a modifiable risk factor in the development of metabolic acidosis in patients with CKD. We investigated the hypothesis that dietary acid load is associated with serum bicarbonate and insulin resistance in 49 nondiabetic patients with CKD stages 3-5 and 15 healthy controls. Net endogenous acid production (NEAP) and potential renal acid load (PRAL) were quantified by 3-day prospective food diaries. Insulin sensitivity was measured by hyperinsulinemic euglycemic clamp. Mean (SD) values for NEAP, PRAL, serum bicarbonate, and insulin sensitivity were 60.5 (30.3) mEq/day, 9.7 (18.3) mEq/day, 24.8 (2.7) mEq/L, and 4.5 (2.2) mg/min glucose per µU/mL insulin, respectively. Both measures of dietary acid intake were significantly associated with serum bicarbonate in analyses adjusted for age, eGFR, sex, race, BMI, and use of diuretics. In NEAP, difference between the lowest (≤ 45.6 mEq/day) and highest (>64.2 mEq/day) tertiles was -2.0 mEq/L (95% CI -4.0, -1.0) of bicarbonate; p = .045 for continuous exposure. In PRAL, difference between the lowest (≤ 2.21 mEq/day) and highest (> 19.1 mEq/day) tertiles was -2.0 mEq (95% CI -3.0, -0.4) of bicarbonate; p <.001 for continuous exposure. Neither NEAP, PRAL, nor serum bicarbonate were significantly associated with insulin sensitivity. These results suggest that modification of dietary acid may be effective in blunting metabolic acidosis in CKD, but may not affect insulin resistance. Abstract 113 Understanding the role of POPDC2 & POPDC3 in colorectal epithelia Suzanne Kissel (Indiana University School of Medicine), Mukal Mittal, Sarah Short, Benjin Facer, Bobak Parang, Vishruth Reddy, Cody Keating, Christopher Williams. Vanderbilt University School of Medicine, Nashville, Tennessee Colorectal cancer (CRC) is the second most common cancer in the United States, striking 140,000 people annually and causing 60,000 deaths. The Popeye domain containing family is a group of homologous proteins with three members identified to date: BVES (POPDC1), POPDC2, and POPDC3. All three proteins contain both a Popeye domain as well as three conserved transmembrane domains. BVES in particular has recently been associated with cellular tight junctions (TJs), and is thought to be involved in regulating the association of TJ components. Dysregulation of TJs is often associated with carcinogenesis and recent studies support a role of TJ integral proteins in the regulation of Epithelial-to-Mesenchymal Transition, a key feature of advanced malignancy. BVES (POPDC1) expression has been shown to be reduced in CRC by promoter hypermethylation but the status of other family members has not been studied. Therefore, the purpose of this research served to define the role of POPDC2 and POPDC3 in the colonic epithelial cell and their role in CRC. First, we analyzed mRNA expression from The Cancer Genome Atlas database to determine expression differences in the POPDC genes between normal and CRC tissues. These data suggest [1][2]that all three genes are downregulated significantly in cancer tissue versus normal. We further measured mRNA expression in a variety of colon cancer cell lines, but POPDC gene expression did not obviously correlate with cell aggressiveness or epithelial morphology. We then utilized immunofluorescent and immunohistochemical (IHC) staining to identify the location of POPDC3 in the cell. Interestingly, this expression appeared to be largely nuclear with varying appearance in the cytoplasm. IHC staining of human CRCs further validated the nuclear expression observed in vitro, yet also suggested that the POPDC3 protein expression is increased in cancer versus normal tissue. This conflicts with mRNA data for POPDC3, yet may be due to possible posttranslational modifications or another event which impacts protein degradation. While much work remains to be done, these studies support a role for POPDC2 and POPDC3 in colon cancer tumorigenesis, which is likely distinct from the role of BVES. Further investigation will be conducted in order to fully identify their function in colonic epithelia and CRC cells. Abstract 114 Mechanisms for Pathway-Selective Insulin Resistance: Implications for Obesity Associated Coronary Heart Disease Eileen A. Wang (Rush Medical College), Brian T. Palmisano, John M. Stafford, Vanderbilt University Obesity and diabetes type 2 greatly increase the risk for coronary heart disease, which is a major cause of morbidity and mortality in Western society. Insulin resistance is associated with a number of risk factors including dyslipidemia, elevated serum glucose and insulin levels, and hypertension, culminating in increased risk of type 2 diabetes and coronary heart disease. The liver is a major contributor in triglyceride production and glucose metabolism. Hepatic lipid metabolism is regulated through de novo lipogenesis, fatty acid esterification, and VLDL secretion. An interesting observation is that with obesity, insulin signaling is impaired for glucose metabolism, yet seems to be intact with regards to promoting lipid synthesis. In hepatic metabolic disease, hepatic glucose and lipid metabolism are altered, causing a dysregulation of insulin dependent glucose metabolism and increased insulin dependent lipid metabolism. This is called pathway selective insulin resistance. It is not known the mechanisms of that lead to this phenomenon. We hypothesize that pathway-selective insulin resistance can be induced in mice fed with a high fat diet through induction of known pathways. C56B1/6 mice that were fed a high fat diet for 8 weeks. Mice were fasted for 5 hours and hyperinsulinemic-euglycemic clamp was performed with a glucose and insulin infusion and a saline infusion. It was found that mice fed a high fat diet were insulin insensitive compared to mice fed a low fat diet. To determine how insulin regulates mRNA targets of known glucose and lipid metabolic pathways in the setting of hepatic insulin resistance in vivo, mRNA of liver cells were extracted and reverse transcribed into cDNA. Relative mRNA expression of known genes in the high fat diet and low fat diet treated with insulin was compared to control saline low fat diet. We found that insulin treatment in high fat mice was unable to repress Pepck-1 and G6pase compared to insulin treatment in the low fat diet mice. Additionally, insulin treatment in high fat diet mice upregulated the expression of Sqs1 and SREBP2, thus inducing the lipogenesis pathway. A high fat diet induces pathway-selective insulin resistance in mice. Deep sequencing data will reveal novel gene targets and pathways of pathway-selective insulin resistance. Our proposal is a systems approach to discovering pathway selective insulin resistance with regards to mRNA target. A better understanding of insulin regulation with mRNA may yield novel insights into insulin regulation of gene expression such as novel transcriptional regulators of insulin sensitivity in the pathway that will suppress hepatic gluconeogenesis and suppress de novo lipogenesis. An understanding of the genes responsible for pathway selective insulin resistance may result in a more effective treatment for diabetes type 2 and a reduction in cardiovascular complications. Abstract 115 The use of family centered programming to improve physical activity for parents and preschool children Bianca Marks (Meharry Medical College School of Medicine), David Maynard, William Heerman MD, Shari Barkin MD, Vanderbilt University Medical Center Childhood obesity affects 1 in 4 children in the state of Tennessee, with Latino children being at a disproportionally higher risk. The GROW trial is an NIH funded, large randomized controlled trial aimed at preventing childhood obesity through family centered and community focused behavioral intervention programming This project fits under the umbrella of the GROW trial, and its specific purpose is to assess how underserved families enrolled in GROW with preschool aged children community centers and programming. It was hypothesized that the undeserved families will use community centers and programming in ways that are most convenient. This specific project was done by conducting ethnographic field work at the two Nashville community centers that participate in the GROW study. The results from this study suggested that programs centered on swim and dance were most popular with preschoolers. Additionally, the peak time of day for preschool attendance was the afternoon. Furthermore, families utilized programming most when they shared strong social networks with other families. Another finding of particular importance is that bilingual classes had higher attendance rates that classes that were solely offered in English. Overall, the results from this study can inform the approach of the GROW trial as a whole as well as Nashville Parks and Recreation. Abstract 116 A mouse model of diabetes-accelerated atherosclerosis exhibits elevated endothelial cell Vcam1, Icam1 and Ccl2 expression: Possible role for hyperglycemia in combination with inflammation Dilreet Rai1, Jenny E. Kanter2 and Karin E. Bornfeldt2 1.University of Missouri-Kansas City School of Medicine, Kansas City, MO, 2. Diabetes and Obesity Center of Excellence, University of Washington School of Medicine, Seattle, WA Diabetes results in an increased risk of cardiovascular disease mediated by increased atherosclerosis. Diabetes accelerates early atherosclerotic lesions, which are primarily composed of myeloid cells. Preliminary data show that freshly isolated aortic endothelial cells from a mouse model of diabetes-accelerated atherosclerosis (streptozotocin-induced diabetes in LDL receptor-deficient mice) express higher levels of adhesion molecules including Vascular Cell Adhesion Molecule 1 (VCAM-1) and Intercellular Adhesion Molecule 1 (ICAM-1) as well as the chemokine CCL-2. Whereas the effects of glucose on Human Umbilical Vein Endothelial cells are known, it is not known if similar changes occur in mouse primary endothelial cells in a model of diabetes-accelerated atherosclerosis. We hypothesize that diabetes, in part, accelerates atherosclerosis by augmenting monocyte adhesion to the endothelium and that the effects on the endothelium is mediated by a combination of elevated glucose and inflammatory mediators. To test the hypothesis that elevated glucose stimulates increased myeloid cell adhesion via increased expression of Vcam1, Ccl2 and Icam1, we isolated primary heart endothelial cells from LDL-receptor deficient mice. Endothelial cells were stimulated in 20 mM glucose plus 5 mM mannitol (osmotic control) or 5.5 mM glucose plus 19.5 mM mannitol in the presence or absence of 20 ng/ml TNF-α for 4 hours. RNA from freshly isolated aortic endothelial cells from steptozotocin-diabetic mice and controls were isolated by flushing the aorta with Qiazol. RNA was isolated, reverse transcribed, and subjected to real-time PCR. As expected, TNF-α stimulated the expression of Vcam1 (8.76±0.8-fold), Ccl2 (34.4±11.7-fold) and Icam1 (12.9±1.7-fold) mRNA compared to unstimulated cells under normal glucose conditions: Vcam1 (1.05±.2-fold), Ccl2 (1.01±0.05-fold), Icam1(1.26±0.5-fold). Interestingly, the increase in TNF-astimulated Vcam1 (14.6±4.6-fold over basal; N=6), Ccl2 (135±73.2-fold; N=5) and Icam1 (37.5±15.8-fold; N=5) expression was further elevated if cells were stimulated in the presence of high (20 mM) glucose. Elevated glucose alone, in the absence of TNF-a, had no effect. Finally, diabetes resulted in similarly increased expression of Vcam1 (2.74 ± 0.1; N=4), Icam1 (2.4 ± 0.4; N=3) and Ccl2 (1.78 ± 0.6; N=4) in aortic endothelial cells from diabetic mice compared to non-diabetic mice; Vcam1 (1.19 ± 0.4-fold; N=4), Icam1 (1.12±0.3-fold; N=4) and Ccl2 (1.05±0.2-fold; N=4). We conclude that diabetes enhances the expression of Vcam1, Icam1 and Ccl2 in the aortic endothelium, and that these changes are mimicked by exposure of isolated endothelial cells to a combination of elevated glucose and TNF-a. Our results suggest that the diabetic effects on the endothelium might be driven by hyperglycemia and an increased inflammatory environment in diabetes. These proinflammatory changes in the vasculature likely play a role in accelerating atherosclerosis in diabetes. Abstract 117 Disruption of Hepatocellular ERBB3 Expression Suppresses Diethylnitrosamine-Induced Hepatocellular Carcinogenesis Michael A. Weintraub (Rutgers - Robert Wood Johnson Medical School), Annam Abbasi, Xiuqi Zhang, Lawrence A. Scheving, William E. Russell, Vanderbilt University Medical Center The liver has an extraordinary ability to regenerate in response to injury. With chronic injury, regeneration can eventually become dysregulated, leading to hepatocellular carcinogenesis. Hepatocellular carcinoma (HCC) is the second leading cause of cancer death with ineffective treatment options and a five-year survival rate of 12%. Members of the ERBB receptor tyrosine kinase family are major regulators of liver regeneration and may have a role in HCC. To test the hypothesis that ERBB3 and EGFR (ERBB1) are important regulators of HCC formation, ERBB3 and EGFR were inactivated in a hepatocellular-specific manner by breeding Cre-recombinase and ERBB3 or EGFR loxP mice. To initiate tumorigenesis, diethylnitrosamine (DEN) was injected into wildtype (WT), ERBB3 hepatocyte-specific knockout (HS-KO) and EGFR HS-KO mouse pups. At 8 months, mouse liver tissue was harvested, tumors were counted and cell proliferation was assessed. It was determined that ERBB3 HS-KO mice, but not EGFR HS-KO mice, had a reduction in hepatic tumors (p<0.01) and lower levels of the two proliferation markers that were evaluated, Ki67 (p=0.05) and cyclin A (p<0.01). ERBB2, normally not expressed in adult mouse liver, was detected in DEN-injected mice. ERBB3 HS-KO mice showed reduced ERBB2 expression (p<0.01), suggesting that the induction of ERBB2 may be partly dependent upon ERBB3. ERBB3 HS-KO resulted in diminished STAT3 activation (p<0.01), but did not affect activation of other downstream signaling molecules AKT, ERK1, or ERK2. These observations suggest that ERBB3 may have a role in hepatocellular carcinogenesis and could become a target in the treatment of HCC. Abstract 118 TRPM7 is a critical regulator of endocrine pancreatic development and islet cell function. Thomas J. Galletta (Northeast Ohio Medical University), Prasanna K. Dadi, David A. Jacobson, Vanderbilt University, Nashville, TN Transient receptor potential melastatin-like 7 (TRPM7) is the most highly expressed TRP channel in the human pancreatic islet. TRPM7 serves many important functional roles, such as modulating Ca2+ influx, apoptotic signaling, neurotransmitter release, and exocrine pancreatic development. While these roles may influence pancreatic islet function, the influence of TRPM7 in the pancreatic islet has not been determined. Here we assessed whether TRPM7 plays a role in the growth and development of the endocrine pancreas, islet cell apoptosis, and glucosestimulated insulin secretion (GSIS). Our approach incorporated mouse models with acute beta cell specific (MIP-cre/ERT-TRPM7fl/fl) and chronic pancreatic specific (Pdx-Cre-TRPM7fl/fl) TRPM7 ablation. First we measured pancreatic mass and total pancreatic insulin from Pdx-CreTRPM7fl/fl mice; pancreatic mass was reduced by 36% (p < 0.05) and total pancreatic insulin was reduced by 47% (p < 0.05) compared to control pancreata. We then evaluated islet cell function by measuring Ca2+ influx and GSIS utilizing islets from both TRPM7 ablation models. TRPM7 activity is modulated by ATP, and in pancreatic beta cells the ATP/ADP ratio increases in response to glucose. Thus, we measured glucose-stimulated beta cell Ca2+ influx and found that it was reduced by 27% (p < 0.001) with ablation of TRPM7. Interestingly, however, islets isolated from both Pdx-Cre-TRPM7fl/fl and MIP-cre/ERT-TRPM7fl/fl mice demonstrated increased GSIS with a 40% increase in Pdx-Cre-TRPM7fl/fl islets (p < 0.01) and a 47% increase in MIPcre/ERT-TRPM7fl/fl islets (p < 0.01). Therefore, TRPM7 may regulate the exocytosis of insulin granules independent of Ca2+ influx. Finally, we measured TRPM7’s effect on islet cell apoptosis because apoptotic signaling has been shown to activate TRPM7 channels. We found that when islet apoptosis was induced via pro-inflammatory cytokine stimulation, glucose-stimulated Ca2+ influx was reduced by 58% in beta cells with acute ablation of TRPM7 (p < 0.001). Additionally, islets with acute beta cell and chronic pancreatic TRPM7 ablation were more sensitive to cytokine-induced apoptosis compared with controls; there was a 25% increase in apoptosis for islets without beta cell TRPM7 (p < 0.01) and a 22% increase in apoptosis for islets without TRPM7 (p < 0.05). Our results indicate that TRPM7 is a critical regulator of endocrine pancreatic development and physiological islet cell function. Furthermore, these results identify an important role for TRPM7 in limiting beta cell destruction during inflammatory conditions associated with diabetic islets. Thus, TRPM7 is a potential therapeutic target for increasing GSIS and reducing beta cell destruction during the pathogenesis of diabetes. Abstract 119 Resting State fMRI and PET Correlations in the Dopamine Reward Pathway in Obese Adults with Type 2 Diabetes Mellitus Le Zhong, B.S. (University of Miami Miller School of Medicine); Waqas Majeed, Ph.D. (Lahore University of Management Sciences); Heidi Silver, Ph.D., R.D. (Vanderbilt University Medical Center); Kevin Niswender, M.D., Ph.D. (Department of Medicine, Vanderbilt University Medical Center); Malcolm J. Avison, Ph.D. (Vanderbilt University Institute of Imaging Science) The dopamine (DA) reward pathway, whose dysfunction is a hallmark of drug addiction, has been hypothesized to exhibit similar alterations in obesity. A recent model of the DA reward pathway suggests that the nucleus accumbens (NAcc) integrates inputs from salience attribution areas including the amygdala, which drive motivated drug and food seeking behaviors, and frontal lobe areas such as medial orbitofrontal cortex (MOFC) that exert cognitive control over this motivational drive. Thus signals from the amygdala have been hypothesized to upregulate the reward pathway, while signals from MOFC have been hypothesized to suppress the cue-driven reward seeking. Using resting state functional magnetic resonance imaging (rsfMRI) and 18F-Fallypride positron emission tomography (PET) data combined with fat-water imaging, biochemical, and demographic data obtained from the 2nd and 6th weeks of a longitudinal study of the impact of insulin therapy on brain DA systems, we examined the impact of adiposity, insulin resistance, NAcc DA D2 receptor availability, and leptin on the relative strengths of the NAcc-Amygdala and NAcc-MOFC connections determined from the strength of the inter-regional temporal correlation (r2) of the resting state fMRI time course. We show that in obese adults with moderate type 2 diabetes mellitus, the ratio of visceral adipose tissue to lean tissue, a direct measure of adiposity, correlated negatively with the non-displaceable binding potential obtained from PET in several brain regions. Treatment with amphetamine (AMPH) led to a significant negative NAcc-MOFC r2 relationship with the total adipose tissue to lean tissue ratio (TAT/LT), but no significant change in the dependence of the NAcc-Amygdala r2 on TAT/LT. Treatment with insulin detemir led to decreased dependence of the NAcc-MOFC r2 and increased dependence of the NAcc-Amygdala r2 on TAT/LT. AMPH administration alone did not significantly change the NAcc-Amygdala r2, while it increased the NAcc-MOFC r2. There were significant correlations between fasting leptin levels, an indirect measure of adiposity, and the NAcc-Amygdala r2 but not the NAcc-MOFC r2. The NAccAmygdala r2 correlation with leptin levels was unaffected by AMPH, but the direction of the correlation was affected by insulin detemir treatment, suggesting an alternate mechanism through which leptin may mediate NAcc-Amygdala connections with modulation from insulin. Our studies provide evidence that adiposity and its correlates play both direct and indirect roles in modulating the DA reward pathway and affecting adjacent cortical regions that govern it. Abstract 120 Regulation of Iron Homeostasis in Macrophages in Response to Helicobacter pylori Infection Cassie Xu (Indiana University School of Medicine), Jennifer Noto, Richard Peek, Vanderbilt University Medical Center Progression to gastric adenocarcinoma from Helicobacter pylori infection, the strongest known risk factor in the development of gastric cancer, is dependent on an interplay between host and pathogen factors. The development of gastric adenocarcinomas induced by H. pylori infection has been observed to be accelerated by iron deficiency; however, the effects on iron levels in infected individuals by H. pylori are largely unknown. As the expression of hepcidin, the major regulator of iron homeostasis which inhibits the release of iron from duodenal enterocytes and macrophages by binding and leading to the degradation of the only known iron exporter, ferroportin, is known to be enhanced by inflammation and infection, we predict that H. pylori infection of macrophages, which play a large role in iron recycling and exhibit strong expression of ferroportin, will lead to an increase of hepcidin expression and a decrease in ferroportin expression. In order to address our prediction, hepcidin and ferroportin expression was assessed in RAW cells, murine macrophages, infected with oncogenic H. pylori cag+ strain 7.13 and its cagA- and cagE- mutants using quantitative real-time PCR. Similarly, hepcidin and ferroportin expression were measured in THP-1 cells, a human monocyte line, that were infected with H. pylori strain 7.13 and concurrently secreted hepcidin protein levels by the infected THP-1 cells were measured using an ELISA assay. In the RAW cells, a moderate increase of hepcidin expression was observed in a cagA- and cagE- independent manner, while ferroportin expression was observed to be decreased in a cagA- and cagE- independent manner. Similar trends in hepcidin and ferroportin expression were observed in the human THP-1 cells, but with a greater induction of hepcidin expression levels. Secreted hepcidin protein levels by infected THP-1 cells were also found to be elevated. This data suggests that hepcidin expression in macrophages is increased upon infection with H. pylori, while ferroportin expression is decreased, demonstrating that H. pylori might have an effect on host iron homeostasis by limiting circulating iron; however, more studies must be done to prove stronger statistical significance. Abstract 121 Technology-Mediated Lifestyle Interventions for Reducing Risk for Type 2 Diabetes: a Systematic Review Rachel Bian (University of Michigan Medical School); Neal Kaufman, MD; Ananda Sen, PhD; Caroline Richardson, MD, University of Michigan Context: Weight loss through lifestyle interventions, such as those delivered through the Diabetes Prevention Program, has been shown to reduce risk for developing type 2 diabetes. Technology-mediation of these interventions can help overcome barriers to program delivery and can be used to disseminate diabetes prevention programs on a larger scale. Methods: In this systematic review, 6 databases were searched to identify studies reporting weight loss that used technology to carry out diet and exercise interventions targeting those at high risk for diabetes. Controlled trials and prospective cohort studies published after January 1, 2002 were included. Results: The search identified 431 citations, and out of those ten studies met inclusion criteria, evaluating a total of 14 technology-mediated intervention arms. Average weight loss ranged from 0.1kg to 6.7kg across the intervention arms. Effects of intervention on diabetes incidence and decreased A1c levels were also noted, and results of technology-mediated interventions were compared to that of standard care and in-person intervention controls. Conclusion: Technology-mediated diabetes prevention programs can result in clinically significant amounts of weight loss and cause significant decreases in prediabetes prevalence and diabetes incidence. Abstract 122 Role of Discoidin Domain Receptor 1 (DDR1) in Mesangial Cells Vivian Joan Ortiz Baez (San Juan Bautista School of Medicine, Caguas PR.), Ambra Pozzi, and Corina Borza. Department of Medicine, Division of Nephrology, Medical Center North, Vanderbilt University, Nashville, TN. Discoidin Domain Receptor 1 (DDR1) is an extracellular matrix and tyrosine kinase receptor that binds to and is activated by collagen. Up-regulation of this receptor has been observed in many fibrotic diseases, including kidney diseases. However, whether up-regulation of this receptor contributes to or counteracts the disease process in unclear. Based on the finding that collagen-mediated activation of DDR1 leads to increased collagen synthesis, and collagen is a key matrix deposited in the course of fibrosis, we hypothesize that activation of DDR1 plays a deleterious effect in kidney fibrosis. Our study was aimed to identify the potential pro-fibrotic pathways activated by DDR1. To do this, we utilized kidney cells either lacking or expressing DDR1 and analyze the activation of three major mediators of fibrosis, namely ß-catenin, STAT3 and c-Jun, upon collagen stimulation. Upon collagen stimulation no differences in total ß-catenin and c-Jun were observed between untreated and collagen treated cells. Interestingly the phosphorylation of STAT3 on Y705 was decreased in cells expressing DDR1 stimulated with collagen compared to collagen-stimulated DDR1-null cells. Although the well-accepted function of activated (e.g., phosphorylated) STAT3 is to translocate the nucleus and activate pro-fibrotic signaling, cytokine mediated-STAT3 phosphorylation has been recently shown to protect certain organs against damage and fibrosis. Thus, it is conceivable that DDR1 might promote collagen synthesis by decreasing the activation of STAT3. More studies are needed to determine whether indeed decreased STAT3 phosphorylation in DDR1 expressing cells stimulated with collagen is required to drive DDR1-mediated production of collagen. Abstract 123 Effects of a clinically relevant dose of immunosuppressant drug, Tacrolimus, on human islet function and survival in vivo Kelsey Eitel (Loyola University Chicago Stritch School of Medicine), Scott Wisniewski, Ana Padgett, Chunhua Dai, Alvin C. Powers, Vanderbilt University Patients undergoing organ transplantation are at an increased risk of developing diabetes mellitus post-transplantation. This is thought to be due to the diabetogenic effects of immunosuppressive drugs, but the mechanism of this effect is poorly understood. To test whether a clinically relevant dose of the immunosuppressive drug Tacrolimus (TAC) impairs human islet function and survival in vivo, we transplanted 1,500 islet equivalents (IEQ) beneath the kidney capsule of immunodeficient, normoglycemic NOD-scid IL2r-γ-null (NSG)-DTR mice and treated them with Diphtheria toxin (DT) to ablate mouse beta cells. Two weeks later the normoglycemic mice were treated with 0.25 mg/kg/day of TAC or saline via a subcutaneous osmotic pump for four weeks. The TAC dose needed to obtain a clinically relevant level was determined by measuring the TAC level in blood. During an intra-peritoneal glucose tolerance test, TAC-treated mice had a slightly elevated fasting blood glucose (TAC 158 mg/dl, n = 3 vs. control 87 mg/dl, n = 3, p = 0.0373) and had impaired glucose clearance with the most significant blood glucose elevation seen at 15 minutes post-IP glucose injection (TAC 392 mg/dl, n = 3 vs. control 182 mg/dl, n = 3, p = 0.0242). TAC treatment did not change human basal insulin levels but did impair glucose/arginine-stimulated human insulin secretion. Human islet grafts in TAC-treated mice and control mice had similar levels of beta cell apoptosis (n = 3/group, p = 0.4226), beta cell proliferation (n = 3/group, p = 0.4514), and beta (n = 3/group, p = 0.0804), alpha (n = 3/group, p = 0.0564), and delta (n = 3/group, p = 0.3343) cell ratios. These results suggest that a clinically relevant dose of TAC impairs human beta cell function, but does not affect beta cell survival, proliferation, or mass. Abstract 124 Designing Targeted Fracture Therapies for Diabetic Patients Sagar S. Deshpande (University of Michigan), Noah S. Nelson, Yekaterina Polyatskaya, Ernestina Schipani, Karl J. Jepsen, Steven A. Goldstein, Steven R. Buchman, University of Michigan. It is clinically well established that diabetic bone fractures heal poorly, often taking twice as long as non-diabetic fractures to achieve consolidation. Despite this fact, there are no current therapies targeted at accelerating bone healing in this specific population. Our laboratory has previously utilized Deferoxamine, an FDA-approved iron chelator, as an osteogenic and angiogenic agent in the setting of irradiated fracture healing. Moreover, Deferoxamine has demonstrated utility in healing diabetic pressure ulcers and other soft-tissue deficits. As such, it our hypothesis that administration of Deferoxamine will create bony unions in a diabetic, criticalsized femoral fracture in a rat model. Currently, this study is in pilot phase, wherein we are evaluating the ideal temporal endpoint for this study. Once we have done so, we plan to move forward to the main arm of the study, in order to assay the efficacy of Deferoxamine in our model. Abstract 125 Evaluating Methods to Enhance Tufts D2d Study Recruitment Pamela Sherwood (Tufts University School of Medicine), Sarah Gunn, MS, Anastassios Pittas, MD, Tufts Medical Center D2d is a multi-center clinical trial to test whether vitamin D supplementation reduces diabetes risk in people at high-risk (pre-diabetes). A major recruiting challenge for D2d is that 9 out of 10 people with pre-diabetes are unaware of their risk. This study addresses two major aims: (1) to assess the highest yield methods of recruitment for the study at the present time and (2) to evaluate the attitudes of current participants toward the use of social networking for clinical trial recruitment. To test the hypothesis that referrals from primary-care physicians were the highest yield method of recruitment, first, a chi square test was performed to assess whether there was a significant difference in the method of recruitment for participants who made it to randomization as opposed to those who did not qualify for randomization into D2d. Next, pairwise comparisons of the main D2d recruitment methods were performed. There was a significant difference (p=0.025) in recruitment methods for those who made it to randomization as compared to those who did not make it to randomization. It was shown by pairwise comparison that referral was the highest yield recruitment method as compared to advertisements (p=0.005), database (p= 0.038), and other (p=0.014). To test the hypothesis that Tufts D2d study participants are active on social networking sites and would be interested in using social networking to learn about clinical trials for which they are eligible, surveys were created and distributed to participants by mail and email. Survey results were analyzed using basic statistics. 60% of survey respondents used social networking and 40% said that they would use or join a social networking site to receive information about the D2d study or other clinical trials. 100% of those who would use a social networking site to site to receive information about the D2d responded that their preferred platform would be Facebook. The results support the conclusion that referrals from primary-care physicians are the highest yield method for recruiting pre-diabetic participants to D2d. More data is needed to fully analyze the results of the social networking survey so no valuable conclusions can yet be made from the data available. Abstract 126
