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5th annual

researchsummit
UNIVERSITY OF WISCONSIN DEPARTMENT OF SURGERY

Wednesday, January 22, 2014


The Marquee at Union South 1308 West Dayton Street Madison, Wisconsin

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Innovation

Welcome
Welcome to the 5th Annual University of Wisconsin Department of Surgery Research Summit.
Our theme this year is Innovation, and we anticipate that the Summit will be an opportunity to showcase the
vigorous creativity and novelty that has always characterized our Departmental research efforts. We are honored
by the presence of our two guest keynote speakers this year, Irving L. Kron, MD, Professor and Chair of the
Department of Surgery, University of Virginia, and Rebecca M. Blank, PhD, Chancellor of the University of
Wisconsin, Madison. Dr. Kron is internationally recognized in Surgery and Cardiovascular Surgery with extensive
expertise in cardiac transplantation and the management of complex aortic aneurysms. Chancellor Blank is
a world-renowned expert in economics and the interrelationships between research innovation and economic
growth. Prior to becoming Chancellor of the University of Wisconsin, Madison, she served as Dean and Professor
of the Gerald R. Ford School of Public Policy at the University of Michigan, and Secretary of the US Department
of Commerce. Our speakers will provide their unique and welcome insights into the impact of innovation in
surgical research and academia.
This year, five awards will be given in honor of five individuals whose iconic contributions have made our own
research enterprise possible. The Layton F. Rikkers, MD Best Oral Presentation Award is named in honor of our
previous Chairman, who established the academic vigor and strength of our Department. The K. Craig Kent, MD
Best Basic Science Poster Award honors our current Chairman, who has led our department to its current standing
as the fifth-highest NIH funded Department of Surgery in the nation. The Charles N. Ford, MD Best TranslationalClinical Poster Award is named after our previous Otolaryngology Division Chair who built a world-class voice and
laryngology enterprise within our Department. The Debra A. Hullet, PhD Best Basic Science Poster Award - Student
honors a beloved colleague whose contribution as a leader and mentor to many in our Department has been
even greater than her own research accomplishments. The Louis C. Bernhardt, MD Best Translational-Clinical
Poster Award Student honors a man whose long-standing mentorship to our medical students and residents
has been legendary.
Thank you for taking time out of your busy schedules to attend this years Summit. We hope that todays
activities will increase your awareness of the innovative research activities that are taking place every day in our
Department of Surgery.
Welcome to the Summit.
Nathan V. Welham, PhD and Clifford S. Cho, MD
Program Co-Chairs

Acknowledgements
We would like to thank the following individuals who served on the organizing committee and were invaluable in
their contributions to the planning of this event.
Herb Chen, MD
Mary Marshall
Kelly Mallon
Mary Nutt
Karen Williams

We would also like to recognize the abstract reviewers who served on our program committee. The following
individuals generously donated their time to review the 144 abstracts that were submitted.
Suresh Agarwal, MD
Shahab Akhter, MD
Amal Alhefdhi, MD
Rebecca Bartlett, MA
Jyoti Brar, MD
Guilherme Campos, MD, PhD
Luis Fernandez, MD
David Foley, MD

Ankush Gosain, MD, PhD


Samuel Gubbels, MD
Lian-Wang Guo, PhD
Michael Hammer, PhD
Deb Hullet, PhD
Gregory Kennedy, MD, PhD
Suzanne King, MA
Ciara Leydon, PhD

Kenneth Meredith, MD
Joshua Mezrich, MD
Sarah Oltman, MD
Amy Reeder, PhD
Madhu Roy, MD, PhD
David Schneider, MD
Lee Wilke, MD

Finally, we would like to acknowledge todays onsite judges whose work is in determining the winners of the
Bernhardt, Ford, Hullett, Kent and Rikkers awards.
Suresh Agarwal, MD
Shahab Akhter, MD
Amal Alhefdhi, MD
Michael Bentz, MD
Herb Chen, MD
Michelle Ciucci, PhD
Luis Fernandez, MD
Luke Funk, MD, MPH
Ankush Gosain, MD, PhD
Samuel Gubbels, MD

Lian-Wang Guo, PhD


Michael Hammer, PhD
Dixon Kaufman, MD, PhD
Gregory Kennedy, MD, PhD
Craig Kent, MD
Irving Kron, MD
Glen Leverson, PhD
Bo Liu, PhD
Tim McCulloch, MD
Kenneth Meredith, MD

Joshua Mezrich, MD
Sarah Oltmann, MD
Madhu Roy, MD, PhD
Sara Sackett, PhD
David Schneider, MD
Rebecca Sippel, MD
Lee Wilke, MD
Dai Yamanouchi, MD, PhD

Program
University of Wisconsin Department of Surgery
5th Annual Research Summit - Innovation
Wednesday, January 22, 2014
AM
7:30

Continental Breakfast and Registration

8:15 Welcome and Opening Remarks, The Marquee



Herbert Chen, MD, Vice-Chairman for Research, Department of Surgery
Clifford Cho, MD, Program Co-Chair
Nathan Welham, PhD, Program Co-Chair
8:30 UW Department of Surgery Research Talks
Four 10-minute talks each followed by 5-minute Q&A
 Clifford Cho, MD
Adoptive Cell Transfer and Checkpoint Blockade as Combinatorial Cancer Immunotherapy
Timothy King, MD, PhD
Developing Novel Treatments to Improve Cutaneous Wound Healing
Takushi Kohmoto, MD, PhD
Resuscitation Potential and Functional Evaluation of Human Donation After Cardiac Death
Donor Hearts Using Ex-Vivo Perfusion Technique
Kenneth Kudsk, MD
Infections, Nutrition, Mucosal Immunity and Neuropeptides
9:45

Invited Speaker, The Marquee

Irving L Kron, MD
Professor and Chair, Department of Surgery
University of Virginia School of Medicine
Development of a Clinical Trials Network
10:30

Oral Poster Viewing and Judging, Varsity Hall

11:30

Lunch & Poster Viewing, Varsity Hall

PM
12:30

Research Update, The Marquee

K. Craig Kent, MD


A.R. Curreri Professor of Surgery
Chairman, Department of Surgery
Professor of Surgery, Division of Vascular Surgery

Program
1:30

(continued)

Invited Speaker, The Marquee

Rebecca M Blank, PhD


Chancellor, University of Wisconsin-Madison
2:15

Coffee and Ice Cream Break

2:45 Top six abstract presentations


5 minutes + 3 minutes Q&A
Christopher Dodgion, MD
Expanding the Use of Free Text in EMR to Study Breast Cancer
Sarah Franco, BS
TGF-beta and Its Signaling Protein, Smad3, Play a Role in the De-Differentiation of
Smooth Muscle Cells
Kun-Tai Hsu, MD
Is Central Lymph Node Dissection Necessary for Parathyroid Carcinoma?
Harry Nayar, MD
The State of Global Health Training in Plastic Surgery Residency: Pragmatic
Considerations and Future Directions
Jennifer Philip, BS
-arrestins Regulate Mitochondrial Oxidative Stress and Collagen Synthesis in Adult
Human Cardiac Fibroblasts
Martynas Ziemelis, BS
Protective Role of Complement Blockade in the Context of Renal Ischemia-Reperfusion
Injury and Kidney Transplantation
3:30 UW Department of Surgery Research Talks
Four 10-minute talks each followed by 5-minute Q&A
Timothy McCulloch, MD
High Resolution Manometry: The Future of Dysphagia Management
Joshua Mezrich, MD
Smoke em If You Got em Turns Out This May Be Bad Advice
Daniel Ostlie, MD
Optimal Pain Management for Pectus Excavatum Repair
Dai Yamanouchi, MD, PhD
Role of Balanced Mineralization in Aortic Disease
4:45

Closing and Announcement of Research Prize Winners

5:15

Reception, the lounge area just outside The Marquee at Union South

Abstracts
GROUP ONE: BASIC SCIENCE
Poster
Number:
0. Semidirect Pathway Regulates Indirect Alloresponse by Inducing Abortive Activation of CD4 T Cells Clones;
Bracamonte-Baran W, Burlingham W
1. Role of monocytes in Collagen Type V peptide specific responses in LDLR-/- atherosclerotic mice; Vrushali Agashe,
Arick Park, Ewa Gan, John Kernien, Daniel Greenspan, William Burlingham
2. Inhibition of Notch signaling Reduces Healing of Splinted Cutaneous Excisional Wounds in Mice;
Marielle Brenner, B.S., Alex Thompson, Taylor J Jaraczewski, B.S., Priya R Pathak, Madhuchhanda Roy, M.D., Ph.D.,
Timothy W. King, M.D., Ph.D., FACS, FAAP
3. Bombesin stimulates Paneth cell antimicrobial enzyme levels and intestinal antibacterial function otherwise lost with
parenteral nutrition.; Rebecca A. Busch, MD; Joseph F. Pierre, PhD; Aaron F. Heneghan, PhD; Xinying Wang, MD;
Kenneth A. Kudsk, MD
4. The Aryl Hydrocarbon Receptor has a Protective Effect in Tumor Cells When Expressed; Carney, Patrick; Megna, Bryant;
Kennedy, Gregory; Geiger, Pete
5. Poly(ethylene glycol)-containing hydrogels promote primary granule release from human blood-derived
polymorphonuclear leukocytes; Hannah Caitlin Cohen, Tyler Jacob Lieberthal, W. John Kao
6. Video-based method of quantifying performance and instrument motion during simulated phonosurgery;
Ketan Surender, M.Sc., Ellen Conroy, Zhixian Geng, Ph.D., Ting Chen, M.D., Seth Dailey, M.D., Jack Jiang, M.D., Ph.D.
7. Thailandepsin A, a Novel HDAC Inhibitor, Decreases Tumor Cell Survival and Activates the Notch Pathway in Carcinoid
Cancer.; Ajitha Dammalapati, April Harrison, Madhuchhanda Roy, Colin Korlesky, Renu Nair, Cheng Wang, Yi-Qiang Cheng,
Renata Jaskula-Sztul and Herbert Chen
8. TGF-beta/Smad3 Stimulates Stem Cell/Developmental Gene Expression and Vascular Smooth Muscle Cell DeDifferentiation; Daniel DiRenzo; Xudong Shi; Lianwang Guo; Sarah Franco; Bowen Wang; Stephen Seedial; K. Craig Kent
9. Mechanical Testing of Modeled Rib Fractures with Plated Fixation on the Internal Face; Elegbede, A., Goodspeed, M.,
Vanderby, R., Agarwal, S.
10. TGF-beta and its Signaling Protein, Smad3, Play a Role in the De-Differentiation of Smooth Muscle Cells;
Sarah Franco, Xu Dong Shi, Lian-Wang Guo, Daniel DiRenzo, Toshio Takayama, Bowen Wang, Bo Liu, and K. Craig Kent
11. Residential Arterial Cells and Circulating Bone Marrow-Derived Cells both Contribute to Intimal Hyperplasia in a Rat
Allograft Carotid Transplantation Model; Yi Si
12. Transgenic Reporter Mice Enable Delineation of Temporal and Spatial Enteric Cholinergic Neurogenesis;
Scott Lee, BS, Christopher S. Erickson, BS, Amanda J. Barlow, PhD, Noah R. Druckenbrod, PhD, Miles L. Epstein, PhD,
Ankush Gosain, MD, PhD
13. Early ultrasonic vocalization dysfunction in a PINK1 knock-out model of prodromal Parkinson disease; LM Grant1,2, KP
Cullen1,2, J Pultorak4, CA Kelm-Nelson1 MR Ciucci1,2,3 1Department of Surgery, Division of Otolaryngology-Head and Neck
Surgery;2 Department of Communication Sciences and Disorders;3 Neuroscience Training Program;4 Department of Zoology
14. A Newly Synthesized Collection of SAHA Analogs Show High Potency and Notch Induction in Carcinoid Cancer;
April D. Harrison, Ajitha Dammalapati, Casi M. Schienebeck, Renu Nair, Weiping Tang, Renata Jaskula-Sztul, Herbert Chen
15. Bombesin Improves Adaptive Immunity of the Salivary Gland During Parenteral Nutrition; Aaron F Heneghan,
Joseph F. Pierre, Xinying Wang, Drew A Ronnenburg, Guy E. Groblewski, and Kenneth A. Kudsk
16. Comparison of straw configurations and their effects on vocal economy using a simulated system of straw phonation
therapy; Terah M. Hennick, Ellen R. Conroy, Emily Olszewski, Jack J. Jiang, M.D., Ph.D.

Abstracts

(continued)

Poster
Number:
17. Targeted Delivery of Thailandepsin A using Unimolecular Micelles to Improve Antitumor Effect in Carcinoids;
Renata Jaskula-Sztul, Wenjin Xu, Ajitha Dammalapati, Guojun Chen, Omer Sadak, April Harrison, Qifeng Zheng, Renu Nair,
Colin Korlesky, Cheng Wang, Yi-Qiang Cheng, Shaoqin Gong and Herbert Chen
18. Environmental Pollutants Accelerate Skin Transplant Rejection and Increase Th17 Differentiation in a Murine Model;
Walker Julliard MD, John Fechner, Joshua Mezrich MD
19. Development of an Optogenetic Sensory Peripheral Nerve Interface; Sahil K. Kapur MD, Thomas J. Richner MS,
Sarah K. Brodnick BS, Justin C. Williams PhD, Samuel O. Poore MD PhD
20. Hybrid Electrical-Optical Functional Stimulation System; Sahil K. Kapur MD, Thomas J. Richner MS, Sarah K. Brodnick BS,
Justin C. Williams PhD, Samuel O. Poore MD PhD
21. Early and progressive oromotor and swallowing dysfunction in a PINK1 knockout model of Parkinson Disease;
CA Kelm-Nelson1 LM Grant1,2 , E Paul1 , MR Ciucci1,2,3 1 Department of Surgery, Division of Otolaryngology-Head and Neck
Surgery; 2 Department of Communication Sciences & Disorders; 3 Neuroscience Training Program
22. Cellular Based Immunodepletion and Tomotherapy for Pre-conditioning of Tolerance Induction in a Rhesus Macaque
Renal Allotransplant Model; Kempton, Steve, MD, Haynes, Lynn, MS, Jankowska-Gan, Ewa, Christensen, Neil, BVSc, Forrest,
Lisa, VMD, Hematti, Peiman, MD, Burlingham, William, PhD, Kaufman, Dixon, MD PhD
23. Treatment of rat vocal fold scar by gp46 silencing; Yo Kishimoto, Alice Wei, Yutaka Toya, Nathan V. Welham
24. Tourniquet Master Training for Junctional and Inguinal Hemorrhage Control (TMT); Peter W. WEYHRAUCH, PhD,
James NIEHAUS, PhD, Max METZGER, BS, Shlomi LAUFER, PhD, Calvin KWAN, BS, Carla PUGH, MD, PhD
25. Selective Induction of the Nrf2-Antioxidant Response Element (ARE) Pathway in Hepatocytes Decreases the
Development of Non-Alcoholic Steatohepatitis and is Associated with Augmented Expression of -Oxidation and
Triglyceride Export Genes in Mice; Lung-Yi Lee, Drew Roenneburg, Li Zhang, Jeffrey A. Johnson, David Foley
26. Overexpression of the Nrf2-Antioxidant Response Element (ARE) pathway in hepatic stellate cells (HSCs) ameliorates the
development of Non-Alcoholic Steatohepatitis (NASH) in mice; Lung-Yi Lee, Li Zhang, Drew Roenneburg,
Jeffrey A. Johnson, David Foley
27. Novel isolation and characterization of porcine vocal fold epithelial cells; Elizabeth Erickson Levendoski, Ciara Leydon, &
Susan L. Thibeault
28. Establishment of human pancreatic extracellular matrix co-culture system for endocrine specification and maturation
from human embryonic stem cells; Xiang Li, Sara Dutton Sackett, Cori OBrien, Jon S. Odorico
29. Utilizing proteomics approaches to better understand tissue engineering process; Qiyao Li, Zhen Chang, Basak E. Uygun,
Changying Lin, Brian L. Frey, Lloyd M. Smith, Nathan V. Welham
30. Murine Allograft Elastase-treated Abdominal Aorta Transplantation for Pathogenesis Research of Abdominal Aortic
Aneurysm; Zhenjie Liu, Qiwei Wang, Stephanie Morgan, Jun Ren, Jasmine Giles, and Bo Liu
31. High Molecular Weight Polyethylene Glycol (PEG) Inhibits Myocardial Ischemia-Reperfusion Injury and Improves Survival
and Ventricular Function; James Lloyd, Xianyao Xu, Jennifer L. Philip, Charlie Muller, and Shahab A. Akhter
32. Nerve Stimulation does not Increase Mitochondrial DNA Deletion Mutations in the Tongue Musculature; Entela Lushaj,
Heidi Kletzien, Besa Jonuzi, Maria Barnes, Alexander Cook, Nadine Connor
33. Quantifying Vocal Fold Fluid Content Using Tissue Dielectric Properties; Allison L. Maytag, Jacob P. Meyer, Fuqiang Gao,
Susan C. Hagness, Jack J. Jiang
34. Traditional Medicine in the Sidama Region of Ethiopia: A global health field experience; Alex McAlvay, Yuli Chen,
Heidi Busse, Girma Tefera

Abstracts

(continued)

Poster
Number:
35. Healthcare access and traditional medicine among the Sidama of Ethiopia; Alex McAlvay, Yuli Chen, Heidi Busse,
and Girma Tefera
36. Permeability of canine vocal fold lamina propria; Jacob P. Meyer, Anton Kvit, Erin Devine, Jack Jiang
37. Macrophage Phenotypes in Murine Abdominal Aortic Aneurysm; Stephanie Morgan, Calvin Harberg, Qiwei Wang, Bo Liu
38. PTF1a dosage influences pancreatic cell fate decisions in mouse embryonic stem cells; Gopika Nair, Joel Alvarez,
Jon Odorico
39. The role of endodermal apoptosis in Duodenal atresia formation; PF Nichol, KM Zaremba, AL Reeder, A Kowalkowski
40. Disruptions in endoderm development can result in intestinal atresia formation: time for a new hypothesis?; PF Nichol,
AL Reeder, A Kowalkowski, CM Vezina, KM Zaremba, R Liebl
41. Epidermal growth factor mediated wound repair in human embryonic stem cell-derived epithelial cells;
Liliana Palencia, BS; Amritava Das, MEng; Sean Palecek, PhD; Susan Thibeault, PhD; Ciara Leydon, PhD
42. Regulation of Cardiac Fibroblast Collagen Synthesis by -Arrestins in Maladaptive Ventricular Remodeling;
Jinju Li, Jennifer L. Philip, Tiju Theccanat, Shahab A. Akhter
43. Regulation of Cellular Oxidative Stress and Cell Survival by G Protein-Coupled Receptor Kinase-2;Tiju Theccanat,
Jennifer L. Philip, Nicholas Ludmer, Jinju Li, Xianyao Xu, and Shahab A. Akhter
44. -arrestins Regulate Mitochondrial Oxidative Stress and Collagen Synthesis in Adult Human Cardiac Fibroblasts;
Jennifer L. Philip, Jinju Li, Tiju Theccanat, Xianyao Xu, Shahab A. Akhter
45. The Role of GRK2 and -Arrestins in Maladaptive Post-Infarction Ventricular Remodeling; Jennifer L. Philip, Jinju Li,
Xianyao Xu, Shahab A. Akhter
46. Andrographolide attenuates experimental abdominal aortic aneurysm by inhibiting proinflammatory cells infiltration
and cytokines production; Jun Ren, Zhenjie Liu, Qiwei Wang, Bo Liu
47. Development of an in vitro organotypic wound model to characterize of Notch expression in healing wounds;
Madhuchhanda Roy, Taylor J Jaraczewski, Priya R Pathak, Timothy W King
48. Effects of age and radiation treatment on function of extrinsic tongue muscles; John A Russell, Nadine P Connor
49. Endocrine specification and maturation of human pluripotent stem cell-derived pancreatic progenitors after
transplantation to immunodeficient mice; Sara Dutton Sackett, Xiang Li, Xiaofang Xu, Gopika Nair, Cori OBrien and
Jon S Odorico
50. Pollution Enhances Th17 Differentiation Through the Aryl Hydrocarbon Receptor; Samantha Knopp, John Fechner,
Walker Julliard MD, Joshua Mezrich MD
51. Role of bi-directional regulation in skin allograft survival; Miwa Satomi
52. The Association of CARM1 Isoform mRNA Expression with Patient Clinical Characteristics and Molecular Subtypes in
Breast Cancer Tumors, and the Differential Localization of CARM1 Protein Isoforms; David Shlensky BS, Wei Xu PhD,
Lee Wilke MD
53. NOTCH3 expression suppresses growth and induces cell cycle arrest in follicular thyroid cancer; Yash R. Somnay, BS,
Xiao-Min Yu, MD, PhD, Herbert Chen, MD, FACS
54. Growth factor delivery from mineral coated devices; Darilis Suarez-Gonzalez, Yan Lu, Alisha Diggs, Brett Nemke,
Scott Hollister, Mark Markel, William L. Murphy
55. Removal of CD39+ T cells Uncovers a Population of Col V Specific, CD39-Responder T cells in Normal, Healthy
Individuals.; Jeremy A Sullivan, Ewa Jankowska-Gan, Subramanya Hedge, Matt Pestrak and William J Burlingham

Abstracts

(continued)

Poster
Number:
56. Hyperglycemia suppressed abdominal aortic aneurysm through inhibition of osteoclastogenesis; Teruyoshi Tanaka,
Koji Kurosawa, Dai Yamanouchi
57. Combination effector and memory T cell immunotherapy; a novel approach for optimal anti-cancer immunity;
Andrew Tatar, Siddhartha Sen, Justin Meyers, Prakrithi Srinand, Amanda Contreras, David Mahvi, Clifford Cho
58. Induction of the Nrf2-Antioxidant Response Element (ARE) Pathway in Cultured Hepatic Stellate Cells (HSCs) Leads to
Increased Markers of HSC Activation; Lindsay Taylor, Lung-Yi Lee, Li Zhang, Jeffrey A. Johnson, David Foley
59. Morphological Changes in Fibroblasts induced by PBMCs from Patients with Type II Diabetic Nephropathy;
Trinh Pham, Austin Eckhoff, Deb Hullett, Hans Sollinger, Bianca Tomasini-Johansson
60. Interspecies comparison of vitamin A-storing vocal fold stellate cells; Yutaka Toya, Napaporn Riabroy, Christopher R. Davis,
Sherry A. Tanumihardjo, Yo Kishimoto, Diane M. Bless, Nathan V. Welham
61. Cell Line Variability in the Differentiation of Human Pluripotent Stem Cells to an Otic Progenitor-like Fate; Parul Trivedi,
Cynthia Chow, Su-Chun Zhang and Samuel Gubbels
62. Receptor Interacting Protein (RIP)-mediated necroptosis as a novel target for abdominal aortic aneurysm; Qiwei Wang,
Zhenjie Liu, Jun Ren, Stephanie Morgan, Bo Liu
63. Age related changes in rat cortical motor area dedicated to tongue and jaw; Julie M. Wenninger, John Russell, Heidi
Kletzien, Allison Schasser, Nagheme Thomas, Jeffrey A. Kleim, Nadine P. Connor

GROUP TWO: CLINICAL TRIALS


64. Clinical Evaluation of Stratagraft Skin Tissue as an Alternative to Autografting Deep Partial-Thickness Burns; Lee D.
Faucher, Michael J. Schurr, Kevin N. Foster, Steven E. Wolf, Booker T. King, Christina Thomas-Virnig, Kelly Van Winkle, Allen R.
Comer, Mary A. Lokuta, James H. Holmes IV, B. Lynn Allen-Hoffmann
65. And I think that we can fix it: A qualitative study of surgeons preoperative discussions before high-risk operations;
Jacqueline M Kehler, MD; Kristen E Pecanac, MS, RN; Karen J. Brasel, MD, MPH; Zara Cooper, MD, MSc; Nicole Steffens, MPH,
Martin McKneally, MD, PhD; Margaret L Schwarze, MD, MPP5

GROUP THREE: OUTCOMES


66. Discordance of histological grade between primary and metastatic neuroendocrine carcinoma; Adesoye, Taiwo; Daleo,
Marie A.; Weber, Sharon M.; Winslow, Emily; Loeffler, Agnes G.; Cho, Clifford
67. Elevated Postoperative PTH levels Prevent Bone Mineral Density Improvements in Patients with Primary
Hyperparathyroidism; Kamal Ahmed, MD, Amal Alhefdhi MD;; David F. Schneider, MD, MS; Herbert Chen MD, FACS; Rebecca
Sippel MD, FACS
68. Recurrent Primary Hyperparathyroidism Occurs More Frequently in Patients with Double Adenomas; Amal Alhefdhi,
David Schneider, Rebecca Sippel, and Hebert Chen
69. Comparison of Patient Outcomes in Laparoscopic and Open Proctocolectomy with Ileal Anal-Pouch Anastomosis; Brown,
A.; Kennedy, G. MD, PhD; Tevis, S. MD
70. Comparison of Laparoscopic and Robotic Proctectomy for Rectal Cancer at the University of Wisconsin; Brown, A.;
Kennedy, G. MD, PhD; Tevis, S. MD
71. Survey of Coding Practices by Laryngologists Who Perform Both Office- and Operating Room-Based Procedures for
Dysphonia; Seth H. Dailey, MD; David A. Hyman, MD; Tamara J. LeCaire, PhD
72. National Trends in Pediatric Blunt Spleen and Liver Injury Management and Potential Benefits of an Abbreviated Bed
Rest Protocol; Dodgion CM, Gosain A, Rogers A, St. Peter SD, Nichol PF, Ostlie DJ

Abstracts

(continued)

Poster
Number:
73. Preoperative Cognitive and/or Functional Impairment is Associated with the Need for Higher Level of Care after Hospital
Discharge; Colin C. Ehlenbach, BS; Sarah E. Tevis, MD; Gregory D. Kennedy, MD, PhD; Sarah C. Oltmann, MD
74. The Age Of Patients With Rib Fractures Is Associated With Higher Complication Rates And Increased Length Of Stay.; A.
Elegbede, M. Molnar, H. Jung, A. ORourke, S. Agarwal
75. Surgical Site Infection after Thyroidectomy: A Rare but Significant Complication; Dawn M. Elfenbein, David F. Schneider,
Herbert Chen, Rebecca S. Sippel
76. Faculty Evaluations of Residents Medical Knowledge: Can they be used to predict ABSITE Performance?; Dawn M.
Elfenbein, Rebecca S. Sippel, Robert McDonald, Tammy Watson, John E. Scarborough, John Migaly
77. Patient Profile at Tikur Anbessa Specialized Hospital Vascular Surgery Clinic,Addis Ababa, Ethiopia; Sean Fling, BA.
Girma Tefera, MD. Nebyou Seyoum, MD. Charles Penn, BS. Rakeb Tibebu
78. Outcomes of Open Versus Laparoscopic Ladds Procedure for Intestinal Malrotation in Adults; Lane Frasier, MD; Glen
Leverson, PhD; Ankush Gosain, MD PhD; Jacob Greenberg, MD EdM
79. Long-term Cost-effectiveness of Medical, Endoscopic, and Surgical Management of Gastroesophageal Reflux Disease;
Luke M. Funk MD MPH, James Y. Zhang BS, Joseph M. Drosdeck MD, W. Scott Melvin MD, Kyle A. Perry MD
80. Bariatric Surgery Outcomes in Medicaid vs. Non-Medicaid Patients: Different Cohorts, Similar Outcomes; Luke M Funk
MD MPH, Andrew Suzo BS, Dean J Mikami MD, Bradley J Needleman
81. Nasolacrimal System Fractures Associated with Epiphora and Dacryocystitis; Ravi K. Garg, Michael J. Hartman, Mark J.
Lucarelli, Glen Leverson, Ahmed M. Afifi, Lindell R. Gentry
82. CT-Related Radiation Exposure in Children Transferred to a Level 1 Pediatric Trauma Center; Adam S. Brinkman MD, Kara
G. Gill MD, Carly M Glarner MD, Jocelyn Burke MD, Andrew Rogers, MD, Mary J. Anderson, RN, Charles M. Leys MD, Daniel J.
Ostlie MD and Ankush Gosain MD, PhD
83. Characterizing readmission in ulcerative colitis patients undergoing restorative proctocolectomy.; Thomas P. Hanzlik,
Sarah E. Tevis, MD, Amy Suwanabol, MD, Bruce Harms, MD, Charles Heise, MD, Eugene F. Foley, MD, and Gregory Kennedy,
MD
84. Paradoxical Vocal Fold Motion: Epidemiological Presentation and Clinical Characterization; N.A. Hartley, K.M. McConville
& S.L. Thibeault
85. Is Central Lymph Node Dissection Necessary for Parathyroid Carcinoma?; Kun-Tai Hsu, M.D., Rebecca S. Sippel, MD,
FACS, Herbert Chen, MD, FACS and David F. Schneider, M.D., M.S.
86. Does Postoperative Drain Amylase Predict Pancreatic Fistula Following Pancreatectomy?; Jacqueline S. Israel MD, Robert
J. Rettammel MA, Glen E. Leverson PhD, Laura R. Hanks
87. The Impact of Comorbidity on Mortality in Trauma Outcomes; Christopher Janowak, Philip Mercier, Suresh Agarwal
88. The Venous Anastomotic Flow-Coupler for Free Flap Monitoring: A Prospective Analysis of 85 Microsurgical Breast
Reconstruction Cases; Kempton, Steve, MD, Poore, Samuel, MD, Chen, Jenny, MD, Afifi, Ahmed, MD
89. The Effect of an Integrated System on the Rate of Duplicate CT Scanning in Transferred Trauma Patients and the Costs
Associated with Care; Amy E. Liepert, MD*; Joseph Bledsoe, MD; Sarah Majercik, MD, MBA; and Mark H. Stevens, MD
90. Short-term Clinical Outcomes after Off-pump CABG at a Single VA Medical Center; Entela Lushaj, MD, PhD, Athanasia
Schreiner, NP, Besa Jonuzi, Lucian Lozonschi, MD
91. Surgical Outcomes of Patients with Locally Advanced Non-Small Cell Lung Cancer; Entela Lushaj, Wassim Aghnatios Abi
Jaoude, Ryan Macke, James Maloney

Abstracts

(continued)

Poster
Number:
92. Anatomic Segmentectomy vs. Lobectomy for Stage I Non-Small Cell Lung Cancer (NSCLC): Extent of Resection is
Important in Determining Preservation of Pulmonary Function.; Macke, Ryan, Schuchert, Matthew J, Odell, David D.,
Landreneau, James R., McCormick, Kristen., Pennathur, Arjun; Luketich, James D; Landreneau, Rodney J
93. Midesophageal and Epiphrenic Diverticula: A 15 Year Experience using Minimally Invasive Approaches for Surgical
Management; Macke, Ryan, Luketich, James D., Landreneau, Rodney J., Pennathur, Arjun, Awais, Omar, Weksler, Benny,
Shende, Manisha R, Christie, Neil A, Schuchert, Matthew J, Nason, Katie S, Levy, Ryan M.
94. Adolescent Shared Decision in Plastic and Reconstructive Surgery; McCarthy, J., Mount, D.
95. Robotic Ivor-Lewis Esophagectomy After Neoadjuvant Chemoradiation Therapy; Smith FO MD, Almhanna K MD, Shridhar R
MD PhD, Hoffe S MD, Karl RC MD, and Meredith KL MD
96. Robotic assisted Ivor Lewis esophagectomy in the elderly patient; Andrea M Abbott MD, Tobin Strom MD, Nadia Saeed, Ravi
Shridhar MD, Sarah Hoffe MD, Khaldoun Almhanna MD, Kenneth Meredith MD
97. The Surgical Apgar Score Correlates with an Increased Risk for Readmission in Emergency Surgery Patients; Gajanthan
Muthuvel, BS, Sarah E. Tevis, MD, Suresh K. Agarwal, MD, Gregory D. Kennedy, MD, PhD
98. Dont Just Tell Somebody They Need This Surgery: A Qualitative Study of Patient Decision Making Preferences in HighStakes Surgical Decisions; MJ Nabozny MD, JM Kehler MD, NM Steffens MPH, KJ Brasel MD, ME Gaines JD, TC Campbell MD,
ML Schwarze MD
99. Are all thyroid ultrasound evaluations created equal? Sonographers specialized in thyroid cancer correctly label clinical
N0 disease in well differentiated thyroid cancer; Sarah C. Oltmann, M.D., David F. Schneider, M.D., M.S., Herbert Chen,
M.D., Rebecca S. Sippel, M.D.
100. Lung Transplant Outcomes for High-Risk Patients (LAS50) with Idiopathic Pulmonary Fibrosis (IPF); Satoru Osaki,
Keith C Meyer, James Maloney, Richard D Cornwell, Nilto C De Oliveira
101. Left Ventricular Assist Device Placement in Very Obese Patients: Proceed with Caution; Satoru Osaki, Maryl R Johnson,
Peter S. Rahko, Margaret A Murray, John H. Blabaum, Entela B. Lushaj, Sue Ulschmid, Lucian Lozonschi, Takushi Kohmoto
102. Expanding the Use of Free Text in EMR to Study Breast Cancer; Christopher M Dodgion MD, MSPH, MBA; Thien Nguyen
BS; Marquita R Decker MD, MPH; Heather B Neuman MD, MS; Wei Jiang MS; Yue-Yung Hu MD, MPH; Anita Karcz MD, MBA;
Stuart R Lipsitz ScD; Leonard DAvolio PhD; Caprice C Greenberg MD, MPH
103. Surgical Coaching to Improve Performance; Caprice C Greenberg MD MPH, Hala N Ghousseini PhD, Carly E Glarner MD
MEd, Sudha R Pavuluri Quamme MD MS, Heather L Beasley BA, Douglas A Wiegmann PhD
104. Institutional Variation in Surgical Care for Breast Cancer at Community Hospitals; Marquita R Decker MD, MPH; Stuart
Lipsitz ScD; Christopher M Dodgion MD, MSPH, MBA; Yue-Yung Hu MD, MPH; Thien Nguyen; Wei Jiang MS; Heather B
Neuman MD; Anita Karcz MD, MBA; Leonard DAvolio PhD; Caprice C Greenberg MD, MPH
105. Outpatient Care Alone is Not Sufficient to Prevent Readmission After Vascular Surgery; R. Scott Saunders, MD; Sara
Fernandes-Taylor, PhD; Jason T. Wiseman, MD; Travis L. Engelbert, MD, Jon S. Matsumura, MD, K. Craig Kent, MD
106. Patient Reported Readiness for Discharge is Associated with Higher Patient Satisfaction and Lower Readmission Rates;
Ryan K. Schmocker, MD, Sara Murry MD, Xia Vang BA, Glen Leverson PhD, Linda Cherney Stafford MPH, Emily R. Winslow, MD
107. Reduction of Surgical Site Infections after Laparoscopic Gastric Bypass with Circular Stapled Gastrojejunostomy;
P Shabino MD, A Elegbede MD, R Schmocker MD, M Nabozny MD, M Khoraki MD, M Garren MD, J Greenberg MD,
GM Campos MD
108. A Nomogram Predicting Post-operative Readmission in General Surgery Patients; Sarah E. Tevis MD,
Sharon M. Weber MD, K. Craig Kent MD, Gregory D. Kennedy MD PhD

Abstracts

(continued)

Poster
Number:
109. Examining the role of cervical ultrasound in detecting thyroid pathology in patients with primary hyperparathyroidism;
Deena Weiss, BS, Herbert Chen, MD, FACS
110. General and Vascular Surgery Readmissions in the Modern Era: A Systematic Review; Jason T. Wiseman MD,
Amanda Guzman, Sara Fernandes-Taylor PhD, Travis L. Engelbert MD, R. Scott Saunders MD, and K. Craig Kent MD

GROUP FOUR: TRANSLATIONAL/CLINICAL


111. Notch Signaling in pancreatic differentiation from embryonic stem cells; Joel Alvarez, Gopika Nair, Jon Odorico
112. De novo donor-specific HLA antibodies increase the risk of ischemic cholangiopathy in donor after cardiac death liver
allografts; O. Andacoglu, A.M. DAlessandro, D.P. Foley, T.M. Ellis, A.J. Powell, G.E. Leverson, M.R. Lucey, A.I. Musat
113. Periadventitial Application of Rapamycin-Loaded Nanoparticles Produces Sustained Inhibition of Vascular Restenosis;
Guojun Chen, Xu Dong Shi, Lian-Wang Guo, Yi Si, Men Zhu, Srikanth Pilla, Bo Liu, Shaoqin Gong*, K. Craig Kent*
114. In-vivo treatment of brain dead rat donors with novel fusion protein improves islet health and increases functional
potency, reducing the islet mass required or successful reversal of diabetic hyperglycemia post-transplant.; Peter J.
Chlebeck, Xiaobo Ma, Juan S. Danobeitia, Luis A. Fernandez
115. Sex Differences in Voice-Related Modulation of Mechanosensory Detection Thresholds in the Human Larynx; Michael J.
Hammer, Mallory A. Krueger, Stephanie A. Palm
116. Evaluating the Tongue-Hold Manuever Using High-Resolution Manometry and Electromyography; Michael J. Hammer,
Corinne A. Jones, Jason D. Mielens, Chloe H. Kim, Timothy M. McCulloch
117. Thirty Year Follow-up of Total Hand Replantation: A Case Report; Jacqueline S. Israel MD, Venkat K. Rao MD and Samuel
O. Poore MD, PhD
118. Endoscopic Laryngopharyngeal Reflux Findings are Poorly Correlated with Combined Multichannel Intraluminal
Impedance pH Monitoring; Marie Jette, MS, Eric Gaumnitz, MD, Martin Birchall, MD, Nathan Welham, PhD, Susan Thibeault,
PhD
119. Using High-Resolution Manometry and Videofluoroscopy to Detect Subtle Swallowing Changes in Early and Mid-Stage
Parkinson Disease; Corinne A. Jones, Michelle R. Ciucci, Timothy M. McCulloch
120. iPS-MSCs reduce incidence of Delayed Graft Function in the Non-human Primate Brain Death and Renal Transplant
Model; Laura Zitur, Juan Danobeitia, Saritha Dsouza, Xiaobo Ma, Anthony DAlessandro, Igor Slukvin, Luis Fernandez
121. In-office biopsy for suspected laryngeal and pharyngeal malignancy: safety, tolerance, subsite analysis, and time to
treatment.; Dylan Lippert, M.D.; Matthew R. Hoffman, Ph.D.; Phat Dang, M.D.; Greg Hartig, M.D.; Tim McCulloch M.D.; Seth
H. Dailey, M.D.
122. Functional evaluation of human DCD donor hearts using a continuous isolated myocardial perfusion technique;
potential for expansion of the cardiac donor population.; Satoru Osaki, MD, PhD, Matthew R. Locher, PhD, Entela B.
Lushaj, PhD, Shahab A. Akhter, MD, and Takushi Kohmoto, MD, PhD
123. Sensor-Based Assessment of Cast Placement and Removal; Anne-Lise D. Maag, MD, Shlomi Laufer, PhD, Calvin Kwan, BS,
Elaine R. Cohen, MEd, Rachel L. Lenhart, MS, Natalie C. Stork, MD, Matthew A. Halanski, MD, Carla M. Pugh, MD, PhD
124. Use of Advanced Sensor and Motion Tracking Technology to Classify Clinical Breast Examination Techniques; Anne-Lise
D. Maag, MD, Shlomi Laufer, PhD, Elaine R. Cohen, MEd, Calvin Kwan, BS, Carla M. Pugh, MD, PhD
125. AKT Expression is Associated with Degree of Pathologic Response in Adenocarcinoma of the Esophagus Treated
with Neoadjuvant Therapy; Nadia Saeed, Maki Yamamoto, Jill Weber, Ravi Shridhar, Sarah Hoffe, Khaldoun Almhanna, and
Kenneth L. Meredith

Abstracts

(continued)

Poster
Number:
126. The State of Global Health Training in Plastic Surgery Residency: Pragmatic Considerations and Future Directions;
Harry S. Nayar, MD, MBE; Delora L. Mount, MD; Michael L. Bentz, MD
127. Elevated PTH After Curative Parathyroidectomy Delays Symptom Improvement; Priya R. Pathak, Sara E. Murray, MD,
Sarah C. Schaefer, NP, Glen Leverson, PhD, Herbert Chen, MD, Rebecca S. Sippel, MD
128. Use of the Ankle Brachial Index (ABI) as a possible diagnostic tool for peripheral artery diseases (PAD) in Diabetic
patients at Tikur Anbessa Specialized Hospital, Ethiopia.; Charles Penn
129. Validation of final ioPTH level and risk of disease persistence and recurrence; Mohammad H. Rajaei, David F. Schneider,
Rebecca S. Sippel, Herbert Chen, Sarah C. Oltmann
130. Magnetic resonance imaging evaluation of lateral pterygoid contributions to Eustachian tube opening; Michael H.
McDonald, M.D.; Thomas R. Richards; John Park, M.D., Matthew R. Hoffman, Ph.D.; Jack J. Jiang, M.D., Ph.D.
131. Cost Analysis of Colon Cancer - Comparing Colorectal Surgeons within the Same Institution; David Rivedal BS, Sarah
Tevis MD, Eugene Foley MD, Bruce Harms MD, Charles Heise MD, Gregory Kennedy MD, PhD
132. The Cross-Training and Detraining Effect of Tongue Exercise in the Cranial Sensorimotor System; Allison J. Schaser &
Nadine P. Connor
133. Preclinical and Clinical Evaluation of a Human Skin Substitute Engineered to Secrete Elevated Levels of a Host
Defense Peptide; Christina Thomas-Virnig, Cathy Rasmussen, Allen Comer, Mary Lokuta, Lee Shaughnessy, Sandy Schlosser,
Colette Johnston, Rebecca Bauer, Thomas Cleven, Nathan Wieczorek, B.Lynn Allen-Hoffmann
134. Pharyngeal Pressure and Timing During Bolus Transit; Chelsea Walczak [1], Timothy McCulloch [2], Corinne Jones [3],
William Bleifuss [4]
135. High-throughput Screening Identifies Idarubicin as a Preferential Inhibitor of Smooth Muscle versus Endothelial
Cell Proliferation; Shakti A. Goel, Lian-Wang Guo, Bowen Wang, Song Guo, Drew Roenneburg, Gene Ananiev, F. Michael
Hoffmann, K. Craig Kent
136. Determination of an Accurate and Consistent Method to Measure the Take-off Angle of Supra-aortic Branch Vessels;
Bradley Wendorff, Taylor Smits, Hillary Alberta, Wendy Meadows, Jon Matsumura, M.D.
137. Measuring listening effort using pupil dilation: implications for management of hearing loss; Matthew Winn, Jan
Edwards, Ruth Litovsky
138. Resolution of Thyroglobulin Antibodies after Total Thyroidectomy for Cancer; Jimmy Xu, B.S., Ryan Bergren, B.S., David
Schneider, M.D., Herbert Chen, M.D., F.A.C.S., Rebecca S. Sippel, M.D., F.A.C.S.
139. Yellow coronal vocal fold scar a unique pattern; William G Young, MD; Seth H Dailey, MD
140. Overexpression of Dysadherin in Thyroid Cancer is Associated with Tumor Aggressive Phenotypes; Xiao-Min Yu, MD,
PhD, Celina Montemayor Garcia, MD, PhD, Eric Weinlander, BA, Ricardo V. Lloyd, MD, PhD, Herbert Chen, MD, FACS
141. Protective Role of Complement Blockade in the Context of Renal Ischemiareperfusion Injury and Kidney
Transplantation.; Martynas Ziemelis, Juan S. Danobeitia, Xiaobo Ma, Gengwen Huang, Drew A. Roenneburg and Luis A.
Fernandez
142. The Evolving Integrated Vascular Surgery Residency Curriculum; Brigitte K. Smith, MD, Jacob A. Greenberg, MD, EdM,
Erica L. Mitchell, MD
143. Learning from developing countries in strengthening health systems: An evaluation of personal, professional, and
institutional impact of global health volunteers; Busse H, Aboneh E, Azazh A, Wubben R, Tupesis J, Tefera G
144. Re-differentiation of human T cell-derived induced pluripotent stem cells into the T cell lineage; Matthew E. Brown,
William J. Burlingham, PhD

G ROUP ONE

Basic Science

0
Semidirect Pathway Regulates Indirect Alloresponse by Inducing Abortive
Activation of CD4 T Cells Clones
Bracamonte-Baran W, Burlingham W, University of Wisconsin

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type

Bracamonte-Baran W
(608)263-7048
bracamonte-baran@surgery.wisc.edu
Research Assistant
Surgery
Dr. Will Burlingham
Basic Science

Semidirect pathway implies allogeneic antigen acquisition (AgAq) of MHC molecules by dendritic cells (DC).
Its role as an amplifier of antiviral immune response had been described. We hypothesize that it might be
associated with the tolerogenic effect induced by maternal-fetal microchimerism (MMc). In vivo MLR were
performed using the NIMA (non-inherited maternal antigen) murine model, in which H2b/b offspring are
exposed to H2d antigens and are prone to develop MMc. In this assay CFSE-labeled CD4 T cells from TEa
TCR transgenic mice (specific for indirect pathway by recognizing IEd peptide-IAb complex) were iv injected
in NIMA mice, which statuses of AgAq (H2Kd) were previously screened by FACS (hemisplenectomy). After
4 days spleen and lymph nodes were harvested and proliferation of TEa CD4 T cells determined by CFSE
dilution method. One third of NIMA mice had detectable H2Kd AgAq on myeloid DCs (CD11c+/B220-)
subset. Mice with undetectable AgAq prior to MLR allow proliferation of TEa cells (8 division cycles), whereas
those with detectable AgAq show regulatory properties, occurring minimal proliferation of TEa cells,
undergoing 1 division cycle and shifting to non-nave intermediate immunophenotype (CD62Llo/CD44lo).
These findings are compatible with induction of abortive activation of allospecific CD4 TEa cells.

1
RoleofmonocytesinCollagenTypeVpeptidespecificresponsesinLDLR/
atheroscleroticmice

VrushaliAgashe,ArickPark,EwaGan,JohnKernien,DanielGreenspan,WilliamBurlingham

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

VrushaliAgashe
(608)2632485
agashe@surgery.wisc.edu
GraduateStudent
Transplantation
WilliamBurlingham
BasicScience
Autoimmunity,Atherosclerosis,Murinemonocytes,Th17cells,CollagentypeV

PreviousexperimentsidentifiedColV1chainasatargetofCD4Tcellandhumoralautoimmunityinpatientswith
coronaryarterydisease(CAD)andinmousemodelsofthedisease.UsingcelldepletionassaysonpatientPBMCs,CD14+
monocyteswerefoundtobecriticalAPCsforTh17cellsinvolvedinColVreactivitywhichdependedonIL17,IL1,IL22
andTNF.AColVspecificresponsewasalsoobservedinsplenocytesofLDLR/(B6background)mice,fedahigh
cholesteroldietfor14weeks.UsingaDTHadoptivetransferassayweanalyzedtheroleofmonocytesinthisresponse.
Bonemarrowmonocytesweremixedina1:5ratiowithsplenicCD4+TcellsandresponsetoColVwasdetermined.Col
VreactivitywasconsistentlyhigherwithisolatedmonocytesandTcellsascomparedwithwholesplenocytes.The
responsedependedonIL1andIL17,butnotonIL22andTNF,withvariableIFNdependence.Theseresultscontrast
withtheColVresponseselicitedbyS.C.immunizationwithantigen+CFAinWTB6mice.Thoseresponseswere
dependentnotonlyonIL1andIL17(IFN),butalsoonIL22andTNF.Ourresultsimplyadifferentmechanismof
responsetoselfantigensinatherosclerosis,perhapsduetothepredominanceofmonocytesasAPCs.

2
Inhibition of Notch signaling Reduces Healing of Splinted Cutaneous Excisional
Wounds in Mice
Marielle Brenner, B.S., Alex Thompson, Taylor J Jaraczewski, B.S., Priya R Pathak, Madhuchhanda Roy, M.D., Ph.D., Timothy W.
King, M.D., Ph.D., FACS, FAAP

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Marielle Brenner
(608) 358-9963
mabrenner@wisc.edu
Medical Student
Plastic & Reconstructive Surgery
King
Basic Science
Wound Healing, Notch, epidermis, stented wound model, keratinocytes

Introduction
Complications due to impaired wound healing affect millions of people throughout the world & management of wounds
in the US alone cost more than $20B/yr. The predominant cell type in cutaneous wound healing are keratinocytes, which
proliferate, migrate across the wound bed, & then differentiate into a multilayered, waterproof barrier. Our lab is
interested in increasing the efficiency of the wound healing process to help negate some of the problems with nonhealing wounds by investigating the underlying mechanisms that regulate this process. Previously, we have shown that
the Notch signaling pathway plays a role in keratinocyte proliferation & migration in vitro. Here, we tested the effects of
inhibition of Notch signaling on re-epithelialization of wounded epidermis in a murine stented wound model that
resembles cutaneous healing by granulation tissue deposition similar to humans. Our hypothesis is that blocking the
Notch pathway will decrease wound healing in otherwise healthy mice.
Methods
6 week old, healthy male mice were anesthetized & after depilation, full-thickness wounds were placed on their backs
extending through the panniculus carnosus (n=9). A 12 mm dia. silicone stent was secured around each wound with
cyanoacrylate glue & interrupted 6-0 nylon sutures to prevent healing by contraction. Wounds were dressed with sterile
N-terface & Tegaderm, then the mice were wrapped with Coban, which was secured with Transpore tape. Dressings
were changed every day after topical application onto the open wound bed of the Notch inhibitor, DAPT (30 uM) or
vehicle (0.1% DMSO) for 10 days. Digital photographs were taken at every dressing change. At the end of the
experiment, the mice were sacrificed & wounds were harvested for histological analysis. Wounds were analyzed using
ImageJ software & expressed as a ratio of wound area to stent area, with scaling normalized to the inner circumference
of the splint as a standard. Wound area was calculated as a percent area of the original wound size. Statistical
significance was defined as p<0.05 using the students t-test.
Results
Partial to complete re-epithelialization was seen in the wounded tissues over the experimental period in both the
control & DAPT treated groups. Rates of wound closure were significantly reduced in DAPT treated wounds compared
to control (p<0.02). No significant local side effects such as increased edema or allergic reaction were noted in the
DAPT-treated mice.
Conclusion
DAPT decreases re-epithelialization of cutaneous wounds in the in vivo murine stented wound healing model, indicating
that Notch signaling plays a crucial role in wound healing in immunocompetent mice. Histological analysis including
markers for keratinocyte migration, proliferation & angiogenesis/vascular density are underway. Based upon our
findings, further study of Notch in wound healing should be conducted which may then lead to better therapeutics for
the wound healing process in patients.

3
Bombesin stimulates Paneth cell antimicrobial enzyme levels and intestinal antibacterial function otherwise lost with parenteral nutrition.
Rebecca A. Busch, MD; Joseph F. Pierre, PhD; Aaron F. Heneghan, PhD; Xinying Wang, MD; Kenneth A. Kudsk, MD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Rebecca Busch
(312) 909-0213
rbusch@uwhealth.org
Resident
General Surgery
Kenneth Kudsk
Basic Science
parenteral nutrition, innate immunity, infection, gastrointestinal, bombesin

Introduction: Parenteral nutrition (PN) increases the risk of infection in critically ill patients when compared to enteral
feeding. Experimentally, impairments in both adaptive and innate mucosal immunity have been implicated in increasing
the infections during PN. In innate immunity, Paneth cells, under cholinergic stimulation, secrete antimicrobial peptides
including sPLA2, lysozyme, and RegIII- into the intestinal lumen to protect mucosa from bacteria. We previously
demonstrated reduced tissue and luminal levels of the Paneth cell antimicrobial protein, sPLA2, following PN. Bombesin
(BBS), a gastrin releasing peptide analogue, has been shown to stimulate gut adaptive but not innate immune function.
Hypothesis: Since PN reduces intestinal sPLA2, we hypothesized PN would decrease other antimicrobial Paneth cell
products (lysozyme, RegIII-). Secondly, we hypothesized the addition of BBS to PN (PN+BBS) would stimulate expression
of antimicrobial Paneth cell products when compared to PN alone restoring bactericidal activity.
Methods: Male ICR mice underwent IV cannulation and were randomized to chow, PN, or PN+ 15 g TID BBS
(n=7/group) for 5 days. Ileal tissue was analyzed for expression of sPLA2, lysozyme, and RegIII- by RT-PCR. Tissue was
also stimulated with the cholinergic agonist, bethanechol (100 M), to measure bactericidal activity against
Pseudomonas.
Results: Compared to Chow tissue, PN significantly decreased expression of sPLA2, lysozyme, and RegIII-. The addition
of BBS to PN significantly increased expression of sPLA2, lysozyme, and RegIII- to levels similar to and not significantly
different from Chow feeding. Functionally, PN demonstrated a significant decrease in bactericidal activity while the
addition of BBS to PN restored bactericidal activity to Chow levels (Table 1).
Table 1. Effects of feeding Chow, PN, and PN+BBS on expression of ileal tissue sPLA2, lysozyme, and RegIII- and
bactericidal activity.
Chow

PN

PN+BBS

sPLA2

.99 + 0.2

0.54 + 0.1*

0.99 + 0.1

lysozyme

1.7 + 0.07

1.3 + 0.07*

2.0 + 0.1

RegIII-

3.4 + 0.09

1.3 + 0.1*

2.7 + 0.6

Bactericidal activity

29.0 + 4.3

14.4 + 1.9*

32.6 + 7.7

*P <0.05 vs. Chow and BBS


Conclusion: The neuropeptide BBS preserves innate immune antimicrobial enzymes and anti-bacterial function that was
lost with PN.

4
The Aryl Hydrocarbon Receptor has a Protective Effect in Tumor Cells
When Expressed
Carney, Patrick; Megna, Bryant; Kennedy, Gregory; Geiger, Pete
Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Patrick Carney
(608) 445-3493
pcarney@wisc.edu
Undergraduate Student
General Surgery
Kennedy Lab
Basic Science
Aryl Hydrocarbon Receptor, Colon and Rectal Cancer, Tumor, Chemotherapy, 5-Fluorouracil

Introduction: The aryl hydrocarbon receptor (AhR) is a protein that is responsible for responses to exogenous and
endogenous compounds in vivo. Recent data indicates that loss of AhR increases susceptibility to colon tumor induction
in animals. However, the role of AhR in human colon cancer has not been explored. The goals of the current study are
to determine the pattern of expression of AhR in human colon cancer. In addition, we want to determine if AhR
activation or suppression has an effect on 5-FUs ability to kill cells in vitro.
Methods: Frozen human colon tumors and matched normal tissue was obtained from an IRB approved human tissue
bank maintained in the department of surgery. AhR and Cyp1a1 mRNA expression was measured by quantitative rt-PCR
in both cancer and normal tissue. Clinical data was extracted from the medical record and levels of AhR was stratified
according to tumor type, disease stage, and other clinical variables.
For the in vitro studies. colorectal adenocarcinoma cells (DLD-1) were propagated in DMEM medium
supplemented with 10% fetal bovine serum, 5 mL non-essential amino acids (100x) and 5 mL Penicillin/Streptomycin at
37 C in a 5% CO2 humidified atmosphere. Cells growing in logarithmic phase were plated in 96-well plates. After 4
hours, the cells were treated accordingly with the AhR agonist 6-formylindolo[3,2-b]carbazole (FICZ), the AhR antagonist
CH-223191, or control. After 2 more hours, the cells were treated with either 1% DMSO or 5-fluoruracil (5-FU), a
chemotherapeutic pyramidine analog. After 20 hours of treatment, an XTT cell viability assay was performed to evaluate
approximate cell death. The Wilcoxon rank sum test was used for statistical analysis.
Results: The levels of AhR were found to be higher in the colon cancer tissue compared to the matched normal tissue
(3.10.1 vs 10.5, p=0.0002). This was consistent with the finding of increased levels of Cyp1a1 in malignant tissues
(37.10.0002 vs 10.0003, p=0.04). After treatment, DLD-1 cells with agonized AhR were found to resist cell death by 5Fluorouracil while those with antagonized AhR were much more susceptible to the drug (Figure 1).
Conclusions: We found that AhR is increased in colon cancer compared to normal tissue. We also found that activation
of AhR decreases cell death in colon cancer cells treated with 5-FU. While this is a relatively small study, it indicates that
AhR antagonists may serve as a supplement to increased susceptibility of colon cancer to 5-FU.
Key Words: Aryl Hrdrocarbon Receptor, Colon and Rectal Cancer, Tumor, Chemotherapy, FICZ, CH-223191, 5Fluorouracil

Cell Survivorship

DLD-1 Tumor Cell Death in Response to


5-FU with Varying AhR Epxression
2.5
2
1.5
1
0.5
0
Control/5-FU (No Tx)

FICZ/5-FU (Agonist)
Treatment

CH223191/5-FU
(Antagonist)

5
Poly(ethyleneglycol)containinghydrogelspromoteprimarygranulerelease
fromhumanbloodderivedpolymorphonuclearleukocytes

HannahCaitlinCohen,TylerJacobLieberthal,W.JohnKao

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

HannahCohen
(703)6263799
hccohen@wisc.edu
GraduateStudent
GeneralSurgery
W.JohnKao
BasicScience
biomaterials,poly(ethyleneglycol),polymorphonuclearleukocytes,neutrophils,
degranulation

Polymorphonuclearleukocytes(PMNs;neutrophils)arerecruitedtositesofbiomaterialimplantsduringacute
inflammationandrespondtobiomaterialsthroughinteractionsbetweencellsurfacereceptorsandadsorbedproteins.
PMNscontainfourgranulesubsets:primarygranules,secondarygranules,tertiarygranulesandsecretoryvesicles.Upon
receptormediatedactivation,PMNscanreleasetheirgranulestorecruitmonocytes(MCs)andmediate
MC/macrophageactivationandfunction,therebypotentiallyinfluencingsubsequentinflammatoryorwoundhealing
eventsintheforeignbodyresponsetoimplantedbiomaterials.Wesoughttoinvestigatethereleaseofmyeloperoxidase
(MPO),aprimarygranulemarker,andmatrixmetalloproteinase9(MMP9),atertiarygranulemarker,fromhuman
bloodderivedPMNsculturedonselectedbiomaterials:poly(ethyleneglycol)(PEG)hydrogels,polydimethylsiloxane
(PDMS),tissueculturepolystyrene(TCPS)andgelatinPEG(GP)hydrogels.Primarygranulescontainhighlytoxic
microbicidalproteinsandproteasessuchashumanneutrophilpeptides13,cathepsinGandelastase,manyofwhich
areimplicatedinchronicinflammation.Therefore,thereleaseofprimarygranulesistightlycontrolledandgenerally
occursonlyafterPMNsreachthesiteofinflammation,whereasPMNsreleasetheirothergranulesubsetsduring
extravasation.PMNsculturedonPEGcontaininghydrogels(i.e.,PEGandGPhydrogels)releasedhigherconcentrations
ofMPOthanPMNsculturedonPDMSorTCPSat2hours,suggestingthatPMNsmayundergoaninflammatoryresponse
toPEGcontaininghydrogels.ThisisnotablebecausePEGhastraditionallybeenconsideredabioinertandnon
immunogenicmaterial.PMNsonallbiomaterialsreleasedcomparablelevelsofMMP9at2hours,indicatingthatPMNs
culturedonPEGcontaininghydrogelshavedifferentmechanismsofreleaseforprimaryandtertiarygranules.Inongoing
studies,weareinvestigatingthesignalingpathwaysinvolvedinbiomaterialmediatedPMNdegranulationtounderstand
howwecandirectdegranulationtoachieveamorefavorableforeignbodyresponsethatlimitsinjurytobystandercells
andminimizesproinflammatoryMC/macrophagepolarization.

6
Videobasedmethodofquantifyingperformanceandinstrumentmotionduring
simulatedphonosurgery

KetanSurender,M.Sc.,EllenConroy,ZhixianGeng,Ph.D.,TingChen,M.D.,SethDailey,M.D.,JackJiang,M.D.,Ph.D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

EllenConroy
(608)3860952
erconroy@wisc.edu
UndergraduateStudent
OtolaryngologyHead&NeckSurgery
JackJiang,MD,PhD
BasicScience
Phonomicrosurgery,surgicalassessment,surgicaltraining,simulatedmodel,VPITS

Objective:ToevaluatetheVideobasedPhonomicrosurgeryInstrumentTrackingSystem,anovelmethodofacquiring
instrumentpositiondataduringsimulatedphonomicrosurgery.Thepositiondatacanbeusedtocomputequantitative
motionmetrics.Weusedthisinstrumenttrackingsystemtodetermineifnovicesubjectmotionmetricsimprovedover
oneweekoftraining.
StudyDesign:Prospectivecohortstudy.
Methods:Tensubjectsperformedsimulatedsurgicaltasksonceperdayforfiveconsecutivedays.Instrumentposition
datawerecollectedandusedtocomputemotionmetrics(pathlength,depthperception,andnetmotionsmoothness).
Datawereanalyzedtodetermineifmotionmetricsimprovedwithpracticetime.Qualityoftaskoutcomewasalso
determinedeachdayandtherelationshipbetweentaskoutcomeandmotionmetricswasusedtoevaluatethevalidity
ofderivedmotionmetricsasindicatorsofsurgicalperformance.
Results:Significantdecreasesovertimewereobservedforpathlength(P<0.001),depthperception(P<0.001),andtask
outcome(P<0.001).Nosignificantchangewasobservedfornetmotionsmoothness.Asignificantrelationshipwas
observedbetweentaskoutcomeandpathlength(P<0.001),depthperception(P<0.001),andmotionsmoothness
(P<0.001).
Conclusion:TheVideobasedPhonomicrosurgeryInstrumentTrackingSystemcanestimateinstrumenttrajectoryand
providequantitativedescriptionsofsurgicalperformance.Itmaybeausefulnewmethodofevaluatingperformance
duringsimulatedphonomicrosurgery.Pathlengthanddepthperceptionmaybeparticularlyusefulindicatorsof
phonomicrosurgicalperformance.

7
ThailandepsinA,aNovelHDACInhibitor,DecreasesTumorCellSurvivaland
ActivatestheNotchPathwayinCarcinoidCancer.

AjithaDammalapati,AprilHarrison,MadhuchhandaRoy,ColinKorlesky,RenuNair,ChengWang,YiQiangCheng,RenataJaskula
SztulandHerbertChen

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AjithaDammalapati
(803)6148182
dammalapati@surgery.wisc.edu
Other
GeneralSurgery
HerbertChen
BasicScience
neuroendocrine(NE)tumors,ThailandepsinA(TDPA),Notchsignaling,HDAC
inhibitor,antitumor

Introduction:Carcinoidsareneuroendocrine(NE)tumorsthatsecretehormonescausingthecarcinoidsyndrome.
Metastaticcarcinoidsarenotamenabletocurativesurgery.OurpreviousresearchhasshownthatNotchsignalingacts
asatumorsuppressorinNEtumors.Inthepresentstudy,wesoughttoexaminethemechanismsofNotchpathway
activationbyThailandepsinA(TDPA),anewlydiscoveredHDACinhibitor,anddelineateantitumorpropertiesofthis
drugingastrointestinal(GI)carcinoidcancercells.
Methods:TDPAisabacterialnaturalproductisolatedfromthefermentationbrothofBurkholderiathailandensisE264.
ThecytotoxicityofTDPAonhumanGIcarcinoidcells(BON)wasmeasuredbydeterminingtheIC50value.Theanti
proliferativeeffectsofTDPAonBONcellgrowthwereassessedbythemethylthiazolyldiphenyltetrazoliumbromide
(MTT)rapidcolorimetricassayandclonogenicassay.Themechanismsofgrowthinhibitionweredeterminedusingcell
cycleandapoptoticmarkersviaWesternblotandflowcytometryanalyses.ExpressionofNotchisoforms13was
assessedatthetranscriptionallevels(quantitativerealtimeRTPCR)andtheproteinlevels(Westernblot)from
untreatedandTDPAtreatedBONcells.FunctionalanalysisoftheNotchIntracellularDomain(NICD)wasdoneby
measuringchangesinluciferaseactivityusingtheCBF1bindingassay.ThemRNAexpressionofHESandHEY(Notch
signalingeffectors)wasquantifiedbyrealtimeRTPCR.Toinvestigatethetranscriptionalregulationspecificallyof
Notch1,weusedluciferasereporterconstructsharboringvarious5deletionsoftheNotch1promoter.EffectsofTDPA
onNEmarkerschromograninA(CgA)andachaetescutecomplexlike1(ASCL1)wereassessedbyWesternblotanalysis.
Results:TDPAtreatmentdecreasedcellproliferationinadoseandtimedependentmannerbelowtheIC50valueof
7nM.Westernblotanalysisandflowcytometryexperimentsindicatedthatthegrowthinhibitionwasduetobothcell
cyclearrest(atG2/Mphase)andapoptosis.TreatmentofBONcellswithTDPAledtoaninductionofNotchisoforms13
inadosedependentmanner.FunctionalanalysisofNotchsignalingdemonstratedanincreaseintheCBF1binding
activityandupregulationoftranscriptionallevelsofHESandHEYfamilies.ThedeletionmappingoftheNotch1promoter
revealedthattheTDPAtargetsarecontainedwithinthe257bp5promoterregionrelativetothetranscriptionstart
site.Moreimportantly,coincidentwithNotchactivation,wenoticedadosedependentreductioninNEmarkersASCL1
andCgA.
Conclusions:WedemonstratedthatTDPAactivatestheNotchpathwayandisapotentinhibitorofcarcinoidcell
proliferationatlownanomolarconcentrations.TheseresultsindicatethatthereinstitutionofNotchexpressionbyTDP
Ahastherapeuticpotentialforcarcinoidcancer.

8
TGFbeta/Smad3StimulatesStemCell/DevelopmentalGeneExpressionand
VascularSmoothMuscleCellDeDifferentiation

DanielDiRenzo;XudongShi;LianwangGuo;SarahFranco;BowenWang;StephenSeedial;K.CraigKent

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DanielDiRenzo
(608)2659823
direnzo@surgery.wisc.edu
PostdoctoralFellow
VascularSurgery
Dr.K.CraigKent
BasicScience
TGFbeta,smoothmusclecells,dedifferentiation,development,genearray

AtheroscleroticassociateddiseasesaretheleadingcauseofdeathintheUnitedStates.Despiterecentprogress,
interventionaltreatmentsforatherosclerosiscanbecomplicatedbyrestenosisresultingfromneointimalhyperplasia.
WehavepreviouslydemonstratedthatTGFbetaanditsdownstreamsignalingproteinSmad3:1)areupregulated
followingvascularinjury,2)togetherdrivesmoothmusclecell(SMC)proliferationandmigrationand3)enhancethe
developmentofintimalhyperplasia.InordertodetermineamechanismthroughwhichTGFbeta/Smad3promotethese
effects,AffymetrixgeneexpressionarrayswereperformedonprimaryratSMCsinfectedwithSmad3andstimulated
withTGFbetaorinfectedwithGFPalone.
Morethan200genesweredifferentiallyexpressed(>2.0foldchange,p<0.05)inTGFbeta/Smad3stimulatedSMCs.We
thenperformedGOtermenrichmentanalysisusingtheDAVIDbioinformaticsdatabaseandfoundthatTGFbeta/Smad3
activatedtheexpressionofmultiplegenesrelatedtoeitherdevelopmentorcelldifferentiation,severalofwhichhave
beenshowntobeassociatedwithmultipotentstemorprogenitorcells.QuantitativerealtimePCRconfirmed
upregulationofseveraldevelopmentalgenesincludingFgf1,Ngf,andWnt11byapproximately2.5,6and7fold,
respectively.TimedependentupregulationofthestemcellassociatedgenesCd34andCxcr4(bothby50fold)wasalso
confirmedbyrealtimePCR.Finally,TGFbeta/Smad3downregulatedtranscriptionofSMCcontractilegenesaswellas
proteinproductionofsmoothmusclealphaactin,calponin,andsmoothmusclemyosinheavychain.Thesecombined
resultssuggestthatTGFbeta/Smad3stimulationdrivesSMCstoaphenotypicallyalteredstateofdedifferentiation
throughtheupregulationofdevelopmentalrelatedgenes.

9
MechanicalTestingofModeledRibFractureswithPlatedFixationontheInternal
Face

Elegbede,A.,Goodspeed,M.,Vanderby,R.,Agarwal,S.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AnuElegbede
(702)3081663
AElegbede@UWHealth.org
Resident
GeneralSurgery
Agarwal
BasicScience
RibFractures,Fixation,Plating,Stress,MechanicalTesting

Introduction:Traditionally,ribfractureshavebeentreatednonoperativelywithpaincontrol,pulmonarytoilet,and
internalpneumaticstabilizationasnecessary.Overthepastdecade,operativefixationofribshasbecomeapopular
treatmentoption;however,thissurgerycarrieswithitsignificantmorbidityanddiscomfort.Anovelsurgicaldrillwith
anarticulatingdriveshafthasbeendevelopedtofacilitatefixationoffracturedrib(s)ontheinternalfaceoftheribvia
minimallyinvasivesurgery.Priortocontinueddevelopmentofthisnoveltool,treatmentefficacyoffracturedribsplated
ontheinternalfacemustbecomparedtofixationontheexternalface.Wehypothesizedthatinternalfacedfixationwill
beatleastequivalentinstiffnessanddurabilitytostandardexternalfacedfixation.
Methods:Fortymodelsawboneribswerecuttosimulateanobliqueribfracture:completeandangulated.Eachribwas
reconstructedbyplatefixationoneithertheinternalorexternalfaceutilizingaUplate(AcuteInnovations)oraflat
platewithlockingscrews(AcuteInnovations)foreachorientationoffixation.Theplatedribswerecycled50,000times
at10Hzwithloadingfrom0to+2Ntosimulatebreathingloads.Stiffnesswasmeasuredandrecordedatintervals
throughoutthetest.
Results:StiffnessfortheribconstructwhentheUplatewasattachedtotheinternalface,1.550.26N/mm,was
greaterthanwhensecuredtotheexternalface,1.150.24N/mm(p=0.04).However,therewasalargerdropin
stiffnessbetweeninitialandfinalloadingwhentheUplatewasattachedtotheinternalfacecomparedtotheexternal
face,2.3%and1.4%respectively(p=0.05).Stiffnessfortheribconstructsecuredusingaflatplatewithlockingscrews
ontheinternalface,1.410.23N/mm,wasgreaterthanwhensecuredtotheexternalface,1.060.10N/mm(p=
0.02).Testingshowedasimilardecreaseinstiffnesswhentheflatplatewasattachedtotheinternalfacecomparedto
theexternalface,1.2%and1.6%respectively.
Conclusions:Internalfaceplacementofosteosyntheticplatesshowedgreaterstiffnessoffixationonmodelribs.In
addition,flatplatessecuredtotheinternalfacehadequivalentdurability(asdefinedbydropinstiffness).Thesetwo
factorssuggestfixationontheinternalfaceoffracturedribsutilizingflatplateswithlockingscrewsismechanically
similartocurrentsurgicalmethodsandsupportsthepotentialforavideoassistedthoracoscopicsurgeryasaminimally
invasivealternativetorepairfracturedribs.

10
TGFbetaanditsSignalingProtein,Smad3,PlayaRoleintheDeDifferentiation
ofSmoothMuscleCells

SarahFranco,XuDongShi,LianWangGuo,DanielDiRenzo,ToshioTakayama,BowenWang,BoLiu,andK.CraigKent

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SarahFranco
(505)8045226
srfranco.77@gmail.com
GraduateStudent
VascularSurgery
K.CraigKent
BasicScience
IntimalHyperplasia,Restenosis,SmoothmusclecellDedifferentiation,
Developmental,Stemcell

Intimalhyperplasia(IH)isacomplexprocessinvolvingthephenotypicswitchofvascularsmoothmusclecells(SMC)from
adifferentiatedstatetoadedifferentiatiedstate,anditisthemaincontributortorestenosis,whichistherenarrowing
ofthevesselaftervascularreconstructiveprocedures.WehavepreviouslyshownthatTGFanditssignalingprotein,
Smad3,playastimulatoryroleinIH.However,themolecularbasisofTGF/Smad3stimulatedSMCdedifferentiation
andIHremainsunclear.Ourpreviousmicroarraystudiesrevealedthattheexpressionofmorethan200geneswas
significantlyalteredbyTGF/Smad3stimulationofrataorticSMCs.Analysisoftheseresultsdemonstratedthatmost
ofthesegenesweredevelopmentalrelated.Whiledifferentiationgenesweredownregulated,dedifferentiationgenes,
includingSOX18,anddevelopmentalrelatedgeneswereupregulated.
Inthisstudy,wefocusedouranalysisonSOX18,whichisageneassociatedwithmultipotentstemcells.RataorticSMCs
wereinfectedwithadenovirusexpressingeitherSmad3orGFP(control),followedbytreatmentwithTGForsolvent
for24h.RNAwasextractedfromthetreatedcellsandquantitativeRTPCRwasperformed.WefoundthatSOX18
expressionwasupregulatedby28foldinTGF/Smad3treatedSMCsincomparisontotheGFPcontrol.Tofurther
analyzetheeffectofTGF/Smad3onSOX18expressioninvivo,weperformedballoonangioplastyinratcarotid
arteries,whereTGFandSmad3levelsareknowntobeelevatedbyinjury.Twoweekspostballooninjury,ratcarotid
arterieswerecollectedandcrosssectionsprepared.ImmunohistochemistryshowedthatSOX18wassubstantially
upregulatedintheneointimalayerofinjuredarteriescomparedtouninjuredcontrol.
OurdatasuggeststhatinjuryinducedupregulationofTGFandSmad3stimulatesSOX18expression,whichmay
contributetoSMCdedifferentiationandthedevelopmentofneointima.Thus,ourstudyprovidesnewinsightsintoTGF
/Smad3mediatedIH.FurtherstudiesarewarrantiedtouncovertheintriguingmechanismswherebyTGF/Smad3
stimulatesIHbyregulatingexpressionofstemcellassociatedgenes.

11
ResidentialArterialCellsandCirculatingBoneMarrowDerivedCellsboth
ContributetoIntimalHyperplasiainaRatAllograftCarotidTransplantation
Model

YiSi

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JasmineGiles
(608)7707659
giles@surgery.wisc.edu
Other
VascularSurgery
BoLiu
BasicScience
Vascular,Transplant,SMC(smoothmusclecell),IH(intimalhyperplasia),BM(bone
marrow)

Introduction:Intimalhyperplasia(IH),animportantfeatureofrestenosisfollowingvascularinterventions,is
characterizedbyaccumulationofsmoothmusclecells(SMCs)orSMClikecellsintheneointima.Bonemarrowderived
progenitorsarebelievedtoplayanimportantroleinIHbasedonmousestudiesusingbonemarrowtransplantation
(BMT).However,aratfemoralinjurymodelcoupledwithBMTrevealedaninsignificantpresenceofbonemarrow
derivedcellsinIH,whichindicatedthatcellularmechanismsunderlyingvascularrepairinratsaredramaticallydifferent
fromthatinmice.ToinvestigatetheoriginofIHinrat,wecreatedanewratIHmodelwhichcombinesballooninjury
withcarotidallografttransplantation.
Methods:AllografttransplantationwasadministratedbetweentransgenicinbredLewisGFPanditsWildType(WT)rats.
Theleftcarotid(57mminlength)ofdonorrats,ballooninjured,withintactsurroundingconnectivetissuewas
harvestedimmediatelyandtransplantedtothehostleftcarotidarterywithendtoendanastomosis.Also,bone
marrowderivedcellswithDiIlabelingfromtheGFPrats7daysfollowingcarotidtransplantationwereautologously
injectedbacktothesameratviatailvein.
Results:Followingallografttransplantationoftheballooninjuredcarotidcommonartery(n=3),smoothmuscleactin
positiveandCD68positivecellswereobviouslyfound14dayslaterintheneointima,wherereendotheliazationwasnot
achievedyet.Themajorityofneointimacellswereapparentfromrecipient(85%)versusdonor(15%)usingGFPand
WTallografttransplantation,whereMCP1/CCR2andSDF1/CXCR4signalingwerebothinvolved.Inaddition,bone
marrowderivedDiI/GFPdoublepositivecellsaccumulatedthroughouttheIHofWTGFPtransplantation.
Conclusions:ThenewangioplastytransplantationhybridmodelallowsexaminationofIHinnonirradiatedrats,inwhich
bonemarrowandpossiblyotherremotesourcesgiverisetomajorityofSMClikecellsintheneointima.

12
Transgenic Reporter Mice Enable Delineation of Temporal and Spatial Enteric
Cholinergic Neurogenesis
Scott Lee, BS, Christopher S. Erickson, BS, Amanda J. Barlow, PhD, Noah R. Druckenbrod, PhD, Miles L. Epstein, PhD, Ankush
Gosain, MD, PhD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Ankush Gosain
(608) 263-9419
gosain@surgery.wisc.edu
Faculty
General Surgery
Gosain
Basic Science
Enteric Nervous System, Cholinergic, Neurogenesis, Transgenic, Embryology

Introduction:
Gastrointestinal motility, luminal secretions and blood flow are regulated by the enteric nervous system (ENS) present
along the bowel wall. The ENS is derived primarily from a population of migratory progenitor neural crest cells (NCC)
that delaminate from the vagal region of the neural tube. Vagal NCC enter the murine foregut at embryonic day 9 (E9)
and migrate proximal to distal, colonizing the entire bowel by E14.5. Normal functioning of the ENS depends upon NCC
differentiation into enteric neurons with the correct complement of neurotransmitter phenotypes that regulate
muscular contraction and relaxation. Postnatally, cholinergic neurons expressing choline acetyltransferase (ChAT)
constitute a large proportion of enteric neurons and are the major excitatory neurons in the bowel. As a first step in
understanding the mechanisms regulating neurotransmitter expression in enteric neurons, we examined the spatial and
temporal differentiation of cholinergic enteric neurons during embryonic and post-natal development.
Methods:
Following IACUC approval, we mated ChAT-GFP reporter mice (green fluorescence) with transgenic animals that
contained ChAT-Cre/R26R tdTomato (red fluorescence) to create progeny that express both GFP and tdTomato in
cholinergic neurons. We obtained ileum and colon from embryos and postnatal (P) animals starting at ages E10.5
through P30. These tissues were fixed and stained with antisera to ChAT and Hu, a pan-neuronal marker. Using
confocal imaging, the localization of GFP, tdTomato, and ChAT immunoreactivity (ChAT-IR) was quantified at each age
studied.
Results:
The earliest ChAT-positive cells were seen at E10.5 in the rostral small intestine approximately one day after the entry of
enteric precursors into the gut. These cells were identified by the presence of ChAT-IR and were also ChAT-GFP positive.
However there was no detectable expression of ChAT-tdTomato at this stage. As development proceeded we found that
cholinergic neurons within the distal small intestine (dSI) and proximal colon were first labeled by ChAT-IR and then
ChAT-GFP, which were followed by a delay of 2-3 days by Chat-tdTomato. Additionally, the percentage of Chat-IR
neurons in the embryonic gut is equivalent to adult levels in the dSI by E13.5 and in the proximal colon by P0. After these
stages the percentages remained relatively constant although the neuronal density underwent dramatic changes.
Conclusions:
We observed a spatial and temporal gradient in the differentiation of enteric cholinergic neurons. These appear first in
the embryonic foregut, later in the hindgut, and their numbers increased with age. The difference between the
proportion of ChAT-IR Hu+ neurons expressing GFP and tdTomato suggests that the ChAT-GFP reporter is a more reliable
indicator of cholinergic differentiation than ChAT-Cre/R26R tdTomato. Using these mice, we can interrogate the
mechanisms regulating neurotransmitter expression in enteric neurons.

13
Early ultrasonic vocalization dysfunction in a PINK1 knock-out model of
prodromal Parkinson disease
LM Grant1,2, KP Cullen1,2, J Pultorak4, CA Kelm-Nelson1 MR Ciucci1,2,3 1Department of Surgery, Division of OtolaryngologyHead and Neck Surgery;2 Department of Communication Sciences and Disorders;3 Neuroscience Training Program;4 Department
of Zoology

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Laura Grant
(608) 262-6122
grant@surgery.wisc.edu
Graduate Student
Otolaryngology-Head & Neck Surgery
Dr. Michelle R Ciucci
Basic Science
Parkinson Disease, Voice Deficits, Ultrasonic Vocalizations, Rat, Transgenic

Introduction: Parkinson disease (PD) is devastating to cranial sensorimotor function and voice deficits include a harsh or
breathy vocal quality and reduced loudness and pitch variability. However, the onset, progression and neural correlates
of voice deficits are poorly understood. To address this, we used a transgenic rat model of PD and hypothesized that
voice deficits would manifest early, be progressive in nature and be related to pathologies in brain structures associated
with cranial sensorimotor function.
Methods: Homozygous and heterozygous PINK1 knock-out rats and wild-type (WT) controls (n=48) were evaluated at 2,
4, 6 and 8 months of age. Ultrasonic vocalizations were analyzed for average bandwidth, peak frequency, duration,
intensity and complexity of frequency modulated calls. Immunohistochemistry was performed at 8 months of age for
tyrosine hydroxylase (dopamine marker) in the substantia nigra and striatum and for -synuclein aggregates in
brainstem and cortical regions important to vocal control. To determine differences between groups, a repeated
measures ANOVA (age x genotype) and Fishers Least Significant Difference tests were used (alpha<0.05).
Results: There was a significant interaction between age and genotype for bandwidth F(6, 110)=3.00, p<0.0003).
Homozygous rats had decreased bandwidth compared to WT at 4 and 6 months, (p<0.05)) and beginning at 4 months,
demonstrated reduced bandwidth compared to 2 months (p<0.05).
There was a significant interaction between age and genotype for peak frequency F(6, 110)=3.00, p<0.0003). Beginning
at 4 months of age, homozygous rats demonstrated decreased peak frequency compared to heterozygous but not WT
rats (p<0.05) and at 6 and 8 months, peak frequency was reduced compared to 2 and 4 months (p<0.05).
For intensity, there were significant main effects for genotype F(2, 45)=24.80, p<0.0001) and age F(3, 110)=4.62,
p=0.0044). Homozygous rats were quieter compared to WT and heterozygous rats (p<0.05), regardless of age. However,
in general, at 4 and 6 months all rats had significantly louder calls compared to 2 months (p<0.05).
Tyrosine hydroxylase levels did not differ between genotypes (F (2, 29)=5.78, p=0.1). However, -synuclein positive
aggregates were present in the substantia nigra and periaqueductal gray regions of the brainstem of multiple
heterozygous and all homozygous rats.
Conclusions: Homozygous PINK1 rats demonstrated reduced intensity beginning at 2 months, while bandwidth and peak
frequency were compromised beginning at 4 months. Although duration, intensity, and percent complex calls generally
increased with age for all genotypes-probably as an effect of size, maturation, and practice-intensity remained impaired
in the homozygous rats compared to heterozygous and WT. Interestingly, heterozygous rats did not show any robust
vocalizations deficits. These data suggest homozygous PINK1 knock-out rats demonstrate robust, early vocalization
deficits that may be progressive in nature, in the absence of nigrostriatal dopamine depletion. However, despite the
lack of dopamine loss at 8 months (primary PD pathology), -synuclein aggregates were found in the substantia nigra
and could represent early degeneration of dopaminergic pathways. Aggregates were also found in the periaqueductal
gray-a region important in vocal control. These data suggest that vocalization deficits may be an early behavioral
biomarker in pre-manifest Parkinsonism.

14
ANewlySynthesizedCollectionofSAHAAnalogsShowHighPotencyandNotch
InductioninCarcinoidCancer

AprilD.Harrison,AjithaDammalapati,CasiM.Schienebeck,RenuNair,WeipingTang,RenataJaskulaSztul,HerbertChen

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AprilHarrison
(608)2653749
harrison@surgery.wisc.edu
Other
GeneralSurgery
HerbertChen
BasicScience
Carcinoid,HDACInhibitor,SAHAAnalog,Notch,Neuroendocrine

Introduction:Carcinoidcancerconsistsofslowgrowingneuroendocrine(NE)tumorswithinthegastrointestinaltract,
whichareoftenmetastaticbythetimeofdiagnosismakingcurativesurgeryinfeasible.Carcinoidtumorscause
excessiveproductionofvariousbioactivehormonesresultinginthecarcinoidsyndromeandapoorqualityoflifefor
patients.TheNotchpathwayhasproventobetumorsuppressiveinNEcancer,resultinginaclinicalneedfor
therapeuticoptionstoactivateNotch.NonselectiveHDACinhibitors,suchasSAHA,havedemonstratedNotch
induction,buthavebeenproventocausesignificantsideeffects.SAHAanalogsAB2,AB3,andAB12wererecently
synthesizedaspotentiallyselectiveHDACinhibitors,whichmayreducenegativesideeffectswhileactingasequally
effectiveNotchpathwayactivators.
Methods:TodeterminetheantiproliferativeeffectofAB2,AB3,andAB12onhumangastrointestinalcarcinoidBONcells
weperformeda3(4,5dimethylthiazol2yl)2,5diphenyltetrazoliumbromide(MTT)assayforIC50values.Similarly,a
MTTassaywasperformedonhumanfibroblastWI38cells,todeterminethedrugseffectonnoncancerouscells.BON
cellsweretreatedwithAB2,3,or12,upto30Mfor48hoursandproteinlysateswereanalyzedviaWesternblotting
forNotch13isoformexpression.AfunctionalanalysisofNotchexpressionattheNotchIntracellularDomain(NICD)
wasobtainedbymeasuringluciferaseactivityviaaCBF1bindingassay.Westernblottingwasutilizedtoevaluate
expressionofNEtumormarkersachaetescutecomplexlike1(ASCL1)andchromograninA(CgA).Apoptoticandcell
cyclemarkerswerealsoevaluatedtodeterminethemechanismofactioninvolved.
Results:DosedependentreductionsincellproliferationresultedfromtreatmentwithAB2,AB3,andAB12,withIC50
valuesof30Morless.EachdrugshowedminimaleffectonWI38cellproliferation.ProteinlysatestestedviaWestern
blottingrevealedeachdrugtreatmentinducedNotch13isoforms.CBF1bindingactivityincreasedinadosedependent
mannerforallthreedrugswithaminimumofathirtyfoldinduction.WesternblotanalysisofNEtumormarkers
revealedasignificantreductioninASCL1andCgAfollowingeachdrugtreatment.Increasedexpressionofapoptoticand
cellcyclemarkerswasobserved.
Conclusion:AB2,AB3,andAB12eachdemonstratedasignificantreductionincarcinoidcellviability,withanegligible
effectonnoncancerouscells.AllthreedrugsarepotentNotchinducers,andeachdrugtreatmentresultedina
substantialdecreaseinNEtumormarkers.TheseresultsindicatethatAB2,AB3,andAB12mayprovetobeasuccessful
collectionofcompoundstoprovidenoveltherapeutictreatmentoptionsforcarcinoidcancer.

15
BombesinImprovesAdaptiveImmunityoftheSalivaryGlandDuringParenteral
Nutrition

AaronFHeneghan,JosephF.Pierre,XinyingWang,DrewARonnenburg,GuyE.Groblewski,andKennethA.Kudsk

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AaronHeneghan
(608)2620170
heneghan@surgery.wisc.edu
Scientist
GeneralSurgery
Kudsk
BasicScience
salivaryglands,parotid,submandibular,sIgA,amylase

Background:Theparotidandsubmandibularsalivaryglandsaregutassociatedlymphoidtissues(GALTs)thatsecrete
immunecompoundsintotheoralcavity.Parenteralnutrition(PN)withoutenteralstimulationdecreasesGALTfunction,
includingintestinallymphocytecountsandsecretoryimmunoglobulinA(sIgA)levels.Sincetheneuropeptidebombesin
(BBS),agastrinreleasingpeptideanalogue,stimulatesintestinalfunctionandrestoresGALTparameters,we
hypothesizedthatPN+BBSwouldstimulateparotidandsalivaryglandIgAlevels,Tlymphocytes,andIgAplasmacell
countscomparedwithPNalone.Methods:Male(InstituteofCancerResearch)ICRmicereceivedintravenouscatheters
andwererandomizedtochowwithsaline,PN,orPN+BBS(15g/tid/mouse)for5days(8/group),2daysafter
cannulation.SalivaryglandswereweighedandeitherfrozenforIgAandamylaseanalysisorfixedforhistologicalanalysis
ofacinarcells,IgA+plasmacells,andTlymphocytes.SmallintestinalwashfluidwascollectedforIgAregressionanalysis
withsalivaryglands.Results:PNreducedorganweight,acinarcellsize,andamylaseactivitycomparedwithchow;BBS
hadnosignificanteffectsontheseparameters.Comparedwithchow,PNsignificantlyreducedsalivaryglandIgAlevels,
IgA+plasmacells,andTlymphocytes.PN+BBSsignificantlyelevatedIgAandrestoredcellularitycomparedwithPN.
SalivaryglandtissuehomogenateIgAlevelssignificantlycorrelatedwithintestinalfluidIgAlevels.Conclusions:
Comparedwithchow,PNresultsinatrophyofthesalivaryglandscharacterizedbyreducedamylase,IgA,andimmune
cellularity.BBShasnoeffectonacinarcellsoramylaseactivitycomparedwithPNbutmaintainstissueIgAandplasma
cellsandTlymphocytenumberscomparedwithchow.

16
Comparison of straw configurations and their effects on vocal economy using a
simulated system of straw phonation therapy
Terah M. Hennick, Ellen R. Conroy, Emily Olszewski, Jack J. Jiang, M.D.,Ph.D.

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Terah Hennick
(817) 739-6265
thennick@wisc.edu
Undergraduate Student
Otolaryngology-Head & Neck Surgery
Jack Jiang, M.D., Ph.D.
Basic Science
Straw Phonation, Excised Larynx, Inertance, Phonation Threshold Pressure, Phonation
Threshold Flow

Purpose: Straw phonation voice therapy employs a semi-occlusion and extension of the vocal tract to increase
supraglottal pressure and acoustic impedance in the vocal tract, thereby improving vocal economy. The advantage of
using straws in semi-occluded voice therapy is the ability to control their dimensions and, theoretically, the level of
impedance and supraglottal pressure. This study used an excised larynx setup to evaluate the effects of varying
combinations of tube semi-occlusion, tube extension, and recently proposed direct airflow input on vocal economy.
We hypothesized that combining extension, semi-occlusion, and direct supraglottal airflow input would result in the
largest decrease in phonation threshold pressure (PTP) and phonation threshold flow (PTF) compared to all other
configurations. Furthermore, we hypothesized that inertance values calculated for each vocal tract configuration
would inversely correlate with PTP.
Methods: Nine conditions were evaluated: control 1 (before treatment), control 2 (after treatment), and seven
combinations of tube semi-occlusion (6.5 mm), tube extension (30 cm), and supraglottal airflow input (200 mL/s).
Outcome measures included PTP, PTF, and inertance values for each condition. Data were collected for twenty-two
excised canine larynges and results from all conditions were compared to each other using a one-way analysis of
variance. PTP values and calculated inertance values were compared using the Pearson product-moment correlation
coefficient.
Results: PTP decreased for each experimental condition compared to control 1, and comparisons between all
configurations revealed a significant difference between PTP means for the following: control 1 vs. extension+airflow,
control 1 vs. airflow, control 1 vs. extension, control 1 vs. semi-occlusion, semi-occlusion+extension vs.
extension+airflow, semi-occlusion+extension vs. airflow, and semi-occlusion+extension+airflow vs. extension+airflow.
The greatest decrease in PTP compared to control 1 was observed for semi-occlusion+airflow. PTF increased slightly for
all but two experimental conditions compared to control 1. PTF values for extension+airflow and semi-occlusion each
showed a statistically significant decrease compared to control 2. The correlation coefficient revealed a weak
correlation between inertance and PTP (r=0.387517822). We observed high interlarynx variability in PTP and PTF
changes.
Conclusions: The observed trend of decreasing PTP for each straw configuration further supports the theory that vocal
tract extensions, semi-occlusions, and direct airflow input improve vocal economy. However, we rejected our
hypothesis that extension+semi-occlusion+airflow would result in the largest decrease in PTP and PTF as semiocclusion+airflow had the largest significant decrease in PTP and largest decrease in PTF, though insignificant. We also
rejected our hypothesis that inertance would inversely correlate with PTP. Future work will include supraglottal
pressure and acoustic data to more accurately calculate inertance and better understand the relationship between
inertance and PTP. As PTF is more sensitive to glottal width than PTP, the insignificant increases in PTF observed for
some conditions may have been caused by changes in glottal width caused by straw insertion and removal. Additional
studies that use varying straw configurations in humans may be beneficial for optimizing straws to yield the greatest
vocal economy improvements for normal and dysphonic patients.

17
TargetedDeliveryofThailandepsinAusingUnimolecularMicellestoImprove
AntitumorEffectinCarcinoids

RenataJaskulaSztul,WenjinXu,AjithaDammalapati,GuojunChen,OmerSadak,AprilHarrison,QifengZheng,RenuNair,Colin
Korlesky,ChengWang,YiQiangCheng,ShaoqinGongandHerbertChen

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

RenataJaskulaSztul
(608)2653749
sztul@surgery.wisc.edu
Scientist
GeneralSurgery
Chen
BasicScience
nanomedicine,carcinoids,Thailandepsin,HDACi,somatostatinreceptors

Introduction:Carcinoidsaregastrointestinal(GI)neuroendocrine(NE)tumorsoccurringthroughouttheGItractwhich
producehormonescausingthecarcinoidsyndrome.AlthoughNEtumorsareslowgrowing,theyarefrequently
metastaticatthetimeoftheirdiscoveryandnolongeramenabletocurativesurgery.Therefore,thereisagreatneedto
developnoveltherapeuticstrategiesbothtoreducetumorburdenandcontrolsymptomsinpatientswithcarcinoid
neoplasms.Toaddressthisneed,wedevelopedandoptimizedafamilyofnovelunimolecularmicellescapableof
deliveringanewlyreportedanticancerdrugThailandepsinA(TDPA).ThesemicellesspecificallytargettheNEcancer
cellswithoverexpressedsomatostatinreceptors(SSTRs)usingoctreotide(OCT)astheactivetumortargetingligand.In
thecurrentstudyweassessedtheantitumoreffectsofTDPAloadedunimolecularmicellesbothinvitroandinvivo.
Methods:Stableunimolecularmicelleswerepreparedinanaqueoussolutionusingmultiarmstaramphiphilic
blockcopolymer.Thehydrophobicpolyestercore(PLA)wasusedtoencapsulatetheTDPAandthepolyethyleneglycol
shell(PEG)wasusedtoprovidewatersolubilityandreduceopsonization.OCTwasconjugatedtothePEGforactive
tumortargeting.AhumanGIcarcinoidcellline(BON)wastreatedwithafamilyofmicelles(TDPAloadedtargetedand
nontargeted,andemptytargetedandnontargeted)ataTDPAconcentrationof5nMfor2or24,48and72hours.The
cellproliferationandtheexpressionofNEmarkers(ASCL1andCgA)wereassessedbyMTTassayandWesternblot,
respectively.TheeffectoftheOCTtargetingligandsonthecellularuptakeofthemicelleswasmeasuredbyflow
cytometryandconfocallaserscanningmicroscope(CLSM).TheantitumorefficacyofTDPAloadedmicelleswas
determinedinBONxenograftsafterfiveintravenousinjectionsperformedevery5dayswithadoseof3.125mg/kgBW.
Results:ThefamilyofpHresponsiveunimolecularmicellesweredevelopedfortargeteddeliveryofTDPAto
carcinoids.TheTDPAloadedandOCTconjugatedtargetedmicelleshadthestrongestinhibitoryeffectonBONcell
proliferationandNEmarkersexpression.Moreover,thetargetedmicellesexhibitedamuchhighercellularuptake(4
foldincrease)thannontargetedmicellesbasedonflowcytometryandCLSManalyses.Additionally,TDPAloaded
targetedmicellesdemonstratedthebestantitumorefficacywiththehighestdecreaseoftumorvolume(76%).
Conclusions:TheOCTtargetingligandsconjugatedontothesurfaceofthemicellessubstantiallyincreasedthe
cellularuptakeofthemicellesinBONcellsandimprovedcytotoxiceffectofTDPAbothinvitroandinvivo.Thus,
targeteddeliveryofanticancerdrugTDPA,specificallytothetumortissuecouldsignificantlyimprovethetherapeutic
outcomesintreatingcarcinoiddiseasewhilereducingsystemictoxicity.

18
EnvironmentalPollutantsAccelerateSkinTransplantRejectionandIncreaseTh17
DifferentiationinaMurineModel

WalkerJulliardMD,JohnFechner,JoshuaMezrichMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

WalkerJulliard
(608)2659147
wjulliard@uwhealth.org
Resident
Transplantation
JoshuaMezrich
BasicScience
Th17cells,ArylHydrocarbonReceptor,Pollutants,Skintransplantation,IL17

Background:Thearylhydrocarbonreceptor(AhR)isacytosolictranscriptionfactorwithnumerousxenobioticand
endogenousligands.Polycyclicaromatichydrocarbons(PAHs)areAHRligandsthatcanbefoundinairpollutants.
RecentevidenceindicatesthatpollutantsandPAHsinparticularmayalterthefunctionofantigenpresentingcells(APCs)
andTcells.OurlaboratoryhasextensivelystudiedIL17producingTcells(Th17cells)andtheirrelationshipwiththeAhR
andpollutants.Th17cellsplayasignificantroleinautoimmunediseasesandtransplantrejection.Thecurrentstudy
examinedtheeffectsofastandardizedlotofatmosphericparticulatematter(UrbanDustParticles;UDP)onTh17cell
polarizationinvitroandtheeffectsofintranasalUDPexposureinamouseskintransplantmodel.Wehypothesizethat
activationoftheAhRwillleadtoanincreaseinTh17polarizationinvitroandtherebyanincreaseintheinflammatory
cytokineIL17.Furthermore,wehypothesizethatinhalationofUDPwillleadtosystemiceffectswhichwillaccelerate
rejectionofskintransplantationinmice.
Methods:ToassesstheeffectofUDPonTcellpolarization,naveTcellsisolatedfromwildtypeB6andAhRnullspleens
werethenculturedinTh17conditionsfor4daysinthepresenceorabsenceofUDP.IL17wasmeasuredinculture
supernatantsbyELISAandthecellswereanalyzedbyflowcytometryforintracellularcytokines,AhR,andcellsurface
markers.Inotherexperiments,B6micewereexposedtoeitherintranasalphosphatebufferedsaline(PBS;control)or
UDPevery3daysfor15days.Onday12,skinfromaBalb/cmousewastransplantedontotheB6miceandthesemice
werethenmonitoredforrejection.Followingrejectionofskingrafts,miceweresacrificedandspleensandlymphnodes
werecollectedandanalyzedbymixedlymphocytereaction(MLR)usingirradiatedallogeneic(Balb/c)orsyngeneic(B6)
stimulatorcells.After4daysofculture,supernatantswereharvestedandanalyzedbyELISAandthecellswerestained
andanalyzedbyflowcytometry.
Results:Th17differentiationandIL17productionwereenhancedwithadditionofUDPincultureinwildtypeTcells.
However,therewasnoenhancementwhentheexperimentwasrepeatedwithAhRnullTcells.Furthermore,skin
transplantrejectionwasacceleratedinmicethatinhaledUDP.OnanalysisoflymphnodesfromUDPtreatedmicepost
graftrejection,therewasashiftintheratioofTh1:Th17cellstowardsagreaterTh17response.
Conclusions:TheseexperimentsdemonstratethatpollutantssuchasUDPcancauseanincreaseinTh17differentiation
andinflammatorycytokineproductioninvitroinanAhRdependentmanner.Furthermore,inhalationofpollution
acceleratedskintransplantrejectioninamousemodel.Whenthesemicewereanalyzed,therewasatrendtowards
moreofaTh17responseasopposedtoaTh1response.Theseresultssuggestthatpollutioncouldplayanimportant
roleintherejectionoftransplantedorgansbyalteringtheimmuneresponse.

19
Development of an Optogenetic Sensory Peripheral Nerve Interface
Sahil K. Kapur MD, Thomas J. Richner MS, Sarah K. Brodnick BS, Justin C. Williams PhD, Samuel O. Poore MD PhD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

SAHIL KAPUR
(607) 592-4436
kapur.sahil@gmail.com
Resident
Plastic & Reconstructive Surgery
Samuel Poore MD/PhD Justin Williams PhD NITRO Lab.
Basic Science
Peripheral Nerve, Optogenetics, Neural Interface, Sensory, Electrocorticography

Purpose:
Improvement in afferent sensory feedback is the necessary next step in the development of functional neuroprostheses.
While electrical stimulation serves as the standard of peripheral nerve manipulation, the use of light sensitive ion
channels in optogenetic models could provide a sensitive and specific alternative for afferent signal generation. In this
study, we demonstrate similarities between the cortical representation of afferent neural signals that have been
generated by electrical and optical stimulation of peripheral nerves. Furthermore, we demonstrate the ability to
generate afferent signals that retain cortical localization following transcutaneous optical and electrical peripheral nerve
stimulation.
Methods:
Flexible thin film microelectrode arrays were implanted over the sensorimotor cortex in optogenetically modified
transgenic mice expressing channelrhodopsin in accordance with IACUC guidelines. Seven days following cortical
implantation, peripheral nerve signals were generated under four different experimental conditions: electrical
stimulation following surgical exposure of the sciatic and median nerves, optical stimulation following surgical exposure
of the sciatic and median nerves; transcutaneous electrical stimulation of the sciatic and median nerves and
transcutaneous optical stimulation of the sciatic and median nerves. Local field potentials were recorded by the
implanted cortical electrode arrays during these sessions.
Results:
Cortical signals recorded by the multielectrode arrays were localized to regions of the hindlimb or forelimb cortex
corresponding to the peripheral nerve being stimulated. Localization of the signal was preserved across both electrical
and optical stimulation modalities. Furthermore, cortical signals maintained their localization during transcutaneous
optical and electrical stimulation. Signal amplitudes varied proportionately with the amplitude and pulse width of
peripheral nerve stimulation. Generation of consistent localized cortical signals via both electrical and optical stimulation
was maintained in experiments carried out over a period of at least 5 days. Control experiments carried out on wild type
mice produced similar results following electrical stimulation but no cortical signals following optical stimulation of the
peripheral nerves.
Conclusion:
Preserved similarity and localization of cortical signals following electrical and optical stimulation of peripheral nerves
implies that ascending sensory information can be reliably transmitted to the brain via either electrical stimulation or
optical pulses applied to peripheral nerves. Furthermore, the robust cortical response to transcutaneus optical and
electrical stimulation permits the non-invasive manipulation of peripheral nerves. These results along with our
previously demonstrated results open the possibility of developing neuroprostheses capable of generating afferent and
efferent neural signals in response to optical stimulation.

20
HybridElectricalOpticalFunctionalStimulationSystem

SahilK.KapurMD,ThomasJ.RichnerMS,SarahK.BrodnickBS,JustinC.WilliamsPhD,SamuelO.PooreMDPhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SAHILKAPUR
(607)5924436
kapur.sahil@gmail.com
Resident
Plastic&ReconstructiveSurgery
SamuelPooreMD/PhDJustinWilliamsPhDNITROLab.
BasicScience
FunctionalElectricalStimulation,Neuroprosthetics,PeripheralNerve,Optogenetics,
Muscle

Purpose:Conventionalneuroprosthesesmakeuseoffunctionalelectricalstimulationasamodalitytoproduceuseful
motion.Whiletherehavebeensignificantadvancesmadeinelectricalstimulationbasedcontrolalgorithms,major
obstaclestothebroadadoptionofthistechnologyhavebeenthehighlevelsofmusclefatigueandtissuedamagedueto
extendedperiodsofelectricalstimulation.
Thefastemergingtechnologyofoptogeneticshasallowedustoopticallystimulateperipheralnervesviatheexpression
oflightsensitivemicrobialopsinsinmammalianneurons.Optogeneticstimulation,however,islimitedbythelevelsof
ionchannelexpressioninaperipheralnervesuchthatthemagnitudeofcompoundmuscleactionpotentialsgenerated
byopticalstimulationaloneissignificantlylowerthantheresponsespossiblewithelectricalstimulation.
Ourobjectiveistocombinebothelectricalandopticalstimulationmodalitiestodevelopahybridstimulationinterface
thathasthecapabilityofproducingbothlessfatigueaswellasahighmagnitudeofmuscleEMGresponse.
Method:Transgenicmiceexpressingbluelightsensitivechannelrhodopsinionchannelswereanesthetizedandtheir
sciaticnerveandgastrocemiusmuscleswereexposedinaccordancewithIACUCguidelines.Simultaneouselectricaland
opticalstimulationofthesciaticnervewascarriedoutusingaconcentricneedleelectrodeandabluelightemitting
diode.Compoundmuscleactionpotentialsweremeasuredusingneedleelectrodesplacedinthegastrocnemiusmuscle.
Results:IsolatedelectricalstimulationgeneratedasignificantlyhigherEMGresponseinthemuscleandmorefatigue
thanisolatedopticalstimulation.Combinedelectricalopticalstimulationwasabletogenerateacomparablemagnitude
ofEMGresponsewithlesselectricalandopticalstimulusthanwhenbothmodalitieswereusedinisolation.Subtwitch
thresholdelectricalstimulationwasamplifiedinthepresenceofsimultaneousopticalexposureofthenervetogenerate
EMGresponseinthegastrocnemiusmuscle.Combinedelectricalopticalstimulationdidnotproducesimilareffectsin
controlmice.
Conclusion:Combinedelectricalopticalstimulationallowsonetousetheadvantagesofbothmodalitiestodevelopan
interfacethathasthecapabilityofproducinglessfatigueaswellasahighmagnitudeofmuscleresponse.Thishybrid
modelcanbeintegratedintoexistingcontrolalgorithmstoimproveperformanceandlongevityoffunctionalelectrical
stimulationdevices.

21
Early and progressive oromotor and swallowing dysfunction in a PINK1 knockout model of Parkinson Disease
CA Kelm-Nelson1 LM Grant1,2 , E Paul1 , MR Ciucci1,2,3 1 Department of Surgery, Division of Otolaryngology-Head and Neck
Surgery; 2 Department of Communication Sciences & Disorders; 3 Neuroscience Training Program

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Cynthia Kelm-Nelson
(608) 262-6122
cakelm@wisc.edu
Postdoctoral Fellow
Otolaryngology-Head & Neck Surgery
Dr. Michelle Ciucci
Basic Science
Parkinson Disease, Oromotor, Swallowing, Rat, Transgenic

Introduction: Parkinson disease (PD) is devastating to oropharyngeal swallowing. However, the onset, progression and
neural correlates of PD-related dysphagia are poorly understood. To address this, we used a transgenic rat model of PD
and hypothesized that deficits would manifest early in the disease process, progress and be related to pathologies in
brain structures associated with cranial sensorimotor function.
Methods: Rats with homozygous and heterozygous knock-out (KO) of PINK1, and WT controls (n=48) were studied. Bite
force and timing characteristics were measured during consumption of pasta at 4 and 8 months. Lingual force and
timing control was measured during a complex licking task at 2, 4, 6 and 8 months. At 8 months, we performed
immunohistochemistry for tyrosine hydroxylase in the substantia nigra and striatum and for alpha-synuclein aggregates
in brainstem and cortical regions important to oromotor control. To determine differences between groups, a KruskalWallis ANOVA based on ranks was used for biting measures and a repeated measures ANOVA with Holm-Sidak
comparisons was used for lingual forces (alpha<0.05).
Results: There was a significant main effect for genotype for the inter-bite interval (F(3, 48)=10.12, p=0.001). The
homozygous rats had significantly longer intervals compared to heterozygous rats (p=0.005) and WT (p=0.001). There
was a significant main effect for age (F (2,48)=6.88, p<0.001); rats at 8 months pressed with more force compared to rats
at 4 months.
There was a significant interaction for age x genotype (F(6, 99)=12.14, p<0.0001) for average force. At 4 months, the
homozygous rats pressed with greater force than the heterozygous (p<0.0001)and WT rats (p=0.005). This persisted at
the 6 and 8 month time points (p< 0.0001 for all comparisons). There were no significant differences between WT and
heterozygous rats.
There was a significant interaction for age x genotype F(6, 99)=4.32, p=0.0006) for variability of force. Homozygous rats
showed an increased variability in the amount of force per press during the session as compared to WT and
heterozygous rats at 2 (p<.0001, p=.0086-uncorrected), 4(p<.0001, p<.0001), 6(p<.0001, p<.0001) and 8 months
(p<.0001, p=.0026). Notably, homozygous rats significantly increased their variability between 2 to 4 (p=.0014) and 4 to
6 months (p<.0001); this plateaued at 6 months. Heterozygous rats also significantly increased their variability from 4 to
6 (p<.0001) and 6 to 8 (p<.0001)months.
There were no differences in tyrosine hydroxylase among genotypes in the substantia nigra or striatum (F (2, 29)=5.78,
p=0.1). Alpha synuclein positive aggregates were present in the substantia nigra, nucleus ambiguus, and dorsal motor
nucleus of the vagus regions of the brainstem in homozygous and to some degree heterozygous rats.
Conclusion: Overall results indicate that transgenic PD rats, especially with homozygous KO of PINK1, demonstrate early
oromotor dysfunction and that lingual deficits regarding fine control and timing of licking behavior is progressively
impaired. Although there was no depletion of nigrostriatal dopamine in PD rats, there were alpha synuclein positive
aggregates in the substantia nigra as well as brainstem areas that control oromotor function in PD rats. Understanding
the pathology of swallowing deficits is vital to developing PD treatments.

22
CellularBasedImmunodepletionandTomotherapyforPreconditioningof
ToleranceInductioninaRhesusMacaqueRenalAllotransplantModel

Kempton,Steve,MD,Haynes,Lynn,MS,JankowskaGan,Ewa,Christensen,Neil,BVSc,Forrest,Lisa,VMD,Hematti,Peiman,MD,
Burlingham,William,PhD,Kaufman,Dixon,MDPhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SteveKempton
(909)5531690
skempton@uwhealth.org
Resident
Transplantation
Burlingham/KaufmanLab.PI:DixonKaufman
BasicScience
Tolerance,Tomotherapy,Chimerism,Rhesus,RenalAllotransplant

INTRODUCTION:Currenttrendsintransplantimmunologyfocusontheutilizationoftolerancemechanismsinherentin
thehumanimmunesystemwiththeintenttocreateahostimmuneenvironmentthatishyporesponsivetothe
transplantedorgan.Todate,donorhematopoieticstemcell(HSC)andTcellinfusiontoachieveimmunetolerancehas
shownpromiseinthecreationofamixedchimerichostinMHCidenticaldonor/recipientpairs.Inordertotestthis
methodacrossdifferentMHCdisparities,wearedevelopinganonhumanprimaterenalallotransplantandHSC/Tcell
transplantmodel.Intheearlystagesofdevelopingsuchamodel,itisimportanttoreportthesafetyprofileandchanges
totheimmunesystemduringandafterprofoundimmunosuppression.
METHODS:Sevenrhesusmacaquemonkeysweredividedinto3separatetreatmentgroups.Allanimalsreceived5days
ofantithymocyteglobulin(ATG),withsomeanimalsreceiving10fractionsof1.2Gyhelicaltomotherapybasedtotal
lymphaticirradiation(TLI),andsomeanimalsreceivingakidneyallotransplantonday0.Group1(n=1)receivedATG
only;Group2(n=3)receivedbothATGandTLI;Group3(n=3)receivedATGandTLIandarenalallograft.Allanimals
underwentregularintervalblooddrawstofollowabsoluteneutrophilandlymphocytecountsaswellaschemistry
panelsforrenalfunctionandelectrolytes.Weeklyflowcytometricanalysiswasperformedtodetectlymphocytesubset
populations.CytomegalovirusreactivationwasmeasuredbyquantitativePCRevery1to2weeks.TrainedVeterinary
staffmonitoredanimalsclinicalconditiondaily.
RESULTS:Allanimalsinitiallydidwellwithnofeverormajorillness.Cellularbasedimmunodepletivetherapyalone
(Group1)didresultinlymphocytedepletion;however,lymphocyterecoverywasnotedafterday7.Thecombineduse
ofATGandTLIinGroups2and3demonstratedadequatelymphocytedepletion,lastingbeyond14days;however,
severeneutropenia(ANC<500)wasnotedinsomeanimalsrequiringantibioticsandtemporarycessationofmaintenance
immunosuppression.AnadjusteddecreaseddoseofTLIwasadministeredto2animals(oneeachingroups2and3)and
alsoresultedinadequatelymphocytedepletion.Lymphocytesubsetsingroups2and3recoveredtopretransplant
levelsby79weekspostinitiationoftreatmentandproportionsofCD4andCD8cellsreturnedtonormalby26weeks.
Allanimalsintherenalallograftgroup(3)developedCMVreactivationrequiringtreatment.
CONCLUSION:AcombinationofATGandhelicaltomotherapyforTLIcansafelybeadministeredtotherhesusmacaque,
providingprofoundimmunosuppressionbeyond2weekswithouttoxicity.ItwasanimportantobservationthatCMV
reactivationoccurredinthissetting(especiallywhenarenalallotransplantwaspresent)andshouldbeanticipatedand
treatedwithprophylacticantiviralmedication.FurtheradjustmentstoTLIdoseanddistributionoftherapymayresult
inlessneutropeniaandpossiblylessCMVreactivation.Thisappearstobeanidealpreconditioningprotocoltoallowfor
posttransplantinfusionandengraftmentofdonorHSCandTcellinfusionstogenerateamicrochimericstatefor
toleranceinduction.

23
Treatmentofratvocalfoldscarbygp46silencing

YoKishimoto,AliceWei,YutakaToya,NathanV.Welham

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

YoKishimoto
(608)2130121
toya@surgery.wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
Welham
BasicScience
vocalfold,scar,gp46,collagen,fibrosis

Purpose:Gp46isacollagenspecificchaperoneproteinresidingintheendoplasmicreticulum.Itisessentialforthe
maturationofcollagenandthusisthoughttoplayanimportantroleintheprogressionoffibrosis/scarring.The
purposeofthisstudywastoevaluatethetherapeuticpotentialofgp46silencingforthetreatmentofchronicvocalfold
scar.
MaterialsandMethods:First,normalandscarratvocalfoldfibroblastsweretransfectedusinggp46siRNA/liposome
complexesatfirstpassage.Transfectionefficiencywasevaluatedusingflowcytometry,fluorescentmicroscopyandqRT
PCR,andeffectoncollagensynthesiswasevaluatedviahydroxyprolineabundance.Next,theeffectofgp46silencingon
chronicallyscarredvocalfoldswasexaminedinvivo.Ratvocalfoldswereinjuredbilaterallyandallowedtohealfor8
weeks.siRNAcomplexeswereinjectedintotheresultingscarredvocalfoldstwiceperweekfor4weeks.Afterthefinal
injection,animalswereeuthanizedandthevocalfoldmucosawasevaluatedforcollagentranscriptionandprotein
abundance.
Results:Gp46knockdownwassuccessfullyachievedviainvitrotransfection,andcorrespondedtoareductionin
collagensynthesisbynormalandscarvocalfoldfibroblasts.Further,invivodeliveryofgp46siRNA/liposome
complexesreversedtheexcessivecollagendepositionassociatedwithchronicvocalfoldscar.
Conclusions:Gp46silencingisapromisingtherapeuticstrategyforthetreatmentofvocalfoldscar.

24
TourniquetMasterTrainingforJunctionalandInguinalHemorrhageControl
(TMT)

PeterW.WEYHRAUCH,PhD,JamesNIEHAUS,PhD,MaxMETZGER,BS,ShlomiLAUFER,PhD,CalvinKWAN,BS,CarlaPUGH,MD,
PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

CalvinKwan
(608)2625241
kwan@surgery.wisc.edu
Scientist
GeneralSurgery
Dr.CarlaPugh
BasicScience
Assessment,Tourniquet,Performance,SensorEnabledManikin,TrainingTool

Junctionalandinguinalbleedingisasignificantandchallengingproblemonthebattlefield.Inventorshavedeveloped
newtypesoftourniquets,includingtheAbdominalAorticTourniquet(AAT)andtheCombatReadyClamp(CRoC)to
addressthesegroinandpelvicinjuries.Whilethesehemorrhagecontroltechnologieshavebeendeveloped,validated,
andapprovedforuse,trainingsystemsthatteachandrefreshskillsrelatedtothesetechnologieshavenotbeen
developed.Trainingsystemsforthesetypesofinjuriesarevitalbecausetheinjuriesarerareonthebattlefieldand
difficulttotrain.Toaddresstheseneeds,asensorenabledmanikinwasdesigned.Usingthesensordata,thedifferent
phasesofapplyingthetourniquetwerecapturedandkeyperformancewasmeasured.

25
Selective Induction of the Nrf2-Antioxidant Response Element (ARE) Pathway in
Hepatocytes Decreases the Development of Non-Alcoholic Steatohepatitis and is
Associated with Augmented Expression of -Oxidation and Triglyceride Export
Genes in Mice
Lung-Yi Lee, Drew Roenneburg, Li Zhang, Jeffrey A. Johnson, David Foley

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Lung-Yi Lee
(608) 556-4601
LLee2@uwhealth.org
Resident
Transplantation
David Foley
Basic Science
Nrf2, NAFLD, NASH, lipid metabolism, transgenic

Background:
Oxidative stress is implicated in the development of non-alcoholic steatohepatitis (NASH). The Nrf2-ARE pathway
protects cells from oxidative damage. We have demonstrated that the absence of Nrf2 exacerbates NASH progression.
The purpose of this study was to determine whether hepatocyte-specific overexpression of Nrf2 mitigates the
progression of NASH.
Methods:
Transgenic mice with hepatocyte-specific overexpression of Nrf2 (AlbCre/caNrf2, n=16) and littermate controls (n=13)
were fed either standard chow or methionine-choline-deficient (MCD) diet. After 28 days, livers and serum were
obtained for analyses. Serum ALT and AST were measured using the IDEXX VetTest Chemistry Analyzer. Oil Red O
staining of liver tissue was performed to evaluate for area of steatosis. Staining was quantified using ImageJ. Hepatic
triglyceride was extracted using the Folch method, and concentration determined enzymatically. qRT-PCR was used to
assess mRNA abundance of Nrf2-dependent and lipid metabolism genes. Comparisons between the groups were
performed with one-way ANOVA followed by Fishers LSD post hoc test. P<0.05 was used for statistical significance.
Results:
Compared with standard chow, both AlbCre/caNrf2 and control mice on MCD diet had elevated serum transaminases,
area of steatosis, and liver tissue triglyceride. However, among the MCD diet groups, AlbCre/caNrf2 animals had
significantly decreased serum transaminases, area of steatosis, and liver tissue triglyceride as compared to the control
animals. There are significant increases in the baseline mRNA abundance of Nrf2-dependent genes in AlbCre/caNrf2
mice as compared to the control mice. When fed MCD diet, Nrf2 induction did not alter the expression of genes involved
in de novo fatty acid synthesis. However, compared to control mice on MCD diet, AlbCre/caNrf2 mice on MCD diet
demonstrated significantly increased expression of CPT2, a critical gene required for -oxidation of fatty acid, and MTTP,
an important gene regulating the assembly of VLDL and export of hepatic triglyceride. In addition, AlbCre/caNrf2 mice
on MCD diet have increased expression of genes involved in fatty acid transport (CD36, FATP1, and FATP4) and
decreased expression of genes involved in intracellular fatty acid binding (FABP1 and FABP5).
Conclusion:
Selective overexpression of Nrf2 in hepatocytes decreases the development of steatosis in a dietary model of NASH.
These findings are associated with increased mRNA abundance of genes involved in fatty acid transport, -oxidation, and
triglyceride export. The current study suggests novel Nrf2-dependent mechanisms involving lipid metabolism in NASH.
Future studies are warranted to elucidate these interactions.

26
Overexpression of the Nrf2-Antioxidant Response Element (ARE) pathway in
hepatic stellate cells (HSCs) ameliorates the development of Non-Alcoholic
Steatohepatitis (NASH) in mice.
Lung-Yi Lee, Li Zhang, Drew Roenneburg, Jeffrey A. Johnson, David Foley

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Lung-Yi Lee
(608) 556-4601
LLee2@uwhealth.org
Resident
Transplantation
David Foley
Basic Science
Nrf2, NASH, Hepatic Stellate Cells, Transgenic, NAFLD

Background:
Oxidative stress is implicated in the development of NASH. The Nrf2-ARE pathway protects cells from oxidative damage.
We have demonstrated that the absence of Nrf2 exacerbates NASH progression, and hepatocyte-specific overexpression
of Nrf2 mitigates the development of steatosis. The purpose of this study was to determine whether overexpression of
Nrf2 in HSCs abrogates the progression of NASH.
Methods:
Transgenic mice with HSC-specific overexpression of Nrf2 were generated using the Cre-Lox technology with the glial
fibrillary acidic protein (GFAP) promoter (GFAP-Cre/caNrf2). HSCs from transgenic animals were isolated and cultured.
Quantitative RT-PCR was used to assess mRNA abundance of Nrf2-dependent genes in cultured HSCs to confirm
phenotype. Transgenic mice (GFAP-Cre/caNrf2) (n=11) and their littermate controls (n=9) were then fed either control or
methionine-choline-deficient (MCD) diet. After 28 days of diet, the livers and serum were obtained for analyses. Serum
ALT was measured using IDEXX VetTest Chemistry Analyzer. Oil Red O staining was performed to evaluate for steatosis.
Hepatic oxidative stress was determined by measuring thiobarbituric acid reactive substances (TBARS).
Immunohistochemistry and immunoblotting with anti--smooth-muscle actin (-SMA) antibody was performed to
assess HSC activation. The mRNA abundance of genes involved with HSC activation and fibrogenesis was determined
from qRT-PCR of liver tissue samples. One-way ANOVA followed by Fishers LSD post hoc test was performed for
statistical analysis. P<0.05 for statistical significance.
Results:
Quantitative RT-PCR demonstrated significant increases in the expression of Nrf2-dependent genes in GFAP-Cre/caNrf2
HSC cultures as compared to controls. After 28 days of MCD diet, the percentage of steatosis was significantly higher in
mice fed MCD vs. control diet. Among the MCD diet groups, both serum ALT and the percentage of steatosis were
significantly lower in the GFAP-Cre/caNrf2 animals as compared to the control animals. This was associated with
significantly decreased TBARS in the livers of GFAP-Cre/caNrf2 animals vs. control animals on MCD diet. The GFAPCre/caNrf2 animals on MCD diet also have significantly decreased mRNA abundance of fibrogenic genes, including
COL1A1 and TIMP1, as compared to the controls on MCD diet. Finally, immunoblot demonstrated a decrease in -SMA
protein levels in GFAP-Cre/caNrf2 animals as compared to the control animals on MCD diet.
Conclusion:
Induction of Nrf2-ARE pathway in HSCs decreases the development of steatosis, oxidative stress, HSC activation, and
fibrogenic gene expression in a dietary model of NASH. Future studies are required to delineate the molecular
mechanisms involved in this novel protective role of Nrf2 induction in HSCs.

27
Novelisolationandcharacterizationofporcinevocalfoldepithelialcells

ElizabethEricksonLevendoski,CiaraLeydon,&SusanL.Thibeault

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ElizabethLevendoski
(219)6710260
levendoski@surgery.wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
SusanL.Thibeault
BasicScience
Vocalfolds,Epithelium,Cellculture,Porcine,Barrierfunction

Introduction:Thevocalfoldsareauniquelylayeredstructureconsistingofepithelium,laminapropria,andmuscle.As
theoutermostlayerofthevocalfolds,theepitheliumformsabarrierthatprotectstheunderlyingconnectivetissue
frominjurywhilebeingnearlycontinuouslyexposedtopotentiallyhazardousinsultsincludingenvironmentalor
systemicbasedirritants,surgicalprocedures,andvibratorytrauma.Injurytothevocalfoldepithelialbarrierhasbeen
implicatedinthepathogenesisofmanyvocalfoldpathologies,bothbenignandmalignant.However,thecellularand
molecularmechanismsunderlyingepithelialcellfunctioninhealth,injury,anddiseasehavebeenlargelyunexplored.A
primaryreasonforthisisthatvocalfoldepithelialcelllinesdonotexist.Compoundingthisproblem,epithelialcellsare
difficulttoaccessandcannotberemovedfromthehealthyhumanlarynxwithoutasignificantrisktovocalfunction.
Animalmodelsarecommoninvocalfoldrelatedresearchandprovideanalternativeforhumanepithelialcellculture.
Theaimofthisstudywastoestablishaprimarycultureofporcinevocalfoldepithelialcells.
Methods:Porcinelaryngeswereobtainedfromlocalabattoirswithin15minutesofanimalsacrifice.Vocalfoldswere
excisedandepithelialcellswereisolatedthroughacombinationofdispaseandtrypsintreatments.Isolatedcellsfrom
eachlarynxwerecountedandplatedincollagencoated6wellplatesortranswellinsertsandculturedfor7daysunder
liquidcoveredconditions.Cellswerecharacterizedmorphologicallyandbymeasuringepithelialmarkersusing
immunocytochemistry.Inordertoidentifythebarrierpropertiesoftheepitheliumformed,invitro,transepithelial
resistance(TER)wasalsorecorded.
Results:Onaverage,4.51x105cellswereisolatedperlarynx(range:6.3x1048.86x105cells).Epithelialcellsisolated
fromporcinevocalfoldsgrewto7090%confluentmonolayerswithin7days.Themajorityofcellsinculturewere
morphologicallyepitheliallikeinappearanceasevidencedbytheformationofdenselypackedclustersofpolygonal
shapedcells.Somemorphologicallyelongatedcells,fibroblastlikeinappearance,werealsonotedinculture.Epithelial
appearingcellswerepositiveforepithelialcellsmarkersincludingpancytokeratinandtheepithelialmembrane
associatedmucinproteinMUC4.TERvaluesrangedfrom<50183cm2.
Conclusions:Wehavesuccessfullyisolatedandculturedprimaryporcinevocalfoldepithelialcells.Comparedto
previouslystudiedcellisolationandculturetechniques,thismethodincreasestheyieldofviableepithelialcells,while
simultaneouslylimitingthegrowthofnonepithelialcellssuchasfibroblasts.Continuingstudieswillincludestainingfor
epithelialcelljunctionsandfunctionalmeasurementsofpermeability.Improvingthebarrierfunctionoftheseprimary
cultureswillalsobeafuturefocus.Inconclusion,ourporcinevocalfoldepithelialprimarycellculturesholdpromisefor
useinfutureresearchthatseekstostudythevocalfoldepithelialbarrierinhealthanddisease.

28
Establishment of human pancreatic extracellular matrix co-culture system for
endocrine specification and maturation from human embryonic stem cells
Xiang Li, Sara Dutton Sackett, Cori OBrien, Jon S. Odorico

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Grace Li
(608) 263-6668
li@surgery.wisc.edu
Scientist
Transplantation
Jon S Odorico
Basic Science
pancreatic extracellular matrix, human embryonic stem cell, endocrine progenitor
cell, decellularization, beta-cell

Objectives: The generation of insulin-producing cells through induced-differentiation of human embryonic stem cells
(ESCs) and induced pluripotent stem cells (iPSCs) has been intensively studied for their potential therapeutic effects in
treating patients with Type 1 diabetes. A variety of protocols have demonstrated the ability to promote the in vitro
differentiation of hESCs/iPSCs into insulin positive cells. However, major limits still remain in the efficiency in generating
enriched, robustly
important role during human embryonic development by actively regulating cell behavior through structural support
and biochemical stimulation. Tissue specific ECM has been proposed for use in a number of regenerative strategies for
tissue and organ replacement. However, few studies have explored the potential of using pancreatic ECM to specifically
-culture of human
enhance the differentiation of human ESCs into functional
ESCs with pancreatic ECM would improve upon the conventional methods regarding the efficiency of the in vitro
differentiation and the in vivo maturation processes. This investigation aims to characterize the biological nature and
Methods: In order to establish the ECM-derived cell culture platform, we developed a method to secure 200m sections
of decellularized human pancreas as a biologic scaffold to Transwell membranes in an in vitro culture system. To obtain
optimal ECM scaffold material, various decellularization protocols employing the use of SDS, TritonX-100, sodium
deoxycholate and combinations therein were examined. HESCs (WA01) were differentiated using a stage-specific and
chemically-defined protocol, and then seeded onto the ECM scaffold for further differentiation. Samples from specific
stages were analyzed by immunohistochemistry for pancreatic lineage markers, such as Foxa2, Pdx1, insulin and
glucagon and compared with cells differentiated in the absence of matrix.
Results: Partially differentiated definitive endoderm cells were observed to adhere to and incorporate into the 3-D
scaffold structure throughout the subsequent differentiation stages. In contrast, undifferentiated hESCs cells exhibited a
significantly lower cell attachment rate to the pancreatic ECM scaffold, unless additional vitronectin was present. This
difference in cell attachment performance demonstrates the affinity between the cells with a committed differentiation
fate and the specific ECM material. At the final stage of the in vitro differentiation, a mixed population of
insulin+/glucagon- cells, glucagon+/insulin- cells, and insulin+/glucagon+ cells were observed in the ECM co-culture
condition.
Conclusions/Future directions: Further quantification analysis of the in vitro observation is ongoing. Differentiated cells
co-cultured with ECM scaffold were also co-transplanted under the kidney capsule of SCID/Beige mice and will be
subjected to metabolic analysis to determine if transplantation of progenitors with ECM is able to accelerate the in vivo
maturation process compared to the transplantation of cells alone. A similar co-culture system utilizing pancreatic ECMderived hydrogel is also currently under examination as an alternative method that aims to enhance the differentiation
and maturation of pancreatic progenitor cells.

29
Utilizingproteomicsapproachestobetterunderstandtissueengineeringprocess

QiyaoLi,ZhenChang,BasakE.Uygun,ChangyingLin,BrianL.Frey,LloydM.Smith,NathanV.Welham

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

QiyaoLi
(608)6952768
qli@chem.wisc.edu
GraduateStudent
OtolaryngologyHead&NeckSurgery
NathanWelham
BasicScience
proteomics,liver,decellularization,vocalfolds,proteinturnover

Apromisingtissueengineering/regenerativemedicineapproachforfunctionalorganreplacementhasemergedinrecent
years.Itinvolvesusingdecellularizedbiologicalscaffolds,withinwhichseededcellsgrowtoformnewtissues.Withthe
decellularizationandrecellularizationstrategy,researchershavecreatedpotentiallyfunctionalorgansincludingheart,
liver,lung,kidney,etc.
Proteomicsisthenextstepinthestudyofbiologicalsystemsaftergenomicsandtranscriptomics.Itisthelargescale
studyofproteinsandisgenerallyperformedwithmassspectrometry(MS).MSmethodsaresensitive(permitting
routinedetectionoffemtomoleamountsofpeptides),rapid(sequencingthousandsofpeptidesinasinglerun)and
readilyautomated(permittinghighthroughputstrategies).Thus,MSprovidesanattractivemethodologyforthe
investigationofcomplexproteinmixturesisolatedfromcells,bodyfluidsandtissues.
Ourworkistoutilizeproteomicsapproachestobetterunderstandthetissueengineeringprocess.Twoaspectsofour
recentcollaborativeworkarepresentedbelow,withoneinvolvingratliverandtheotherinvolvinghumanvocalfolds.
i)Evaluationofratliverdecellularizationeffectiveness:Thereisevidencethatremnantcellularmaterialsmaycontribute
toadversehostresponsefollowingimplantation.Therefore,thegoaloforgandecellularizationistoeffectivelyremove
cellularmaterialswhilepreservingthecompositionandbiomechanicalpropertiesoftheECM.Currentquantitative
criteriaofdecellularizationaresolelyrelatedtonucleicmaterialssincetheyweredirectlycorrelatedtoadversehost
responsesandtheywereeasytoquantify.However,ithasalsobeenrecognizedthatothercellularmaterialssuchas
thoseassociatedwithcytoplasmormembranealsohaveeffectsonhostreactions.Toprovideanassessmentmethod
complementarytothenucleibasedcriteria,weappliedaproteomicsapproachtoanalyzingdecellularizedliverforboth
ECMproteinsandremnantcellularproteins,whicharepotentiallyantigenicuponfuturetissueimplantation.
ii)Determinationofthecontributionofrepopulatinghumanvocalfoldfibroblasts(VFFs)toreconstitutionandturnover
ofdecellularizedvocalfold(VF)ECMscaffolds:Thestepfollowingdecellularizationisrecellularization.Inthecaseof
vocalfoldsengineering,thecellsusedforrecellularizationcouldbeVFFsandVFepithelials.VFFsareresponsibleforECM
turnoverandmaintenance.Proteomictechnologieshaveintroducedthepossibilityofdeterminingtheturnoverratesof
multipleproteinssimultaneously.WeusedtheSILAC(stableisotopelabelingbyaminoacidsincellculture)approachto
differentiatebetweenproteinsthatarealreadyintheECMscaffoldsandthenewlysynthesizedones.Byunderstanding
theextentandratetowhichrepopulatingcellsturnoverthedecellularizedECM,wewilloptimizethistissueengineering
approachforfutureclinicalapplications.

30
MurineAllograftElastasetreatedAbdominalAortaTransplantationfor
PathogenesisResearchofAbdominalAorticAneurysm

ZhenjieLiu,QiweiWang,StephanieMorgan,JunRen,JasmineGiles,andBoLiu

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ZhenjieLiu
(608)7727134
lawson3001@gmail.com
PostdoctoralFellow
VascularSurgery
BoLiu
BasicScience
Abdominalaorticaneurysm,Elastase,Atherosclerosis,Transplantation,Inflammation

Background:Syngeneicheterotopictransplantationofaorticarchhasbeenusedforatherosclerosisstudies.However,
therearenosimilarmurinemodelsforpathogenesisresearchofabdominalaorticaneurysm(AAA).MurineelastaseAAA
modelisoneofthemostpopularmodelsforpathogenesisstudies.Murineallograftelastasetreatedabdominalaorta
transplantationfromgeneticallymodifiedmicetowildtyperecipientsorviceversa,isusefulforstudiesonAAA.
Methods:Theinfrarenalaortafromthedonormousewastreatedbyelastase(0.45U/Ml)5min,andanastomosedtothe
recipientsaortaatthesameposition.Orthotopicaortictransplantationwasperformedin45mice.Themicewere
sacrificedatday7or14.InflammatorycytokineswereevaluatedbyqPCR,AndHE,elastindegradationgrade,apoptosis,
andinflammatorycellswereevaluated.
Results:Theoverallsurvivalratewas51.1%.Mostoftheelastasedtreatedtransplantedaortasstilldevelopedinto
aneurysm(95.7%).Theelastindegradationgrade,apoptosisindex,CD68/CD3postivecellratiosweresimilarbetween
thetransplantedgroupandelastasegroup.AndthereisnosignificantdifferenceofCCL2,CCL5,TNF,INFbetween
thesetwogroups.
Conclusions:Theelastasetreatedtransplantedabdominalaortastilldevelopedaneurysm.Thisallograftelastasetreated
abdominalaortatransplantationmodelcouldbeusedasanalternativeofbonemarrowtransplantationinthe
pathogenesisstudyofAAA.

31
HighMolecularWeightPolyethyleneGlycol(PEG)InhibitsMyocardialIschemia
ReperfusionInjuryandImprovesSurvivalandVentricularFunction

JamesLloyd,XianyaoXu,JenniferL.Philip,CharlieMuller,andShahabA.Akhter

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JamesLloyd
(608)2636551
akhter@surgery.wisc.edu
MedicalStudent
CardiothoracicSurgery
ShahabA.Akhter,MD
BasicScience
Ischemiareperfusioninjury,Highmolecularweightpolyethyleneglycol,myocardial
infarction,cellsurvivalsignaling,oxidativestress

Introduction:Cardiacischemiareperfusion(IR)injuryremainsasignificantclinicalproblemastherearenotherapies
availabletominimizethedamageresultingfromnecroticandapoptoticcelldeaththatmayleadtoimpairedfunction
andheartfailure.WehavepreviouslyshownthathighmolecularweightPEG(1520kD)canprotectcardiacmyocytesin
vitrofromhypoxiareoxygenationinjury.Themechanismincludespreservationofplasmamembranelipidraft
architecture,decreasedoxidativestress,andupregulationofcellsurvivalsignaling.Inthisstudy,wetranslatedthesein
vitrofindingstoasmallanimalmodelofmyocardialischemiareperfusioninjurytoinvestigatethepotentialprotective
effectsofPEGinvivo.
Methods:AdultmaleSpragueDawleyratsunderwentleftthoracotomyandLADocclusionfor60minutesfollowedby
either48hoursor4weeksofreperfusion.Onemlof10%PEGsolutionorsalinecontrol(n=10ineachgroup)was
administeredviatheinternaljugularveinjustpriortoreperfusion.SurvivalwasmeasuredbyKaplanMeieranalysis.
Cardiacfunctionwasstudiedbyechocardiography.Othermethodsincludedproteinimmunoblotting,andhistological
stainingforfibrosis.StatisticalanalysiswasperformedusingStudentsttest.
Results:FluoresceinlabeledPEGwaswellvisualizedinthemyocardiumafterintravenousdeliveryandwasnotdifferent
thandirectintracoronarydelivery.Survivalat4weekspostIRinjurywasgreaterinthePEGgroupwith86%survival
comparedtoonly60%survivalinthecontrols.ThePEGgrouphadfullrecoveryofLVfunctionat4weeksvs.a25%
declinefrombaselineinthePBSgroupasquantitatedbyejectionfraction(761%vs.642%,PEGvs.PBS;p<0.001)and
fractionalshortening(0.460.01vs.0.360.02;p<0.001).At4weeks,LVendsystolicdimensionswerepreservedinthe
PEGgroupcomparedtoPBSwheretherewasa1.5foldincrease(3.80.3vs.4.60.2mm;p=0.05).Therewasa50%
declineinLVfibrosisinthePEGgroupat4weekspostIRinjuryvs.PBScontrolwithsignificantlysmallerinfarctand
remoteterritoryfibrosisasmeasuredbytrichrome(p<0.01)andpicrosiriusred(p<0.003)histologicalstaining.Cell
survivalsignalingwasalsoupregulatedinthePEGgroupwithincreasedAkt(3fold,p<0.005)andERK(4fold,p<0.05)
phosphorylationcomparedtoPBScontrolsat48hourspostIRinjury.
Conclusions:HighmolecularweightPEGappearstohaveasignificantprotectiveeffectfromIRinjuryintheheartwhen
administeredintravenouslyjustpriortoreperfusion.Thismayhaveimportantclinicalimplicationsinthesettingofacute
coronarysyndrome,myocardialprotectionincardiacsurgery,andorganpreservationfortransplantation.Futurestudies
willexploreadditionalmechanismsofthisbeneficialeffectaswellasusinglargeranimalmodels.

32
NerveStimulationdoesnotIncreaseMitochondrialDNADeletionMutationsin
theTongueMusculature

EntelaLushaj,HeidiKletzien,BesaJonuzi,MariaBarnes,AlexanderCook,NadineConnor

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

EntelaLushaj
(608)2635221
lushaj@surgery.wisc.edu
Scientist
OtolaryngologyHead&NeckSurgery
NadineConnor
BasicScience
Mitochondria,Mutations,Tongue,Exercise,DNA

Introduction
TheabundanceofmitochondrialDNA(mtDNA)deletionmutationsintonguemusclesmayincreasewithageand
contributetomuscledysfunctionassociatedwithagerelatedswallowingdeficits.Itiscontroversialwhetherexercise
effectsgenerationofmtDNAdeletionmutationsinskeletalmuscle.Weexaminedtheeffectsofageandneuromuscular
electricalstimulation(NMES),aclinicaltreatmentfororopharyngealdysphagia,onmtDNAdeletionmutationsandtheir
associatedelectrontransportsystem(ETS)abnormalitiesinthetonguemusculature.
Methods
YoungAdult(9monthold;n=15),middleaged(24monthold;n=12),andold(32monthold;n=16)ratsweredivided
intogroupsthatreceivedhypoglossalnervestimulationat10Hzor100Hz(5days/week).ThepresenceofmtDNA
deletionsandETSabnormalitieswereanalyzedafter8weeksinthegenioglossus(GG),hyoglossus(HG),andstyloglossus
(SG)muscles.
Results
MtDNAdeletionsofthemajorarcwereidentifiedinallthreemusclesafterNMES.Statisticalanalysisdidnotshowa
significantdifferenceinthepresenceofmtDNAdeletionsbetweenyoungandoldortheintensityofexercise.Deletions
rangedfrom110kilobasesinsizeasshownbystandardPCR.AprescreeninghistologicalanalysisdidnotshowETS
abnormalmusclefibersinoldnormalorstimulatedstyloglossusmuscle.
Conclusions
OurdatashowthatNMESdoesnotpromotemtDNAdeletionformationinthetonguemusculature.

33
QuantifyingVocalFoldFluidContentUsingTissueDielectricProperties

AllisonL.Maytag,JacobP.Meyer,FuqiangGao,SusanC.Hagness,JackJ.Jiang

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AllisonMaytag
(414)5813504
maytag@surgery.wisc.edu
Scientist
OtolaryngologyHead&NeckSurgery
JackJ.Jiang
BasicScience
vocalfold,excisedlarynx,hydration,tissuedielectricproperties,fluidcontent

Introduction:Hydrationhasasignificantimpactonthemechanicalpropertiesofthevocalfolds.Asthevocalfolds
vibrateduringphonation,stressesareexertedonthetissuecomponents.Undercircumstancesofextremeloudnessand
duration,thestressescancauseextracellularandcellulardamagewhichinitiatethewoundhealingcascadeinthevocal
folds.Oneimportantcomponentofthehealingcascadeistheinfluxoffluidassociatedwithproteinsandimmunocytes,
resultinginlocalizededema.Theadditionallocalmassofthefluidcanincreasetheimpactstressintheinjuredareaand
thesameareaonthecorrespondingfold,creatingacycleofinjuryandfluidinfluxthatmayeventuallyleadtopathology
development.However,thereiscurrentlynomethodforquantifyingvocalfoldfluidcontentinvivo.Wethereforeapply
themicrowavefrequencydielectricpropertiesoftissue(TDP)tothevocalfoldsasanobjectivemeasureoffluidcontent.
Methods:ToestablishacorrelationbetweenvocalfoldfluidcontentandTDPparameters(relativepermittivityand
effectiveconductivity),thedielectricpropertiesofexcisedvocalfoldswereevaluated.Foreachmeasurement,a2.0
mmdiameteropenendedcoaxialprobewasappliedtotheblottedsurfaceofavocalfold.Therelativepermittivityand
effectiveconductivityweretakenfirstinthreeplacesin20vocalfoldsstoredin0.9%salinetoestablishnormalvalues.
Subsequentmeasurementsweretakenat1mgchangesinvocalfoldmassuntilsaturationortotaldehydration;10vocal
foldswerehydratedindeionizedwaterand10weredehydratedinadryingoven.ColeColemodelfitsareusedto
validatetheintegrityofeachmeasurement.
Results:Thelevelofhydrationwascloselycorrelatedtomeasurementsoftissuedielectricproperties.Therelative
permittivityandeffectiveconductivityofvocalfoldsincreasedwithincreasinglevelsofhydrationanddecreasedwith
dehydration.
Conclusion:Theseexperimentsestablishastandardcorrelationbetweenabsolutevocalfoldfluidcontentandthe
microwavefrequencydielectricpropertiesofvocalfoldtissue.Measurementsofvocalfolddielectricpropertiescan
thenbeusedinfuturestudiesofvocalfoldedemaandpathologydevelopment.

34
TraditionalMedicineintheSidamaRegionofEthiopia:Aglobalhealthfield
experience

AlexMcAlvay,YuliChen,HeidiBusse,GirmaTefera

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AlexMcAlvay
(206)2716295
mcalvay@wisc.edu
GraduateStudent
VascularSurgery
GirmaTefera
BasicScience
TraditionalMedicine,AccesstoMedicine,Healthsovereignty,Traditionaldisease
classification,HerbDrugInteractions

Acknowledgements:ThisstudywasmadepossiblethroughthegeneroussupportbytheDivisionofVascularSurgeryat
theUniversityofWisconsinSchoolofMedicineandPublicHealth.
BackgroundandObjectives:Morethan80%ofEthiopiaspopulationdependsontraditionalmedicineforprimarycare.
TheWHOdefinestraditionalmedicineashealthpractices,approaches,knowledgeandbeliefsincorporatingplant,
animal,andmineralbasedmedicines,spiritualtherapies,manualtechniquesandexercises,appliedsingularlyorin
combinationtotreat,diagnose,andpreventillnessandmaintainwellbeing.Theobjectivesofthisstudywereto:(1)
DocumentmedicinalplantuseamongtheSidamapeople,(2)Understandbarrierstohealthcareaccess,(3)Identify
autochthonousdiseaseclassifications,and4)Developahandbookofplantsandtheirtraditionalmedicinaluses.
Methods:ThisstudywasconductedinAletaWondo,locatedintheSidamaregionofsouthernEthiopia.Semistructured
interviewsandguidedfieldwalksconductedthroughatranslatorwerecarriedoutwithknowledgeableinformants
chosenthroughsnowballsampling.Barrierstoaccessinghealthservicesandresourceswereanalyzedthrough
communitymappingwithlocalelders.Medicinalplantspecimenswerecollected,photographedandspecimensthat
werenotreadilyidentifiablewerebroughttotheNationalHerbariumatAddisAbabaUniversityforconfirmation.The
informationsheetincludednameoftheplantinthelocallanguageandAmharic,pharmacologicalproperties,medicinal
usageandetc.
Results:TheSidamause47medicinalplantspeciesfrom30plantfamilies.Thesespeciesareusedtotreat36
distinctdiseases,includingfourlocallyacknowledgedconditionslackingdirectequivalentsinWesternbiomedicine.The
categoryofillnesswiththehighestproportionofplanttreatmentswasgastrointestinalcomplaints(40%).Followup
researchfound91.8%oftheplantspeciesmentionedtohavedocumentedpharmacologicalproperties,40.9%ofwhich
matchedthediseasetreatedwiththeplant(excludingdiseaseswithnoWesternequivalent).5specieswerefoundto
havepreviouslyidentifiedpotentialherbdruginteractions.Theplantsandtraditionalknowledgerecordedweremade
availabletothecommunityina40pageAmhariclanguagehandbooktosupportthecontinuationofthiscultural
knowledgeandsharefindingswithhealthcareproviders.
Conclusions:Preservationoftraditionalbotanicalknowledgeisimportanttoensurecontinuedaccesstomedicineby
underservedcommunities,preservingculture,reducingbiodiversityloss,andpromotingenvironmentalhealth.More
examplesareneededofwaysinwhichWesternmedicalresearchersandpractitionerscancollaboratewithindigenous
peoplestobuildcommonknowledgeaboutplantsandtheirmedicinaluses.Tothebestoftheauthorsknowledge,this
studywasthefirstofthistypetobecarriedoutwiththeSidamapeopleinhabitingtheSouthernNationsand
NationalitiesRegion(SNNPR)ofEthiopia.

35
Healthcare access and traditional medicine among the Sidama of Ethiopia
Alex McAlvay, Yuli Chen, Heidi Busse, and Girma Tefera

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Alex McAlvay
(206) 271-6295
mcalvay@wisc.edu
Graduate Student
Vascular Surgery
Girma Tefera
Basic Science
Traditional Medicine, Health Sovereignty, Healthcare Access, Disease classification,
Herbal Contraindications

Background: More than 80% of Ethiopias population depends on traditional medicine for primary care. The WHO
defines traditional medicine as health practices, approaches, knowledge and beliefs incorporating plant, animal, and
mineral based medicines, spiritual therapies, manual techniques and exercises, applied singularly or in combination to
treat, diagnose, and prevent illness and maintain well-being. Understanding local medicinal plant use and disease
constructs is an important step toward providing culturally appropriate health care to underserved communities
worldwide. Documentation of traditional plant medicine also plays a vital role in the conservation of biological and
cultural diversity. To the best of the authors knowledge, this is the first study of this type to be carried out with the
Sidama people inhabiting the Southern Nations and Nationalities Region (SNNPR) of Ethiopia.
Objectives: This study was conducted with three primary objectives: (1) Documenting medicinal plant use among the
Sidama people, (2) Understanding barriers to healthcare access, and (3) Identifying autochthonous disease
classifications.
Methods: Semi-structured interviews and guided field walks conducted through a translator were carried out with
knowledgeable informants chosen through snowball sampling. Barriers to medical resource access were analyzed
through community mapping with local elders. Medicinal plant specimens were collected, and specimens that were not
readily identifiable were brought to the National Herbarium of Addis Ababa University for confirmation.
Results: The Sidama use 47 medicinal plant species from 30 plant families. These species are used to treat 36
distinct diseases, including 4 locally acknowledged conditions lacking direct equivalents in Western biomedicine. The
category of illness with the highest proportion of plant treatments was gastrointestinal complaints (40%). Follow-up
research found 91.8% of the plant species mentioned to have documented pharmacological properties, 40.9% of which
matched the disease treated with the plant (excluding diseases with no Western equivalent). 5 species were found to
have previously identified potential herb-drug interactions.
Conclusion: More examples are needed of ways in which Western medical researchers and practitioners can collaborate
with indigenous peoples to build common knowledge about plants and their traditional medicinal uses. Preservation of
traditional medical knowledge is important in ensuring continued access to medicine by underserved communities,
preserving culture, reducing biodiversity loss, and promoting environmental health. This knowledge may help the
Sidama and other underserved groups achieve the health sovereignty they are entitled to by the United Nations
Declaration on the Rights of Indigenous Peoples. Our findings could be used in the by government and international
health care personnel to become aware of potential herb-drug interactions and culturally appropriate practice with the
Sidama. Plant usage data can further provide information for biodiversity conservation by drawing attention to possible
overexploitation of non-timber forest products, and mapping species distributions. The traditional knowledge recorded
will be made available to the community in a 40 page Amharic language handbook to ensure continuity in cultural
knowledge.
Funding: This project was generously funded by the UW-Madison Division of Vascular Surgery.

36
Permeabilityofcaninevocalfoldlaminapropria

JacobP.Meyer,AntonKvit,ErinDevine,JackJiang,

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JacobMeyer
(920)6061876
jpmeyer3@wisc.edu
MedicalStudent
OtolaryngologyHead&NeckSurgery
JackJiang,M.D.,Ph.D.
BasicScience
vocalfold,permeability,strain,laminapropria,Darcy'sLaw

Introduction:Theinertialmovementofinterstitialfluidinvocalfoldlaminapropriaduringandaftervibrationmay
exposethetissuetolocalizedstressconcentrationsthatleadtodamage.Thismovementispartiallygovernedbythe
connectivetissuespermeabilitytofluidflow.Thepermeabilityvaluesofthelaminapropriaarethereforenecessaryto
describevocalfoldfluiddynamicsthathaveconsequencesforcomputationalmodelsandpathologicalinterventions.
Methods:AformulationofDarcy'sLawwasappliedtocalculatethepermeabilityofexcisedcaninevocalfoldlamina
propriabymeansofameasuredfluidflow.Flowwasfoundusinganapparatuswithasalinecolumnofknownheightto
applyconstantdifferentialpressuresof5and20kPaacrosssamplesinthedeepsuperficialdirection.Additionally,fluid
flowthroughtissuesplacedunder10%,20%,and30%strainat20kPawasmeasuredtodeterminepermeabilityduring
straindeformation.
Results:Tissuesamplesunder5kPaand20kPawerefoundtohavemeanpermeabilityvalues(SD)of1.02x1013
m3s/kg(2.99x1014)and1.41x1013m3s/kg(3.98x1014),respectively.Permeabilitywasthenmeasuredforlamina
propriaplacedunder10%,20%,and30%strains,andresultedinsignificantincreasesforthe20%(p<0.05)and30%
(p<0.05)straingroups.
Conclusions:Thesepermeabilityresultscanbeusedforevaluatingfluidmovementincaninevocalfoldlaminapropria,
andsuggestsuchdynamicschangewithsubstantialtissuedeformation.

37
MacrophagePhenotypesinMurineAbdominalAorticAneurysm

StephanieMorgan,CalvinHarberg,QiweiWang,BoLiu

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

StephanieMorgan
(608)2157088
morgan@surgery.wisc.edu
PostdoctoralFellow
VascularSurgery
BoLiu
BasicScience
VascularDisease,Aneurysm,MouseModels,Macrophage,Inflammation

RATIONALE
Macrophageinfiltrationisawellestablishedcharacteristicofabdominalaorticaneurysm(AAA),however,relevanceof
macrophagesubsetshavenotbeenexplored.
OBJECTIVE
Wehypothesizedthatthemicroevironmentinaneurysmaltissuesskewsmacrophagedifferentiationtowardthe
inflammatory(M1)subtype.Strategiesthatenhancealternativelyactivated(M2)macrophagepopulationsmay
attenuatesmallaneurysmgrowth.
METHODSANDRESULTS
BothM1andM2macrophagesweredetectedinhumanandexperimentalaneurysm.Inmice,thenumberoftotal
macrophagesfoundinarteriestreatedwithelastaseorheatinactivatedelastasewascomparable.However,the
percentageofM1macrohpageswassignificantlyhigherinelastasetreatedtissues.WhenadministeredafterAAA
induction,theperoxisomeproliferationactivationreceptorgamma(PPAR)agonistRosiglitazone(RGZ)significantly
increasedtheM2population,decreasedM1/M2ratiointheaorticwall,andinhibitedaorticdilation(RGZ:82.813.2%
vs.DMSO:145.718.87%).
CONCLUSIONS
AnM1skewedmacrophagepopulationisassociatedwithaneurysmdevelopment.Alteringorinhibitingthispro
inflammatorybiasmaybeaneffectivetherapeuticstrategytoslowaneurysmprogresssion.

38
PTF1adosageinfluencespancreaticcellfatedecisionsinmouseembryonicstem
cells

GopikaNair,JoelAlvarez,JonOdorico

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

GopikaNair
(608)7724192
gopika.gn@gmail.com
GraduateStudent
Transplantation
JonOdorico
BasicScience
StemCells,Pancreas,Insulin,Diabetes,Endocrine

Pancreatictranscriptionfactor1a(PTF1a)isrequiredforpancreasspecificationandexocrinedifferentiation.Recent
observationsinzebrafishembryossuggestaroleforPTF1ainexocrinevsendocrinefatechoices.Partialfunctional
mutantsandmorpholinobasedknockdownexperimentsshowedthatlowdosesofPTF1afavoredanendocrine
commitmentwhilehighdosesinhibitedendocrinedifferentiationandpromotedexocrinefateinzebrafish.Suchstudies
arenoteasilyfeasibleinmouseembryos.Todetermineifasimilarmolecularcontroloverfatechoicesexistsinmouse
development,weutilizedatetracyclineinduciblePTF1amouseembryonicstemcellline(tetptf1amESCs)thatwas
establishedasarobustmodelofpancreaticdevelopmentinourpreviousstudies.OninducingdifferentdosesofPTF1a
byvaryingdoxycycline(dox)concentration,weobserveddifferencesintranscriptlevelsofmanypancreaticfactorsand
thenumbersofNGN3+cells,Insulin+cellsandAmylase+cells.LowdosesofPTF1agaverisemoreendocrineprogenitors
(NGN3+cells)andhadsignificantlyhighergeneexpressionofproendocrinemarkers(neurog3,nkx6.1)whencompared
toculturesthatreceivedhighdosesofdox.Lowdoseculturessubsequentlygeneratedmoreinsulin+cellsthanhigh
dosecultures.TheseculturesalsohadsignificantlyhigherCpeptidecontentasdeterminedbyacidethanolextraction
followedbyELISA.Incontrast,cultureswithhighPTF1alevelshadmoreexocrinegeneexpression(amy2,ela1,cpa1)and
amylase+cellsthancultureswithlowPTF1adoses.WealsoobservedanincreaseintranscriptlevelsofRBPJL,oneofthe
bindingpartnersofPTF1athatisimplicatedinexocrinecommitmentwithincreasingdosesofPTF1a,whileanother
partnerRBPJremainsmoreorlessconstant.ThisindicatesapotentialreplacementofRBPJbyRBPJL,andhenceafate
shift.TheseresultssuggestthatPTF1aexertsadosagedependantcontroloverendocrinevsexocrinecommitment
decisionsinmammaliansystemsaswell.

39
TheroleofendodermalapoptosisinDuodenalatresiaformation

PFNichol,KMZaremba,ALReeder,AKowalkowski

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

PeterNichol
(608)5126357
nichol@surgery.wisc.edu
Faculty
Administration
NicholLab
BasicScience
Fgfr2IIIb,mouse,duodenum,atresia,apoptosis

Purpose:WepreviouslyreportedthatformationofduodenalatresiainFgfr2IIIb/mouseembryosoccursinthe
absenceofaformationofanendodermalplug,contrarytoastandinghypothesisthatendodermalplugsareacauseof
intestinalatresia.Precedingthedefectthereisanincreaseinapoptosisintheregionwheretheatresiawillform(the
atreticprecursor).Becausetheincidenceofduodenalatresiaisonly3845%withinthesemutants,theFgfr2IIIbrec/rec
mousemodel(phenotypicallyequivalenttoFgfr2IIIb/mice)isidealfordeterminingtheroleofapoptosisinthe
formationofthisdefect.Wehypothesizedthatapoptosisisacriticaleventinduodenalatresiaformation.Therefore,
apoptosisshouldoccurintheatreticprecursorofroughlyhalfofthemutantembryos.Totestthis,weharvesteda
numberofthesemutantsandlittermatecontrolsandassessedapoptosisintheatreticprecursorregion.
Methods:Approvalofouranimalresearchprotocolwasobtainedfromourinstitution.Fgfr2IIIbrec/recembryoswere
generatedbybreedingFgfr2IIIbrec/+;HprtCre/+toFgfr2IIIbflox/floxmales.EmbryoswereharvestedatEmbryonicage
(E)9.5andE10.5,E18.5.GenotypeswereconfirmedbyPCR.Normal(Fgf2IIIb+/+)littermatesmicewereusedas
controls.Embryoswerefixedovernight,dehydrated,embeddedinparaffin,andsectionedat5mbeforebeingfloated
ontoslides.SectionswerethenprocessedusingTUNELstainingtovisualizeapoptosis.Comparativeanalyseswere
performedatmatchedembryonicstagesforFgfr2IIIbrec/rec,heterozygous,andwildtypeembryos.Sectionswere
photographedat200XmagnificationandreconstructedinAdobePhotoshop.
Results:Muchtooursurprise,weobservedendodermalapoptosisintheduodenalatreticprecursorofallmutants(5/5)
atE10.5,butitsintensitywasfarlessthanthatseeninthecolon(anareawhichwillalsofromanatresia).Thelumenof
theduodenalatreticprecursorappearedverynarrow.ThischangeinstructuresuggestedthatbyE10.5thisregionwas
inthepostapoptoticphaseofatresiaformation.
WhenrepeatingthisinE9.5embryos,weobservedmassiveapoptosisinallmutants(7/7)throughouttheentire
intestinalendoderm.TherewasalackofvariationintheanatomicdistributionofTUNELstainingbetweenmutants.
Embryosheterozygousforthemutationhadalower,butstillsignificant,increaseinapoptosisoverthelevelseenin
littermatecontrols.
Conclusions:Theseresultschallengethehypothesisthatendodermalapoptosisistheprimarycriticaleventinatresia
formation.WhileapoptosisisevidentintheatreticprecursorregionatE10.5,itisaglobalintestinalendodermeventat
E9.5withinthesemutants.Despitethisglobalphenomenon,duodenalatresiaratesdonotreach100%.Thenext
obviousstepwillbetoblockapoptosisinhomozygousmutantsandobservetheeffectsontheincidenceofatresiaand
severityoftissuelossinthismousemodel.

40
Disruptions in endoderm development can result in intestinal atresia formation:
time for a new hypothesis?
PF Nichol, AL Reeder, A Kowalkowski, CM Vezina, KM Zaremba, R Liebl

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Peter Nichol
(608) 512-6357
nichol@surgery.wisc.edu
Faculty
Administration
Nichol Lab
Basic Science
Fgfr2iiib, atresia, endoderm, mechanism, mouse

Purpose: Intestinal atresia is a well-recognized congenital defect whose cause remains unknown. Historically, proposed
mechanisms of atresia formation included disruptions in the mesoderm-derived vasculature (called vascular accidents)
and an occlusion of lumen, following excessive endoderm proliferation, leading to solid core of tissue. These
mechanisms were hypothesized prior to data derived from genetic animal models.
The Fibroblast growth factor receptor 2IIIb (Fgfr2IIIb) is a tyrosine kinase trans-membrane receptor. In mice, a
homozygous Fgfr2IIIb null mutation at conception results in colonic atresia. The formation of this defect is preceded by
an increase in endodermal apoptosis and a reduction in endodermal cell proliferation at Embryonic (E) day 10.5. The
location of these events suggests that a loss of Fgfr2IIIb receptor function within the endoderm is mechanistically central
for the formation of colonic atresias. However, at this time there is only one published piece of data on Fgfr2IIIb
expression in the mouse intestine at E10.5.
To test whether a tissue specific Fgfr2IIIb loss would result in intestinal atresias, we selectively mutated Fgfr2IIIb within
the endoderm utilizing the endoderm-specific Cre recombinase Shh EGFPCre to recombine Fgfr2IIIb floxed alleles.
Resulting embryos were assessed for colonic atresias, lineage analyses were carried out to visualize Cre expression
patterns, and in situ hybridization was used to confirm Fgfr2IIIb recombination.
Methods: Experiments utilized mouse lines on a C57B6 background. Female mice Fgfr2IIIb flox/flox or Fgfr2IIIb flox/flox;
26Rosa lacZ/LacZ were bred to Fgfr2IIIb rec/+; Shh cre/+ males. Embryos were harvested at E10.5, E11.5, E13.5, and
E18.5. Expression of Fgfr2IIIb was assessed by in situ hybridization in both controls and mutant embryos at E11.5 and
E13.5. Rates of cell proliferation and apoptosis were examined on E10.5 embryos by BrdU labeling and TUNEL staining,
respectively. Colonic atresia incidence was quantified on E18.5 embryos. Lineage analysis by LacZ staining was
performed on embryos between E10.5 and E13.5.
Results: LacZ staining revealed that Shh cre/+ activity was limited specifically and exclusively to the endoderm of the
intestine, with robust staining present in the small intestine and colon. In embryos homozygous for the conditional
Fgfr2IIIb allele, ShhCre activity eliminated expression of Fgfr2IIIb in the endoderm. This was demonstrated by an absence
of mRNA expression visualized using in situ hybridization.
The lack of endodermal Fgfr2IIIb expression was associated with a pan-colonic increase in apoptosis and a decrease in
cell proliferation. By E13.5, , an absence of the endoderm in the distal colon was observed . All Fgfr2IIIb rec/rec; Shh
cre/+ embryos developed a colonic atresia by E18.5, whereas littermate control embryos showed phenotypically normal
colons.
Conclusions: These results indicate that specific disruptions in endoderm development (such as loss of Fgfr2IIIb
expression) can cause intestinal atresias. The Fgfr2IIIb rec/rec mutation lead to substantial apoptosis and a significant
decrease in endodermal cell proliferation. Colonic atresias reliably followed this aberrant expression of Fgfr2IIIb in the
endoderm during early development. These data suggests that intestinal atresias can arise through a mechanism
altogether different from the previously suggested vascular accidents or solid core of tissue due to hyper-proliferation
of the endoderm.

41
Epidermalgrowthfactormediatedwoundrepairinhumanembryonicstemcell
derivedepithelialcells

LilianaPalencia,BS;AmritavaDas,MEng;SeanPalecek,PhD;SusanThibeault,PhD;CiaraLeydon,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

LilianaPalencia
(414)5316687
lpalencia@wisc.edu
MedicalStudent
OtolaryngologyHead&NeckSurgery
ThibeaultLab(SusanThibeaultandCiaraLeydon)
BasicScience
Proliferation,Epithelial,Wound,Repair,EGF

Vocalfoldinjuryduetoblunttrauma,iatrogenicinjuries,surgicalintubation,orlaryngealinfectionsandinflammation
canresultinthelossofthevibratoryfunctionofvocalfolds,leadingtodysphonia.Followinginjury,thefundamental
goalofwoundhealingisthereconstitutionoffunctionalvocalfoldtissue,includingtheepithelium.Themechanisms
underlyingreepithelialization,anearlyandnecessarypartofvocalfoldwoundrepair,arenotknown.Physical
inaccessibilityandethicalconstraintsprecludestudyofhumanvocalfoldsinvivo.Theaimofthisstudywastoexplore
howepidermalgrowthfactor(EGF)anditsreceptor,theepidermalgrowthfactorreceptor(EGFR),mediate
reepithelializationbycellproliferationinaninvitromodel.
Toassessthewoundrepairresponsetoinjury,wecreatedascratchwoundinathreedimensionalmodelofvocalfold
mucosaofhumanorigindevelopedinourlaboratory.Themodelconsistsofhumanembryonicstemcellderivedsimple
epithelialcellscoculturedwithprimaryvocalfoldfibroblasts.Rateandextentofwoundhealing,EGFRactivation,and
cellproliferationpostinjurywereanalyzedwithandwithoutapplicationofbothexogenousEGFandanEGFRinhibitor,
Gefitinib.Extentofwoundrepairwascalculatedasapercentageoftheoriginalwound.EGFRactivationwasvisualized
viaimmunocytochemistryusinganantityrosine1068antibody,whilecellproliferationwasvisualizedusinganantiki67
antibody.
Resultsindicatethatinvivovocalfoldepithelialwoundhealingcanbesuccessfullyreplicatedinourthreedimensional
modelofvocalfoldmucosa.IncreaseddensityandheterogeneousdistributionofKi67intheepitheliumwasobserved
postinjuryintheareasurroundingthescratchwound,indicativeofproliferationandreepithelializationinthevocalfold
mucosamodel.WoundrepairafterinjurywasstimulatedbyEGFbutdepressedwithconcurrentadditionofGefitinib.
TheseresultsindicatethatEGFstimulateswoundhealinginanEGFRdependentmanner.Thesefindingsprovideinsight
intohowEGFandEGFRmayregulatethewoundrepairresponseofstemcellderivedepithelialcellsinathree
dimensionalmodelofvocalfoldmucosa.Thisstudydemonstratesthatournovelmodelofwoundhealinginvocalfold
mucosacanbeusedsuccessfullytogaininsightintothemechanismsthatenableandregulateepithelialrepairfollowing
injury.

42
Regulation of Cardiac Fibroblast Collagen Synthesis by -Arrestins in Maladaptive
Ventricular Remodeling
Jinju Li, Jennifer L. Philip, Tiju Theccanat, Shahab A. Akhter

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Jennifer Philip
(773) 251-7737
jlphilip@uchicago.edu
Medical Student
Cardiothoracic Surgery
Shahab A. Akhter, MD
Basic Science
Heart Failure, Receptors, Signal Transduction, Myocardial Fibrosis, Fibroblasts

Introduction: Cardiac fibroblasts (CF) produce and degrade the myocardial extracellular matrix and are critical in
maladaptive ventricular remodeling that can result in heart failure (HF). -arrestins are important signaling molecules
involved in -adrenergic receptor (-AR) desensitization and have receptor-independent functions including activation of
downstream signaling pathways that may be involved in CF-mediated fibrosis. We hypothesize that -arrestins play an
important role in the regulation of adult human CF biology with regard to myofibroblast transformation, increased
collagen synthesis, and myocardial fibrosis which are important in the development of HF.
Methods: CF were isolated from normal and failing adult human left ventricles. -arrestin expression was manipulated
using adenoviral-mediated overexpression or siRNA-mediated knockdown. Smad2/3 expression was inhibited by siRNA
mediated knockdown. Collagen synthesis was determined by [3H]Proline incorporation. CF were treated with either
TGF-, -agonist Isoproterenol (Iso), or ERK-inhibitor PD 98059.
Results: -arrestin1 and 2 expression are upregulated 3.3 and 2.1-fold respectively in adult human CF isolated from
failing left ventricles [p<0.03] and -AR signaling is uncoupled with 80% decrease in Iso stimulated cAMP production
[p<0.001] and loss of -agonist-mediated inhibition of collagen synthesis which is characteristic of normal control CF
[4158157 vs. 169260 cmp/mg protein, HF vs. Control, p<0.001]. Knockdown of either -arrestin1 or 2 decreased
myofibroblast transformation as measured by 21% and 32% decreases in -SMA expression compared to scramble
control [p<0.01]. In addition, -arrestin1 or 2 knockdown, restored -AR signaling and -agonist inhibition of collagen
synthesis [2128200 & 1794105 vs. 284155 cmp/mg protein, p<0.01] and decreased collagen I expression over 30%
compared to scrambled control. In contrast, overexpression of -arrestins in normal CF led to a failing phenotype with
increased baseline collagen expression, impaired -AR signaling, and loss of -agonist-mediated inhibition of collagen
synthesis [7669203 & 6297341 vs. 354319 cmp/mg protein, p<0.01]. -arrestin knockdown in failing CF diminished
TGF- stimulated collagen synthesis and decreased collagen I and III expression over 35% [p<0.05] and also inhibited ERK
phosphorylation compared to scramble control [501 & 394 vs. 1051 relative densitometric units, p<0.03].
Overexpression of -arrestins in normal CF increased basal ERK1/2 and Smad2/3 phosphorylation 1.4-fold and 2.7-fold
respectively [p<0.03] and enhanced TGF--stimulated collagen synthesis vs. null adenovirus control [13150409 &
10730606 vs. 6344663 cmp/mg protein, p<0.02]. These increases in collagen synthesis were prevented by pretreatment with ERK-inhibitor PD 98059 or siRNA knockdown of Smad2/3.
Conclusions: In summary, our results demonstrate that -arrestins play an important role in the regulation of collagen
synthesis and degradation by CF in normal and failing human ventricles. Enhanced -arrestin signaling appears to be
deleterious in CF by promoting a pro-fibrotic phenotype via uncoupling of -AR signaling as well as potentiating ERK and
Smad signaling. Targeted inhibition of -arrestins in CF may represent a novel therapeutic strategy to prevent
maladaptive myocardial fibrosis and ventricular remodeling.

43
Regulation of Cellular Oxidative Stress and Cell Survival by G Protein-Coupled
Receptor Kinase-2
Tiju Theccanat, Jennifer L. Philip, Nicholas Ludmer, Jinju Li, Xianyao Xu, and Shahab A. Akhter

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Jennifer Philip
(773) 251-7737
jlphilip@uchicago.edu
Medical Student
Cardiothoracic Surgery
Shahab A. Akhter, MD
Basic Science
G protein-coupled receptor kinases, oxidative stress, NADPH oxidase, heart failure,
cardiac myocytes

Introduction: Cardiac myocyte oxidative stress and apoptosis are important mechanisms for the development of heart
failure (HF). Chronic HF is characterized by increased levels of circulating catecholamines to augment cardiac output.
Long-term stimulation of myocardial -adrenergic receptors (-ARs) is known to be deleterious in cardiac myocytes,
however, the potential mechanisms underlying increased cell death are unclear. We hypothesize that GRK2, a critical
regulator of myocardial -AR signaling and function, plays an important role in mediating cellular oxidative stress and
apoptotic cell death in response to -agonist stimulation.
Methods: Experiments were carried out in rat heart derived H9c2 cells. Total cellular oxidative stress was quantitated by
CellRox fluorescence and mitochondrial oxidative stress by MitoSox. GRK2, GRK2 inhibitor (GRK2ct), and -arrestin were
overexpressed using adenoviruses. NADPH oxidase-4 (Nox4) was inhibited by siRNA mediated knockdown. Osmotic
pumps that delivered either PBS, angiotensin II (AngII) or isoproterenol were implanted in 6 week old C57BL-6N males
mice for 10 days.
Results: Stimulation of H9c2 cells with a non-selective -agonist, isoproterenol (Iso) led to a 3-fold increase in oxidative
stress [p<0.001] and increased apoptosis demonstrated by 1.8-fold increase in 8-oxo-dg staining [p<0.05] and 1.6-fold
increase in caspase-3 activity [p<0.01]. There was also a 2.5-fold increase in Nox4 expression [p=0.01] but no change in
Nox2, the primary NADPH isoforms and major sources of ROS generation in cardiac myocytes. Iso stimulation led to a 3fold increase in GRK2 activity [p<0.05], similar to the myocardial GRK2 activity present in chronic HF. A 3-fold adenoviralmediated overexpression of GRK2 led to similar increases in ROS production and apoptosis as seen with Iso stimulation.
The increases in oxidative stress by either Iso stimulation or GRK2 overexpression were abolished by pre-treatment with
non-specific Nox inhibitor, apocynin, or siRNA knockdown of Nox4. Direct activation of adenylyl cyclase with forskolin,
did not lead to increased ROS production suggesting that -agonist-induced mitochondrial oxidative stress is likely
mediated by activation of GRK2 and increased Nox4 activity and not cAMP/PKA. Adenoviral-mediated expression of a
GRK2 inhibitor prevented ROS production and apoptosis in response to Iso stimulation. -arrestins are signaling proteins
that function downstream of GRK2 in -AR uncoupling. Adenoviral-mediated overexpression of -arrestins led to a 3.5fold increase in ROS production [p<0.03] as well as a nearly 4-fold increase in Nox4 expression [p<0.05]. Chronic agonist stimulation in mice lead to a 2-fold increase in Nox4 expression [p<0.05] and increased apoptosis as
demonstrated by 1.6-fold increase caspase 3 activity [p<0.02] and increased in TUNEL staining compared to PBS or AngII
treatment.
Conclusions: These data demonstrate that -agonist stimulation of cardiac myocytes leads to a robust upregulation of
Nox4, mitochondrial oxidative stress, and apoptotic cell death. Overexpression of GRK2 had similar whereas inhibition of
GRK2 activity abolished this -agonist stimulated increase in Nox4 expression and ROS production and also led to
decreased apoptotic signaling. This appears to be an additional novel mechanism for the deleterious effects of increased
myocardial GRK2 activity and provides further evidence for the therapeutic potential of inhibiting GRK2 to prevent the
development of HF.

44
-arrestins Regulate Mitochondrial Oxidative Stress and Collagen Synthesis in
Adult Human Cardiac Fibroblasts
Jennifer L. Philip, Jinju Li, Tiju Theccanat, Xianyao Xu, Shahab A. Akhter

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Jennifer Philip
(773) 251-7737
jlphilip@uchicago.edu
Medical Student
Cardiothoracic Surgery
Shahab A. Akhter, MD
Basic Science
NADPH Oxidases, Oxidative stress, Heart Failure, Myocardial Fibrosis, Cardiac
Fibroblasts

Introduction: Oxidative stress in cardiac fibroblasts (CF) promotes transformation to myofibroblasts and collagen
synthesis which can lead to myocardial fibrosis, a precursor to heart failure (HF). NADPH oxidase 4 (Nox4) is a major
source of cardiac reactive oxygen species (ROS), however, mechanisms of Nox4 regulation are unclear. -arrestins are
scaffold proteins which signal in G protein-dependent and independent manners including ERK activation. We
hypothesize that -arrestins can regulate oxidative stress in CF in a Nox4-dependent manner and increase collagen
synthesis in the setting of HF.
Methods: CF were isolated from normal and failing adult human left ventricles. Mitochondrial oxidative stress was
quantitated by MitoSox fluorescence. -arrestin expression was manipulated using adenoviral-mediated overexpression
or siRNA-mediated knockdown. Nox4 expression was inhibited by siRNA mediated knockdown. CF were treated with
either TGF-, Nox-inhibitor apocynin, or ERK-inhibitor PD 98059.
Results: Mitochondrial oxidative stress is significantly upregulated in failing CF versus normal controls [2.30.16 vs.
1.00.04 relative fluorescence units, p<0.001]. Nox4 expression is increased 3.5-fold in the failing CF relative to control
[p<0.05]. Treatment with the Nox-inhibitor, apocynin, decreased collagen synthesis in failing CF under basal conditions
[3891366 vs. 84891165 cmp/g protein, p<0.01] and following TGF- stimulation [5452532 vs. 112541525 cmp/g
protein, p=0.01]. Knockdown of Nox4 in failing CF resulted in a 42% decrease in mitochondrial oxidative stress [p<0.01]
as well as decreased collagen synthesis relative to scramble control [2436741 vs. 5681689 cmp/g protein, p<0.02]
and a 56% decrease in -SMA expression, a marker of myofibroblast differentiation, [p<0.02]. Similar results for
apocynin treatment and Nox4 knockdown were seen in control CF. CF -arrestin1 and 2 expression are increased over 4fold in HF versus control [p<0.05]. Knockdown of -arrestin1 or 2 in failing CF decreased ROS by 57% [p<0.04] and Nox4
expression by 60% [p<0.04]. Overexpression of -arrestin1 or 2 in normal CF increased both Nox4 expression and ROS
production over 2-fold [p<0.01]. These increases in oxidative stress were inhibited by pre-treatment with the Noxinhibitor, apocynin, giving evidence that -arrestin mediated increases in oxidative stress are Nox4 mediated. ERK
phosphorylation is increased 4.6-fold in failing versus normal CF [p<0.05]. Pre-treatment with the ERK inhibitor PD98059
led to a 74% decrease in basal and a 66% decrease in TGF- stimulated Nox4 expression in failing CF [p<0.01]. Treatment
with the ERK inhibitor decreased both basal and TGF- stimulated mitochondrial oxidative stress over 50% in the setting
of -arrestin1 or 2 overexpression in control CF [p<0.05].
Conclusions: Mitochondrial oxidative stress is significantly increased in CF isolated from failing adult human left
ventricles versus normal controls. Upregulation of Nox4 appears to be a primary mechanism for the increase in ROS
production, which stimulates collagen deposition. -arrestin expression is also upregulated in failing CF and appears to
play an important and novel role in the regulation of mitochondrial ROS production via Nox4. The specific mechanism
for this effect on Nox4 is potentially ERK mediated and requires further investigation. Inhibition of -arrestin signaling in
CF may inhibit pathological cardiac fibrosis and adverse remodeling in the setting of increased myocardial oxidative
stress.

45
The Role of GRK2 and -Arrestins in Maladaptive Post-Infarction Ventricular
Remodeling
Jennifer L. Philip, Jinju Li, Xianyao Xu, Shahab A. Akhter

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Jennifer Philip
(773) 251-7737
jlphilip@uchicago.edu
Medical Student
Cardiothoracic Surgery
Shahab A. Akhter, MD
Basic Science
G Protein-coupled receptor kinases, Cardiac Remodeling, Myocardial Infarction, Heart
Failure, Cardiac Fibroblasts

Introduction: Remote territory fibrosis following myocardial infarction (MI) is an important mechanism for the
development of heart failure (HF). G protein-coupled receptor kinase-2 (GRK2) plays a key role in this process through
uncoupling -adrenergic receptor (-AR) signaling in failing human cardiac fibroblasts (CF). -arrestins are involved in AR uncoupling and have receptor-independent functions including activation of downstream signaling pathways that
may be involved in CF-mediated fibrosis. This study investigates the role of GRK2 and -arrestin in regulating CF biology
in post-MI remodeling.
Methods: Adult Sprague-Dawley rats underwent left anterior descending coronary ligation to induce MI. Left ventricular
(LV) function was assessed by echocardiography. Myocardial fibrosis was quantitated by histologic staining. LV CF were
isolated and cultured. In cultured CF, -arrestin expression was inhibited by siRNA-mediated knockdown and GRK2
activity was inhibited using adenoviral overexpression of GRK2 inhibitor, GRK2ct (Ad-GRK2ct). GRK2 activity was
inhibited in vivo by intra-aortic injection of Ad-GRK2ct immediately following LAD ligation.
Results: There was a significant decline in LV function at 2 weeks post-MI which was present through 12 weeks
[Fractional shortening: 0.350.01 vs. 0.520.01, p<0.01]. Remote territory (non-infarct area) fibrosis increased by 2
weeks post-MI [61% vs. 21% fibrosis, p<0.01] progressing by 12 weeks to 12% fibrosis [p<0.01], consistent with
adverse remodeling. Collagen synthesis was significantly upregulated in isolated CF as early as 2 weeks post-MI
[3132115 vs. 1082130 cmp/mg protein, p<0.01] which persisted through 12 weeks post-MI [2735856 cmp/mg
protein, p=0.02]. -arrestin expression was increased 2-fold by 8 weeks post-MI and GRK2 activity was increased 1.4-fold
by 12 weeks post-MI. These were associated with a 42% decrease in intracellular cAMP [p<0.05] and loss of -AR agonist
stimulated inhibition of collagen synthesis characteristic of normal CF [39691058 vs. 70895 cmp/mg protein, p<0.01].
Knockdown of -arrestin in CF isolated 8 weeks post-MI decreased transformation to activated myofibroblasts as
measured by a 50% decrease in -SMA expression [p<0.04] and inhibited collagen synthesis compared to scramble
siRNA controls [1985400 vs. 343151 cmp/mg protein, p=0.01]. Adenoviral mediated overexpression of GRK2ct, an
inhibitor of GRK2, in CF isolated 12 week post-MI significantly decreased collagen expression and synthesis compared to
null adenovirus (Ad-Null) control [1928126 vs. 2611213 cmp/mg protein, p=0.02]. Intra-aortic injection of Ad-GRK2ct
immediately following LAD ligation significantly decreased post-MI cardiac dysfunction; as measured by improved
fractional shortening [0.420.01 vs. 0.300.02, p<0.01] and ejection fraction [721% vs. 572%, p<0.01] at 12 weeks
post-MI vs. Ad-Null controls. Ad-GRK2ct also led to significantly inhibition of myocardial fibrosis at 12 weeks post-MI in
both the infarct area [41% vs. 111% fibrosis, p<0.001] and the remote territory [41% vs. 121% fibrosis, p<0.001].
Conclusions: Remote territory fibrosis occurs by 2 weeks post-MI and increases over time. Uncoupling of -AR signaling
via increased GRK2 and -arrestin appears to be an important mechanism of increased myocardial fibrosis, as
intracellular cAMP is known to inhibit collagen synthesis. Targeted inhibition of -arrestin and/or GRK2 and restoration
of -AR signaling in CF may represent novel approaches to inhibiting pathological fibrosis and adverse remodeling postMI.

46
Andrographolideattenuatesexperimentalabdominalaorticaneurysmby
inhibitingproinflammatorycellsinfiltrationandcytokinesproduction

JunRen,ZhenjieLiu,QiweiWang,BoLiu

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JunRen
(847)8685856
jren9@wisc.edu
GraduateStudent
VascularSurgery
BoLiu
BasicScience
Abdominalaorticaneurysm,Andrographolide,Proinflammatorycells,Cytokines,NF
B

Background:
Abdominalaorticaneurysm(AAA),characterizedbyexuberantinflammation,isacommonvasculardiseaseassociated
withhighmortalityrate.ThereiscurrentlynoestablishedtherapytotreatAAAbyalteringtherateofaneurysmal
expansion.Andrographolide(Andro),amajorbioactivechemicalfoundintheplantofAndrographispaniculata,iswidely
usedinTraditionalChineseMedicine.Inseveraldiseasemodels,Androisfoundtoexhibitpotentantiinflammatory
propertybyinhibitingNFBactivity.ThepurposeofthisstudyistotesttheeffectofAndroondevelopmentand
progressionofAAA.
MethodsandResults:
AAAwasinducedinC57BL/6Jmicethroughtransientperfusionoftheabdominalaortawithelastase(0.45U/mL,5min).
Andro(5mg/kg/i.p.injection)orsolventwasadministratedtomicebeforeorafterAAAinduction..Theaorticdiameter
wasmeasuredbeforeelastaseperfusion(day0)andatharvest(day14).DailyadministrationofAndropriortoelatase
inductioncompletelypreventeddevelopmentofAAA.Inaddition,administrationofAndrotomicewithestablished
aneurysmsignificantlyattenuatedprogressionofAAA.ImmunohistochemistryanalysesrevealedthatAndrodecreased
monocytesandmacrophagesinfiltrationandrecruitmentofTcells.Cytokinesproduction(MCP1,CCL7,CXCL10,TNF
andIFN)andiNOSofaortictissuewereattenuatedbyAndrotreatmentcomparedwithcontrolasanalyzedbyqPCR.In
vitro,AndroblockedMCP1expressionupregulatedbyproteinkinaseCdelta(PKC),acriticalmediatorduring
aneurysmformation,inprimaryvascularsmoothmusclecells.
Conclusion:
TheseresultsindicatethatAndrosuppressesdevelopmentandprogressionofAAAthroughinhibitionof
proinflammatorycellsrecruitmentandcytokinesproduction.Andromayofferatherapeuticstrategytoslowdisease
progressioninpatientswithsmallaneurysm.

47
DevelopmentofaninvitroorganotypicwoundmodeltocharacterizeofNotch
expressioninhealingwounds

MadhuchhandaRoy,TaylorJJaraczewski,PriyaRPathak,TimothyWKing

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MadhuRoy
(608)2653749
miroy@wisc.edu
Scientist
Plastic&ReconstructiveSurgery
TimothyKing
BasicScience
Woundhealing,keratinocytes,Notch,JAG1,DLL1

Woundhealingaffectsmillionsofpeopleannually.Followinginjury,keratinocytesfromthewoundedgeproliferate,
migrate&differentiatetorecapitulatethe3Dstructureneededtoprovideabarrierfunction.Weareinterestedin
discoveringnovelstrategiestoenhancethewoundhealingprocess.Humankeratinocytes(NIKS)weregrownin
organotypicculturescomposedofhumandermalfibroblastsembeddedintypeIcollagen.Cultureswerekept
submergedfor5days,thenraisedtotheairinterface,&maintainedforanadditional15daysallowingformationof
fullystratifiedsquamousepithelia.Afullthicknesswoundwascreatedintheorganotypicculture&allowedtohealover
14days.Theorganotypicculturesexpressdifferentiationmarkers&Notchcomparabletohumantissueviaindirect
immunoflorescence(IIF).Notch1&2wereexpressedatthebasallayers,whileNotch3expressionextendedtocellsin
thegranularlayeroftheepidermis,indicatingapossibleroleofNotch1&2inproliferation&migrationofbasal
keratinocytes&Notch3indifferentiatingkeratinocytes.NotchligandJAG1wasexpressedthroughouttheepidermis,
JAG2localizedtothebasallayerwhileDLL1localizedtothesuprabasal&granularlayeragainindicatingadifferential
role.Ourhypothesisisfurthersupportedintheorganotypicwoundmodelwehavedeveloped.Theexpressionpatterns
oftheNotchisoforms&ligandsattheleadingedgeofthehealingwoundwereobservedthroughDay14.Whilethe
areaawayfromthewoundededgeresembledtheexpressionpatternsforNotch&itsligandtothatofstaticcultures,
wenoticedarobustexpressionofNotch&JAG1atthehyperproliferativewoundedge&alongtheleadingedge.DLL1
wasnotdetected.Weareusingthisnovelwoundhealingmodeltoinvestigatetheroleofnotchinwoundhealing.
Broaderapplicationsofthismodeltoinvestigatenewtherapeuticinterventionsforwoundhealingarealsobeing
pursued.

48
Effectsofageandradiationtreatmentonfunctionofextrinsictonguemuscles

JohnARussell,NadinePConnor

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JohnRussell
(608)2633839
russell@surgery.wisc.edu
GraduateStudent
OtolaryngologyHead&NeckSurgery
NadineConnor
BasicScience
radiation,aging,muscle,tongue,swallow

Purpose:Agingandheadandneckradiotherapyindependentlyimpactswallowingfunction,whichmaybedueinpartto
tonguemuscleweakness.Whenagingandradiotherapyinteract,itmaybeexpectedthattherewillbeevengreater
deviationsinfunction.However,therehavebeennoinvestigationsofhowagingandheadandneckcancerradiation
treatmentcombinetoimpactthemusclesofthetongue.
Methods:Twentyyoungadult(9months)andeighteenold(32months)Fischer344/BrownNorwayratsreceivedeither
externalbeamradiationtotheheadorwerenonradiationcontrols.Radiatedratswereexposedto2fractionsof13Gy
overtwoconsecutivedays.Tonguemusclecontractilepropertieselicitedbywholehypoglossalnervestimulationwere
examined12weeksfollowingradiationexposure.
Results:Radiationsignificantlyreducedtonguetetanicandtwitchforcesinboththeyoungadultandoldrats.There
weresignificantinteractioneffectsofageandradiationonmusclecontractiontimewitholdradiationratshavingthe
slowestcontractiontime.Inaddition,therewasaninteractioneffectofradiationandageonmuscledecaytimewiththe
oldradiationratshavingtheslowestdecaytime
Conclusion:Tongueforceisreducedfollowingradiationexposure.Tonguemusclecontractionspeedsarealsoaltered
followingradiationexposurewithgreatesteffectbeingonagedmuscle.Thus,tonguecontractionspeedmayalso
contributetoswallowingproblemsassociatedwithagedindividualsreceivingradiationtreatmentforheadandneck
cancer.

49
Endocrinespecificationandmaturationofhumanpluripotentstemcellderived
pancreaticprogenitorsaftertransplantationtoimmunodeficientmice

SaraDuttonSackett,XiangLi,XiaofangXu,GopikaNair,CoriOBrienandJonSOdorico

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SaraSackett
(608)2630628
sackett@surgery.wisc.edu
Scientist
Transplantation
JonSOdorico
BasicScience
humanembryonicstemcell,pancreaticprogenitors,transplantation,endocrine
specification,betacell

Objective:Diabetesisachronicmetabolicdisorderresultingfrominsufficientproductionofinsulinfrompancreatic
cells.Stemcellderivedlikecellscouldfunctionasasourceoftransplantabletissuewhendonortissueisnotavailable.
Thedifferentiationofcellsfromhumanembryonicstemcells(hESCs)isamultistepprocess,whichincludesthe
sequentialformationofprimitivestreak,definitiveendoderm,posteriorforegut,pancreaticepitheliumandendocrine
cells.CurrentESCdifferentiationprotocolsrequirealongperiodofdifferentiationinvivobeforeachievingfunctional
levelsofinsulinproduction,whichmaymakeitdifficulttotranslatethisworktotheclinic.Toaddressthis,weare
developingnovelapproachestoimprovetheinvitrodifferentiationprocessandaredeterminingifthesepancreatic
progenitorsarecapableofdifferentiatingintoahomogenouspopulationofmature,functionalbetacellsinvivoina
shortenedtimeperiod.Wehypothesizethatthedifferentiationblockinderivingfunctionalendocrinecellsfromhuman
pluripotentstemcellsinvitrocanbeovercomebymanipulatingtheprotocolandbytransplantingcellsatearlierstages
inthedifferentiationprocess.Thus,shorteningtheinvivomaturationanddifferentiationperiodrequiredtocure
diabeticanimals.
Methods:UndifferentiatedhESCsaretreatedwithourprotocolsalongwithseveralpublishedprotocolsascomparisons
toinducedifferentiationthroughadefinitiveendodermstagetopancreaticprogenitors.Inthesevariousprotocols,cells
areeithergrownonMatrigel,insuspension,oronTranswellplateswithMatrigel.AttheendofstageIIIandstageIV,25
x10^6cellsaretransplantedtothekidneysubcapsulecompartmentofimmunodeficientmiceforfurtherdifferentiation
andmaturation.Fasting,stimulatedandrandomfedCpeptidelevels,asameasureofsecretedinsulin,andblood
glucosevaluesareanalyzedat8,12,16,20,25,28and32weeksandmetabolicstudies(GTT,foodchallenge)are
performed.Controlmicearetransplantedwithhumanislets(10002000IEQ)orhumanfetalpancreasrudiments.
Results:Wehaveshownthatourdifferentiationprotocolresultsindifferentiationofcellstodefinitiveendodermand
thatthesecellsgoontofurtherdifferentiateintoPdx1+cellswhichalsoexpressFoxA2,HNF6,Sox9,HNF1bandNkx6.1.
Wefindthatwhilemanyofthecellsarepolyhormonalforinsulinandglucagonattheendofstage4,manyofthesecells
becomemonohormonalduringinvivodifferentiationasshownbyimmunofluorescentstaining,whereasinother
protocolstheyremainpolyhormonal.ELISAanalysesshowdetectablelevelsofCpeptide(>120pMby20weeks).GTT
showimprovedglucoseclearancebeginningasearlyas12weeksandincreasedCpeptidefollowingglucosestimulation.
Conclusions:Wehavedevelopedauniqueprotocolforpromotingthedifferentiationofpancreaticprogenitorcells,
whichhavefunctionalendocrinepotentialaftertransplantationintomice.Althoughtheprecisemechanismofthein
vivomaturationremainsundefined,itisclearthatsomeinvitroderivedcellpopulationshavesignificantlygreater
potentialfordifferentiatingintoinsulinsecretingbetacellsthanothers.

50
PollutionEnhancesTh17DifferentiationThroughtheArylHydrocarbonReceptor

SamanthaKnopp,JohnFechner,WalkerJulliardMD,JoshuaMezrichMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

KnoppSamantha
(608)2659147
slknopp@wisc.edu
MedicalStudent
Transplantation
JoshuaMezrich
BasicScience
Pollution,Th17,IL17,IL22,ArylHydrocarbonReceptor

Introduction:Thearylhydrocarbonreceptor(AHR)isacytosolicreceptorwiththeabilitytoinfluenceTcell
differentiationtoaTregulatoryorThelper17(Th17)response.Itisknownthatairpollutionisariskfactorforasthma
andotherairwaydiseases,andIL17associatedwithincreasedTh17differentiationcanexacerbateairwaydisease.
Polycyclicaromatichydrocarbons(PAHs),whichareligandsoftheAHRandpresentinairpollutioninvarying
compositionandconcentration,areapossibleexacerbatingfactorofairwaydisease.Thisstudyexaminestheeffectsof
varioussourcesofPAHsonTcelldifferentiationtoaTh17effectorresponse.Wehypothesizethatairpollutionsamples
willincreaseTh17differentiationviatheAHR,andthatmorehighlypollutedenvironmentalairsampleswillhavemore
significanteffectsonTcelldifferentiationandAHRactivationinvitrothanwillsampleswithlowerlevelsofpollution.
Methods:WeusedmagneticbeadseparationtoisolatenaveCD4+CD62L+TcellsfromthespleensofwildtypeC57BL/6
(B6)andAHR/mice.ThesenaveTcellswerethenplacedincultureforfourdaysunderTh17conditions(antibodiesto
CD3andCD28,aswellasIL6andtransforminggrowthfactor;antibodiestointerferonandIL4wereincludedin
someexperiments)andexposedtovaryingconcentrationsofurbandustparticles(UDP)orenvironmentalairsamples
obtainedfromtheWisconsinStateLaboratoryofHygiene.Weusedintracellularcytokinestainingfollowedbyflow
cytometrytodeterminecelldifferentiation.LevelsofcytokinesinculturesupernatantwereassessedusingELISA.
Results:MurineB6naveTcellsculturedinthepresenceofUDPunderTh17conditionsresultedinenhancedTh17
differentiation,asshownbyELISAandintracellularcytokinestaining.MurineAHR/naveTcellculturesshowedno
significantincreaseinIL17atanyconcentrationofUDP.FlowcytometricanalysisofmurineB6naveTcellscultured
underTh17conditionsalsoshowedthattherewasincreasedexpressionoftheAHRinIL17positivecellsascomparedto
IL17negativecells.IntracellularcytokinestainingandELISAshowedincreasedIL17expressionwithadditionofdiesel
environmentalairsampletomurineB6naveTcellsculturedunderTh17conditions.
Conclusion:ThesepreliminaryresultsshowthatTh17differentiationinvitroisenhancedwithadditionofUDPordiesel
environmentalairsampletocultureconditionsandUDPmediatedTh17differentiationinvitroisAHRdependent,
suggestingamechanismofimmunemodulationbyairpollutantswithpossibleclinicalimplicationsforairwaydisease.

51
Roleofbidirectionalregulationinskinallograftsurvival

MiwaSatomi

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MiwaSatomi
(608)2632485
satomi@surgery.wisc.edu
Faculty
Transplantation
WilliamJBurlingham
BasicScience
bidirectionalregulation,skinallograft,antiCD40Lmonoclonalantibody,donor
specificsplenocytetransfusion,transvivodelayedtypehypersensitivity(tvDTH)
assay

Background;Wepreviouslyfoundthatthepretransplantimmunestatusoforgandonoraswellasorganrecipient
stronglyinfluenceallograftoutcomeinkidney.Wefoundthathaploidenticalpatientdonorpairswithunidirectionalor
nonregulationstatushadsignificantlyincreasedacuterejection,antidonorantibodyandgraftloss.Incontrast,
bidirectionalregulationkeptlongtermgraftsurvivalwithfewrejections.Thesurvivalofskingraftisalsorelatedtothe
donorrecipientimmunologicalinteraction.Donorpassengerleukocytes,includingskinTcellsandDCmayinfluence
graftoutcome.
Methods;WechosetousetoleranceinducedbyantiCD40Lmonoclonalantibody(mAb)anddonorspecificsplenocyte
transfusion(DST)andrecipientspecifictransfusion(RST)+antiCD40Lasameanstotestthefollowinghypothesis:Pre
transplant,bidirectionalregulationprolongsmouseskinallograftsurvival.B6miceskinallograftswereplacedonthe
backofCBAmice.Wemade3types(nave,tolerance,sensitized)ofB6andCBAmicerespectively.Wecheckedthe
immunestatusbyusingtransvivodelayedtypehypersensitivity(tvDTH)assay.Weexaminedskintransplantof9
patterns,andscoredtheskingraftconditioneveryday.
Results;Skingraftexperimentswerestillinprogressatthetimeofwriting.

52
The Association of CARM1 Isoform mRNA Expression with Patient Clinical
Characteristics and Molecular Subtypes in Breast Cancer Tumors, and the
Differential Localization of CARM1 Protein Isoforms
David Shlensky BS, Wei Xu PhD, Lee Wilke MD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

David Shlensky
(414) 801-7821
dshlensky@wisc.edu
Medical Student
General Surgery
Dr. Lee Wilke
Basic Science
Breast Cancer, Molecular Expression, Clinical Characteristics, Protein Localization,
Oncology

Introduction: Breast cancers are classified by differences in several receptors, including the estrogen (ER) receptor.
Molecular differences between normal and malignant cells are targetable for treatment, yet alternative mRNA splicing
events that contribute to malignant transitions may also offer targets for therapy. Coactivator-associated arginine
methyltransferase 1 (CARM1) is a coactivator for ER and cancer-relevant transcription factors, and can methylate
diverse cellular targets including histones. Most full-length CARM1 (CARM1FL) is automethylated at R551 in Exon 15, but
the alternatively spliced CARM1E15 lacks this site. CARM1FL and CARM1E15 function differently in transcriptional
regulation, protein methylation, and mediation of pre-mRNA splicing in cellular models. Recently, CARM1E15 was
found to be dominant in ER- human breast cancer cell lines, and may therefore be an oncogenic form. We hypothesized
that the RNA ratio of CARM1E15 to CARM1FL and the absolute level of the short isoform would be higher in ERcancers compared to ER+ cancers and benign samples. We also hypothesized that each protein isoform might have a
different subcellular localization that contributes to their functionality
Methods: With IRB approval, twelve flash frozen human breast cancer samples and three benign fibroadenoma tumors
were processed to quantify each mRNA transcript by qRT-PCR. Further studies to localize CARM1 protein isoforms were
peformed on MB-MDA-231 ER- breast cancer cells and BG-1 ER+ ovarian cancer cells. Immunofluorescent labeling was
completed using E15 antibody, which recognizes CARM1FL, and E16 antibody, which recognizes both isoforms. DAPI and
phalloidin were used to stain nuclear and cytoplasmic compartments. Data were analyzed by ANOVA and relationships
were determined between continuous variables with bivariate correlations.
Results: Human breast cancers did not have significantly higher expression of CARM1FL or CARM1E15 (p=0.21 and
0.31) compared to fibroadenomas. Triple-negative tumors, ER+/PR+ tumors, and fibroadenomas did not differ in
CARM1FL or CARM1E15 expression (p=0.39 and 0.30). Tumors from patients with lymph node involvement did not
have higher expression of CARM1FL (p=0.94) nor CARM1E15 (p=0.67). In MDA-MB-231, E15 localized chiefly to the
nucleus versus the cytoplasm, with a significantly higher pixel intensity of 11.73 and 0.44, respectively (p = 0.001). E16
also localized to nucleus and cytoplasm, with a significantly higher intensity of 28.16 and 5.92, respectively (p<0.005).
Finally, the percentage of cytoplasmic to total fluorescence was higher for total CARM1 (15.75 %) than CARM1FL (3.39%;
p < 0.005). BG-1 cells displayed a similar CARM1 localization pattern: the cytoplasmic to total fluorescence was higher
for total CARM1 (16.73%) than CARM1FL alone (3.8%; p <0.005).
Conclusions: In breast tumors, CARM1 isoform mRNA levels are not associated with patient clinical characteristics or
molecular subtypes. Recent studies, however, have shown that high CARM1 protein expression correlates with poor
prognosis. Our results imply that CARM1FL is likely nuclear, whereas CARM1E15 is cytoplasmic and nuclear. Studies are
needed to validate this localization difference in human breast cancer samples.

53
NOTCH3expressionsuppressesgrowthandinducescellcyclearrestinfollicular
thyroidcancer

YashR.Somnay,BS,XiaoMinYu,MD,PhD,HerbertChen,MD,FACS

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

YashSomnay
(248)9530631
somnay@wisc.edu
MedicalStudent
GeneralSurgery
HerbertChen
BasicScience
ThyroidCancer,Notch,Metastasis,CellCycle,Apoptosis

INTRODUCTION:TheNotchproteinfamily(Notch14)regulatecellproliferation,survival,andstemandprogenitorcell
hierarchiesinbothnormalandcanceroustissues.Notably,Notch3proteinisdistinctinstructurefromtheconserved
Notchfamilyhomology,andhasnarrowerexpressionspecificitytoproliferatingneuroepithelium.Furthermore,Notch3
hasbeendemonstratedtoplayanoncogenicroleinhumanovarianandbreastcancersaswellasmurineTcell
differentiationandleukemogenesis.However,theroleofNotch3inthyroidcancerhasyettobedescribed.Inthisstudy,
wesoughttocharacterizetheroleofNotch3activityinfollicularthyroidcancerproliferationandphenotype.
METHODS:FTC236cellsweretransfectedwithplasmidpRevTetOn(Clontech)containingtheTetresponsive
transcriptionalactivatorusingLipofectamine2000(Invitrogen).FTCTetoncloneswerescreenedfordoxycycline
dependentinducibilityofthereportergeneluciferasebytransienttransfectionofthepRevTRELucvectorintoFTCTet
onclones.CloneswereselectedandusedfortransfectionwiththeresponseplasmidcontainingtheNOTCH3gene
(pRevTRENOTCH3)ortheemptyvector(pRevTRE).Notch3expressionfollowingdoxycyclinetreatmentwasdetermined
usingWesternanalysisforNotch3,Hes1,andcmyc.Cellgrowthwasassessedusingamethyltetrazoliumbromide
(MTT)rapidcolorimetricassay.Expressionlevelsofvariouscellcyclearrestmarkerswerealsodetermined.Finally,flow
cytometryanalysisusingpropidiumiodidestainingwasperformed.
RESULTS:TreatmentofFTCNOTCH3cellswithdoxycyclinedosedependentlyinducedNOTCH3expressionwith
correspondingincreasesinHes1andcmyc,twodownstreamNotchfamilytargets.Importantly,Notch3wasabsentin
uninducedFTC236cells.CellsstablytransfectedwithanemptyvectorexperiencednoeffectonNotch3expressionor
cellviability.Notch3overexpressionledtoadosedependentdecreaseinFTC236cellgrowthover4days.Increased
expressionofNotch3ledtoanincreaseinp21,p27,andareductionincyclinfamilyproteinsanddependentkinases.
FlowcytometryanalysisrevealedanincreaseinG1/SarrestedcellswithincreasingNOTCH3induction.
CONCLUSIONS:Notch3activationinfollicularthyroidcancercellsdosedependentlysuppressedcellproliferationviacell
cyclearrest.ThesearethefirstfindingstosuggestthatNotch3activationmaybeapotentialtherapeutictargetof
activationratherthansuppressionforaggressivefollicularthyroidcancersininoperableandmedicallyintractablecases.

54
Growthfactordeliveryfrommineralcoateddevices

DarilisSuarezGonzalez,YanLu,AlishaDiggs,BrettNemke,ScottHollister,MarkMarkel,WilliamL.Murphy

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DarilisSuarezGonzalez
(608)2655933
suarezgonzalez@surgery.wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
WilliamMurphy
BasicScience
growthfactor,tissueengineering,sustainedrelease,mineralcoatings,medicaldevices

Theobjectiveofthisworkistopresentasimplemechanismforthereleaseofgrowthfactorsfrommineralcoatings
formedondifferentdevices,specificallymicroparticlesandscaffoldmaterials.Itisknownthatgrowthfactorsarekey
playersduringtissueregeneration;however,thereremainsaneedforsimple,clinicallyrelevantdeliveryapproaches.
Wehypothesizedthatmineralcoatingscouldbeusedasaplatformforcontrolledincorporationandreleaseofsingleor
multiplegrowthfactors.Inaddition,wehypothesizedthatthecharacteristicsofnanostructuredmineralcoatingswould
allowforoptimizedproteinstability,anduniquelyhighbiologicalactivityofreleasedgrowthfactorsrelativetoexisting
growthfactordeliveryapproaches.Betatricalciumphosphate(TCP)scaffoldsandmicroparticlesweremineralcoated
usingabiomimeticapproach.Mineralcoateddevicesweredippedinagrowthfactorsolution.Wecharacterizedbinding,
release,andbiologicaleffectsofaseriesofgrowthfactors,includingrecombinanthumanvascularendothelialgrowth
factor(rhVEGF),amodular,mineralbindingversionofbonemorphogeneticprotein2(mBMP2),andbasicfibroblast
growthfactor(bFGF).Ourresultsdemonstratethatwecouldformmineralcoatingsonvariousdevices.Inaddition,we
showedsustainedreleaseofmBMP2,andrhVEGF,andbFGF.Thebiologicalactivityofthegrowthfactorsreleasedfrom
differentdeviceswascharacterizedinvitroandinvivo.ScaffoldsreleasingrhVEGFwereimplantedinanintramuscular
sheepmodelandevaluatedat2weeksforbloodvesselingrowth.Basedonhistologicalevaluation,therewasan
increaseinbloodvesselingrowthinadosedependentmanneringroupsreleasingrhVEGF.Microparticlesreleasing
bFGFwereincorporatedinamodifiedhyaluronicacidhydrogelwithhumanvocalfoldfibroblasts.Sustainedreleaseof
bFGFresultedinincreasedmetabolicactivityofhVFFsuggestinganincreaseinproliferation.Alltogether,thesestudies
demonstratethesecoatingshaveauniqueabilitytomaintainbiologicalactivityofgrowthfactors.Inaddition,this
approachdemonstratesamodulardesignapproach,inwhichacontrollablebiologicscarrierisintegratedintoa
structuraldevice(i.e.ascaffold)asathinsurfacecoating,withoutnegativelyimpactingthebulkphysicalpropertiesof
thedevice.

55
RemovalofCD39+TcellsUncoversaPopulationofColVSpecific,CD39
ResponderTcellsinNormal,HealthyIndividuals.

JeremyASullivan,EwaJankowskaGan,SubramanyaHedge,MattPestrakandWilliamJBurlingham

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JeremySullivan
(608)2634875
SULLIVAN@SURGERY.WISC.EDU
Scientist
Transplantation
Burlingham
BasicScience
autoimmunity,Th17cells,CD39,selfantigen,CD4

WeandothershavereportedthatautoimmunitytotheselfantigenCollagenV(ColV)afterlungtransplantation,is
stronglycorrelatedwiththedevelopmentofBronchiolitisObliteransSyndrome,theclinicalmanifestationofobliterative
bronchiolitisandthemaincauseofmortalityandgraftloss.Inaddition,wehaveshownthatColVautoimmunityisa
Th17dependentprocess,asantibodytoIL17andCD4,butnottoIFNorCD8,blockedtheresponse.Innormal
individuals,approximately50%ofcirculatingTregsexpresstheextracellularATPaseCD39.Thissubsetisthoughttoplay
akeyroleinmaintainingselftolerancebysuppressingTh17associatedautoimmunity.Withthisinmind,wesetoutto
investigateifColVspecificTh17cellsarepresentinnormal,healthyindividuals.WecollectedPBMCsforuseintrans
vivoDTHandintracellularcytokinestainingassays.Results:ColVreactivityinwholePBMCofhealthysubjectswaslow,
butsignificantlyhigherthantheresponsetoCollagentypeI.WefoundthattheremovalofCD39+Tcellsfromnormal
PBMCrevealedstrongColVreactivity.Inaddition,theremovalofCD39+TcellsfromTcellmonocytecocultures
uncoveredaColVinducedproductionofIL17,albeitatlowerlevels/CD4TcellascomparedtoColVsensitizedlung
transplantrecipients.Furthermore,theTh17naturalresponseinCD39Tcellswasalsofoundtooccurselectivelyin
responsetoktubulin,HSP65andvimentin,butnotinresponsetocardiacmyosin,CollagentypeIV,myelinbasic
proteinorprostaticacidphosphatase.OurdatastronglyimpliesthatCD39+TregsrestrainColVautoimmunityinnormal,
healthyindividuals.ThenaturalColVreactiveTh17likecellsmayrepresentanovelclassofautoreactiveTcellthatis
releasedfromCD39+Tregcontrolafterlungtransplant,compromisinggraftsurvival.

56
Hyperglycemia suppressed abdominal aortic aneurysm through inhibition of
osteoclastogenesis
Teruyoshi Tanaka, Koji Kurosawa, Dai Yamanouchi

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Teruyoshi Tanaka
(608) 556-4358
tanaka@surgery.wisc.edu
Postdoctoral Fellow
Vascular Surgery
Dr. Yamanouchi Lab.
Basic Science
abdominal aortic aneurysm, diabetes mellitus, osteoclastogenesis, hyperglycemia,
osteoclast

Background
Abdominal aortic aneurysm (AAA) is the 15th leading cause of death in men over the age of 55. Diabetes mellitus (DM)
is also a significant medical condition which is listed as the 7th leading cause of death in adult age over 65. Although AAA
shares most of the risk factors with other cardiovascular disease, many large-scale clinical data analyses reported that
DM is an independent negative risk factor for AAA.
Increasing evidence suggests that arterial calcification may be the result of highly organized processes resembling those
seen in osteogenesis in bone, which relies upon a delicate balance between mineral deposition and resorption by
osteoblasts and osteoclasts, respectively. We have previously shown the involvement of osteoclast-like cell in AAA
through pathological absorption of vascular calcification. Since DM are well known to be associated with severe vascular
calcification, we hypothesized that DM induced more vascular calcification but less aneurysmal degeneration through
the inhibition of osteoclastogenesis.
Materials and Methods
The effect of hyperglycemia on osteoclastogenesis was tested by in vitro osteoclastogenesis assay. We stimulate
RAW264.7 cell with receptor activator of nuclear factor-B ligand (RANKL) for 5 days to induce osteoclast confirmed by
the number of enzymatic tartrate-resistant acid phosphatase (TRAP) staining positive multinucleated cells. Cells were
treated under normoglycemic (5.5 mM glucose), normoglycemic with high osmotic pressure (2.5 mM mannitol), and
hyperglycemic condition (30.5 mM glucose). Secreted matrix metalloproteinase-9 (MMP-9) levels in the media and
nuclear factor of activated T-cells (NFAT) c1 expression from cell lysate were detected by Western blotting.
To test the effect of hyperglycemia on aneurysm in vivo, we introduced type I diabetes in C57Bl6 mice by 150 mg/ml of
streptozotocin (STZ) or control vehicle. Calcium Phosphate (CaPO4) induced aneurysm was then surgically induced and
the maximum diameter of the carotid artery was measured before and after 4 weeks from surgery.
Protein expression standardized by alpha-tubulin was quantified by densitometry. Fold increase of the maximum
diameter of the artery was used for in vivo aneurysm quantification. Data are presented as means standard deviation
(SD). One-way analysis of variance followed by post-hoc Tukeys or students t-test was used to examine differences
between the each group. P<0.05 was considered as significant.
Results
Osteoclastogenesis was significantly suppressed under hyperglycemic condition (167.023.4 vs 25.21.8 cells/well,
normoglycemic vs hyperglycemic, P<0.01) in vitro. MMP-9 and NFATc1 expression also was significantly suppressed
(5.50.3 vs 3.10.1, and 3.60.2 vs 1.20.1, P<0.05). CaPO4 induced aneurysm was significantly suppressed in STZ
induced DM mice compared to control mice (1.80.2 vs 1.50.2, control vs DM, P<0.05). Immunohistochemical analysis
showed suppression of osteoclastogenesis in diabetic mice.
Conclusion
In vitro osteoclastogenesis assay showed inhibitory effect of hyperglycemia on osteoclastogenesis. We have also shown
the inhibitory effect of DM on mouse aneurysm with suppression of osteoclastogenesis in the arterial wall. Taken
together, our results indicated the inhibitory effect of DM on AAA through the inhibition of osteoclastogenesis. Further
research to elucidate underlying mechanisms is warranted and could lead to the novel therapeutic approach to AAA.

57
CombinationeffectorandmemoryTcellimmunotherapy;anovelapproachfor
optimalanticancerimmunity

AndrewTatar,SiddharthaSen,JustinMeyers,PrakrithiSrinand,AmandaContreras,DavidMahvi,CliffordCho

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AndrewTatar
(608)2631401
tatar@surgery.wisc.edu
Other
GeneralSurgery
Dr.CliffordCho
BasicScience
Melanoma,Immunotherapy,Tcells,murine,adoptivecelltransfer

Background.Severalnewimmunotherapiesarecurrentlybeinginvestigatedinthetreatmentofmelanoma.One
experimentalstrategyutilizesadoptivecelltransfer(ACT)ofmelanomaspecificCD8+cytotoxicTlymphocytes(CTL)
harvestedfromtumorinfiltratinglymphocytes(TIL).WhileautologoustumorreactiveeffectorTcells(TEFF)recognize
andcleartumorcellsinvitro,previousworkfromourlaboratoryhasshownthatTEFFarehighlysusceptibletoapoptotic
celldeathinthepresenceofmelanoma.Incontrast,wehavefoundthatmemoryTcells(TMEM)areresistantto
melanomainducedcelldeath;moreover,wehaveshownthatTMEMbasedACTresultsinmoredurabletumor
suppressionthantraditionalTEFFbasedACT.WetestedthehypothesisthatcombinationACTusingbothTEFFand
TMEMwouldhaveadditiveeffectsonmelanomacomparedwithTEFForTMEMbasedACTalone.
Methods.WeusedamurinemodelofmelanomaACTpreviouslyestablishedinourlaboratory.Immunocompetent
C57BL/6micewereinoculatedviasubcutaneousinjectionswithB16F10melanomacellstransfectedtoexpressverylow
levelsofthelymphocyticchoriomeningitisvirus(LCMV)peptideGP33(B16GP33).CD8+GP33specificTcellswere
harvestedfromthespleensofC57BL/6mice8days(TEFF)or>50days(TMEM)afterLCMVinfection.Sevendaysafter
tumorinoculation,miceweretreatedwithnoACT(control),ACTusingCD8+GP33specificTEFFcells,ACTusingCD8+
GP33specificTMEMcells,orACTusingCD8+GP33specificTEFFandCD8+GP33specificTMEMcellsincombination.At
varioustimepointsaftertumorinoculation,localandsystemicTcellresponsestomelanomawereassessedbyanalyzing
TILandsplenocytes,respectively,usingflowcytometry;tumorvolumesweremeasuredevery3days.
Results.Consistentwithourpreviousfindings,TMEMACTresultedinmoredurablesuppressionofB16GP33melanoma
growththanTEFFACT.TumorgrowthwasmaximallyinhibitedinmicetreatedwithcombinationTEFF+TMEMACT.
TMEMACTinducedlargerpopulationsofCD8+TILthanTEFFACT.However,circulatingpopulationsofmelanoma
specificCD8+TcellswereonlyobservedinmicetreatedwithTEFF+TMEMACT.
Conclusions.OurdataidentifyanovelapproachtoimprovingTcellbasedmelanomaimmunotherapy.TMEMACT
inducesastrongerlocalimmuneresponsethanTEFFACT.TheabilityofcombinationACTusingbothTEFFandTMEMto
inducebothalocalandsystemicantitumorimmuneresponsemaybethebasisforitssuperiorefficacy.Future
experimentswillfocusonunderstandingthemechanismsthatmayexplaintheseresults.

58
Induction of the Nrf2-Antioxidant Response Element (ARE) Pathway in Cultured
Hepatic Stellate Cells (HSCs) Leads to Increased Markers of HSC Activation
Lindsay Taylor, Lung-Yi Lee, Li Zhang, Jeffrey A. Johnson, David Foley

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Lindsay Taylor
(414) 364-0623
lntaylor@wisc.edu
Medical Student
Transplantation
Dr. David Foley
Basic Science
Nrf2, Hepatic Stellate Cells, TGFb1, fibrosis, in vitro

INTRODUCTION
Oxidative stress is a major stimulus of hepatic stellate cell (HSC) activation, a process that leads to the development of
hepatic fibrosis. The Nrf2-ARE pathway is an endogenous antioxidant pathway that protects cells from oxidative
damage. The relationship between the Nrf2-ARE pathway and HSC activation remains poorly understood. The purpose
of this in vitro study was to determine whether induction of Nrf2-ARE pathway in HSCs could decrease both HSC
activation and the expression of fibrogenic genes.
METHODS
Primary murine HSCs were isolated from Nrf2 wild-type (WT) and Nrf2 knockout (KO) mice using both in situ and ex vivo
pronase/collagenase digestion followed by density centrifugation in Percoll. Transforming growth factor beta 1 (TGF-1)
was used to activate HSCs. Nrf2 WT and KO HSCs were grown to confluence in 6-well plates and then treated with or
without TGF-1 (3.0ng/mL). After 24 h, RNA and protein were isolated from cultured cells. Quantitative RT-PCR was used
to quantify the mRNA abundance of Nrf2-dependent genes and fibrogenic genes that are recognized as markers of HSC
activation (SMA, COL1A1, TIMP1, MMP2, and MMP9). Western blot was performed to determine SMA protein
expression. To evaluate the effect of pharmacologic induction of Nrf2, WT HSCs were treated with vehicle or tertButylhydroquinone (tBHQ), a known inducer of the Nrf2-ARE pathway, for 24 hours prior to TGF-1 treatment. After 24
h, RNA and protein were isolated and the same end points were evaluated. Statistical analysis was performed with one
way-ANOVA followed by Fishers LSD post hoc test, or Students t-test as appropriate. P<0.05 indicates statistical
significance.
RESULTS
Treatment of Nrf2 WT and KO HSCs with TGF- 1 resulted in significant increases in gene expression of SMA, TIMP1,
MMP2, and MMP9 compared to vehicle-treated cells. When compared to Nrf2 WT cells, Nrf2 KO cells demonstrated
significantly lower mRNA abundance of SMA, COL1A1, and TIMP1, but increased MMP2 and MMP9 gene expression
both at baseline and with TGF-1 treatment. Western blot demonstrated decreased SMA protein in Nrf2 KO HSCs as
compared to the WT cells. Treatment of WT HSCs with tBHQ resulted in significant increases in the expression of Nrf2dependent genes. Treatment of HSCs with tBHQ followed by TGF-1 resulted in significantly higher expression of SMA,
MMP2, and MMP9 as compared to HSCs treated with TGF-1 in the absence of tBHQ.
CONCLUSIONS
Treatment of HSCs in vitro with TGF-1 leads to increased HSC activation and increased expression of fibrogenic genes:
SMA, TIMP1, MMP2 and MMP9. Contrary to our hypothesis, the absence of Nrf2 in HSCs leads to decreases in markers
of HSC activation, and pharmacologic induction of Nrf2 in HSCs with tBHQ leads to increases in HSC activation at baseline
and in response to TGF-1. Further studies are needed to clarify the role of the Nrf2-mediated antioxidant response in
HSC activation.

59
MorphologicalChangesinFibroblastsinducedbyPBMCsfromPatientswithType
IIDiabeticNephropathy

TrinhPham,AustinEckhoff,DebHullett,HansSollinger,BiancaTomasiniJohansson

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

BiancaTomasiniJohansson
(608)2631366
tomasini@surgery.wisc.edu
Scientist
Transplantation
Sollinger
BasicScience
diabetes,PBMC,fibroblasts,fibrosis,nephropathy

Diabeticnephropathy(DN)isdefinedasrenaldeficiencyassociatedwithTypeIIdiabetes;itisthemostcommoncause
ofendstagerenaldisease,andhasbeentermedamedicalcatastropheofworldwidedimensions.Theincreasing
prevalenceofDNmakesitimperativetosearchformechanismsleadingtofibrotickidneydisease,characterizedby
increasedinfiltrationofleukocytesanddepositionofextracellularmarixproteins,suchascollagensandfibronectin.
Previousfindingsfromourlabhaveshownthatperipheralbloodmononuclearcells(PBMCs)frompatientswith
nephropathycanaltermorphologyinrenalepithelialcellsinvitro.Becauseactivatedfibroblasts,orsocalled
myofibroblasts,arethelikelygeneratorsofextracellularmatrixproteins(fibrosis,wheninexcess),weinvestigated
whetherthereareprofibroticchangesassociatedwithfibroblastswhencoculturedwithPBMCsfromTypeIIDN
patients.
PBMCswereisolatedfrombloodofhealthyvolunteersorpatientsatvariousstagesofDN,asdefinedbyurinary
albumincontent,andcryopreservedpriortococulturewithfibroblasts.Foreskinfibroblastsfromanimmortalline
(AH1F)wereculturedinDMEM,10%FBS,thenplatedina24wellplateongelatincoatedcoverslipsfor24hin0.2%fatty
acidfreeBSAinDMEM.PBMCs(1X10^6percoverslip)wereaddedinRPMI,1%FBSinthepresenceorabsenceof
fibronectin(FN)assemblyinhibitorFUD(500nM)or,asacontrolformyofibroblastictransformation,TGF(20ng/ml)in
RPMI.Mixedcultureswereincubatedfor24or48h,washedwithPBS,fixed,permeabilized,blockedwithBSAand
incubatedwithantibodiestomatrixproteins,smoothmuscleactin,severalmononuclearcellsurfacemarkers,orwith
phalloidinforactinstaining.Analysesincludedphasecontrastandimmunofluorescence(IF)microscopyimaging.
PhasecontrastrevealedthatfibroblastsundergomorphologychangesinthepresenceofPBMCsfrompatientsat
variousstagesofdiabeticdiseaseprogression.Fibroblastswerehypertrophicanderraticindirectionalityrelativeto
thosetreatedwithnormalPBMCs.FibroblastdensityseemedmarkedlyreducedinpresenceofdiseasedPBMCs,
however,AH1Fcellcounts(DAPI)suggestotherwise.IFfindingsforactinstressfibersparalleledmorphologicalfindings
asseenbyphasecontrast.ConsistentchangesinFNorsmoothmuscleactinwerenotapparentbyqualitativeIF
assessment.However,FUDdecreasedadherenceofPBMCs(asdetectedbycountingantiCD45+cells)tothefibroblast
monolayer.Mostofthecellsadheredtothefibroblastmonolayersfromdiabeticpatientsweremonocyticinorigin.
ThechangeindirectionalityandsizeoffibroblastspromotedinthepresenceofPBMCsfrompatientswithdiabetes,
comparedtothoseofnormalsubjects,suggeststheinductionofamigratoryphenotype.Thischangeinphenotypemay
representamechanismviawhichdiseasedPBMCsmaypromotetheinitialstagesoffibroblastconversionto
myofibroblastsandthusthebeginningstagesoffibrosis.ChangesobservedwithFUDsuggesttheinvolvementofFNin
theinteractionsbetweenPBMCsandfibroblasts.Furtherexperimentsneedtobecarriedouttoidentifyquantifiable
markersoftheobservedchangesinducedbydiseasedPBMCstoconfirmthesefindingsandelucidatetheunderlying
mechanisms.

60
InterspeciescomparisonofvitaminAstoringvocalfoldstellatecells

YutakaToya,NapapornRiabroy,ChristopherR.Davis,SherryA.Tanumihardjo,YoKishimoto,DianeM.Bless,NathanV.Welham

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

yutakatoya
(608)2130121
toya@surgery.wisc.edu
Scientist
OtolaryngologyHead&NeckSurgery
Welham
BasicScience
vocalfold,stellatecell,vitaminA,larynx,HPLC

Purpose:
Previousreportsdescribedavocalfoldstellatecell(VFSC)populationinhumanvocalfoldmucosa.Thesecells,localized
tothemaculaflavae(MF),appeartobealocalrepositoryofvitaminA;theirexactroleinvocalfoldbiologyisunclear
andcannotbedeterminedfromhumanstudies.Experimentalresearchinthisareawouldbenefitfromasuitableanimal
model;therefore,wepursuedinterspeciescomparisonstoidentifythemostappropriatemodelforfutureinvivo
experimentalVFSCresearch.

DesignandMethod:
Weconductedhematoxylinandeosin(H&E;generalmorphology),oilredO(OR;lipiddroplet),andgoldchloride(GC;
vitaminA)stainingofserialfrozenvocalfold(VF)sectionsfromfivespecies:human,dog,pig,rabbitandrat(n=23per
species).WeimmunostainedsectionsforthestellatecellmarkerGFAP.Finally,weperformedHPLCbasedanalysisto
obtainretinolandretinylesterprofiles.
SummaryofResults:
HumanandratVFmucosaexhibitedaclearsubpopulationofOR+GC+GFAP+cellswithintheanteriorandposteriorMF.
HPLCanalysisconfirmedthepresenceofretinolandretinylestersinhumansandrats.However,VFSCsandMFwerenot
identifiedindog,pigandrabbitspecimens.
Conclusions:
OurdataindicatethatratVFSCandMFaremostsimilartohuman,suggestingtheratmodelisbestsuitedtoinvivo
experimentalVFSCresearch.

61
CellLineVariabilityintheDifferentiationofHumanPluripotentStemCellstoan
OticProgenitorlikeFate

ParulTrivedi,CynthiaChow,SuChunZhangandSamuelGubbels

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ParulTrivedi
(608)8902673
trivedi@wisc.edu
Other
OtolaryngologyHead&NeckSurgery
Dr.SamuelGubbels
BasicScience
HumanPluripotentStemcells,OticlikeProgenitorcells,Celllinevariability,
Differentiation,geneexpression

Background:Pluripotentstemcellshavethepotentialtogenerateanycelltypeinthebody.Giventhisquality,human
pluripotentstemcellscanbeusedfordevelopmentalstudies,diseasemodeling,drugdiscoveryandpotentiallyfor
therapeuticpurposes.Notableprogresshasbeenmadeinapplyinghumanpluripotentstemcelltechnologytowards
thesegoalsinorgansystemssuchastheretina,centralnervoussystem,peripheralnervoussystemandthe
cardiovascularsystem.Recently,severalmethodsforthegenerationofinnerearhaircellsfrommouseandhuman
pluripotentstemcellshavebeendescribed.Continuedprogressinthisareaofauditoryresearchwillrequirerefinement
ofdifferentiationprotocolstoenrichtheresultantoticprogenitorandhaircellpopulations.Onesourceofvariabilityin
humanpluripotentstemcelldifferentiationcanbeaninherentpredilectionofeachstemcelllinetodifferentiate
towardscertaingermlayerandprogenitorcellderivatives.Wehypothesizethatdifferenthumanpluripotentstemcell
lineswillhavevariablepotencyforgeneratingoticprogenitorlikecellsupondifferentiationunderidenticalconditions.
Methods:Threecommonlyusedandfederallyapprovedhumanpluripotentstemcelllinesweredifferentiatedtowards
anoticprogenitorlikelineageusingadifferentiationprotocolwehavedevelopedthroughmodificationofanembryoid
bodybasedneuraldifferentiationprotocol.Wecomparedhumanpluripotentstemcelllinesusingastepwise,
longitudinalanalysisoftheexpressionofearlyoticgeneandproteinmarkersduringthedifferentiationprocessusingRT
PCR,qPCRandimmunocytochemistry.
Results:Individualhumanpluripotentstemcelllinesdemonstratedvariabilityintheirabilitytodifferentiatetowardsan
oticprogenitorlikefate.ThetemporalpatternandlevelofexpressionofoticprogenitormarkerssuchasPax2,Pax8,
Six1,Eya1andDlx5variedbetweencelllinesupondifferentiationunderidenticalconditions.Individualcelllines
demonstratedahighlevelofconsistencyintheexpressionofoticprogenitormarkertranscriptsandproteins.
Conclusions:Humanpluripotentstemcelllinesappeartofollowasimilartranscriptionalprogramuponotic
differentiation;howeverthereareimportantdifferencesintheefficiencyofindividuallinesindoingso.Ourfindings
highlighttheneedtoconsidertheeffectofindividualcelllinevariabilityinstudiesofHuPSCdifferentiationtowardsan
oticprogenitorandhaircelllikefate.

62
ReceptorInteractingProtein(RIP)mediatednecroptosisasanoveltargetfor
abdominalaorticaneurysm

QiweiWang,ZhenjieLiu,JunRen,StephanieMorgan,BoLiu

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

QiweiWang
(847)9102137
qiwei.mark@gmail.com
GraduateStudent
VascularSurgery
BoLiu
BasicScience
abdominalaorticaneurysm,necroptosis,ReceptorInteractingProtein,necrostatin1,
inflammation

Abdominalaorticaneurysm(AAA)isahighlylethalvasculardiseasewithnoeffectivepharmacologicaltreatment.
Althoughdepletionofarterialsmoothmusclecell(SMC)hasbeennotedinhumanandexperimentalAAAs,howthecells
dieremainselusive.Emergingstudieshavedemonstratedaregulatedformofnecrosistermednecroptosis,whichcould
resultinthereleaseofhighlyimmunogeniccelldeathassociatedmolecularpatterns.RIP1andRIP3arekeyregulatorsof
necroptosis.Here,wetestedthehypothesisthatRIPmediatednecroptosiscontributestocelldeathandinflammationin
AAAthereforeinhibitingRIPmayattenuatethesepathologicalprocessesanddiseaseprogression.
WefoundhumanAAAscontainedanupregulationofRIP3thatwasessentiallyundetectableinnormalarteries.
ImmunostainingandquantitativePCRanalysisdemonstratedsimilarlyelevatedRIP3inamousemodelofAAAthatwas
inducedbyporcinepancreaticelastase.ThesignificanceofnecroptosisinAAAwasdemonstratedbyourresultthat
RIP3dificientmicewereprotectedfromdevelopingAAA.Inourefforttodevelopapharmacologicaltherapy,wetested
whetherinhibitionofRIP1kinasewithNecrostatin1(Nec1)oranoptimizedformofNec1calledNec1scouldprevent
smallaneurysmsfromadvancing.Indeed,existinganeurysmsineitherNec1orNec1streatedmiceshowednofurther
expansionandevenshowedregressioninthecaseofNec1s.Incorrelationtoitsinhibitoryeffectsonaneurysmgrowth,
Nec1profoundlyreducedcellnecrosisandinflammationintheinjuredaortas.Furthermore,wedemonstratedthat
proteinkinaseCdeltaisrequiredfornecroptosislikelythroughregulationofRIP3function.InhibitionofPKCdeltaY311
phosphorylationconferrednecroptosisresistancetoaSMCcellline.
Takentogether,ourdataidentifiesRIPmediatednecroptosisasanunderlyingpathologicalprocessofAAAandmay
serveasanoveltargetforthisdevastatinghumandisease.

63
Agerelatedchangesinratcorticalmotorareadedicatedtotongueandjaw

JulieM.Wenninger,JohnRussell,HeidiKletzien,AllisonSchasser,NaghemeThomas,JeffreyA.Kleim,NadineP.Connor

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JulieWenninger
(608)2659130
wenninger@surgery.wisc.edu
Scientist
OtolaryngologyHead&NeckSurgery
NadineConnor
BasicScience
tongue,jaw,intracorticalmicrostimulation,age,brainmap

Agerelatedchangesinsensorimotoractionsforspeech,voice,andswallowingmaybeduetoalterationswithincranial
muscles,neuromuscularjunctions(NMJ),andthecentralnervoussystem.Withinthetongueandhypoglossalnucleus
(HN),wehavefoundreductionsinevokedandvoluntarymuscleforces,alterationsinNMJmorphology,alterationsin
serotonergicinputstotheHN,andareductioninprimarydendritesofhypoglossalmotorneurons.
However,itisunknowniftheagerelateddeclineinmotorfunctioninvolvescorticalmapchanges.Andifso,arethere
potentialchangeswithexercise?Totestthehypothesisthatcorticalmotorareasdedicatedtojawandtonguewould
decreaseasafunctionofaging,weperformedintracorticalmicrostimulationmappingexperimentsinyoungadult(9mo,
n=7)andold(36mo,n=7)Fischer344/BrownNorwaymalerats.Inasecondgroupofrats,totestthehypothesisthat
exercisewouldincreasethecorticalareasdedicatedtojawandtongue,youngadult(n=8;4exercisetrained,4
untrained)andold(n=8;4exercisetrained,4untrained)ratsunderwenttargetedtongueexercisetrainingorno
exercisetraining.Following8weeksofexercisetrainingornotraining,animalswereanesthetizedandplacedina
stereotaxicframe.Acranialwindowwasmadeextending35mmrostraland12mmcaudaltoBregma.
Aphotographofthecorticalsurfacewastakenandadigitalgrid(squaresof250m)wasoverlaid.Sitesofstimulationat
adepthof1550(5)mweremadeatgridintersectionsusingaglassmicroelectrodefilledwith3NNaClandplatinum
wire.Currentwasincreaseduntilmovementwasevoked(maximumstimulus100A).Meanareasoftongueandjaw
movementwerecalculated.Wefoundthatoldratshadalargertonguemovementrepresentationsthanyoungrats
(0.8890.125and0.2040.093mm2,respectively;p=0.001).
Inthefirstgroupof(untrained)rats,therewasnodifferenceinthesizeofjawmovementrepresentationsbetweenold
andyoungrats(1.080.334and0.6660.104mm2,respectively;p=0.28).Additionally,only2ofthe7youngratshada
tongueareagreaterthan0.06mm2.Thus,theseresultsdidnotallowustoacceptourhypothesis.Inthesecondgroup
ofrats(exercisetrainedorunexercisedtrained),preliminaryresultssuggestthatexerciseincreasecorticalareainthe
tonguebutnotjawinbothyoungadultandoldrats.Whileitappearsthatcorticalareadedicatedtotongueincreases
withageinmalerats,agerelatedcorticalthinningmayhaveoccurred(inuntrained)rats.Becausestimulationwas
evokedataconstantcorticaldepth,furtherinvestigationoftotalcorticalvolumededicatedtothetongueisneeded.
Additionally,preliminaryfindingssuggestthattargetedtongueexerciseincreasedthecorticalareadedicatedtotongue
butdidnotaffectthecorticalareadedicatedtojaw,suggestingthatourtargetedexerciseisspecifictotongue.
However,otherformsofexercise(hypoglossalnervestimulationorincreasedventilatorydrive)aswellasinvestigation
ofpotentialcorticaldepthchangesinexercisedanimalswouldalsoproveinsightful.

G ROUP TWO

Clinical Trials

64
CLINICALEVALUATIONOFSTRATAGRAFTSKINTISSUEASANALTERNATIVETO
AUTOGRAFTINGDEEPPARTIALTHICKNESSBURNS

LeeD.Faucher,MichaelJ.Schurr,KevinN.Foster,StevenE.Wolf,BookerT.King,ChristinaThomasVirnig,KellyVanWinkle,Allen
R.Comer,MaryA.Lokuta,JamesH.HolmesIV,B.LynnAllenHoffmann

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

LeeFaucher
(608)2659574
faucher@surgery.wisc.edu
Faculty
GeneralSurgery
LynnAllenHoffmann
ClinicalTrials
skinsubstitute,deeppartialthicknessburns,clinicaltrial,tissueengineering,wound
therapeutic

Introduction:Standardofcarefordeeppartialthickness(DPT)burnsissurgicalexcisionandautografting.However,
becauseautograftinggeneratespainfuldonorsitewoundspronetoinfectionandscarring,alternativestoautografting
areurgentlyneeded.
Objective:ThisstudyisdesignedtoevaluatethesafetyandefficacyofStrataGrafthumanskinsubstituteasan
alternativetoautografting.
Methods:AmulticenterclinicaltrialtoexaminethesafetyandefficacyofStrataGrafttissueinpromotingthehealingof
DPTburnswithoutautograftingisbeingperformedat6regionalburncenters.Burnsof349%TBSAwereincluded.
PatientshadtwocomparableDPTburnsrandomizedtoautograftorStrataGrafttissue.Intwodosecohortsof10
patientseach,patientsreceivedupto220cm^2and440cm^2ofrefrigeratedStrataGrafttissue,respectively.
Enrollmentinathirdcohortof10patientsconsistedofpatientswhoreceivedupto440cm^2ofcryopreserved
StrataGrafttissue.PrimaryendpointsarewoundclosureatthreemonthsandthepercentageofStrataGrafttreatedarea
requiringautograftingbyday28.Otherassessmentsincludesafetyevaluations,cosmesis,donorsitepainimmunological
responses,andpersistenceofallogeneicDNA.
Results:Enrollmentiscomplete.TherehasbeennosafetysignalrelatedtotheuseofStrataGrafttissue.Ofthe30
subjectsenrolledacrossthethreecohorts,nonerequiredautograftingbyday28althoughonesubjectswounds
convertedtofullthicknessandrequiredtemporarygraftingofbothstudyandnonstudysitesbyday28.Thusfar,18of
19perprotocoltreatmentsiteswerecompletelyhealedby3months.Noevidenceofimmunologicalresponsesto
StrataGrafthasbeenseen.DNAfromtheskinsubstitutehasnotbeendetectedafter3months.Patientfollowupfor
cohort3isongoing.

Conclusions:EarlyresultssuggestthatStrataGrafttissueisabletopromotethehealingofDPTburnsbythepatients
owncellsandisgraduallyreplacedbyautologoustissueasthewoundheals.Ongoingpatientfollowupwilldetermine
whetherthesepromisingearlyresultsarereflectedinthefinalstudyoutcomes.

65
And I think that we can fix it: A qualitative study of surgeons preoperative
discussions before high-risk operations
Jacqueline M Kehler, MD; Kristen E Pecanac, MS, RN; Karen J. Brasel, MD, MPH; Zara Cooper, MD, MSc; Nicole Steffens, MPH,
Martin McKneally, MD, PhD; Margaret L Schwarze, MD, MPP5

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Jacky Kehler
(651) 260-5542
jkehler@uwhealth.org
Resident
Vascular Surgery
Gretchen Schwarze
Clinical Trials
medical decision making, informed consent, communication, high-risk surgery, shared
decision making

Background
The fix-it model of medical decision making describes disease as an acute abnormality that can be readily restored to
normal form or function through targeted medical intervention. This model is particularly ineffective for chronic
conditions and interventions that do not completely return patients to normalcy. It is unknown how the fix-it model is
used by physicians while discussing complex medical decisions such as high-risk surgery.
Objective
To characterize how surgeons communicate with patients during decisions about high-risk operations.
Design
We conducted an observational study of surgeons during preoperative clinic visits. We audio-recorded and transcribed
48 conversations and analyzed the data using qualitative content analysis.
Setting and Participants
We used purposive sampling to select 9 surgeons who routinely perform high-risk surgery at three hospitals in the
United States (2) and Canada (1).
Results
Surgeons commonly used fix-it to describe disease and operations in an effort to facilitate understanding. This model
was also used as the decision making framework in 3 of the 48 conversations. However, after employing fix-it
language to foster understanding, most surgeons explicitly discussed limitations of the fix-it model as an independent
rationale for operating.
Limitations
We did not evaluate non-verbal communication or events that occurred outside the audio-recorded clinic visit. Our
ability to capture the patients understanding of the surgical decision was limited, as the conversations were universally
surgeon-led.
Conclusions
Surgeons deliberate about the decision to operate by highlighting the oversimplification of the fix-it model, suggesting
to patients that the rationale for choosing surgery is substantially more complex. Whether patients are able to make
this distinction between fix-it as an explanatory model of disease and fix-it as a decision making framework is
unclear.

G ROUP THREE

Outcomes

66
Discordanceofhistologicalgradebetweenprimaryandmetastatic
neuroendocrinecarcinoma

Adesoye,Taiwo;Daleo,MarieA.;Weber,SharonM.;Winslow,Emily;Loeffler,AgnesG.;Cho,Clifford

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

TaiwoAdesoye
(405)4107760
TAdesoye@uwhealth.org
Resident
GeneralSurgery
DrCliffordCho
Outcomes
Neuroendocrine,Grade,Discordance,metastatic,tumor

Introduction.Theprognosisandmanagementofneuroendocrinecarcinomaislargelydrivenbyhistologicalgradeas
assessedbymitoticactivityorKi67expression.Wereviewedourinstitutionalexperiencetodetermineifthehistological
gradeofneuroendocrinecarcinomachangesbetweenprimaryandmetastatictumors.
Methods.Wereviewedourinstitutionaldatabasetoidentifypatientswithmetastaticneuroendocrinecarcinoma.
Specimenswereindependentlyreviewedandprimaryandmetastatictumorswerecategorizedaslow,intermediate,or
highgradeasdeterminedbymitoticcountorKi67immunohistochemicalstaining.
Results.Weidentified18patientswithmetastaticneuroendocrinecarcinomatreatedatourinstitutionbetween1997
and2013forwhomcompletepathologicalreviewofprimaryandmetastatictumorswaspossible.Primarylesionswere
foundinthesmallintestine(n=9),pancreas(n=6),ampulla(n=1),stomach(n=1)andrectum(n=1).Metastaticlesions
werehepatic(n=17)orperitoneal(n=1).Timingofmetastasiswassynchronousin13casesandmetachronousin5cases.
Histologicalgradewasconcordantbetweenprimaryandmetastatictumorsin7cases,anddiscordantin11cases.
Amongthediscordantcases,7hadahighermetastaticgradethanprimarygrade,and4hadalowermetastaticgrade
thanprimarygrade.Metachronouspresentationwasassociatedwithahigherlikelihoodofgradediscordance(p=0.036);
inthisseries,thehistologicalgradeofallmetachronousmetastasesdifferedfromthatofprimarytumors.
Conclusions.Thereisahighprevalenceofhistologicalgradediscordancebetweenprimaryandmetastaticfociof
neuroendocrinecarcinoma,particularlyamongpatientswithmetachronousmetastaticpresentation.Giventhe
importanceofhistologicalgradeindiseaseprognosticationandtreatmentplanning,thisfindingmaybeinformativein
themanagementofpatientswithmetastaticneuroendocrinecarcinoma.

67
ElevatedPostoperativePTHlevelsPreventBoneMineralDensityImprovements
inPatientswithPrimaryHyperparathyroidism

KamalAhmed,MD,AmalAlhefdhiMD;;DavidF.Schneider,MD,MS;HerbertChenMD,FACS;RebeccaSippelMD,FACS

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

kamalahmed
(608)6205041
ahmedk@surgery.wisc.edu
PostdoctoralFellow
GeneralSurgery
HerbertChen
Outcomes
PostoperativePTH,BoneMineralDensity,patient,surgery,improvement

Objective:Theaimofthisstudyistoevaluatetheimpactofpostoperative(PTH)levelsonBMDinPHPTpatients.
Background:Primaryhyperparathyroidism(PHPT)isassociatedwithdecreasesinbonemineraldensity(BMD)which
improveafterasuccessfulparathyroidectomy.Cureafterparathyroidectomyisdefinedaseucalcemia,butupto30%of
patientsaftercurativeparathyroidectomywillhavepersistentlyelevatedPTHlevels.
Methods:AretrospectivereviewofallpatientswhounderwentparathyroidectomyforPHPTatasingleinstitutionfrom
July2001April2012.PatientswhohadpreandpostoperativeBMDscanreadings,normalpostoperativevitaminD
levels,andeucalcemiafor>6monthswereincluded.Patientsweredividedintotwogroups;Group1patientshada
normalpostoperativePTHlevelandGroup2patientshadanelevatedpostoperativePTHlevel(>65pg/mL).
Results:Therewere284patients,44(15%)patientsinGroup2and240(85%)inGroup1.Therewerenodifferences
betweenthetwogroupsinformofage,gender,preoperativecalciumorPTHlevel,orpostoperativevitaminDlevels.
Group1patientsweresignificantlymorelikelytohaveanimprovementintheirBMDincomparisontoGroup2patients
(86.3%vs.18.2%,p>0.001).Interestingly,Group2patientswere8timesmorelikelytohaveadeclineintheirBMD
postoperatively(72.7%ofGroup2vs.9.1%ofGroup1,p>0.001).
Conclusion:Ourdatademonstratesthatforpatientswithbonedisease,curativeparathyroidectomyshouldbedefined
notonlybynormalpostoperativecalciumbutalsoanormalpostoperativePTHlevel.

68
RecurrentPrimaryHyperparathyroidismOccursMoreFrequentlyinPatientswith
DoubleAdenomas

AmalAlhefdhi,DavidSchneider,RebeccaSippel,andHebertChen

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AmalAlhefdhi
(202)7589254
alhefdhi@surgery.wisc.edu
PostdoctoralFellow
GeneralSurgery
HebertChen
Outcomes
PrimaryHyperparathyroidism,Recurrent,persistent,doubleadenoma,hyperplasia

Introduction:Theincidenceofrecurrentprimaryhyperparathyroidism(PHPT)hadbeenreportedtobebetween1and
10%.Thepurposeofthisstudywastoexamineifpatientswithmultiglanddiseasehaveadifferentrecurrencerate.
Methodology:AretrospectiveanalysisofaprospectivelycollecteddatabasewasperformedonpatientswithPHPTwho
underwentparathyroidectomyatoneinstitutionbetween2001and2013.Patientswhounderwentinitial
parathyroidectomywithatleast6monthsoffollowupwereincluded,andweredividedintothreegroupsaccordingto
operationnotes:singleadenoma(SA),doubleadenoma(DA),andhyperplasia(H).Anelevatedpostoperativeserum
calciumlevelwithin6monthsofsurgerywasdefinedasapersistentdiseasewhileanelevatedcalciumafter6months
wasdefinedasarecurrence.
Results:Intotal,1402patientsmetinclusioncriteria,andthesuccessrateofparathyroidectomywas98.4%.Themean
agewas60.414yearsand78.5%werefemale.Amongthem1097patients(78%)hadSA,124patients(9%)hadDA,
and181patientshadH(13%).TherateofpersistentPHPTwashigheramongstpatientswithDA(4%),SA(1.3%),andH
(2.2%)(p=0.0049).Moreover,therecurrenceratewashigheramongpatientswithDA(7.3%)vs.SA(1.7%)andH(4.4%)
(p=0.0005)withidenticalmedianfollowuptime.Themedianofthefollowupwas11forpatintswithSA,12.5for
patientswithDA,and12forpatientswithH(p=0.1603).
Conclusion:RecurrentandpersistentPHPToccurmorefrequentlyinpatientswithDA.ThesedatasuggestthatDAin
somecasescouldrepresentasymmetricorasynchronoushyperplasia.Therefore,patientswithDAmaywarrantmore
rigorousmonitoringafterparathyroidectomy.

69
Comparison of Patient Outcomes in Laparoscopic and Open Proctocolectomy
with Ileal Anal-Pouch Anastomosis
Brown, A.; Kennedy, G. MD, PhD; Tevis, S. MD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Allison Brown
(608) 469-9593
akbrown@wisc.edu
Medical Student
General Surgery
Dr. Gregory Kennedy
Outcomes
Outcome, Proctocolectomy, Laparoscopic, Complication, Readmission

Introduction:
Restorative proctocolectomy with ileal pouch-anal anastomosis is the standard procedure in the surgical management of
patients suffering from ulcerative colitis (UC) and familial polyposis syndromes (FAP). With the advent of laparoscopic
techniques, questions have arisen about benefit of these techniques over traditional open methods. In 2008, Dr. Heise
et al. compared the outcomes of patients who underwent open and laparoscopic procedures between the years 2003 to
2007 at a large research university. Since then the institutional database has grown to include twice the number of
procedures. Our objective is to build upon the work of Dr. Heise et al. by examining the outcomes of patients who
underwent open and laparoscopic proctocolectomies to understand if there is benefit to the newer laparoscopic
techniques.
Methods:
Three hundred twenty patients were identified from our institutional database who underwent restorative
proctocolectomy with ileal pouch-anal anastomosis between the years 2003 and 2013. Patients were classified as having
undergone open, laparoscopic assisted, or completely laparoscopic procedures. Laparoscopic assisted and completely
laparoscopic procedures were grouped together. Patients in both arms were characterized based on diagnosis (UC or
FAP), gender, age, BMI, and other co-morbid conditions. The primary intra-operative outcomes of interest were
operative time and estimated blood loss. Post-operative outcomes such as length of hospital stay and return of bowel
function were examined. Hospital readmission rates, return to surgery, and complications were measured for patients
within 30 days of initial discharge. Univariate analysis was performed using Fishers exact test and ANOVA. Multivariate
analysis was performed on variables considered significant on univariate analysis (p 0.10) with binary logistic
regression and univariate linear models. All statistical analyses were performed with SPSS v. 22 and p values 0.05 were
considered significant.
Results:
Of the 320 patients sampled, 61% (n=195) underwent laparoscopic procedures. There were significant differences found
between the procedures in terms of age, BMI, and other co-morbidities. These were adjusted for on multivariate
analysis. The laparoscopic approach took longer to perform than the open approach (516 min vs. 373 min; p 0.000).
However, length of hospital stay (6.8 days vs. 8.7 days; p 0.016), time to ostomy output (2.6 days vs. 4.4 days; p
0.000), and time to tolerance of a liquid (3.6 days vs. 4.9 days; p 0.014) and solid diet (5.1 days vs. 6.4 days; p 0.023)
were shorter in the laparoscopic group. Importantly, there was no difference in complication rate between the
techniques (p 0.194), but patients who underwent open procedures were more likely to be readmitted within 30 days
of discharge.
Conclusions:
Newer laparoscopic approaches to restorative proctocolectomy are associated with longer operative times as compared
to open techniques. However, length of hospital stay and time to return of bowel function were shorter in the
laparoscopic group. Readmission rates were higher in the open group, but there was no difference in complication rates
between the groups. Further studies are needed to consider the monetary implications of shorter hospital stays and
lower rates of readmission for laparoscopic procedures.

70
Comparison of Laparoscopic and Robotic Proctectomy for Rectal Cancer at the
University of Wisconsin
Brown, A.; Kennedy, G. MD, PhD; Tevis, S. MD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Allison Brown
(608) 469-9593
akbrown@wisc.edu
Medical Student
General Surgery
Dr. Gregory Kennedy
Outcomes
Laparoscopic, Robotic, Proctectomy, Outcomes, Rectal Cancer

Introduction:
Robotic proctectomy has been found to be feasible and safe in patients with rectal cancer. Studies have demonstrated
that robotic surgery has definite benefits in the confines of the pelvis as it improves dexterity, ergonomics and allows for
3-dimensional viewing. However, outcomes after robotic proctectomy have not been well documented. Concerns about
longer operative times and increased cost have limited its use. The aim of this study is to evaluate post-operative
outcomes in patients who underwent robotic and laparoscopic proctectomy for rectal cancer between 2011 and 2013.
Methods:
Patients who underwent laparoscopic or robotic proctectomy between 2011 and 2013 were identified from the
prospectively maintained, rectal cancer database at the University of Wisconsin. Univariate analysis was performed
using Fishers exact test and ANOVA. Multivariate analysis was performed using binary logistic regression and linear
models. Statistical analyses were carried out with SPSS v. 22 and p-values 0.05 were considered significant.
Results:
Thirty-five patients were identified from our institutional rectal cancer database. Robotic proctectomy was performed
in 51% of patients. Patients who underwent robotic procedures were similar to patients who underwent laparoscopic
procedures in terms of age, gender, ASA classification, and Body Mass Index. Other pre-operative conditions such as
ultrasound staging, neoadjuvant chemotherapy or tumor height were not significant between the two groups.
Analysis of intra-operative measures showed no difference in mean estimated blood loss (235ml vs. 161ml, p = 0.473) or
conversion rate to an open procedure (12% vs. 6%, p = 0.603). However, mean operative time was significantly longer in
robotic cases (220 minutes vs. 318 minutes, p = 0.001).
There was also no significant difference in oncologic measures. The number of lymph nodes retrieved (15 vs. 20 p =
0.222) and the number of positive lymph nodes (1 vs. 0.72, p = 0.723) was comparable between the two groups. The
distance of the resection margin from the radial margin of the tumor was also similar (17.8 mm vs. 16.6 mm, p = 0.889).
Post-operative outcomes demonstrated no significant difference between the techniques. These measures included
length of hospital stay (4.6 days vs. 4.8 days, p = 0.766) and transfusion within 30 days (6% vs. 0%, p = 0.468). There was
no significant difference in the rates of complications (12% vs. 17%, p = 1.000) or readmissions (6% vs 6%, p = 1.000)
between the groups. Multivariate analysis was performed on post-operative variables adjusting for total time of the
procedure. This did not cause significant changes from the results of the univariate analysis: no significant difference
exists in post-operative outcomes between laparoscopic and robotic proctectomy.
Conclusion:
Operative times are significantly longer using robotic methods as compared to laparoscopic methods when performing
proctecomies for rectal cancer. Although we did not identify any significant differences between the two groups in
terms of post-operative outcomes, our small sample size limits the power of our data. Further studies are needed to
assess the long-term patient morbidity, mortality, and oncologic outcomes of these procedures in addition to the shortterm outcomes analyzed here.

71
SurveyofCodingPracticesbyLaryngologistsWhoPerformBothOfficeand
OperatingRoomBasedProceduresforDysphonia

SethH.Dailey,MD;DavidA.Hyman,MD;TamaraJ.LeCaire,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SethDailey
(608)2630192
dailey@surgery.wisc.edu
Faculty
OtolaryngologyHead&NeckSurgery
SethDailey
Outcomes
laryngology,dysphonia,officebased,surgery,utilization

Voicedisordersarefoundinapproximately7.5millionAmericans,havemultipleetiologiesandoccurthroughouta
personslifespan.Approximatelyonethirdofadultsexperiencedysphoniaalteredvocalquality,pitch,loudness,or
vocaleffortthatimpairscommunicationorreducesvoicerelatedqualityoflifeatsomepointduringtheirlifetime.
However,voicedisordersareespeciallyimportantintheagingU.S.workforcewhere65%ofallU.S.jobsare
communicationbased.Personsaged65yearsandolderareespeciallyatriskfordysphoniaresultingfromvocalfold
atrophy,premalignantormalignantlaryngeallesions,andvocalfoldparalysis.
Thefieldofsurgicalvoicecarehasevolvedsuchthatofficebasedoptions,includinglasertreatmentsandvocalfold
injections,havebecomemuchmorecommoninthepastdecade.Themagnitudeofthissettingchangeisuncleardueto
thelackofinformationdescribingtrendsinnationalutilizationandpracticepatternsforsurgicalvoiceprocedures.This
gapinknowledgeiscompoundedbecauseInternationalClassificationofDiseases(ICD)andCurrentProcedural
Terminology(CPT)codesareassignedtosomebutnotallvoicedisordersandtreatments.Thesubsequentambiguityof
proceduralcoding,especiallyintheofficesetting,maycontributetocodingvariationsacrosstheU.S.Whiledatabases
suchastheChronicConditionsDataWarehouse,a5%sampleofMedicareandMedicaidclaimsdata,offerthe
opportunitytoinvestigatepatternsofutilization,quality,safety,andoutcomesofsurgicalvoicecare,alistofcodesused
acrossthecountryforsurgicalvoiceproceduresisfirstrequired.
Thegoalofthecurrent,inprogressinvestigationistodescribethecodingpracticesusedbyasampleofU.S.
Otolaryngologistsspecializinginlaryngologywhoperformoperatingroomandofficebasedsurgicalproceduresfora
definedsetofvoicedisorders.Codingpracticeswillbedeterminedbyqueryingprovidersinordertodefinecode
identifiablevoicedisordersandprocedures.Fiftylaryngologistsfromacrossthecountrywillbeinvitedtoparticipateina
surveyontheirtraining,practiceandcodingfortreatmentofpathologiesknowntoaffectthevoiceincluding
premalignantandbenignvocalfoldlesions,laryngealmalignancyandunilateralvocalfoldparalysis.Descriptiveanalyses
willincludetheoverallfrequencyandvariationofcodeusage.Effectsofphysiciancharacteristics,suchasfellowship
training,experience,practicetypeandsetting,aswellastheextensivenessoftheirofficebasedpractice,willbe
capturedinordertodescribevariationincodingpracticesacrossproviders.Thisworkwillprovidepreliminary
informationwithwhichtoseekfundingtoaddressthedescribedgapsinknowledgeonbothregionalandnational
patternsofsurgicalvoicecareandtheiroutcomes.

72
NationalTrendsinPediatricBluntSpleenandLiverInjuryManagementand
PotentialBenefitsofanAbbreviatedBedRestProtocol

DodgionCM,GosainA,RogersA,St.PeterSD,NicholPF,OstlieDJ

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ChrisDodgion
(435)6406226
cdodgion@uwhealth.org
Resident
GeneralSurgery
DanOstlie
Outcomes
bluntspleen,bluntliverinjury,pediatrictrauma,bedrestprotocol,costsavings

Purpose
Recentreportssuggestthatanabbreviatedbedrestprotocol(ABRP)maysafelyreducelengthofstay(LOS)andresource
utilizationinpediatricbluntspleenandliverinjury(BSLI)patients.ThisstudyevaluatesnationaltemporaltrendsinBLSI
managementandestimatesnationalreductioninLOSusinganABRP.
Methods
Pediatricpatients(<=17yo)sustainingBLSIwereidentifiedintheKidsInpatientDatabasefrom20002009.Yearlyrates
ofinjuryandoperativeinterventionwereexaminedandstratifiedbytypeofinjury.Temporaltrendswereevaluatedfor
significanceusingMantelHaenszelchisquare,accountingforclusteringwithinhospitals.APSAguidelinesandthe
reportedABRPwereappliedbasedonabbreviatedinjuryscore(AIS)andcomparedwithactualLOS.
Results
22,153patients(meanage=12.0y(+/4.7),meaninjuryseverityscore(ISS)=16.0(+/11.5))wereidentified.Overthestudy
period,operativeratesforspleenandliverinjuries,andoverallmortalitysignificantlydeclined.LOS=3.1d(+/1.6)and
2.7d(+/1.9)forspleenandliver,respectively.IfAPSAguidelineswerefollowed,LOS=3.7d(+/1.1)and3.4d(+/0.7),
respectively.ApplicationoftheABRPwouldresultinLOS=1.3d(+/0.5)forallBSLIpatients.AnABRPcouldpotentially
save1.7hospitaldays/patient,or34,724patienthospitaldaysnationally.Thisrepresentsacostsavingsofover$19
millionannually.

Conclusion
OurstudyconfirmsasignificantnationaldecreaseinoperativeinterventionandoverallmortalityinpatientswithBSLI.
Additionally,itappearsthatashorterobservationperiodthantheAPSAguidelinesisbeingutilized.Theimplementation
ofABRPholdspotentialinfurtherreducingLOSandresourceutilization.

73
PreoperativeCognitiveand/orFunctionalImpairmentisAssociatedwiththe
NeedforHigherLevelofCareafterHospitalDischarge

ColinC.Ehlenbach,BS;SarahE.Tevis,MD;GregoryD.Kennedy,MD,PhD;SarahC.Oltmann,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ColinEhlenbach
(414)5341457
ccehlenbach@wisc.edu
MedicalStudent
GeneralSurgery
GregoryKennedy
Outcomes
geriatrics,postoperativecomplications,caretransitions,hospitalreadmission,
outcomes

Objective:Toidentifyvalidatedcognitiveandfunctionalcapacitymeasurementsthatcorrelatewithdischargelevelof
careinolderadultpatientsundergoinggeneralsurgery.
Background:Preoperativecomprehensivegeriatricassessment(CGA)hasbeenfoundtopredictmorbidityandmortality
inpatientsundergoingelectivesurgery.Preoperativeriskfactorshavebeenshowntoleadtopostdischarge
institutionalization.Anassociationbetweenpreoperativeriskfactors,preoperativelevelofrequiredcare,anddischarge
tohigherlevelsofcarehasnotpreviouslybeendemonstrated.
Methods:Aretrospectivereviewofgeneralsurgerypatientsfromanacademic,tertiaryreferralcenterACSNSQIP
databasewasperformed.Patientsundergoinginpatient,nonemergent,generalsurgeryprocedureswithcomplete
electronicmedicalrecordswereincluded.Patientswithunknowndischargedestination,inhospitalmortality,or30day
mortalitywereexcluded.Initialunivariateanalysiswasperformedbetweenagegroupsoflessthan65yearsand65
yearsandolder.Univariateandmultivariateanalyseswerethenperformedinpatients65yearsandoldertoidentify
correlationsbetweendischargelevelofcare,30dayreadmission,andpostoperativecomplicationswithcognitive,
functional,anddemographicriskfactors.
Results:Between2006and2012,3369patientswereincludedintheinstitutionalACSNSQIPdatabase,ofwhich2012
hadcompleteelectronicmedicalrecordsandmetinclusioncriteria.Ofthe2012patients,1429(71.0%)werelessthan
65yearsold,while583(29.0%)were65yearsorolder.Ofpatientslessthan65yearsold,1.9%weredischargedtoa
higherlevelofcare,while12.2%ofpatients65yearsandolderweredischargedtoahigherlevelofcare.LowMini
MentalStateExaminationscore(OR4.627,95%CI1.66012.898),lowLawtonInstrumentalActivitiesofDailyLivingScale
score(OR2.026,95%CI1.1193.669),advancedage(OR4.676,95%CI2.6528.246),highAmericanSocietyof
Anesthesiologistsphysicalstatusclass(OR1.939,95%CI1.0273.660),andlonghospitallengthofstay(OR5.570,95%CI
3.1249.933)werefoundtobeassociatedwithpostoperativedischargetoahigherlevelofcareinpatients65yearsand
older.Longhospitallengthofstaywasfoundtobeassociatedwithoneormorepostoperativecomplications(OR5.571,
95%CI3.6908.412)and30dayhospitalreadmission(OR3.820,95%CI2.1916.662)inpatients65yearsandolder.
Conclusions:Cognitiveandfunctionalcapacityscoringcanbeusedassimplewaystopredictdischargetoahigherlevel
ofcareforolderadults.Preoperativecounselinginhighriskolderadultsneedstoincludethelikelihoodfordischargeto
ahigherlevelofcare,andifnecessary,atimelyreferraltosocialworkcanbeplacedtofacilitatepostoperativedischarge
planning.

74
TheAgeOfPatientsWithRibFracturesIsAssociatedWithHigherComplication
RatesAndIncreasedLengthOfStay.

A.Elegbede,M.Molnar,H.Jung,A.O'Rourke,S.Agarwal

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AnuElegbede
(702)3081663
AElegbede@UWHealth.org
Resident
GeneralSurgery
Agarwal
Outcomes
Trauma,RibFractures,Outcomes,Age,Morbidity

Introduction:Ageinpatientswithribfractureshavebeenassociatedwithincreasedmortality;however,theimpactof
ageandribfracturesuponotherpostoperativefactors,suchascomplicationsandlengthofstayhasnotbeenstudied.
Methods:Aretrospectivereviewofaprospectivelycollectedandvalidateddatabaseatalarge,university,LevelItrauma
centerwasreviewedfrom2008to2012.Fivecommoncomplications;atelectasis,pleuraleffusion,pneumonia,
respiratoryfailure,andpulmonaryedema,wereexaminedinthreegroups:Ages1829,Ages3059,andAges60and
above.Furthermore,hospitallengthofstaywasexaminedforeachofthesecohorts.Results:Overthefiveyearperiod,
atotalof1,500ribfracturepatientswerefound.Althoughtrendstowardsincreasingatelectasisandpneumonia,were
found,particularlyintheoldestgroup,thesedidnotreachstatisticalsignificance.However,olderpatients(Ages60and
above)werefoundtohaveastatisticallysignificantincreaseinpleuraleffusion(p=0.02),respiratoryfailure(p<0.01),
andpulmonaryedema(p<0.01).Furthermore,olderpatients(Ages60andabove)werefoundtohavelongerhospital
staysthanboththeyounger(Ages1829)andmiddleaged(Ages3059)patientswithribfractures.Conclusions:Rib
fracturescontributegreatlytomorbidityandcomplicationinelderlypatients.Furthermore,elderlypatientsutilize
greateracutehospitalcarethantheiryoungercounterparts.Futureeffortsshouldemphasizepreventionsofrib
fracturesinallpatients,butparticularlytheelderly.Aggressivemanagementofolderpatientswithribfracturesshould
beimplementedinordertoattempttodecreasetherateofcomplications.

75
SurgicalSiteInfectionafterThyroidectomy:ARarebutSignificantComplication

DawnM.Elfenbein,DavidF.Schneider,HerbertChen,RebeccaS.Sippel

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DawnElfenbein
(608)2635214
elfenbein@surgery.wisc.edu
Other
GeneralSurgery
Chen
Outcomes
Thyroidectomy,SurgicalSiteInfection,Prophylacticantibiotics,.,.

Background:Surgicalsiteinfections(SSI)afterthyroidectomyarearareoccurrence,butwhentheyhappen,theycan
havesignificantconsequencesforthepatient.Cervicalendocrineoperationsareclassifiedascleancasesandthe
practicepatternforuseofprophylacticantibioticsvarieswidely.Wehypothesizedthatcertainpatientfactorsand
characteristicsoftheoperationmaypredictahigherriskofsurgicalsiteinfection(SSI),thusidentifyingahighrisk
population.Furthermore,wehypothesizethatSSIareassociatedwithothercomplicationswhichcanleadtoincreased
resourceutilization.
Methods:DatafromtheAmericanCollegeofSurgeonsNationalSurgicalQualityImprovementProgram(ACSNSQIP)
datasetforpatientwhounderwentthyroidectomythroughacervicalincisionfrom20052011wereincluded.Patients
whounderwentasternotomyoratransthoracicthyroidectomywereexcluded.Univariateanalysisusingttests(for
continuousvariables)andchisquare(forcategoricalvariables)wereperformed,andvariableswithp<0.2were
consideredforinclusioninamultivariatelogisticregressionmodel.
Results:49,326patientsunderwentcervicalthyroidectomyfrom20052011and179(0.36%)hadaSSI.Onunivariate
analysis,higherbodymassindex(BMI),presenceofdiabetes,higherAmericanSocietyforAnesthesiology(ASA)class,
steroiduse,>10%weightloss,alcoholuse,historyofchronicobstructivepulmonarydisease(COPD),diagnosisof
disseminatedcancerandlongeroperativetime(mean157+112vs.117+62minutes)wereassociatedwithincreased
riskofSSI.Surprisingly,theextentofoperation,gender,age,smokingstatusanddialysisstatuswerenotassociatedwith
anincreasedriskofSSI.Inamultivariatelogisticregressionmodel,SSIremainedstatisticallyassociatedwiththepre
operativepatientfactorsofobesity(BMI35.0140comparedtonormalBMI)(OR=2.1;95%CI,1.33.5),steroiduse(OR
=2.1;95%CI,1.04.4),andhistoryofsevereCOPD(OR=2.0;95%CI,1.04.1).Operativetimeremainedsignificantinthe
multivariatemodel(p<0.001).PatientswithSSIweremorelikelytorequirereintubation,haveapostoperative
pneumonia,andhavedifficultyweaningfromtheventilator.Theywerealsomorelikelytobereadmitted,returntothe
operatingroom,andhadalongerlengthofstay(3.22+5.7vs1.27+5.1days).
Conclusions:Thoughrare,woundinfectionsafterthyroidectomyareassociatedwithotherpostoperativecomplications,
readmission,andsignificantlyincreasedlengthofstay.Wehaveidentifiedpatientfactorsandoperativefactorsthatare
associatedwithSSI,andthismayhelpidentifyhighriskpatientswhomaybenefitfromselectiveuseofprophylactic
antibiotics.

76
FacultyEvaluationsofResidentsMedicalKnowledge:Cantheybeusedtopredict
ABSITEPerformance?

DawnM.Elfenbein,RebeccaS.Sippel,RobertMcDonald,TammyWatson,JohnE.Scarborough,JohnMigaly

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DawnElfenbein
(607)2635214
elfenbein@surgery.wisc.edu
Other
GeneralSurgery
Sippel
Outcomes
SurgicalEducation,ResidentEvaluations,StandardizedTesting,CoreCompetencies,.

Objective:Althoughimperfect,theABSITEexamoffersoneobjectiveassessmentofresidentmedicalknowledge.
Programdirectorsusethisscoretoidentifyunderperformingresidentsforremediation.TheABSITEisadministered
annually,yetearlyrecognitionofknowledgedeficitsandfrequenttrackingofprogressshouldbeagoal.Wesoughtto
determineifendofrotationevaluationsbyfacultyofresidentsmedicalknowledgecorrelatewithABSITEperformance,
andstudiedtwoevaluationtoolsfromdifferentinstitutions.
Methods:Retrospectivecrosssectionalstudyofresidentsovera3yearperiodfromtwogeneralsurgeryresidency
programs.Facultyfromeachinstitutionratedresidentsclinicalknowledgeaspartofaglobalsummativeevaluationat
theconclusionofeachrotation(12monthslong).Theinterraterreliabilityofclinicalknowledgeratingswasassessed
bycalculatingtheintraclasscorrelationcoefficient(ICC),andconvergentvaliditybetweenfacultyevaluationsand
ABSITEperformancewereassessedforeachinstitution.
Results:1105facultyevaluationswerecompletedabout77residentsfrominstitutionA(mean14.4,min6max24),
and457evaluationsfor70residentswerecompletedfrominstitutionB(mean6.5,min2max15).Therewaslow
agreementamongfacultyforeachresident,withICCcoefficientsof0.17and0.11,respectively.Spearmanscorrelation
coefficientwascalculatedforevaluationsandbothpercentcorrectandnationalpercentileontheABSITE.Forinstitution
A,rho=0.42and0.35(p<0.01),respectively,indicatingaweakcorrelationbetweenevaluationsandABSITEscores,
butnocorrelationwasfoundatinstitutionB.Finally,evaluationsacrossquartilesofresidentABSITEscoreswere
examinedandshownocorrelation.
Conclusions:FacultyevaluationsofresidentmedicalknowledgecorrelatepoorlywithresidentABSITEperformance,and
shouldnotbeusedasanongoingpredictivetool.Asthenewgeneralsurgerymilestonesareimplemented,programs
shouldexamineandupdatecurrentevaluationtoolstoprovidemorereliableongoingassessmentofresidentsmedical
knowledge.

77
PatientProfileatTikurAnbessaSpecializedHospitalVascularSurgeryClinic,
AddisAbaba,Ethiopia

SeanFling,BA.GirmaTefera,MD.NebyouSeyoum,MD.CharlesPenn,BS.RakebTibebu

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SeanFling
(757)8488006
sfling@wisc.edu
MedicalStudent
VascularSurgery
Dr.GirmaTefera
Outcomes
patientprofile,Ethiopia,database,vascularhealth,TASH

BACKGROUND:Whiletheravagingeffectsofinfectiousdiseasesarebeingcurtailedinternationally,chronicdiseaseis
becomingincreasinglyprevalentworldwide.Cardiovasculardisease(CVD)isespeciallyafflictingcountriesglobally.
Althoughthereisbountifulinformationrelatingtotheprevalenceofchronicdiseaseinmoreprosperouscountries,
thereislittleinformationthatshowsthesetrendsindevelopingcountriessuchasEthiopia.TikurAnbessaSpecialized
Hospital(TASH)isthelargesthospitalinEthiopiaandislocatedinthecapitalcity,AddisAbaba.Ithasarelativelynew
vascularsurgeryclinic.Theobjectiveofthisstudywastodevelopadatabaseforthisnewclinicandunderstandthe
significanceofapatientprofile.TASHwasthoughttoserveasagoodsourceforcreatingapatientprofileonEthiopian
cardiovascularpatientsasthelargestpublichospitalinAddisAbaba.
METHODS:AnextensivedatasheetwascreatedusingMicrosoftExcelsoftware.Papercopieswereusedtorecord
patientsdemographicsincludinghistory,physicalexam,vascularriskfactors,andlaboratoryinformation.Ankle
BrachialIndexwasalsocalculatedusingratioofAnklesystolicpressuretobestbrachialsystolicpressure(normalvalues
rangebetween.9and1.3).Thistestwasusedtoidentifypatientswithsymptomaticandasymptomaticvasculardisease.
WeinitiallyassessedapplicabilityofthedatabaseintheTASHsetting.
RESULTS:Initially,anextensivedatasheetwasusedtoensurethatallpertinentdatawascollected.However,this
methodposedtobeverydifficultandtimeconsuming.Overthecourseofthesummer,amorecomprehensivedata
sheetwasdevelopedworkingwiththelocalexperts.ThiscurrentdatasheetincludesMR#,height,weight,calculated
BMI,chiefcomplaint,pastmedicalhistory,labvalues,imagingstudiesordered,riskfactors,previousvasculardisease,
finaldiagnosis,anddisposition.Thisnewdatasheetiscurrentlybeingusedtocollectinformation.Asforfutureplans,
thedatasheetthathasbeenmodifiedwillbeputintoanelectronicformat.
CONCLUSIONS:Thisprojectwasverychallenginggiventheconstraintsoftimeandthefeasibilityofdatacollectionby
physiciansandnursesthatarealreadyseverelyoverworked.Thenewdatasheetiscurrentlybeingtestedforfeasibility,
anditisanticipatedthattheelectronicversionwillresultinthecompilationofapatientprofilerepresentativeof
thousandsofEthiopianpatients.Thisispresentlytheonlyvasculardatabaseinthecountry.

78
Outcomes of Open Versus Laparoscopic Ladd's Procedure for Intestinal
Malrotation in Adults
Lane Frasier, MD; Glen Leverson, PhD; Ankush Gosain, MD PhD; Jacob Greenberg, MD EdM

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Lane Frasier
(517) 282-3314
lfrasier@uwhealth.org
Resident
General Surgery
Jacob Greenberg
Outcomes
Malrotation, Laparoscopy, Outcomes, Adults, Surgery

Introduction: The purpose of this study was to investigate outcomes for adult patients undergoing laparoscopic vs open
surgical repair for intestinal malrotation. Intestinal malrotation results from errors in fetal intestinal rotation and fixation
beginning in the first trimester of pregnancy. Most patients are diagnosed in childhood, typically before two years of
age; however, some patients remain undiagnosed into adulthood. Laparoscopic repair is now an accepted alternative to
laparotomy in children but has not been well-studied in adults.
Methods and Procedures: After IRB approval, a retrospective chart review was performed for all patients 18 years of
age diagnosed with intestinal malrotation presenting to a single tertiary-care institution from 2002 - 2013. Data
collected included: age at diagnosis; pre-operative diagnosis; details of pre-operative imaging, surgical management,
and complications; rates of re-operation, and symptom resolution.
Patients were evaluated on an intent-to-treat basis. Categorical data were analyzed using Chi-squared or Fishers exact
test. Continuous data were analyzed using Students T-test. A two-tailed p-value 0.05 was considered statistically
significant.
Results: Twenty-two patients were identified (age range 18 - 63). Fifteen patients were diagnosed pre-operatively; 4
received an intra-operative diagnosis of malrotation during an elective procedure for an alternate diagnosis. Twenty
patients had some type of pre-operative imaging (data not available for one patient), with CT being the most common
imaging modality employed (77.3%). Four patients had volvulus.
Six patients underwent successful laparoscopic repair; an additional three began laparoscopically but converted to
laparotomy. There were no significant differences in age, operative time, or estimated blood loss between the two
groups. There was a trend toward reduced need for nasogastric (NG) tube in the laparoscopic group (1.86 2.7d vs 5.67
4.1 d, p = 0.07), and a statistically significant difference in length of stay (5.0 2.5 d vs 11.6 8.1 d, p = 0.0148).
Three patients required re-operation: 2 underwent side-to-side duodeno-duodenostomy; 1 underwent a re-do Ladds
procedure after an initial open procedure. Ultimately, 4 patients (2 laparoscopic, 2 open) had persistent symptoms:
bloating (n=2), constipation (n=2), and/or pain (n=2)
Table 1. Comparison of Adult Patients Undergoing Laparoscopic vs Open Repair for Intestinal Malrotation
Number of patients
Pre-operative Diagnosis of Malrotation n (%)
Age (years)
Operative Time (minutes)
Estimated Blood Loss (cc)
NG Tube Decompression (days)
Length of Stay (days)
Re-Operation n(%)

Laparoscopic
9
8 (88.9)
34.9 9.5
174.6 69.7
102.8 191.6
1.9 2.7
5.0 2.5
1 (11.1)

Open
13
7 (53.8)
41.2 14.4
200.5 89.6
99.2 125.5
5.7 4.1
11.6 8.1
2 (15.4)

P value
-0.5630
0.2686
0.4807
0.9585
0.0701
0.0148
1.00

Conclusions: Laparoscopic Ladds repair appears to be safe and effective in adults. While small numbers in our study
limit our ability to identify important differences, there was a statistically significant decrease in LOS and a trend toward
decreased need for postoperative nasogastric decompression. There were no significant differences in major or minor
complication rates, need for re-operation, or persistence of symptoms between the two groups.

79
LongtermCosteffectivenessofMedical,Endoscopic,andSurgicalManagement
ofGastroesophagealRefluxDisease

LukeM.FunkMDMPH,JamesY.ZhangBS,JosephM.DrosdeckMD,W.ScottMelvinMD,KyleA.PerryMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

LukeFunk
(617)6718869
funk@surgery.wisc.edu
Faculty
GeneralSurgery
LukeFunk
Outcomes
gastroesophagealrefluxdisease,antirefluxsurgery,minimallyinvasivesurgery,cost
effectiveness,Markovmodel

Context:Thelongtermcosteffectivenessofmedical,endoscopic,andsurgicaltreatmentforadultswith
gastroesophagealrefluxdiseaseisunclear.Cliniciansthereforehavelimiteddatawhenchoosingmedicalmanagement
versusproceduralinterventionforthesepatients.
Objective:Toestimatethecosteffectivenessofmedical,endoscopicandsurgicaltreatmentforadultswith
gastroesophagealrefluxdiseasewhorequiredailyprotonpumpinhibitortherapy
Design,Setting,Participants:AMarkovmodelwasgeneratedfromthepayersperspectiveusingasixmonthcycleand
30yeartimehorizon.Thebasecasepatientwasa45yearoldmalewithsymptomaticgastroesophagealrefluxdisease
taking20mgofomeprazoletwicedaily.Fourtreatmentstrategieswereincludedinthemodel:protonpumpinhibitor
therapy,transoralincisionlessfundoplication,radiofrequencyenergyapplicationtotheloweresophagealsphincterand
laparoscopicNissenfundoplication.Datasourcesforthemodelparametersincludedthepublishedliteratureand
institutionalbillingdata.
MainOutcomeMeasures:Incrementalcosteffectivenessratios(costperqualityadjustedlifeyeargained)
Results:Inthebasecaseanalysis,whichassumedaprotonpumpinhibitorcostof$234oversixmonths($39permonth),
radiofrequencyenergyapplicationtotheloweresophagealsphincterandlaparoscopicNissenfundoplicationwerethe
mostcosteffectiveoptionsovera30yeartimeperiod($2,471and$5,579perQALYgained,respectively).Sensitivity
analysisdemonstratedthatifthecostofprotonpumpinhibitortherapyexceeded$491.95permonthoverfiveyears,
$201.42permonthovertenyears,or$90.63permonthover30years,laparoscopicNissenfundoplicationbecamethe
dominanttreatmentoption(moreeffectiveandlesscostly).Transoralincisionlessfundoplicationwasdominatedby
bothradiofrequencyenergyapplicationandlaparoscopicNissenfundoplicationatallpointsintime.
ConclusionsandRelevance:Inexpensiveprotonpumpinhibitorsarecosteffectivemedicationsforpatientswith
gastroesophagealrefluxdiseasewhorequiredailymedicaltherapy.However,asthecostofprotonpumpinhibitor
treatmentincreases,laparoscopicNissenfundoplicationbecomesamorecosteffectiveoptionoverthelongterm.
SurgicalreferralshouldbeconsideredforGERDpatientswhoareunabletoobtainsymptomaticrelieffromprotonpump
inhibitorsatarelativelylowprice.

80
BariatricSurgeryOutcomesinMedicaidvs.NonMedicaidPatients:Different
Cohorts,SimilarOutcomes

LukeMFunkMDMPH,AndrewSuzoBS,DeanJMikamiMD,BradleyJNeedleman

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

LukeFunk
(617)6718869
funk@surgery.wisc.edu
Faculty
GeneralSurgery
LukeFunk
Outcomes
bariatricsurgery,outcomesresearch,minimallyinvasivesurgery,healthdisparities,
insurancecoverage

Background:Medicaidpatientshavehighratesofmorbidobesity,butbariatricsurgeryisnotroutinelycoveredby
Medicaidinsomestates.Incontrast,bariatricsurgeryiscoveredbyMedicareandmanyprivateinsurers.Reasonsfor
thisdisparityareunclearbutsomehavereportedconcernsthatMedicaidpatientshavepooreroutcomesfollowing
bariatricsurgery.Weperformedasingleinstitutionretrospectiveanalysistoexaminewhethertherearedifferencesin
shortorlongtermoutcomesbetweenMedicaidandnonMedicaidbariatricsurgerypatients.

Methods:AllpatientswhounderwentalaparoscopicRouxenYgastricbypassatTheOhioStateUniversityWexner
MedicalCenterfrom20082010wereidentified.Ofthese609patients,30wereMedicaidpatients.Theywereselected
foranalysisandcomparedto90randomlyselectednonMedicaidpatients(1:3casecontroldesign).Preoperativedata
andpostoperativeoutcomeswereobtainedfromtheelectronicmedicalrecord.Thesevariablesincludeddemographic
data,weightlossattwoyears(%excessbodyweightloss),comorbidityresolution(diabetes,hyperlipidemia,
hypertension,obstructivesleepapnea,andgastroesophagealrefluxdisease),complicationsat90daysandtwoyears
(woundcomplications,acuterenalfailure,pulmonaryembolus,anastomoticleak,deepvenousthrombosis,pneumonia,
urinarytractinfection,mortality,lengthofstay,andERvisitspostoperatively.ChisquaredandstudentsTtestswere
usedtocomparecategoricalandcontinuousvariables,respectively.
Results:ComparedtothenonMedicaidcohort,MedicaidpatientshadsignificantlyhigherpreoperativeBMIs(58.4vs.
49.5;p<.001)andhigherratesofcomorbiditiessuchasobstructivesleepapnea(83.3%vs.51.1%;p=.002)anddiabetes
(73.3%vs44.4%;p=.006).Duringthefirst90daysaftersurgery,MedicaidpatientshadhigherratesofERvisits(33.3%
vs.10.0%;p=.007)butsimilaroverallcomplicationrates(30.0%vs.17.8%;p=.15)andlengthsofstay(median3days).
Bothcohortshadmorethan50%excessbodyweightlosstwoyearsaftersurgery(53.8%forMedicaidvs.64.6%fornon
Medicaid;p=.02).Atamedianoftwoyearsoffollowup,therewerenosignificantdifferencesincomorbidityresolution,
anastomoticcomplicationsormortality.

Conclusion:Despitebeingahigherriskcohort,Medicaidpatientshadsimilarshortandlongtermoutcomescomparedto
nonMedicaidpatients.ConcernforpoorsurgicaloutcomesmaynotbeavalidreasonfordenyingMedicaidpatients
bariatricsurgerycoverage.

81
NasolacrimalSystemFracturesAssociatedwithEpiphoraandDacryocystitis

RaviK.Garg,MichaelJ.Hartman,MarkJ.Lucarelli,GlenLeverson,AhmedM.Afifi,LindellR.Gentry

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

RaviGarg
(805)5019682
2ravigarg@gmail.com
Resident
Plastic&ReconstructiveSurgery
LindellGentry
Outcomes
nasolacrimalsystem,lacrimalfossa,facialfractures,epiphora,dacryocystorhinostomy

Background:Fracturesofthenasolacrimalsystem(NLS),includingthelacrimalfossaandduct,havenotbeen
comprehensivelydescribedinpatientswithfacialtrauma.Characterizationoftheseinjuriesmayhelpfacialtrauma
surgeonsbetterpredictwhichpatientswilldeveloplacrimaloutflowobstructionsymptomsincludingepiphoraand
dacryocystitisandwhomayeventuallyneedlacrimalsurgery.
Methods:CTimagesforallpatientsseenattheUniversityofWisconsinHospitalandClinicsforcraniofacialtraumawere
reviewedfromJanuary2001toDecember2005.PatientswereincludediftheyhadaNLSfractureandatleast1yearof
followup.Fracturepatternsweredescribedandcorrelatedwithclinicaloutcomesdocumentedinthemedicalrecord.
Outcomes,includingthedevelopmentofepiphoraordacryocystitis,ortheneedforlacrimalsurgerywereanalyzed
usingFischersexacttest.
Results:Weidentified104patientswithNLSfracturesamong1,980patientswithcraniofacialtraumawhohadatleast1
yearoffollowup.Elevenpatients(10.6percent)developedepiphoraordacryocystitis,and2patients(1.9percent)
requiredexternaldacryocystorhinostomy(DCR).Tenradiographicinjurypatternswerecharacterized.Avulsionofthe
lacrimalcrest,bonefragmentinthelacrimalfossaorduct,ductcompressiongreaterthan50percent,andnasomaxillary
buttressdisplacementweresignificantlyassociatedwiththedevelopmentofepiphoraordacryocystitis(p<0.05).
NasomaxillarybuttressdisplacementwassignificantlyassociatedwiththeeventualneedforexternalDCR(p=0.03).
Conclusions:PatientswithradiographicevidenceofNLSfracturehaveanapproximately10%riskofdevelopingepiphora
ordacryocystitis.Wedescribe5NLSfracturepatternsthataresignificantlyassociatedwiththedevelopmentoflacrimal
outflowobstruction.Thepresenceofnasomaxillarybuttressfractureanddisplacementsuggestsasignificantlyhigher
riskofeventuallyneedinglacrimalsurgery.

82
CTRELATEDRADIATIONEXPOSUREINCHILDRENTRANSFERREDTOALEVEL1
PEDIATRICTRAUMACENTER

AdamS.BrinkmanMD,KaraG.GillMD,CarlyMGlarnerMD,JocelynBurkeMD,AndrewRogers,MD,MaryJ.Anderson,RN,
CharlesM.LeysMD,DanielJ.OstlieMDandAnkushGosainMD,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AnkushGosain
(608)2639419
gosain@surgery.wisc.edu
Faculty
GeneralSurgery
Gosain
Outcomes
Trauma,Pediatric,ComputedTomography,IonizingRadiation,Radiology

Purpose:
PediatrictraumapatientspresentingtoReferringFacilities(RF)oftenundergocomputedtomographyscans(CT)to
identifyinjuriesbeforetransfertoaLevel1PediatricTraumaCenter(PTC).ThepurposeofourstudywastoevaluateRF
compliancewiththeAmericanCollegeofRadiology(ACR)guidelinestominimizeionizingradiationexposureinpediatric
traumapatientsandtodeterminethefrequencyofadditionalorrepeatCTimagingaftertransfer.
Methods:
AfterIRBapproval,retrospectivereviewofallpediatrictraumaadmissionsfromJanuary2010December2011atour
AmericanCollegeofSurgeonsverifiedPTCwasperformed.Patientdemographicsandmeansofarrivalwereanalyzed.
PatientswhounderwentCTweregroupedbymeansofarrival:thosethatweretransferredfromaRFversusthosethat
presentedprimarilytothePTC.CompliancewithACRguidelinesandneedforadditionalorrepeatCTscanswere
assessedforbothgroups.
Results:697children(<18yo)wereidentifiedwithameanageof9.5years.321(46%)patientspresentedprimarilyto
thePTC.376(54%)weretransferredfromaRF,ofwhich90(24%)patientsunderwentCTpriortotransfer.CTradiation
dosinginformationwasavailablefor79/90patients(88%);61/79(77%)receiveddosesofradiationaboveACR
recommendedlevels.Aftertransfer,9%(8/90)ofchildrenimagedataRFrequiredadditional/repeatCTscans.In
comparison,98%(314/321)ofpatientswhopresentedprimarilytothePTCandunderwentCTreceivedappropriate
pediatricradiationdosing.MeanradiationdoseatPTCwasapproximatelyhalfofthatatRF(p<0.01).
Conclusions:PediatrictraumapatientstransferredfromRFoftenundergoCTscanningwithhigherthanrecommended
radiationdoses,potentiallyplacingthemathighercarcinogenicrisk.Fortunately,fewRFpatientsrequiredadditionalCT
scansafterPTCtransfer.Finally,compliancewithACRradiationdoselimitguidelinesisbetterachievedataPTC
comparedtoaRF.

83
Characterizingreadmissioninulcerativecolitispatientsundergoingrestorative
proctocolectomy.

ThomasP.Hanzlik,SarahE.Tevis,MD,AmySuwanabol,MD,BruceHarms,MD,CharlesHeise,MD,EugeneF.Foley,MD,and
GregoryKennedy,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ThomasHanzlik
(608)7726676
hanzlik@wisc.edu
MedicalStudent
GeneralSurgery
Kennedy
Outcomes
readmission,ulcerativecolitis,restorativeproctocolectomy,,ilealpouchanal
anstomosis,complications

Purpose:Postoperativereadmissionsincreasecostsandaffectpatientqualityoflife.Ulcerativecolitispatientsatour
institutionareatahighriskforpostoperativereadmissionfollowingrestorativeproctocolectomy.Weaimto
characterizepatientsundergoingrestorativeproctocolectomywithrespectto1)thetimeframeofreadmission,2)the
reasonforreadmissionand3)theriskfactorsassociatedwithreadmission.
Methods:Ulcerativecolitispatientswhounderwentrestorativeproctocolectomywereidentifiedfromtheprospectively
maintainedUniversityofWisconsinColorectalSurgerydatabase.Weevaluated533patientswhometourinclusion
criteria.Restorativeproctocolectomyatourinstitutionisroutinelyperformedasa2stageoperation.Thefirststage
involvesaproctocolectomywithilealpouchcreationanddivertingloopileostomy.Thesecondstageinvolvestakedown
oftheloopileostomy.Postoperativereadmissionratesandreasonsforreadmissionwereexaminedfollowingboth
stages.Univariateandmultivariateanalyseswereperformedtoevaluateforriskfactorsassociatedwith30day
readmissionfollowingstageI.
Results:FollowingstageIofrestorativeproctocolectomy,18.2%(n=97)ofpatientswerereadmittedwithin30days
while22.7%(n=121)werereadmittedwithin90days.Youngerpatientage(OR1.825,95%CI1.1392.957),laparoscopic
approach(OR1.943,95%CI1.2173.104)andincreasedlengthofinitialstay(OR1.155,95%CI1.0901.225)wereall
associatedwith30dayreadmission.Themostcommonreasonforreadmissionwasileus/partialbowelobstruction,with
10%ofallpatientsreadmittedforthisreasonwithin30daysofstageI.
Conclusions:Patientsundergoingrestorativeproctocolectomyareathighriskforreadmission,particularlyfollowingthe
firststageoftheoperation.Noveltreatmentpathwaystopreventileusandsubsequentdehydrationasanoutpatient
maydecreasetheratesofreadmissionfollowingrestorativeproctocolectomy.

84
ParadoxicalVocalFoldMotion:EpidemiologicalPresentationandClinical
Characterization

N.A.Hartley,K.M.McConville&S.L.Thibeault

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

NaomiHartley
(608)6094094
hartley@surgery.wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
Thibeault
Outcomes
Voice,PVFM,Epidemiology,ClinicalCharacteristics,vocalfoldfunction

Background&Objectives:Paradoxicalvocalfoldmotion(PVFM)isincreasinglyrecognizedasadisorderthatcanresultin
considerablepatientdistressandmorbidity,yettheunderlyingnatureofthedisorderisnotwellunderstoodandit
remainsmisdiagnosedandsubsequentlyinappropriatelytreatedinclinicalpractice.Theaimofthisstudywastoenrich
thescientificliteratureonPVFMthroughdescriptionofthedemographicandclinicalcharacteristicsofpatientsreferred
toamultidisciplinaryvoiceandswallowingclinicinatertiarycarefacility.
Method:RetrospectiveanalysisofoutcomesdatafromtheUniversityofWisconsinVoiceandSwallowingdatabase
spanningadmissionsOct2006January2012.Participantswereidentifiedusingspecificcriteria,includingICD9code
478.5,SLPandMDimpressionsattimeofassessment.Recordofdemographicinformation,prior,currentand
subsequentdiagnosesandmedications,comorbidities,presentationandqualityofsymptoms,othermedicalhistory,
VoiceHandicapIndex(VHI),refluxsymptomindex(RSI),andthe7itemGeneralizedAnxietyDisorderAssessment(GAD
7)werecollectedforanalysis.
Results:96casesmettheinclusionandexclusioncriteria.Themajorityofcaseswerefemale(n=70),nonsmokers,aged
between3069years(range=1692years)whowerecurrentlyworkingfulltime.Presentingsymptomsweremost
commonlytightnessinthethroat(52%)andchest(46%),with>20%ofcasescitingexacerbatingfactorsofwalking,cold
air,stressandweeklyexercise.Asthmahistorywaspositivein65%ofcases,with61%currentlytakingasthmarelated
medications.Ofthese,83%reportedthattheircurrentbreathingproblemdifferedfromasthma.Othercomorbidities
includedallergies(50%),reflux(37%),anxiety(36%),dysphonia(12%)anddysphagia(7%).Anxiolytics(44%)and
antireflux(43%)medicationswerecommon.
ElevatedRSIscoreswerepresentin60%ofcases,whilepatientratingsoftheVHIindicatedminimalinfluenceofvoice
problemsondailylivingin77%ofpatients.Elevatedvoicehandicapratingswerefoundtobesignificantly(p<.02)
associatedwithmoderatetosevereanxietyscores(GAD7)andrefluxdisease(p<.001).
Conclusion:ThisstudyhighlightscommonpresentingandclinicalfeaturesassociatedwithPVFMinindividualsreferred
forvoiceand/orswallowingexamination.Theseresultsarevaluableforbothfutureresearchandclinicalapplications.

85
IsCentralLymphNodeDissectionNecessaryforParathyroidCarcinoma?

KunTaiHsu,M.D.,RebeccaS.Sippel,MD,FACS,HerbertChen,MD,FACSandDavidF.Schneider,M.D.,M.S.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

KunTaiHsu
(608)7725168
hsu@surgery.wisc.edu
PostdoctoralFellow
GeneralSurgery
Dr.DavidSchneider(withDr.ChenandDr.Sippel)
Outcomes
parathyroidcarcinoma,lymphnodes,lymphadenectomy,diseasespecificsurvival,
SEER

Background
Parathyroidcarcinomahasafiveyearmortalityrateof9%to31%.Unlikeothermorecommonmalignancies,the
significanceoflymphnode(LN)statusremainscontroversialinparathyroidcarcinoma.Currentsurgicalguidelines
recommendenblocresectionoftheparathyroidtumor,theipsilateralthyroidlobe,andipsilateralcentralcompartment
LNdissection.ThepurposeofthisstudywastodeterminetherelativeimportanceofLNmetastasesindiseasespecific
survival(DSS).
Methods
ThiswasaretrospectivereviewusingtheSurveillance,Epidemiology,andEndResult(SEER)databaseofparathyroid
carcinomacasesdiagnosedbetween1988and2010.KaplanMeiersurvivalestimationandCoxproportionalhazards
modelswereusedtoevaluatefactorsaffectingDSS.LogisticregressionwasusedtoidentifypredictorsofLN
metastases.
Results
405parathyroidcarcinomapatientswereidentifiedfromtheSEERregistry.Themedianageatdiagnosiswas56years
(range2089),and212patients(52.3%)weremale.Among114patientswhoseLNswereexaminedatsurgery,only12
(10.5%)hadpositiveLNs.Weperformedsensitivityanalysisandfoundatumorsizethresholdof3cmbestdividedthe
cohortbyDSS.Tumorsize3cm(HR3.67;p=0.03),positiveLN(HR5.63;p=0.02)anddistantmetastasis(HR69.55;
p<0.001)weresignificantadversepredictorsofDSSonunivariateanalysis.Onlytumor3cm(HR4.40;p=0.01)and
distantmetastasis(HR2.89;p=0.004)remainedsignificantonmultivariateanalysis.Notably,LNmetastasesdidnot
independentlypredictDSS(HR0.82;p=0.53).Furthermore,therewasnosignificantrelationshipbetweenlocalinvasion
andpositiveLNs(p=0.40).WhenexaminingfactorsassociatedwithLNstatus,onlytumor3cmpredictedLNmetastasis
(OR40.03;p=0.02).LNmetastaseswere7.5timesmorelikelyinpatientswithtumors3cmthanthosewithtumors
<3cm(21%vs.2.8%;p=0.02)
Conclusions
PositiveLNstatuswasnotassociatedwithDSSforparathyroidcarcinoma.Therefore,centralcompartmentLN
dissectionmaybeunnecessaryinthetreatmentofparathyroidcarcinomaexceptforpatientswithlargetumors(3cm).

86
DoesPostoperativeDrainAmylasePredictPancreaticFistulaFollowing
Pancreatectomy?

JacquelineS.IsraelMD,RobertJ.RettammelMA,GlenE.LeversonPhD,LauraR.HanksBA,CliffordS.ChoMD,FACS,EmilyR.
WinslowMD,SharonM.WeberMD,FACS

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JacquelineIsrael
(715)2109959
jisrael@uwhealth.org
Resident
GeneralSurgery
SharonWeberMD,SurgicalOncology
Outcomes
PancreaticFistula,Amylase,Drain,Pancreatectomy,ISGPF

Background:Previousstudiessuggestthatfollowingpancreatectomy,adrainfluidamylaseobtainedonpostoperative
day1(DFA1)>5000U/Lcorrelateswiththedevelopmentofpostoperativepancreaticfistula(PF).Wesoughttovalidate
whetherDFA1isaclinicallyusefulpredictorofPF.Inaddition,weaimedtoassesswhetherDFA1correlateswithPF
severity.
StudyDesign:Usingasingleinstitutionprospectivedatabase,wereviewedrecordsfrompatientswhounderwenteither
pancreaticoduodenectomy(PD)orleftpancreatectomy(LP)betweenAugust2010andAugust2012.Allpatientshad
oneortwosurgicaldrainsplacedintraoperativelynearthepancreaticandbiliaryanastomoses.Thepresenceandgrade
ofPFwasdeterminedusingtheconsensusguidelinesfromtheInternationalStudyGrouponPancreaticFistula(ISGPF).
Results:Ninetyeightpatientsunderwentmajorpancreatectomyduringthestudytimeperiod.Ofthese,63patientshad
adocumentedDFA1.PFoccurredin27(43%)ofthe63patientsonwhomaDFA1wasobtained.OfthosewithPF,2(7%)
weregradeA,18gradeB(67%),and7gradeC(26%).ThemedianDFA1inpatientsdiagnosedwithPF(4590U/L,range
3216,900)wassignificantlyhigherthanthemedianinthosewithout(45U/L,range25840;p<0.001).WhenDFA1
wasanalyzedatvaryingcutoffvalues,thecorrelationofDFA1withPFwashigh,withanareaunderthereceiver
operatingcharacteristic(ROC)curveof0.903,CI0.8300.976(p<0.0001).Thesensitivity,specificity,positivepredictive
value(PPV),andnegativepredictivevalue(NPV)wereassessedatvaryinglevelsofDFA1.Thehighestsensitivity(96%)
andnegativepredictivevalue(96%)wereobtainedwithacutoffDFA1of<100.Therewasnostatisticallysignificant
relationshipbetweenDFA1levelandISGPFgrade.
Conclusions:Inpatientsundergoingpancreaticresection,althoughDFA1>5000wasassociatedwithhighspecificityand
PPV,thesensitivitywaslow,decreasingitsclinicalusefulness.Importantly,acutoffofDFA1<100resultedinahigh
sensitivityandNPV.Therefore,inpatientswithDFA1<100,earlydrainremovalmaybesafe.Furtherstudiesare
recommendedtovalidatetheroleofDFA1inexcludingPF,potentiallyanticipatingPFseverity,andassistinginthe
managementofsurgicaldrains.

87
TheImpactofComorbidityonMortalityinTraumaOutcomes

ChristopherJanowak,PhilipMercier,SureshAgarwal

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ChristopherJanowak
(608)7744979
cjanowak@uwhealth.org
Resident
GeneralSurgery
SureshAgarwal
Outcomes
Trauma,InjurySeverity,Outcome,Comorbidity,Mortality

Introduction:Traumaticinjurycontinuestobealeadingcauseofhospitalization.Overthepastfivedecades,several
indiceshavebeendevelopedtoevaluatetheprognosisoftraumaticinjurybytakingintoaccountvaryingparameters
abouttheindividual,theinjury,andtheindividual'sresponsetothatinjury.Thesepredictivemethodssufferfrommajor
flawsinthattheyareunreliable,toocumbersomeforbedsidecalculation,ordonotaddressaspectsofthethree
aforementionedparameters.Oftentheindividualisleftoutoftheassessment,whenitappearsintuitivethatan
otherwiseyounghealthypatientwouldbebetterabletosustainthesamecalibertraumaticinjuryasahighlycomorbid
patient.
Methods:UsingaprospectivelykepttraumadatabaseataLevelItraumacenter,weevaluatedtraumaticallyinjured
patientsforchangesinmortalityagainstthenumberofcomorbidconditionspresentaspertheNationalTrauma
Databankguidelines.Werevieweddatafrom1443consecutiveadmissionsin2013forbasicdemographics,injury
severityscore,comorbidconditions,anddisposition.
Results:Ofthe1443patients894patientshadcompleterecordsforanalysisandofthese,60%weremalewithan
averageageof50(range18103).Ofthesepatients,855(95.6%)weredischargedaliveand39(4.3%)died.Thenumber
ofpatientssustainingtraumaticinjurydecreasedasthenumberofcomorbidconditionsincreased;466patientswith
onecomorbidity,211withtwocomorbidconditions,decreasingtojust8patientswithsixcomorbidconditions.The
percentageofpatientsaliveatdischargefollowinginjuryalsodecreasedwithincreasingnumberofcomorbidconditions
from98.5%withonecomorbidcondition,84.9%withfourcomorbidities,and75%withsixcomorbidities.An
exponentialregressionlineforthisdatahadanRsquaredvalueof0.80,suggestingagoodfit.Wealsoevaluatedthe
injuryseverityscore(ISS)ofpatientsversusnumberofcomorbidconditionsandfoundthat,althoughthepatientswho
diedasaresultoftheirinjurieshadanaveragehigherISS,therewasrelativelylittlechangeinISSpernumberof
comorbidconditions.
Conclusions:Ourfindingssuggestthatthenumberofcomorbiditiesperpatientintraumaticallyinjuredpatients
inverselycorrelateswithpercentagealiveatdischarge.Certainlydifferentcomorbiditieshavedifferenthealthimpacts
onindividualsanduniformweightingofcomorbidconditionsmaynotaccuratelyreflecttheindividual.However,our
findingssuggestthatfurtherevaluationofcomorbidconditionswithrespecttotheiroutcomesintraumaticallyinjured
patientsisapromisingavenueandmayhelpimprovecareofthesepatients.Validatingthesefindingswithprospective
observationsmayhelpjustifychangingtheInjuryScoringsystemsthatarecurrentlyavailable.

88
TheVenousAnastomoticFlowCouplerforFreeFlapMonitoring:AProspective
Analysisof85MicrosurgicalBreastReconstructionCases

Kempton,Steve,MD,Poore,Samuel,MD,Chen,Jenny,MD,Afifi,Ahmed,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SteveKempton
(909)5531690
skempton@uwhealth.org
Resident
Plastic&ReconstructiveSurgery
SamuelPoore
Outcomes
FreeFlap,BreastReconstruction,Anastomosis,Monitoring,Doppler

Purpose:Thevenousanastomoticflowcouplerhasrecentlybeendevelopedforclinicaluse,contributingtoamultitude
offlapmonitoringdevicesandtechniques.Todate,onlyonepublishedsmallretrospectiveseries(19patients)reported
thisdevicetobebothreliableandaccurateforuseinheadandneckreconstruction;however,nodataexistsinthe
settingofabdominalbasedfreeflapsforbreastreconstruction.Theauthorspresentaprospectiveanalysisofthe
venousanastomoticflowcouplerin85microsurgicalbreastreconstructioncases.
Methods:Prospectivedatawascollectedonpatientsundergoingpostmastectomyfreeflapbreastreconstructionfrom
May2012toMay2013.Dataobtainedincludedpatientage,BMI,flaptype(DIEP,MSTRAM,SIEA),flowcouplersize,
incidenceofintraoperativeandpostoperativesignalloss,anastomoticproblems,couplerproblems,flaptakeback,and
flapfailure.Proportiondatawascompiledandanalyzed.
Results:EightyfiveconsecutiveabdominalbasedfreeflapsforbreastreconstructionwereperformedfromMay2012to
May2013bytwocosurgeonsattheUniversityofWisconsinHospital.Theaveragepatientagewas49.3yearsand
averageBMIwas28.4.Therewere53MSTRAM,31DIEP,and1SIEAflapsperformed.Thevenousanastomoticflow
coupler(FC)wasusedinallcases.Theoverallflapfailureratewas4.7%andflaptakebackratewas7.1%.Theflow
couplerwasanalyzedinbothintraoperativeandpostoperativesettings.Figure1andFigure2summarizethe
intraoperativeandpostoperativeflowcouplerdatarespectively.Flaptypeandflowcouplersizewerenotfoundtobe
associatedwithflowcouplerproblems.Theintraoperativeandpostoperativesensitivityoftheflowcouplerwasfound
tobe100%(signalpresencecorrelatedwellwithflapviability).However,intraoperatively,theflowcouplerhadafalse
positiverateof75%andapositivepredictivevalueof0.25.Postoperatively,theflowcouplerhada36%falsepositive
rateandapositivepredictivevalueof0.64.
Conclusions:IftheflowcouplerDopplersignalisaudible,itbecomesextremelyfast,reliable,andefficienttoconfirm
patencyofamicrosurgicalanastomosis.However,thereisahighfalsepositiverateinboththeintraoperativeand
postoperativesetting.Thisledtoahighincidenceofintraoperativemaneuverstodiagnoseandamendthecauseof
signalloss.In13%ofcases,theflowcouplerDopplersignalwascompletelyignoredandflapsweremonitoredmore
traditionallywithexternaldopplerorclinicalexam.

89
The Effect of an Integrated System on the Rate of Duplicate CT Scanning in
Transferred Trauma Patients and the Costs Associated with Care
Amy E. Liepert, MD*; Joseph Bledsoe, MD; Sarah Majercik, MD, MBA; and Mark H. Stevens, MD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Amy Liepert
(801) 865-5552
liepert@surgery.wisc.edu
Faculty
General Surgery
Amy Liepert
Outcomes
CT scan utilization, cost accounting, vertical system, trauma, transferred patients

Introduction:
Duplication of Computed Tomography (CT) scanning in transferred trauma patients is a source of quality waste in
healthcare and it is potentially harmful to the patient. This study reports the rates of duplicate CT scan utilization and
cost implications for trauma patients transferred to an ACS-verified Level 1 Trauma Center (TC) in a vertically integrated
healthcare system (Intermountain Health Care) from out-of-network and in-network. The vertically integrated system
provides for immediate viewing of radiology studies via a connected PACs system as well as access to the full electronic
medical record. This system utilizes true COST accounting.
Objectives:
Evaluate the rate of duplicate CT scanning in a transferred trauma population within a vertically integrated healthcare
system both from within the system and from outside of the system.
Compare costs between transfers from OON and IN of patients who had duplicated CT scans.
Methods:
Patients transferred to the ACS-verified Level 1 Trauma Center were prospectively identified at the time of transfer as
either IN or OON. The Level 1 Trauma Center serves as the referral center in a vertically integrated network of 22
hospitals, as well as 20 hospitals that are not directly part of the integrated system. We compared rates of duplicated CT
scanning, and radiology and hospital costs. Duplicated scans were defined as scans that patients received at both the
referring and receiving (Level 1 Center) facilities.
Comparisons were made using chi-squared and Wilcoxon.
Results:
342 patients were included in the study with 252 from within the IMC network (IN) and 90 from out of the IMC network
(OON). 163 (47.7%) patients had at least one duplicated CT scan - of those, 26 (10.3%) were IN and 37 (41.1%) were
OON. The total radiology costs for the CT images were $552 IN and $878 OON.
Discussion:
Median total radiology costs, total hospital costs, and total costs for day one of the trauma center hospitalization were
significantly lower for IN patients as compared to OON patients. The greatest cost differential was in those patients
with an ISS < 14. This applied to both total radiology costs as well as to total day one hospital costs.
This difference may be attributable to technological challenges in the transfer of patient data between two different
hospital systems. The duplication rate for the patients transferred within the vertically integrated system was markedly
less, as were the associated costs.
The lack of a significant cost difference in higher Injury Severity Scores likely is due to the critical nature of this
population. These patients often warrant repeat scans for additional information or for change in clinical status.
This study provides more evidence for improvements in data transfer of electronic patient data especially in less
critically injured patients.
Conclusion:
The rate of repeat CT scanning, radiology costs and total costs for day 1 of hospitalization are significantly lower for
trauma patients transferred within a vertically integrated care system, while patients transferred from out of network
incur greater costs attributable to duplication of CT scans.

90
ShorttermClinicalOutcomesafterOffpumpCABGataSingleVAMedicalCenter

EntelaLushaj,MD,PhD,AthanasiaSchreiner,NP,BesaJonuzi,LucianLozonschi,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

EntelaLushaj
(608)2635221
lushaj@surgery.wisc.edu
Scientist
CardiothoracicSurgery
Cardiothoracic
Outcomes
OffPump,CABG,Outcomes,VA,Onpump

Background:Coronaryarterybypassgraft(CABG)procedurehasbeenthemosteffectivetreatmentforischemicheart
disease,traditionallywithcardiopulmonarybypass(CPB)andcardioplegic.ToavoidthesideeffectsofCPBandreduce
theriskofstroke,offpumpCABGtechniquehasevolvedasanalternativestrategytorevascularizetheischemicheart.
Methods:Weretrospectivelystudied260consecutivepatientsattheWilliamS.MiddletonMemorialVeteransHospital,
betweenOctober2007,andSeptember2010.Demographics,shorttermclinicaloutcomesincluding30dayoperative
mortalityandperioperativemorbiditywereevaluated.
Results:Onehundredseventyeight(178)patientsunderwentanoffpumpCABGand82patientsunderwentanon
pumpCABGprocedure.ThereweremorepatientswithCOPD(p=0.04),LVEFof<35%(p=0.02)andsmokers(p=0.01)in
theoffpumpgroup.Comparedtoonpumppatients,noneofouroffpumppatientsrequiredanewmechanicalsupport
device(p=0.01)orhadastroke(p=0.04).OffpumpCABGpatientshadaverylowconversionratetoonpumpsurgeryof
0.6%.Othershorttermendpointssuchas,30daymortality,numberofpatientshavingcardiacarrest,havinginfections
orrepeatcardiacsurgerywasnotsignificantbetweenthe2groups.
Conclusions:Theonpumpconversionrateinourstudyisoneofthelowesteverreportedinaconsecutivepatientseries
orrandomizedcontrolledtrial.VApatientsmaybenefitfromalowerstrokerateifanoffpumpapproachisusedwhile
higherriskpatientsmayavoidthepredictedadverseoutcomeswithonpumpCABG.

91
SurgicalOutcomesofPatientswithLocallyAdvancedNonSmallCellLungCancer

EntelaLushaj,WassimAghnatiosAbiJaoude,RyanMacke,JamesMaloney

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

EntelaLushaj
(608)2635221
lushaj@surgery.wisc.edu
Scientist
CardiothoracicSurgery
ShahabAkhter
Outcomes
Outcomes,Lung,Cancer,Survival,Therapy

Background:ControversycontinuesregardingtheoptimaltherapyforstageIIIanonsmallcelllungcancer(NSCLCa).
Improvedsurvivalhasbeenshowninpatientsundergoingmultimodalitytherapythatincludessurgicalintervention.Our
goalwastoassesstheimpactoflocallyadvanceddiseaseonoperativeoutcomesbasedonSTScriteria.
Methods:WeretrospectivelyreviewedprospectivelycollectedinstitutionalSTSdata.74patientswithstageIIIaNSCLCa
weretreatedsurgicallybetween08/2006and11/2012.Thecohortwasdividedintopatientshavingsurgeryastheir
initialinterventionfollowedbyadjuvanttherapy,andpatientshavingneoadjuvanttherapyfollowedbysurgery.
Demographics,posttreatmentsurvival,complicationsandsurvivalrateswereassessed.
Results:Meanageathospitaladmissionwas64years.VideoAssistedThoracoscopicSurgery(VATS)wasperformedin
17.5%(n=13)ofpatients.Seventythreepercent(73%;n=54)ofpatientshadsurgeryastheinitialtreatment;27%(n=20)
hadneoadjuvanttherapypriortosurgery.Lengthofhospitalstay(LOS)ofpatientshavingneoadjuvanttherapywas
significantlyhigher(p=0.04;mean=53.4days;median=5days)comparedtoLOSofpatientshavingonlysurgery
(mean=3.73days;median=3days).BothgroupshadsimilarpulmonaryfunctionasshownbyFEV1(8021%vs.8321
%;p=0.59)andDLCOvalues(7521%vs.7222%;p=0.58).Nodifferencewasfoundinprolonged(>5days)airleak(15%
inneoadjuvantvs.9.2%insurgerygroup,respectively;p=0.46),reoperationrate(15%vs.4%inneoadjuvantandsurgery
grouprespectively;p=0.11),pneumonia(0%ofpatientsineachgroup)orUTIrates(5%vs.1.8%inneoadjuvantand
surgerygrouprespectively;p=0.45).30dayreadmissionratewas9%(n=5)forsurgeryand10%(n=2)forneoadjuvant
groups(p=0.89).30daysurvivalwas100%inbothgroups.
Conclusions:Pulmonaryresectionasaninitialtherapyorfollowingneoadjuvantradiationandchemotherapyissafefor
patientswithstageIIIaNSCLCa.ThoughLOSwasincreasedintheneoadjuvantgroupcomparedtoinitialsurgical
therapy,locallyadvanceddiseasedoesnotconferincreasedriskofmorbidityormortalityinourstudypopulation.

92
AnatomicSegmentectomyvs.LobectomyforStageINonSmallCellLungCancer
(NSCLC):ExtentofResectionisImportantinDeterminingPreservationof
PulmonaryFunction.

Macke,Ryan,Schuchert,MatthewJ,Odell,DavidD.,Landreneau,JamesR.,McCormick,Kristen.,Pennathur,Arjun;Luketich,
JamesD;Landreneau,RodneyJ

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

RyanMacke
(608)2635215
macke@surgery.wisc.edu
Faculty
CardiothoracicSurgery
JamesMaloney
Outcomes
Lung,Resection,Cancer,Pulmonary,Function

Purpose:AsuggestedbenefitofsublobarresectionforstageInonsmallcelllungcancer(NSCLC)comparedto
lobectomyisarelativepreservationofpulmonaryfunction.Verylittleobjectivedataexistsupportingthissupposition.
Weevaluatetherelativeimpactofbothanatomicsegmentalandlobarresectiononpulmonaryfunctionatoneyearin
patientswithresectedclinicalstageINSCLC.
Methods:Retrospectivereviewof179patients,diseasefreeatoneyear,followinganatomicsegmentectomy(n=94)or
lobectomy(n=85)forclinicalstageINSCLC.TheprimaryoutcomevariablewasthechangeinFEV1andDLCOoneyear
followingsegmentectomyorlobectomy.Inaddition,comparisonswerealsomadebasedonextentofresection(2
segmentsVS.35segmentsORlobectomy)[Table].ComparativeanalysisofPFTresults,includingpreoperativeand
postoperativevalues,aswellasabsoluteandpercentchangewasperformedusingtheWilcoxansignedranktestand
theMannWhitneyUtest.ThestatisticalpackageSTATA,version11.2(CollegeStation,TX)wasusedfortheanalyses.
Results:Overall,anatomicsegmentectomywasassociatedwithasmallerreductioninpulmonaryfunctioncompared
withlobectomy(FEV14.9%vs.7.8%,DLCO3.9%vs.6.5%,FVC5.0%vs.5.7%)atoneyear.Thesefindingswere
primarilyaffectedbythepreservationinfunctionseenamongpatientsinwhom2ORLESSanatomicsegmentswere
removedCOMPAREDTOthosepatientsundergoingresectionof35segmentsORlobectomy.
Conclusions:Pulmonaryfunctionmaybebetterpreservedat1yearwhensegmentectomyislimitedto(2orless)
anatomicsegments.Largertumorsrequiringresectionofgreaterthan3anatomicsegmentsareprobablybesttreated
bylobectomytoachieveadequatesurgicalmarginswhilefulfillingtheobservedoncologicsuperiorityoflobectomyin
theseclinicalcircumstances.

93
MidesophagealandEpiphrenicDiverticula:A15YearExperienceusingMinimally
InvasiveApproachesforSurgicalManagement

:Macke,Ryan,Luketich,JamesD.,Landreneau,RodneyJ.,Pennathur,Arjun,Awais,Omar,Weksler,Benny,Shende,ManishaR,
Christie,NeilA,Schuchert,MatthewJ,Nason,KatieS,Levy,RyanM.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

RyanMacke
(608)2635215
macke@surgery.wisc.edu
Faculty
CardiothoracicSurgery
JamesMaloney
Outcomes
MinimallyInvasive,esophageal,diverticula,Outcomes,Surgical

Purpose:Midesophagealandepiphrenicesophagealdiverticulaareuncommon.Unfortunately,littleconsensusexists
regardingthemanagementofthesepatients.Indicationsforrepair,optimalapproach,andproceduresincludedarea
fewofthecontroversiesthatexistregardingthesurgicalmanagementofthesepatients.Wecurrentlyfavoraminimally
invasiveapproachforpatientswithsymptomaticdiverticula.However,avarietyofapproacheshavebeenused.The
purposeofthecurrentstudywastoreviewtheresultsofarelativelylargecohortofpatientswiththeserarediverticula
thatweretreatedsurgically.
Methods:Sixtypatientsovera15yearperiodunderwentsurgicaltreatmentofsymptomaticdiverticula.Medianagewas
70.2years.Diverticulawerelocatedinthedistalthirdoftheesophagus(53/60),midesophagus(6/60),andboth
locations(1/60).Ofthosepatientswiththenecessaryinformation,96%(43/45)hadsomeformofesophagealmotility
disorder.Themostcommonsymptomsweredysphagia(46/60),regurgitation(37/60),andheartburn(23/60).
Results:ApproachesincludedVATS(34/60),laparoscopic(19/60),andcombinedlaparoscopic/VATS(7/60).Conversion
toanopenprocedureoccurredin4patients(1laparotomy,3thoracotmies).Proceduresperformedincluded
diverticulectomy/myotomy(29/60),diverticulectomy/myotomy/fundoplication(23/60),anddiverticulectomyalone
(7/60).Complicationsoccurredin23patients(38%).Postoperativeleaksoccurredin12patients,withonly2requiring
operativeintervention.Inhospital/30daymortalitywas1.7%(1/60).
Conclusions:Minimallyinvasiveapproachestomidesophagealandepiphrenicdiverticulaarefeasibleinthehandsof
experiencedesophagealsurgeons.However,theseoperationsarenotwithoutrisk.Theoptimalapproachand
proceduresperformedshouldbedeterminedonanindividualizedbasisandrequiresathoroughpreoperative
investigationtobestunderstandthecauseofthediverticula.

94
AdolescentSharedDecisioninPlasticandReconstructiveSurgery

McCarthy,J.,Mount,D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

jamesmccarthy
(802)2998774
jmccarthy@uwhealth.org
Resident
Plastic&ReconstructiveSurgery
DelMount
Outcomes
shareddecisionmaking,pediatric,decisionaid,plasticsurgery,evidencebased
medicine

Intro:Shareddecisionmakingistheprocessbywhichpatientsnavigatecomplextradeoffsinmakingmedicaldecisions
withtheirprovider.Thereisapaucityofliteratureonhowadolescentsparticipateintheshareddecisionmaking
process.Theobjectivesofthisstudyweretoinvestigateshareddecisionmakinginadolescentsandhowthisprocess
changeswithincreasingagebyanalysisofdecisionalvalues,decisionalconflict,anddecisionalselfefficacy.
Methods:Adolescentsbetweentheagesof1218wererecruitedfromoutpatientplasticsurgeryclinicsforallproviders
inthedivisionofplasticandreconstructivesurgerytocompletethreevalidatedsurveysfromtheOttowaDecision
SupportFramework.Summaryresponseswerethenrecordedinadditiontodifferencesbetweengenderandage;atwo
samplestudent'sttestwasusedtoassessforsignificantdifferencesbetweengenderanyounger(<15years)andolder
(>15years)adolescents.
Results:Atotalof14participantsrespondedtosurveysbetween2012present.Theaverageagewas14.3yearswith
50%male.Thevastmajorityofrespondents(92%)wereunsureabouttheirdecisionwhenanalyzingthedecisional
conflictscale(score>37.5);onerespondent(8%)hadnodecisionalconflict(score<25).Theaverageselfefficacy
("confidence")inmakingdecisionwas83(+/3.9)with0havingextremelylowconfidenceindecisionand100having
extremelyhighconfidence.Decisionalvaluesdemonstratethatphysiciancharacteristics,riskofinfectionandbenefitof
potentiallyimprovedoverallappearancewherethehighestrankedvalues,respectively.Therewerenosignificant
differencesinanydomainbetweengenderoragecategory.
Conclusion:Adolescentspresentingforelectivesurgicalproceduresdemonstratehighdecisionalconflict,butproceed
withgreatconfidenceabouttheirabilitytomakedecisions,andmostgreatlyvaluephysicianpersonalcharacteristics.
Therearenosignificantdifferencesintheshareddecisionmakingprocessbetweengenderorageinadolescent
development.

95
RoboticIvorLewisEsophagectomyAfterNeoadjuvantChemoradiationTherapy

SmithFOMD,AlmhannaKMD,ShridharRMDPhD,HoffeSMD,KarlRCMD,andMeredithKLMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

KennethMeredith
(608)2622025
mereditk@surgery.wisc.edu
Faculty
GeneralSurgery
SurgicalOncology
Outcomes
Robotic,Esophagectomy,Neoadjuvanttherapy,IvorLewis,Outcomes

Background:Neoadjuvantchemoradiationtherapy(NCR)hasbecomestandardofcareforpatientswithlocallyadvanced
esophagealcancer.Inselectedpatients,roboticassistedIvorLewisEsophagectomy(RAIL)isasafeandfeasible
operativestrategyinthemanagementofesophagealcancer.Thisstudywasdesignedtodeterminepotentialdifferences
inperioperativemorbidityandshorttermoutcomesinpatientswithesophagealcancertreatedwithRAILwithor
withoutNCR.
Methods:Aretrospectivereviewwasconductedconsistingofconsecutivepatientswithesophagealcancerwho
underwentRAILesophagectomybetweenOctober2010andJune2012withandwithoutNCR.Clinicalandpathological
variableswereanalyzedwithtwosidedstudentttestassumingequalvariance.Datawereconsideredsignificantatap
value<0.05.
Results:EightyninepatientsunderwentRAILduringthestudyperiod.Seventysevenpatients(87%)receivedNCRand
twentytwopatientsdidnot(13%).Themedianagewas66yearsandthemedianBMIwas28kg/m2.Allpatientshada
R0resection.Therewerenodifferencesinthemeanestimatedbloodloss(149vs.153mL;p=0.52)andmeanoperative
times(434vs.427minutes;p=1.0).Therewerenodifferencesintheincidenceofpneumoniaoratrialfibrillation,
lengthsofstayintheICU,orlengthofhospitalization.Intotal,therewere2(2.3%)anastomoticleaksandoneleakfrom
thegastricconduit.TheanastomoticleaksoccurredinthegroupthatdidnotreceiveNCRandthegastricconduitleak
occurredinthegroupthatreceivedNCR.Therewerenomortalitiesineithergroup.Thecompletepathologicalresponse
rateinthegrouptreatedwithRAILwas31%.Therewasnodifferenceinthemeannumberoflymphnodesharvestedin
theNCRgroup(2211vs.208,p=0.41).
Conclusions:RoboticassistedIvorLewisEsophagectomycanbesafelyperformedfollowingneoadjuvantchemoradiation
therapy.Inthisseriesthereweresimilarperioperativemorbidityandshorttermmortalityoutcomesinpatientswho
receivedNCRcomparedwithroboticassistedIvorLewisalone.Additionalfollowupisrequiredinordertodetermine
longtermoncologicoutcome.

96
RoboticassistedIvorLewisesophagectomyintheelderlypatient

AndreaMAbbottMD,TobinStromMD,NadiaSaeed,RaviShridharMD,SarahHoffeMD,KhaldounAlmhannaMD,Kenneth
MeredithMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

KennethMeredith
(608)2622025
MeredithK@surgery.wisc.edu
Faculty
GeneralSurgery
SurgicalOncology
Outcomes
Robotic,Esophagectomy,Elderlypatient,Outcomes,IvorLewis

Esophagealcancercontinuestoincreaseinincidenceworldwidewiththeageofdiagnosiscontinuingtomovetowards
anolderonset.Roboticassistedapproachestoesophagectomyhavedemonstrateddecreasedcomplicationsandlength
ofhospitalization(LOH).Wesoughttoexaminetheimpactofageonoutcomesinpatientsundergoingroboticassisted
esophagectomy(RAIL).

From20092013,weidentifiedpatientsundergoingroboticassistedIvorLewisesophagectomy.Patientswerethen
stratifiedaccordingto3agegroups.Cohort1,agelessthen50,cohort2,age5070,andcohort3>70.Statistical
comparisonsbetweenLOH,operativetime(OT),estimatedbloodloss(EBL),adverseevents(AE)andmortalitywere
madewithfisherexacttest.
Weidentified134patientswhounderwentRAIL:cohort1(n=12),cohort2(n=67),andcohort3(n=55).Weidentified
nostatisticallysignificantdifferencebetweenthethreecohortsforOT,LOH,daysspentinintensivecare,AEor
mortality.TherewasadifferenceinEBLwithhighermedianbloodloss(150cc)seenincohort1(50600cc)and3(50
400cc)comparedtocohort2(100cc,(25400cc)),p<0.01.TheoverallAEratewas22%.Themostcommon
complicationswereatrialfibrillation(AF)n=13(9.7%)andpneumonian=10(7.5%)butthiswasnotsignificantlydifferent
betweenthecohorts.TheAEratewasn=1,8.3%(cohort1),n=20.9%(cohort2),andn=14,34.5%(cohort3),p=0.14.
Eightofthe14AEincohort3wasAFcomparedtocohort2,n=5,andcohort1,n=0.Therewere4leaks(p=0.38)and2
deaths(0.90)intheentirecohort.Aseparateanalysiswasdonetocompareelderly(>70)tothenonelderly(<70).
MedianEBLwashigherintheelderlycohort(100cc(25600)vs150cc(50400),p<0.01).Therewasatrendtowards
longerLOHintheelderly(9(435)vs11(638)days,p=0.06).AEandmortalitywerenotsignificantlydifferent,although
therewasatrendtowardincreasedAE(p=0.07)intheelderly,withAFbeingthemostcommonAE.
RAILisasafesurgicaltechniqueforuseinanagingpatientpopulation.Wedemonstratedtherewasnoincreasedriskof
LOH,AEordeathintheelderlypatientscomparedtotheiryoungercohort.Thereisanassociatedincreasedriskfor
atrialfibrillationinthepatientsovertheageof70,howeverthisincidenceislowerthenhistoricalseries.

97
TheSurgicalApgarScoreCorrelateswithanIncreasedRiskforReadmissionin
EmergencySurgeryPatients

GajanthanMuthuvel,BS,SarahE.Tevis,MD,SureshK.Agarwal,MD,GregoryD.Kennedy,MD,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

GajanthanMuthuvel
(920)5271249
gmuthuvel@wisc.edu
MedicalStudent
GeneralSurgery
GregoryD.Kennedy
Outcomes
outcomes,emergencysurgery,readmissions,riskfactors,surgicalApgarscore

Introduction:Preventablereadmissionhasbecomeanationalfocus.Itisclearthatsurgicalpatientspresentspecific
challengestothoseinterestedinpreventingreadmission.Patientsundergoingemergencysurgicalproceduresareat
particularlyhighriskforreadmissionandverylittleisknownaboutthiscomplexoutcomeinthispatientpopulation.We
areinterestedintryingtodetermineifwecanusereadilyavailabledatavariablestopredictriskofreadmission.The
surgicalApgarscore(SAS)iscalculatedfromintraoperativevariablesthatarereadilyavailable,andhasbeenstudiedin
thenonemergentsettingandhasbeenshowntobepredictiveofpostoperativemortality.Theobjectivesofthisstudy
weretocharacterize30dayreadmissionsinemergentgeneralsurgeryandtodeterminewhethercertainvariablesor
measureswereassociatedwithreadmissions.WehypothesizedthattheSAScorrelateswithriskforreadmissioninthe
emergencygeneralsurgerypatientpopulation.
Methods:VariablesofinterestwereobtainedfromretrospectiveanalysisoftheUniversityofWisconsinNSQIPdatabase
inadditiontotheelectronicmedicalrecord.Weidentifiedadultgeneralsurgerypatientswhounderwentanemergency
procedurefrom20062012.Univariateanalysisidentifiedfactorsassociatedwith30dayreadmission.Factorswith
p<0.1wereincludedinmultivariateanalysistorevealpotentialriskfactorsforreadmission.
Results:Ascomparedwithnonemergencysurgerypatients,patientswithemergentprocedureshadahigherrateof
readmission(11.1%vs.15.2%,p=0.004).TheSAS(OR3.297,CI1.07410.121,p=0.037),andthecombinedvariableof
theAmericanSocietyofAnesthesiologistsPhysicalStatusClassification(ASAclass)andhospitallengthofstay(OR4.370,
CI2.2518.486,p<0.001),werehighlyassociatedwith30dayreadmissionsinadultemergencygeneralsurgerypatients.
Conclusions:Giventhesignificanceofreadmissionafteremergencysurgery,itisimportanttoestablishmeasuresthat
identifypatientsathighriskforreadmissionbeforetheyaredischarged.Wehaveidentifiedobjectivemeasuresthat
allowforthestratificationofpatientsintolowandhighriskgroups.Patientsfoundtobehighriskcouldbegiven
modifiedpostoperativecarethatmayincludeadditionaleducation,dischargetoadestinationwithahigherstandardof
care,orcloserfollowup.Furthermore,thestratificationofpatientswillenablethestudyofprospectiveinterventions
designedtodecreaseunplannedreadmissionsinemergencysurgerypatients.

98
DONTJUSTTELLSOMEBODYTHEYNEEDTHISSURGERY:AQUALITATIVE
STUDYOFPATIENTDECISIONMAKINGPREFERENCESINHIGHSTAKESSURGICAL
DECISIONS

MJNaboznyMD,JMKehlerMD,NMSteffensMPH,KJBraselMD,MEGainesJD,TCCampbellMD,MLSchwarzeMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MichaelNabozny
(518)9296126
mnabozny@uwhealth.org
Resident
GeneralSurgery
Schwarze
Outcomes
HighRiskSurgery,Decisionmakingpreferences,Palliativecare,Geriatrics,Vascular
Surgery

PURPOSE:OlderAmericanswouldoftenprefertolimitaggressivecareattheendoflife;howeverlittleisknownabout
howpatientswouldprefertomakesurgicaldecisionsinalifethreateningsituation.Theobjectiveofthisstudywasto
evaluatehowolderpatientsevaluatetheoptionofhighrisksurgerywhenthealternativeispalliativecare.
METHODS:Wepurposefullysampledfourfocusgroupsofsociodemographicallydiverseseniors(age60orgreater)in
Wisconsinwhohadbeenfacedwithadifficultmedicaldecisionforthemselvesoralovedone(n=37.)Participants
watchedasevenminutevideoclipshowingasurgeoncommunicatingtreatmentoptionstoafrail,elderlywomanwitha
tenderbutnotrupturedthoracoabdominalaneurysm.Thesurgeonusedadecisionsupporttooltohelpthepatient
understandthechoicebetweensurgeryandpalliativecare.Aprofessionalmoderatorfacilitatedeachgrouptoelicit
understandingandcontrolpreferencesforsurgicaldecisionswithgraveconsequences.Weusedinductivecontent
analysistoqualitativelyanalyzethetranscribedaudiorecordingofeachfocusgroup.
RESULTS:Seniorsreportedthatchoosingbetweensurgeryandsupportivecarewasextraordinarilydifficultandsucha
decisionwouldrequiretimeandassistancefromfamilyandotherhealthcareproviders.Seniorsexpressedshockand
particulardistastethatbothsurgeryandpalliativecarehadpooroutcomeswhichledthemtodistrustthefactual
informationprovided.Seniorsgenerallywishedtohavecontroloverdying,Ithinkthatdeathisnaturalforeverybody
everyonewouldliketohavesomecontroloverhowthey'regoingtodieandtobegivenachoiceaboutwhat
treatmentstopursuewhendeathwaslikely.Seniorsuniversallyreportedthatqualityoflifewasparamountand
overwhelminglyendorsedthatlongtermnursinghomeresidence,orlossofindependence,wasanoutcomeworsethan
death.Nonetheless,severalseniorspreferredtochoselife(surgery)notingthatitwouldbebettertodietryingand
havefaithinGodandyourdoctor.
CONCLUSION:Seniorswanttoassertcontroloverhowtheydiebutstruggletobelievethatmajorsurgeryinafrail
elderlypatientmighthaveonlyundesirableoutcomes.Althoughqualityoflifewasinvokedastheguidingprinciplefor
treatment,seniorswerebotheredthatpalliativecare,andcertaindeath,wasevenofferedasachoice.Thisstudy
highlightsthechallengessurgeonshavehelpingpatientsmakeacutehighstakesdecisionsaboutaggressivetreatment
withfrailelderlypatients.

99
Areallthyroidultrasoundevaluationscreatedequal?Sonographersspecialized
inthyroidcancercorrectlylabelclinicalN0diseaseinwelldifferentiatedthyroid
cancer

SarahC.Oltmann,M.D.,DavidF.Schneider,M.D.,M.S.,HerbertChen,M.D.,RebeccaS.Sippel,M.D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SarahOltmann
(214)4181413
oltmann@surgery.wisc.edu
PostdoctoralFellow
GeneralSurgery
Sippel
Outcomes
ThyroidCancer,Recurrence,Ultrasound,Outcomes,LymphNodeStatus

Background
Cervicalultrasound(US)isastandardpartofpreoperativeevaluationofthyroidcancerpatients.USabilitytoidentify
lymphnode(LN)diseaseinthecentralneckisdebated,leadingsomesurgeonstoutilizeprophylacticcentraldissection
topreventearlydiseaserecurrence.TheaimofthisstudywastoevaluateifUSevaluationbyadedicatedsonographer,
specializinginthyroiddiseasecouldcorrectlydetermineclinicalN0status.
Methods
RetrospectiveanalysisofaprospectivedatabaseofcN0thyroidcancerpatients.Exclusioncriteriaincludedpatients
undergoingLNdissectionattimeofthyroidectomyandthosewithoutdocumentedpreoperativeUS.Patient
demographicsandoutcomeswerereviewed.PatientswerecategorizedbywhoperformedthethyroidUS(asingle
thyroidsurgeonvs.nonsurgeonsonographer).Recurrencewasneedforadditionalradioactiveiodinetreatmentsor
subsequentpositivepathology.
Results
From2005to2012,178patientsmetcriteria.48patientshadsurgeonperformedUSversus130patientswithnon
surgeonperformedUS.Groupswereequivalentinage,gender,andtumorsize.46%ofallpatientshadapreoperative
diagnosisofcancer,whiletheremainingpatientshadbenign(19%)orindeterminate(35%)diagnoses.SurgeonUS
assessedLNstatusmorefrequently(69%vs.20%,p<0.01).Radioactiveiodinetreatmentanddosingwereequivalent.
RadioactiveiodineuptakefollowingwaslowerinsurgeonUS(0.02%0.02vs.0.15%0.03,p<0.01).Recurrencerates
werehigherinnonsurgeonUS(11%vs.0%,p=0.01).USevaluationincludingLNsversusnoLNevaluationdidnotresult
indifferingrecurrencerates(p=0.67).Mediantimetorecurrencewas13months.Meanfollowupwaslongerinthe
nonsurgeonUS(242monthsvs.402,p<0.01),howeverallfollowupencompassedthetimeatwhichallrecurrences
occurred.
Conclusions
PerformanceofanUSbyasonographerspecializinginthyroidcancerensuresoptimumassessmentandcorrectly
identifiespatientsasN0whichmayeliminatetheneedforprophylacticLNdissectionandnotincreasetheriskofearly
recurrence.Asnotallthyroidcancersarediagnosedpreoperatively,USexaminationofthethyroidshouldinclude
routineevaluationofthecervicalLNstations.

100
Lung Transplant Outcomes for High-Risk Patients (LAS50) with Idiopathic
Pulmonary Fibrosis (IPF)
Satoru Osaki, Keith C Meyer, James Maloney, Richard D Cornwell, Nilto C De Oliveira

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Satoru Osaki
(608) 263-0439
osaki@surgery.wisc.edu
Faculty
Cardiothoracic Surgery
Shahab Akhter
Outcomes
Lung, Transplant, IPF, Outcomes, Survival

Purpose:
To evaluate our institutional experience in lung transplantation for high-risk IPF patients as defined by high LAS scores.
Methods:
We retrospectively reviewed 91 consecutive IPF patients who underwent lung transplantation (LTX) at our institution
between May 2005 and October 2012. High risk IPF patients with LAS50 (high-LAS, n=17) were compared to patients
with LAS<50 (low-LAS, n=74).
Results:
High-risk IPF patients had more severe pulmonary dysfunction (%FVC; 37 [high-LAS] vs. 52 [low-LAS], and %FEV1; 42 vs.
55%, p<0.01), required more supplemental oxygen (10 vs. 4 L/min, p<0.01), and 4 (24%) of them were intubated. The
median LAS was significantly higher (73.7 [range, 54.1 - 94.2] vs. 40.7 [32.1 - 49.6], p<0.01) and the waiting time was
shorter (29 vs. 73 days, p=0.03) in the high-LAS group. The incidence of post-LTX pulmonary complications was higher
for the high-LAS group (length of mechanical ventilator support; 3 vs. 1 days, p<0.01, re-intubation; 35 vs. 12%, p=0.03;
tracheostomy; 29 vs. 3%, p<0.01). However, 30-day mortality (0 vs. 4%, p=0.99), hospital mortality (6 vs. 4%, p=0.57),
and overall post-LTX survival did not differ between the groups (Figure), although more early deaths occurred in the
high-risk group.
Conclusions:
Lung transplantation in high-risk IPF patients is associated with increased post-LTX pulmonary complications versus IPF
patients who are less ill (as reflected by lower LAS values). However, long-term, post-LTX survival for carefully selected,
high-risk patients was equivalent to that for lower-risk recipients transplanted at our center. However, management of
high-risk IPF patients is quite challenging both before and after LTX.

101
LeftVentricularAssistDevicePlacementinVeryObesePatients:Proceedwith
Caution

SatoruOsaki,MarylRJohnson,PeterS.Rahko,MargaretAMurray,JohnH.Blabaum,EntelaB.Lushaj,SueUlschmid,Lucian
Lozonschi,TakushiKohmoto

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SatoruOsaki
(608)2630439
osaki@surgery.wisc.edu
Faculty
CardiothoracicSurgery
ShahabAkhter
Outcomes
LVAD,Obese,Outcomes,Complications,Survival

Purpose:
Toevaluateourexperienceinleftventricularassistdevice(LVAD)placementinobesepatients(PTS).
Methods:
Weretrospectivelyreviewed148consecutivePTSwhounderwentLVADplacementatourinstitutionAug2003toSep
2012.Thecohortwasdividedintothenonobese(bodymassindex[BMI]<30,n=89)andtheobese(BMI30,n=59)
groups.Demographicsandoutcomeswereassessed.
Results:
ThemeanweightandBMIwere79(nonobese)vs.106kg(obese)and25vs.34kg/m2,respectively.PTage,diagnosis,
cardiac,pulmonaryandrenalfunctions,anddevicetypedidnotdifferbetweenthegroups.However,obesePTShad
higherprevalenceofdiabetes(58vs.38%,p=0.02),preoperativecereberovascularaccident(14vs.3%,p=0.03),and
moredestinationtherapy(37vs16%,p<0.01).Lengthofhospitalstay(11vs.10days,p=0.59),reoperation(6vs.7%,
p=0.77),andlengthofdevicesupport(184vs.164days,p=0.44)weresimilarinnonobesevsobesePTS.However,
obesePTShadhigherincidenceofVADinfection(29vs.14%,p=0.02),sepsis(17vs.6%,p=0.03),andVADfailure(25vs.
12%,p=0.04).Patientsurvivalwassimilarinthegroups(p=0.50,Figure).However,2/4(50%)severelyobese(BMI>40%)
LVADPTSrequiredLVADexchangeforfailureandexpiredin3monthsafterLVADplacement.
Conclusions:
OurresultsdemonstratethatthesurvivalinobesepatientswithLVADsupportwasequivalenttothatofnonobese
patients,however,obesepatientshadhigherincidenceofinfectiouscomplicationsandfailureofLVAD.Obesepatients
shouldbecarefullyselectedbeforeLVADplacement,especiallyseverelyobesepatientsfordestinationtherapy.

102
Expanding the Use of Free Text in EMR to Study Breast Cancer
Christopher M Dodgion MD, MSPH, MBA; Thien Nguyen BS; Marquita R Decker MD, MPH; Heather B Neuman MD, MS; Wei Jiang MS;
Yue-Yung Hu MD, MPH; Anita Karcz MD, MBA; Stuart R Lipsitz ScD; Leonard DAvolio PhD; Caprice C Greenberg MD, MPH

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Christopher M Dodgion
(608) 263-3076
pavuluri@surgery.wisc.edu
Other
General Surgery
Caprice C Greenberg
Outcomes
Free Text Documents, Electronic Medical Record, Breast Cancer, Machine Learning,
Community Hospitals

Introduction:
Data sources to study breast cancer care in large populations are limited by access to cancer-specific variables. We
sought to address this gap by developing a validated, reusable approach to extract cancer-specific variables from the
free text of electronic medical records (EMR) in a large cohort of US community hospitals.
Study Design:
Free text documents were received as a single XML file per patient, which was parsed into individual documents
(N=103,106). Randomly selected documents (N=1,597) were reviewed and classified by two independent surgical
reviewers, with a breast surgical oncologist adjudicating disagreement. A portion of the adjudicated data set was used to
train an open-source machine learning-based software to identify all breast cancer related operative reports, clinic
notes, and pathology reports. The software was also used to further classify operative reports as diagnostic or
therapeutic. Extraction of cancer-related variables from those documents was performed using a rules-based approach.
Training sets of 200 pathology and 150 clinic notes, and test sets of 50 of each, were randomly selected. Two
independent reviewers coded 8 concept-level variables with a similar adjudication process. Inter-rater reliability (IRR)
was assessed using Cohens kappa. Computer performance was assessed using recall, precision, and f-measure.
Study Population:
An analytic cohort of 5,194 patients (11,022 encounters) from a nationwide cohort of 59 community hospitals was
identified. Each patient underwent a breast procedure between January, 2006 and December, 2009 and had an available
electronic operative report.
Results:
Classification of breast cancer-related document: The abstractor IRR was 0.96 for identification of pathology reports,
0.86 for clinic notes, and 0.91 for operative notes. Computer performance was strong with recall of 0.95, 0.96, and 0.91;
precision of 0.95, 0.88, and 0.85; and f-measure of 0.94, 0.91, and 0.87 for pathology, clinic, and operative notes
respectively. A total of 2,055 pathology reports, 25,714 clinic notes, and 15,131 operative reports were ultimately
identified. The computer identification of therapeutic operative notes showed similar high performance (0.96 recall,
0.93 precision, 0.94 f-measure).
Concept-level classification: IRR between manual reviewers for clinic notes ranged from 0.94 for tumor (T) and nodal (N)
stage to 0.97 for tumor grade and 1.0 for estrogen receptor (ER) status with similar performance for pathology reports.
Computer performance was more variable. Recall was high (1.0) for T and N stage, and grade, but lower for AJCC stage
(0.95), ER (0.86), PR (0.90), and Her-2-neu (0.84) status. Precision was high (1.0) for T stage, PR, and Her-2-neu status,
but lower for N stage (0.92), AJCC stage (0.88), and grade (0.75). F-measures ranged from 0.86 for grade to 1.0 for T
stage.
Conclusions:
We have demonstrated the ability to reliably abstract several cancer-specific variables from free text of EMR notes in a
cohort of US community hospitals. With further refinement, we anticipate improvement in performance for the others.
This new data source can be used for future comparative effectiveness and other health services research in breast
cancer. Given our large, diverse set of hospitals, the approach is scalable, generalizable, and can be modified for future
research questions.

103
SurgicalCoachingtoImprovePerformance

CapriceCGreenbergMDMPH,HalaNGhousseiniPhD,CarlyEGlarnerMDMEd,SudhaRPavuluriQuammeMDMS,HeatherL
BeasleyBA,DouglasAWiegmannPhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SudhaPavuluriQuamme
(608)2633076
pavuluri@surgery.wisc.edu
Other
GeneralSurgery
CapriceGreenberg/WiSOR
Outcomes
SurgicalCoaching,ProfessionalDevleopment,PracticeImprovement,Education,
Framework

INTRODUCTION
Thecurrentapproachtocontinuousprofessionaldevelopmentandpracticeimprovementinhealthcareislimited.
Principlesofadultlearningtheoryandprovenapproachestocontinuedperformanceimprovement,suchasdeliberate
practice,arenotapplied.Toaddresstheselimitations,thefieldisincreasinglyturningtocoaching.Todate,coachinghas
mainlybeenutilizedinprimarycare,focusingonteamdynamics.However,procedurebaseddisciplinesmaybeeven
moreamenabletothisapproach.Thegoalofthisresearchwastodevelopageneralizableconceptualmodeland
frameworkforcoachinginprocedurebasedhealthcare.
METHODS
Acomparativecasestudyidentifiedfeaturesofcoachinginothertechnicaldisciplinesathletics,music,education
thatemploycoachestopromotecontinuedperformanceimprovement.Semistructuredinterviewswereconducted
withcoachesineachdiscipline,andfieldorvideobasedobservationofcoachingsessionswasperformed.Areviewof
theresearchandprofessionalcoachingliteraturewasalsoconducted.
RESULTS
ASurgicalCoachingFrameworkwascreatedthatincorporatesthecoreprinciplesofcoachingandcaninformthe
developmentofcoachinginterventionsinsurgeryandotherprocedurebasedhealthcare.Thesurgeonsexperience,skill
level,anddispositionandthesettinginwhichtheyoperate(context)mustbeconsideredindeterminingtheoptimal
approachtocoaching.Thereisremarkableagreementacrossdisciplinesregardingthecharacteristicsofaneffective
coachthatshouldguideselection.Interpersonalskillsarecritical,specificallyinrelationtoadaptabilityanda
cooperativecommunicationstyle.Coachesmustbewellrespectedbytheindividualswithwhomtheyareworkingin
ordertobeeffective.Thecontentofeachsessionshouldfocusonthethreebasicdomainsofsurgicalperformance:
technicalskill,nontechnicalskill,andcognitiveskill.Thesessionsshouldincorporateadefinedsetofactivities,including
settinggoals,encouraging/motivating,anddeveloping/guiding.Finally,thecoachingcontext(videovs.live,peervs.
expert)mustbedesignedtooptimizetheinteractionbetweenthecoachandthesurgeon.
CONCLUSIONS
TheAmericanBoardofSurgeryhasincludedlifelonglearningandselfassessmentandevaluationofperformancein
practiceasrequirementsformaintenanceofcertification,yetevidencebasedapproachestosuchactivitiesremain
elusive.Surgicalcoaching,asdescribedhere,providesoneinterventionthatcouldmeettheserequirements.

104
Institutional Variation in Surgical Care for Breast Cancer at Community Hospitals
Marquita R Decker MD, MPH; Stuart Lipsitz ScD; Christopher M Dodgion MD, MSPH, MBA; Yue-Yung Hu MD, MPH; Thien Nguyen;
Wei Jiang MS; Heather B Neuman MD; Anita Karcz MD, MBA; Leonard DAvolio PhD; Caprice C Greenberg MD, MPH

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Sudha Pavuluri Quamme


(608) 263-3076
pavuluri@surgery.wisc.edu
Other
General Surgery
Caprice Greenberg/WiSOR
Outcomes
Breast Cancer, Institutional Variation, Surgical Care, Community Hospitals, Electronic
Medical Records

INTRODUCTION
Variation in breast cancer care can reflect patient characteristics, access to care, as well as patient and provider
preferences. We used a novel data set which allows for the automated abstraction of data elements from the free text
in electronic medical records to investigate variation in the surgical treatment at community hospitals, an under-studied
sector of the healthcare system where most breast cancer care is provided.
METHODS
Data on patients who underwent breast cancer surgery from 2006 to 2009 in a large, geographically diverse national
cohort was obtained from the Institute for Health Metrics. Cancer-specific variables were abstracted from pathology and
clinic notes using computer-based algorithms for the natural language processing of free text. Patient zip code was used
to link to U.S. Census Data and to calculate the distance traveled to the closest radiation facility based on the address of
all radiation facilities in the United States. The National Provider Identifier file, the
Area Resource File, and data from the American Hospital Association were used to identify structural characteristics for
each institution and density of providers in each health service area. Polymotous logistic regression with multi-level
outcomes (breast conserving surgery, mastectomy, or mastectomy with reconstruction) was used to identify
associations between these variables and surgical treatment.
RESULTS
At 59 community hospitals, 5,194 patients underwent breast conserving surgery (BCS), mastectomy, or mastectomy with
reconstruction. Patients from rural areas, when compared to patients from metropolitan areas, were less likely to
undergo reconstruction (p=0.005). Patients with advanced nodal stage were more likely to undergo mastectomy without
reconstruction, compared to patients with N0 disease who were more likely to undergo BCS (p<0.001). Estrogen and
progesterone receptor negative patients, as well as patients with Her2neu postive disease, were significantly more likely
to undergo mastectomy or mastectomy with reconstruction rather than BCS (ER negative: p< 0.001; PR negative:
p=0.001; HER2neu positive: p = 0.01). Patients underwent reconstruction more frequently when there were more than 4
reconstructive surgeons per 100,000 people in the health service area (p= 0.03). Patients underwent BCS more
frequently when radiation oncology services were available within the index hospital or health system (p=0.04).
CONCLUSIONS
Disease characteristics as well as the local availability of services influence the type of surgery that patients undergo for
treatment of breast cancer. This study illustrates the multi-faceted and complex decision-making required for the
surgical treatment of breast cancer. These factors must be considered to distinguish warranted variation from unwanted
and unwarranted variation and ensure appropriate care for all patients. Increasingly sophisticated data sources, as
demonstrated here, are critical for such studies.

105
OutpatientCareAloneisNotSufficienttoPreventReadmissionAfterVascular
Surgery

R.ScottSaunders,MD;SaraFernandesTaylor,PhD;JasonT.Wiseman,MD;TravisL.Engelbert,MD,JonS.Matsumura,MD,K.
CraigKent,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ScottSaunders
(847)2179030
saunders@surgery.wisc.edu
PostdoctoralFellow
VascularSurgery
K.CraigKent
Outcomes
vascularsurgery,readmission,rehospitalization,followupvisits,caretransitions

Purpose:
Reducingearlyhospitalreadmissionisamajorfocusofcurrenthealthcarereform.Healthcareprovidersand
administratorsarepursuingclinicalinterventionstoreducereadmissionrates.Earlyoutpatientfollowup,apost
dischargeintervention,hasdemonstratedreducedriskof30dayreadmissioninmedicalpatients,buthasnotbeen
evaluatedinanysurgicalpopulation.Ourstudyexaminestheassociationbetweenoccurrenceofameaningful
outpatientclinicvisitandsubsequentreadmissionfollowinginpatientvascularsurgicalprocedures.
Methods:
Wequeriedthemedicalrecordatalarge,tertiarycareinstitutionfromJuly2008throughDecember2012todetermine
ratesofoutpatientfollowupand30dayreadmissionforadult(18yearsoldorgreater)patientswhounderwentany
inpatientvascularsurgicalprocedure.
Results:
2,480admissionsforvascularsurgeryoccurredfrom20082012,and244(9.8%)resultedinreadmissiontothesame
institutionwithin30days.Overall,1,895(76.4%)inpatientencounterswerefollowedbyameaningfuloutpatientvisitat
thesameinstitutionwithin30daysfollowinginitialdischarge.73.4%(179of244)ofencountersleadingtoreadmission
receivedoutpatientcarefollowingdischargeandpriortoreadmission,while76.7%(1716of2236)ofencountersnot
requiringreadmissionexperiencedoutpatientfollowupwithin30daysofinitialdischarge(Pvalue=.24).Additionally,
wecomparedthetimebetweeninitialhospitaldischargeandthefirstoutpatientvisit(timetofollowup)forinpatient
encounterswithandwithoutsubsequentreadmission.Timetofollowupwassignificantlyshorterforinpatient
encountersresultinginreadmission10.1+/6.2daysversus12.5+/6.5days(Pvalue<.001).
Conclusions:
Outpatientcareoccurredfollowingthevastmajorityofinpatientvascularsurgeryencountersresultinginearlyhospital
readmission.Thissuggeststhatpatientswhoexperienceapostoperativeeventutilizetheoutpatientclinictoseek
furthercarepriortorehospitalization.Consideringthatthetimetofollowupissignificantlyshorterpreceding
readmissionsandthatthereisnodifferenceintherateofutilizationoftheseresources,ourdatasuggestthataccessto
earlyoutpatientcarealoneisnotsufficienttopreventreadmission.

106
PatientReportedReadinessforDischargeisAssociatedwithHigherPatient
SatisfactionandLowerReadmissionRates

RyanK.Schmocker,MD,SaraMurryMD,XiaVangBA,GlenLeversonPhD,LindaCherneyStaffordMPH,EmilyR.Winslow,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

RyanSchmocker
(312)3209024
rkschmocker@gmail.com
Resident
GeneralSurgery
EmilyWinslow
Outcomes
PatientSatisfaction,Readmission,HCAHPS,HospitalSatisfaction,Physician
Satisfaction

Background:WiththeintroductionoftheAffordableCareAct,patientreportedoutcomeshavebeenincreasingly
emphasized.Further,newfinancialpenaltieshaveencouragedeffortstodecreasetherateofreadmission.Current
effortstolimitreadmissionhaveprimarilyaimedtoidentifyhighriskgroupsbasedonclinicalandsocialfactors.Previous
reportshaveshownthatanursingassessmentofapatientsreadinessfordischargecorrelateswithreadmissionrates;
thereforewesoughttodetermineifpatientsselfreportedreadinessfordischargewouldbeusefulasapatient
reportedoutcomeineffortstodecreasereadmission,anddeterminehowreadinessfordischargeimpactspatient
satisfaction.
Methods:Usingapreviouslyestablisheddatabasecontainingbothclinicalinformationandhospitaladministrative
satisfactionsurveydata,aconveniencesamplewasconstructed.Patientshadpreviouslyundergonedetailed
retrospectivechartreviewforalternatestudies.AllpatientsincludedreturnedboththeHospitalConsumerAssessment
ofHealthcareProvidersandSystems(HCAHPS)andthePressGaney(PG)surveysbetween2009and2012.Twogroups
wereconstructedbasedontheresponsetoPGD1:Didyoufeelreadyfordischarge?Thosewhorespondedwiththe
topbox(TB)weretermedreadyfordischarge(RFD)andtheremaindergroupedandtermednotreadyfordischarge
(NRFD).30dayreadmissionratesandHCAHPSsatisfactionscoresforoverallhospitalratingandphysician
communicationdomainswerethenexaminedasafunctionofreadinessfordischarge.StandardHCAHPSmethodswere
usedforTBandcompositescorecalculations.
Results:Of320patients,45%werefemaleand95.5%wereCaucasian.Themeanagewas6215yearsandmeanBMI
was287.Meanlengthofstaywas1716days.DiagnosesincludedSBO(69%),hematologicdisorders(10%),GI
complaints(4%),andtrauma(4%).Theoverall30dayreadmissionratewas14.4%.Whenaskedabouttheextentof
dischargereadiness,55%ofpatientsindicatedtheywereRFD,withtheremainingpatientsindicatinglesserdegreesof
readiness(NRFD).Thesegroupswereotherwisesimilarintermsofbasicdemographics,includingage,sexandrace.
ThoseNRFDhadfewerTBresponsesforoverallhospitalsatisfaction(64%TBvs82%TB;p<0.001)andlowercomposite
scoresforphysiciancommunication(1.81.2vs2.50.8;p<0.001).IntheNRFDgroup,thereadmissionratewashigher
thatofthoseintheRFDgroup(17.9%vs11.4%;p=0.099).
Conclusions:Readinessfordischargeappearstobeanimportantcomponentinbothhospitalandpatientsatisfaction.In
addition,readinessfordischargeappearstobeausefulpatientreportedmetric.Thosewhoreportreadinessfor
dischargehavehighersatisfactionwithbothhospitalsandphysicians,andtrendtowardlowerreadmissionrates.Further
attentiontopatientreadinessfordischargeshouldbeemphasizedduetoitsimpactonpatientsatisfaction,and
additionalinvestigationintotargetedinterventionsforpatientsnotreadyfordischargeshouldbeundertaken.

107
REDUCTION OF SURGICAL SITE INFECTIONS AFTER LAPAROSCOPIC GASTRIC
BYPASS WITH CIRCULAR STAPLED GASTROJEJUNOSTOMY
P Shabino MD, A Elegbede MD, R Schmocker MD, M Nabozny MD, M Khoraki MD, M Garren MD, J Greenberg MD, GM Campos
MD.

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Patrick Shabino
(715) 218-1984
pshabino@uwhealth.org
Resident
General Surgery
Bariatric Surgery - Guilherme Campos
Outcomes
Surgical site infection, Bariatric, Gastric bypass, Gastrojejunostomy, Circular stapled

Purpose: Three techniques are most commonly used for the gastrojejunostomy (GJ) during laparoscopic gastric bypass
(LGB): the circular stapled, linear or hand-sewn technique. The circular stapled is favored in many centers, given its ease
of use, reproducibility and shorter operative times. However circular stapled GJ has been associated with higher rates of
surgical site infections (SSI); with reported rates being in between 5 to 30%. As part of a quality improvement (QI)
project to reduce SSI after Bariatric Surgery, we incorporated standardized technical modifications after LGB in which a
circular stapled GJ was used, while closing primarily all wounds. The purpose of this study is to compare the rates of SSI
after LGB with a circular stapled GJ, before and after the introduction of these technical modifications.
Methods: Patients were studied in two groups; Group 1 (G1) and Group 2 (G2), before and after the introduction of
the technical modifications respectively. G1 consisted of consecutive patients who underwent primary LGB with circular
stapled GJ from 5/2010 to 12/2012, in chronologic order without other exclusion criteria. Data was obtained by
retrospective electronic chart review. G2 consisted of consecutive patients who underwent primary LGB after the start
of the QI initiative in 1/2013. Data was collected prospectively in G2. SSI was defined as documentation of purulent
drainage from wound, symptomatic wounds treated with opening the wound, and any wounds identified as infected by
the surgical team. Technical modifications included the use a modified stapler cover sealed around the tip of the circular
stapler and antibiotic wound irrigation. Patients characteristics, operative details and wound infection rates before and
after the introduction of the technical modifications were compared. Numbers are mean and SEM, unless otherwise
stated.
Results: There were 200 patients in G1 and 43 patients in G2. Groups had similar demographic characteristics
(Age/years: G1 = 48+0.9 vs. G2 = 48+1.9, p=0.87; Female: G1 = 81% vs. G2 = 86%, p=0.52; BMI: G1 = 46.9+0.5 vs. G2 =
46.7+1.1, p=0.84), as well and prevalence of obesity associated diseases including type 2 diabetes, hypertension and
degenerative joint disease. Patients in G2 had a higher prevalence of obstructive sleep apnea (G1 = 59% vs. G2 = 78%,
p=0.03). LGB technical details such as use of trans-oral (TO) or trans-abdominal (TA) anvil insertion were similar (G1: TO
= 36% / TA = 64% vs. G2: TO = 30% / TA = 70%, p=0.59). There was a higher proportion of patients in which a 21mm
circular stapler was used (as opposed to 25mm) in G2 (21mm: G1 = 11%; G2 = 28%, p<0.01). There was a significant
reduction in SSI in G2 (SSI Rates: G1= 30 patients, 15% vs. G2 = 1 patient, 2.3%; p=0.02).
Conclusion: Technical modifications of standard circular stapled GJ for LGB, without leaving the wound open,
significantly reduced SSI rates. Using this technique, SSI rates are comparable to the rates obtained using linear or handsewn techniques.

108
ANomogramPredictingPostoperativeReadmissioninGeneralSurgeryPatients

SarahE.TevisMD,SharonM.WeberMD,K.CraigKentMD,GregoryD.KennedyMDPhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

SarahTevis
(608)6920836
samend@gmail.com
Resident
GeneralSurgery
Dr.GregoryKennedy
Outcomes
Predictive,Nomogram,Postoperative,Readmission,GeneralSurgery

Background:Aspartofthehospitalreadmissionreductionprogram,CentersforMedicareandMedicaidServices(CMS)
plantoimplementpenaltiesforhospitalswithaboveaveragepostoperativereadmissionrates.Easytousealgorithms
foridentifyingpatientsathighriskforreadmissionareneededtoidentifywhichpatientswilllikelybereadmittedatthe
timeofdischarge.Theaimofthisstudywastoidentifyobjectiveperioperativevariableswhichplacepatientsatriskfor
readmission,developapredictivenomogramforreadmissionandvalidatethenomogram.
StudyDesign:Inpatient,generalsurgerypatientswhounderwentelectivesurgeryfrom20062012wereidentifiedfrom
aninstitutionalAmericanCollegeofSurgeonsNationalSurgeryQualityImprovementProgram(ACSNSQIP).Thestudy
populationofpatientsusedtoidentifyriskfactorsforreadmissionanddevelopapredictivenomogramconsistedof
2,799patients,whilethevalidationgroupwasmadeupof350patients.Chisquareanalysiswasperformedtoevaluate
potentialriskfactorsforreadmission.Logisticregressionanalysiswasthenusedtoidentifyindependentpredictorsof
readmissionwithin30daysfromdischarge.Anomogramwasdevelopedusingthelinearpredictormethod.Areaunder
thecurve,positivepredictivevalueandnegativepredictivevaluewerecalculatedtovalidatethenomogram.All
statisticswereperformedinSPSSv.21andpvalues<0.05wereconsideredsignificant.
Results:Preoperativesteroids(OR1.543,95%CI1.0282.316),bleedingdisorder(OR2.591,95%CI1.5874.228),long
operativetime(OR1.623,95%CI1.2062.184)andprolongedlengthofstay(LOS)orinhospitalcomplication(OR2.032,
95%CI1.5442.674)independentlypredictedreadmission.Apredictivenomogramwasdevelopedforreadmission
within30daysofdischargeanditwasfoundtohavemodestpredictiveability(cstatistic0.668).
Conclusions:Developmentofanonlinecalculatorutilizingthispredictivemodelwillallowustoeasilyidentifypatientsat
highriskforreadmissionatthetimeofdischarge.Patientsatincreasedriskforreadmissionmaybenefitfrommore
intensivepostoperativefollowupintheoutpatientsetting.

109
Examiningtheroleofcervicalultrasoundindetectingthyroidpathologyin
patientswithprimaryhyperparathyroidism

DeenaWeiss,BS,HerbertChen,MD,FACS

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DeenaWeiss
(262)9454862
dmweiss2@gmail.com
MedicalStudent
GeneralSurgery
HerbertChen,MD,FACS
Outcomes
ultrasound,thyroid,hyperparathyroidism,endocrine,imaging

Background:
Minimallyinvasiveparathyroidectomyforprimaryhyperparathyroidismismadepossiblewithaccuratepreoperative
imaging.Inadditiontothedetectionofparathyroidadenomas,cervicalultrasoundalsoprovidesconcomitant
assessmentofthethyroidgland,andmanysurgeonsbelievethatitisessential.However,theincidentalidentificationof
thyroidnodulesmaythensubjectpatientstofurtherworkupandpotentiallyinvasivethyroidprocedures.Wesoughtto
determinethelongtermconsequenceofomittingpreoperativeultrasoundonthedevelopmentofthyroidpathology
andcancer.
StudyDesign:
Atourinstitution,222patientswithprimaryhyperparathyroidismunderwentparathyroidectomywithoutpreoperative
cervicalultrasoundfrom1990to2001.Thyroidpathologydiscoveredbyfollowupafterparathyroidectomy,aswellas
subsequentbiopsyandsurgicalinterventionswereanalyzed.
Results:

Ofthe222patientswhounderwentparathyroidectomy,themeanagewas551yearsand149(67%)werefemale.In
thecourseoftheirfollowupafterparathyroidectomy,13(6%)patientsreceivedacervicalultrasound,and7ofthe13
(3%)underwentfineneedleaspirationofathyroidnodule.Only1ofthe7(0.4%ofallpatients)wasultimately
diagnosedwiththyroidcancer.Fouradditionalpatientswerediscoveredtohavethyroidmalignanciesasaresultof
intraoperativedecisionmaking.Allfivepatientsarecurrentlyalive,withanaveragefollowuptimeof14.91.6years.
Nopatientsinthisserieshadanunnecessarythyroidintervention.
Conclusions:
Inpatientswhoundergoparathyroidectomywithoutapreoperativeultrasound,onlyasmallnumber(0.4%)
subsequentlywerediagnosedwiththyroidcancer.Furthermore,omissionofultrasoundduringthelocalizationof
parathyroidglandsdoesnothaveanegativeimpactonthediagnosisofthyroidpathologyasallpatientswhohad
thyroidcancerhadgoodoutcomes,andinfact,maypreventunnecessarythyroidinterventions.Therefore,theuseof
cervicalultrasoundforparathyroidlocalizationshouldbeconsideredoptionalratherthanessential.

110
GeneralandVascularSurgeryReadmissionsintheModernEra:ASystematic
Review

JasonT.WisemanMD,AmandaGuzman,SaraFernandesTaylorPhD,TravisL.EngelbertMD,R.ScottSaundersMD,andK.Craig
KentMD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JasonWiseman
(781)2234901
wiseman@surgery.wisc.edu
PostdoctoralFellow
VascularSurgery
Dr.Kent
Outcomes
Readmissions,Review,Quality,vascularsurgery,generalsurgery

Objective:Todeterminethecurrentlandscapeofgeneralandvascularsurgeryhospitalreadmissions.
SummaryBackgroundData:Higherthanpredictedratesofhospitalreadmissionareincreasinglyunderscrutinyasthey
arecostlyandthoughttorepresentpoorhealthcarequality.Whereasreadmissionsfollowinghospitalizationforan
acutemedicalconditionhavebeensubjectedtoongoingresearchandpolicyinitiativesforanumberofyears,the
descriptionofthisprobleminthesurgicalspecialtiesisstillinitsinfancyandlackssummativeanalysis.
Methods:AliteraturesearchwasperformedutilizingPubMedfromJanuary1,2009toJuly1,2013toidentifyarticles
relatedtohospitalreadmissionsfollowingvascular,general,bariatric,andcolorectalsurgicalprocedures.Thefollowing
endpointswereevaluated:readmissiondiagnosesandrates,predictorsofreadmission,andmortalityinreadmitted
patients.
Results:Theinitialliteraturesearchyielded619articles.Aftersystematicapplicationofexclusioncriteria,therewere41
qualifyingarticles(vascular=10,general=10,bariatric=5,colorectal=16).The30daymedianreadmissionratefor
vascularsurgerywas18.5%(n=9,range[11.9,24.4%]);generalsurgery:8.7%(n=6,range[5.3,12.1%]);bariatricsurgery:
6.8%(n=5,range:[3.7,9.3%]);andcolorectalsurgery:12.8%(n=13,range:[8.3,32.5%]).Themostfrequentlyreported
readmissiondiagnosesincludedwoundcomplications.Themostcommonlyreportedpredictorsofreadmissionwere
postoperativecomplicationssuchaswoundinfectionsandcardiaccomplications,andprolongedlengthofstay.Arisk
predictionmodelwasreportedinonlyonestudy.Readmissionwascharacterizedinthemajorityofthearticlesfromthe
timeofdischarge(n=26,63%)asopposedtothetimeofprocedure(n=12,29%).In3outofthe4studiesthatreported
mortalityratesofreadmittedpatients,therespectiveauthorsdemonstratedasignificantincreaseriskofmortalitywith
readmission.
Conclusions:Earlyreadmissionsarecommonacrosssurgicalspecialties,lendingimpetustoimproveourunderstanding
ofcausesandconsequences.Futureeffortsshouldfocusonstandardizingdefinitionsofreadmissionandreporting
criteria,whichwillultimatelyprovidethefoundationfordecreasingreadmissionsandimprovingthequalityofsurgical
care.

G ROUP FOUR

Translational/Clinical

111
NotchSignalinginpancreaticdifferentiationfromembryonicstemcells

JoelAlvarez,GopikaNair,JonOdorico

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JoelAlvarez
(262)7587710
jdalvarez@wisc.edu
UndergraduateStudent
GeneralSurgery
Odorico
Translational/Clinical
stemcell,pancreas,notchsignaling,tetptf1a,betacell

Notchsignalingisknowntoplayaroleinendocrinedifferentiationinpancreaticdevelopment.However,studieshave
cometodivergentconclusions.Here,wesoughttotestwhethernotchsignalingoritsinhibitionwouldpromoteor
inhibitformationofinsulinproducingcellsfrommouseembryonicstemcells(ESC)usingaPTF1adriveninvitro
pancreaticmodelsystem.Theaimofthisstudywastodefineanintervalofinhibitionofnotchsignalingthatledtoa
moreendocrinerichpopulation.WeusedthegammasecretaseinhibitorN[N(3,5difluorophenacetyl)1alanyl]S
phenylglycinetbutylester(DAPT)toinhibitendogenousnotchsignalinginatetracyclineinduciblePtf1aESClineand
various48hourinhibitionperiodswereanalyzedwithregardtotheeffectsonendocrinedifferentiation.Ourfindings
suggestnotchsignalingisrequiredduringandshortlyafterptf1ainductiontopromoteamoreendocrinerich
population.InhibitionofnotchfromEB7+11EB7+13ledtoanincreaseinthenumberofinsulinproducingcellswhen
analyzedatEB7+28andanincreaseincellspositiveforthebetacellprogenitormarkerneurogenin3(ngn3)when
analyzedatEB7+14whencomparedtovehiclecontrols.Thereisadoserelationshipinthiseffectasaconcentrationof
1MofDAPTresultsinmorecellspositiveforinsulinandendocrinehormoneswhereasa5Mconcentration
decreasedordidnotsignificantlyalterendocrinedifferentiation.Incontrast,inhibitingnotchsignalingatEB7+7EB
7+9orEB7+9EB7+11doesnotsignificantlychangethenumberofinsulinproducingcellsorngn3expressionwhen
comparedwithvehiclecontrol.Thus,ourfindingsindicatethatnotchsignalingisrequiredatearlystagesandhastobe
inhibitedatlaterstagesofdifferentiationofESCstocells.Thisknowledgecouldbeusedtoadvancemoreefficient
generationofnewinsulinproducingglucoseresponsivecellsfromEScellswhichcouldbeusedfortransplantation
purposesinpatientswiththeautoimmunediseaseType1diabeteswhohaveadeficiencyofviablebetacells.

112
DenovodonorspecificHLAantibodiesincreasetheriskofischemic
cholangiopathyindonoraftercardiacdeathliverallografts

O.Andacoglu,A.M.D'Alessandro,D.P.Foley,T.M.Ellis,A.J.Powell,G.E.Leverson,M.R.Lucey,A.I.Musat

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

OyaAndacoglu
(608)2286968
oandacoglu@uwhealth.org
Resident
GeneralSurgery
AlexandruMusat
Translational/Clinical
:livertransplantation,denovoDSA,,ischemiccholangiopathy,c4d,DCD

Donorshortagehasledtoanincreasedutilizationofdonationaftercardiacdeath(DCD)livergrafts.Incontrastto
donationafterbraindeath(DBD)grafts,DCDlivershaveinferiorgraftfunctionandsurvival,mostlyrelatedto
consequencesofischemiccholangiopathyIC)resultingfromanexaggeratedischemiareperfusioninjurytothe
peribiliarymicrovasculature.Asthemainpathogenicmechanismsofhumoralalloreactivityinvolvedamageof
endothelialcellsinmicrovascularbeds,wehypothesizedthathumoralalloreactivityduetodenovodonorspecific
antibodies(dnDSA)generatedafterlivertransplantationasareactiontothegraftmightfurtherpromotedamageof
endothelialcellsinperibiliaryvascularplexusandincreasetheriskofICinDCDgrafts..Therefore,wecorrelatedbiliary
integritywiththedevelopmentofdnDSAagainstHLAclassI(A,B,C)andII(DR,DQ,DP)in19consecutiveprimaryliver
transplantrecipientsthatreceivedanABOcompatibleDCDgraftbetween5/2009and2/2012.Sevenpatients
developedIC;5/7(71%)patientswithIChaddnDSA.Coxproportionalhazardsmodelshowedthatpatientswho
developeddnDSAhada6timeshigherriskofIC(p=0.03).MultivariableanalysisindicatedthatdnDSAandcoldischemia
timewereindependentriskfactorsforIC.ThisisthefirststudytosuggestthatdevelopmentofdnDSAafterDCDliver
transplantationisanindependentriskfactorforIC.Ourobservationsraisethepossibilitythataddressingthegeneration
ofdnDSA,apotentiallymodifiableriskfactor,mayimprovetheDCDlivergraftfunctionandsurvival.

113
PeriadventitialApplicationofRapamycinLoadedNanoparticlesProduces
SustainedInhibitionofVascularRestenosis

GuojunChen,XuDongShi,LianWangGuo,YiSi,MenZhu,SrikanthPilla,BoLiu,ShaoqinGong*,K.CraigKent*

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

GuojunChen
(608)5566964
gchen55@wisc.edu
GraduateStudent
VascularSurgery
ShaoqinGongandCraigKent
Translational/Clinical
Intimalhyperplasia,poly(lactidecoglycolide),nanoparticles,rapamycin,sustained
drugdelivery

Openvascularreconstructionsfrequentlyfailduetothedevelopmentofrecurrentdiseaseorintimalhyperplasia(IH).
Thispaperreportsanoveldrugdeliverymethodusingarapamycinloadedpoly(lactidecoglycolide)(PLGA)
nanoparticles(NPs)/pluronicgelsystemthatcanbeappliedperiadventitiallyaroundthecarotidarteryimmediately
followingtheopensurgery.Invitrostudiesrevealedthatrapamycindispersedinpluronicgelwasrapidlyreleasedover3
dayswhereasreleaseofrapamycinfromrapamycinloadedPLGANPsembeddedinpluronicgelwasmoregradualover4
weeks.Inculturedratvascularsmoothmusclecells(SMCs),rapamycinloadedNPsproduceddurable(14daysversus3
daysforfreerapamycin)inhibitionofphosphorylationofS6kinase(S6K1),adownstreamtargetinthemTORpathway.
Inaratballooninjurymodel,periadventitialdeliveryofrapamycinloadedNPsproducedinhibitionofphosphoS6K114
daysafterballooninjury.ImmunostainingrevealedthatrapamycinloadedNPsreducedSMCproliferationatboth14
and28dayswhereasrapamycinalonesuppressedproliferationatday14only.Moreover,rapamycinloadedNPs
sustainablysuppressedIHforatleast28daysfollowingtreatment,whereasrapamycinaloneproducedsuppressionon
day14withreboundofIHbyday28.Sincerapamycin,PLGA,andpluronicgelhaveallbeenapprovedbytheFDAfor
otherhumantherapies,thisdrugdeliverymethodcouldpotentiallybetranslatedintohumanusequicklytoprevent
failureofopenvascularreconstructions.

114
In-vivo treatment of brain dead rat donors with novel fusion protein improves
islet health and increases functional potency, reducing the islet mass required
for successful reversal of diabetic hyperglycemia post-transplant.
Peter J. Chlebeck, Xiaobo Ma, Juan S. Danobeitia, Luis A. Fernandez

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Peter Chlebeck
(608) 262-8459
chlebeck@surgery.wisc.edu
Faculty
Transplantation
Fernandez
Translational/Clinical
brain death, transplant, islets, fusion protein, organ preservation

BACKGROUND
Current medical understanding indicates that organs from brain dead donors experience severe oxidative stress in-situ
that negatively impacts organ quality and graft survival following transplantation. Pancreatic beta cells present
increased susceptibility to damage from this oxidative stress due to lower levels of endogenous anti-oxidant enzymes.
As a result, therapeutic islet transplantation to cure diabetes-mellitus may require multiple infusions of islets in large
quantities over the course of a patients life. MTS-EndoIII-TAT is a mitochondrially targeted fusion protein with a potent
DNA-repair enzyme specific to the primary DNA base-lesion caused by reactive oxygen and nitrogen species and
implicated in the pathogenesis of diabetes-mellitus. We previously found that in-vitro treatment of isolated pancreatic
islets with MTS-EndoIII-TAT abrogates the negative effects of pro-oxidant cytokines in culture. In this study, we
investigated the effect of MTS-EndoIII-TAT on the health and functional potency of transplanted islets from brain dead
donors treated in-vivo.
METHODS
To simulate the clinical transplant environment, we induced brain death in donor male Lewis rats via an established
protocol and sustained the animals on a ventilator with hemodynamic monitoring. Donors were treated with an IV
bolus injection of vehicle or MTS-EndoIII-TAT 30-minutes after brain death, and euthanized after 6-hours. Non-brain
dead donors were used as a control. Pancreatic islets cells were isolated and cultured for 24-hours. Islet quality was
assessed by flow cytometry measuring viability (FDA), apoptosis (Annexin-V) and mitochondrial membrane polarity (JC1). Islet functional potency was assessed in-vitro by glucose stimulated insulin secretion (GSIS) and in-vivo by
transplanting a minimum 250 islet equivalents (IEQ) into induced-diabetic immunodeficient mice. Intraperitoneal
glucose tolerance testing (IPGTT) was performed 30-days after transplant.
RESULTS
Islet cells from brain-dead donors treated with MTS-EndoIII-TAT exhibited higher MMP than cells from control and
vehicle-treated brain-dead donors by flow cytometry (93.242.5% compared to 86.87%5.4% and 85.997.8%
respectively), and lower levels of necrosis and apoptosis. Islets from treated donors exhibit a significant increase in
insulin secretion response to in-vitro glucose challenge (GSIS) when compared to islets from control and vehicle-treated
brain-dead donors (stimulation index: 18.61.2 vs. 8.33.0 vs. 13.03.1 respectively, p<0.01). In-vivo assessment of
functional potency showed that as few as 250 IEQ from donors treated with MTS-EndoIII-TAT can return diabetic mouse
recipients to sustained normoglycemia within 48-hours of transplantation. In contrast, mice transplanted with only 250IEQ from control and vehicle-treated brain-dead donors failed to achieve normoglycemic control after 30-days. IPGTT
results showed a greater area under the curve (AUC) for insulin release and a faster normalization of blood glucose in
recipients of donor islets treated with MTS-EndoIII-TAT.
CONCLUSION
These results indicate that in-vivo treatment of brain-dead donors with MTS-EndoIII-TAT can improve islet viability postisolation, maintain mitochondrial membrane polarity, enhance islet functional potency and reduce the number of islets
required to reverse diabetes in transplant recipients. We consider MTS-EndoIII-TAT as a promising cyto-protective agent
to improve islet quality prior to transplantation, and as a potential strategy to enhance organ preservation and improve
outcomes for transplant recipients not limited to islets, but including other transplantable organs as well.

115
Sex Differences in Voice-Related Modulation of Mechanosensory Detection
Thresholds in the Human Larynx
Michael J. Hammer, Mallory A. Krueger, Stephanie A. Palm

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Michael Hammer
(608) 807-8566
hammer@surgery.wisc.edu
Scientist
Otolaryngology-Head & Neck Surgery
Hammer Lab
Translational/Clinical
larynx, sensory, sex, efference, afference

I. BACKGROUND: Rapidly adapting mechanoreceptors within the laryngeal mucosa provide the central nervous system
with perceptual and proprioceptive afference for a variety of essential yet diverse human functions including voice
sound production and airway protection. It is unknown why mechanosensory information that yields a defensive
response when an individual breathes may go largely unnoticed when the individual voices. Therefore, a central
question is whether there is voice-related modulation of laryngeal mechanosensory detection. Such modulation would
be consistent with current models of afferent laryngeal control, and may be important tomaintain fluent voice in the
presence of potentially distracting sensory input. Therefore, we employed endoscopic assessment of laryngeal
mechanosensory detection thresholds inten healthy adults during tidal breathing and a voice task. Therefore, the
purpose of this study was to measure and compare laryngeal mechanosensory detection thresholds during a baseline
condition of tidal breathing and an experimental voice condition. We hypothesized that thresholds would be
significantly higher during the voice condition. Given the higher prevalence of voice sensorimotor disturbances in
women, we also hypothesized that women may exhibit less voice related modulation.
II. METHODS: (a) Participants. Data were collected from 10 healthy participants (5 women, 5 men) between 1823 years
old. (b) Laryngeal Mechanosensory Assessment. We employed an endoscopic stimulus delivery paradigm to present a
pressurecalibrated burst of air to the laryngeal mucosa to determine the threshold pressure at which each participant
could detect the stimulus. During tidal breathing, stimulus presentation was triggered by the initiation of the expiratory
phase of respiration. During the voice task, stimulus presentation was similarly triggered after an initial 1.5 s of sustained
voice.
III. STATISTICAL ANALYSIS: We utilized a mixed model ANOVA to examine differences in mechanosensory detection
between baseline tidal breathing and voice conditions and to test for differences between women and men. We
hypothesized that thresholds would be significantly higher during the voice condition.We also hypothesized that women
may exhibit less voice related modulation than men.
V. RESULTS: We observed significant main effects for task [p = 0.005] and sex [p = 0.044]. Sensory thresholds were
elevated for each participant during the voicetask by an average of 3.44 mmHg, representing more than a 3-fold
increase (p = 0.005). Men exhibited a greater 5-fold
increase in threshold during the voice task; women exhibited a more modest 1.39-fold increase (p = 0.003).
VI. DISCUSSION: Our findings suggest that the laryngeal sensorium may modulate mechanosensory afference to
attenuate the potentially distracting influence of sensory input during voice. The finding that women maintain a greater
sensitivity during the voice task than men (lower thresholds) may have important implications for the higher prevalence
of sensorimotor voice disturbances in women. Our results are consistent with the presence of mechanosensory
modulation in other motor systems and with observed sensory differences between women and men. Such modulation
has important implications for understanding the underlying neural mechanisms of laryngeal control and how these
mechanisms may operate in individuals with laryngeal disturbances.

116
Evaluating the Tongue-Hold Manuever Using High-Resolution Manometry and
Electromyography
Michael J. Hammer, Corinne A. Jones, Jason D. Mielens, Chloe H. Kim, Timothy M. McCulloch

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Michael Hammer
(608) 262-6018
hammer@surgery.wisc.edu
Scientist
Otolaryngology-Head & Neck Surgery
McCulloch Lab & Hammer Lab
Translational/Clinical
swallow, muscle, treatment, pressure, electromyography

BACKGROUND: The tongue-hold maneuver was developed as a therapeutic swallow maneuver to restrain retraction of
the tongue and to isolate movement of the posterior pharyngeal wall. To perform the tongue-hold maneuver,
instructions are to protrude the tongue maximally but comfortably, holding it between the central incisors and to
maintain this position during a saliva swallow. Given the anatomical interconnection between the muscles of the tongue
and the posterior pharyngeal wall, the anterior bulge observed during the tongue-hold maneuver may result from
increased contraction of the superior pharyngeal constrictor. However, the immediate and swallow-related effects of
the tongue-hold maneuver remain unknown.
HYPOTHESIS: Therefore, we tested the hypothesis that the amplitude and duration of submental, genioglossus, superior
pharyngeal constrictor, and cricopharyngeal muscle activity would increase during the tongue-hold maneuver, but that
the amplitude and duration of pharyngeal pressure would remain relatively unchanged.
METHODOLOGY: We used simultaneous high-resolution (36-channel) manometry (HRM), fine-wire intramuscular
electromyography (EMG) of the superior pharyngeal constrictor, genioglossus, cricopharyngeus, and surface EMG of the
submental region with a group (N=8) of healthy participants. We tested participants under three conditions: (a) Saliva
swallow (no manuever), (b) Saliva swallow with tongue tip at the lip, (c) Saliva swallow with maximum tongue
protrusion.
ANALYSIS AND STATISTICAL APPROACH: Pressure and EMG data were analyzed using a customized MATLAB program
(MathWorks, Inc., Natick, MA). EMG signals were rectified and normalized so that data could be pooled across
participants. The highest peak from the rectified signal for a given muscle and participant was set to equal 1. Therefore,
all other data points were computed to be between 0 and 1. Task differences were analyzed using repeated measures
analysis of variance (ANOVA) using a criterion significance level of alpha=0.05. We hypothesized that the amplitude and
duration of submental, genioglossus, superior pharyngeal constrictor, and cricopharyngeal muscle activity would
increase during the experimental tasks, but that the amplitude and duration of pharyngeal pressure would remain
relatively unchanged.
RESULTS: Our findings generally supported our experimental hypothesis. BEFORE the swallow, there was a significant
main effect of task as the amplitude of submental [p<0.001], genioglossus [p=0.005], superior pharyngeal constrictor
[p=0.012], and cricopharyngeus [p<0.001] muscle activity increased. DURING the swallow, there was a significant main
effect of task as the amplitude of submental [p<0.001], genioglossus [p=0.038], and superior pharyngeal pharyngeal
constrictor [p<0.001] muscle activity, as well as on the duration of submental [p<0.001], genioglossus [p=0.003], and
superior pharyngeal pharyngeal constrictor [p=0.030] muscle activity increased. Amplitude and duration of
cricopharyngeus muscle EMG activity were unchanged during the swallow (p>0.05). For manometric measures we found
that before the swallow, manometric pressures remained unchanged; during the swallow, pressure magnitude and
duration also remained unchanged (p>0.05).
DISCUSSION: Our preliminary results suggest that the tongue-hold facilitates increased tongue, pharyngeal wall, and
floor of mouth muscle contraction. Therefore, the tongue-hold during the preparatory and oral phases of swallow alters
the central patterning of the pharyngeal swallow and may be utilized to change the pattern of pharyngeal phase swallow
movements and is a unique example of motor-equivalent swallow behavior.

117
ThirtyYearFollowupofTotalHandReplantation:ACaseReport

JacquelineS.IsraelMD,VenkatK.RaoMDandSamuelO.PooreMD,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JackieIsrael
(608)2652535
jisrael@uwhealth.org
Faculty
Plastic&ReconstructiveSurgery
SamuelO.Poore
Translational/Clinical
Hand,Replantation,Transplantation,Reconstructive,microsurgery

Traumatichandinjuriesoccasionallyresultinamputation,andthedecisiontoreplanttheaffectedextremitymustbe
onethatiscarefullyconsidered.Wepresentacaseofatotalhandreplantationperformed29yearsago,aswellasa
reviewofthesignificanceoflongtermfollowupinpatientsundergoingreconstructionofanamputatedupper
extremity.Ourpatient,a21yearoldmale,sustainedaworkrelatedcircularsawamputationoftheleft,nondominant
handin1984.Heunderwentemergentreplantationatthelevelofthecarpus,andexperiencedasatisfactory
postoperativecourse.Heparticipatedinregularoutpatienttherapy,andat12monthsfollowingtheinitialoperation,
subjectivelydeniedpain,anddemonstratedwristactiverangeofmotion(aROM)approximately50%intheinjured
comparedtononinjuredhand.Gripstrengthintheaffectedhandwas19%thatoftheoppositehand.Thepatient
returnedtoourclinicalmost360monthsafterhisinitialoperation.HewasevaluatedutilizingtheMichiganHand
OutcomesQuestionnaire(MHQ).Subjectively,thepatientexpressedsatisfactionwithhisreplantedhand,withno
impairmentsinactivitiesofdailyliving.CombinedtotalMHQscoreintheinjuredhandwas85/100,comparedto99in
theunaffected.TotalaROMoftheleftwristwasapproximately65%thatoftheright,andbothgripstrengthandlateral
pinchinthelefthandwere58%thatoftheright.Twopointsensationofeachfingertipontheaffectedhandwasan
averageof10mm,comparedto5mmontheoppositeside.Overallhandfunctionwasfoundtobesignificantlyimproved
comparedto1yearfollowup.Thiscaseexemplifieshowupperextremityreplantationissuccessfulinthepurposefully
selectedpatient.Theoutcomesobservedinthispatientremindusoftheutilityoflongtermfollowupinall
postoperativepatients,includingthoseundergoinghandtransplantbothnowandinthefuture.

118
ENDOSCOPICLARYNGOPHARYNGEALREFLUXFINDINGSAREPOORLY
CORRELATEDWITHCOMBINEDMULTICHANNELINTRALUMINALIMPEDANCEPH
MONITORING

MarieJette,MS,EricGaumnitz,MD,MartinBirchall,MD,NathanWelham,PhD,SusanThibeault,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MarieJette
(617)5990456
jette@surgery.wisc.edu
GraduateStudent
OtolaryngologyHead&NeckSurgery
Thibeault
Translational/Clinical
chroniclaryngitis,laryngopharyngealreflux,measurement,pHmonitoring,impedance
monitoring

Whileprobablecausativeagentshavebeenidentified(e.g.,refluxatecomponents,tobaccosmoke),thedefinitive
mechanismforlaryngealmucosaldamageremainselusive.MultichannelintraluminalimpedancecombinedwithpH
monitoring(pH/MII)hasemergedhasasensitivetoolfordiagnosisandcharacterizationofgastroesophagealreflux
diseasewithlaryngopharyngealmanifestations.TodeterminetherelationshipbetweenlaryngealsignsandpH/MIIwe
examinedcorrelationsbetweenRefluxFindingScore(RFS)ratingsof142videostroboscopiclaryngealexaminations
averagedacross8trainedcliniciansandfindingsfromcorrespondingpH/MIIacquiredfromparticipantsmedically
untreatedforreflux.TherelationshipbetweenRFSandpH/MIIfindingsanddemographicinformationwasalsoassessed.
WefoundseveralstatisticallysignificantcorrelationsbetweenRFSratings,pH/MIIfindings,anddemographicdata,
includingassociationsbetweenage,smokingstatus,andgender,thoughallrelationshipswereclinicallynegligibleor
weak(0.30<R<0.30).Resultssuggestthattheremaybealternatebiologicalexplanationsfortheinflammatoryclinical
signscommonlyascribedtoreflux.

119
UsingHighResolutionManometryandVideofluoroscopytoDetectSubtle
SwallowingChangesinEarlyandMidStageParkinsonDisease

CorinneA.Jones,MichelleR.Ciucci,TimothyM.McCulloch

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

CorinneJones
(608)2655439
jones@surgery.wisc.edu
GraduateStudent
OtolaryngologyHead&NeckSurgery
McCulloch
Translational/Clinical
dysphagia,highresolutionmanometry,videofluoroscopy,Parkinson'sDisease,
deglutition

Purpose:DysphagiasecondarytoParkinsonDisease(PD)canhavedevastatingeffectsonhealthandqualityoflife.
However,standardmeasurementtechniquesmaynotbesensitiveenoughtodetectearlyandsubtlechangesin
swallowingfunction.WehypothesizedthatpersonswithPDwouldshowdifferencesincoordinationandprecisionof
swallowingeventsonhighresolutionmanometry.
Methods:SimultaneousHRMandvideofluoroscopy(VF)wasperformedtoexamine22personswithearlytomidstage
PDand10normallyagingcontrols.TheModifiedBariumSwallowImpairmentProfile(MBSImP)wasusedtoquantifyVF
parameters,andtheSydneySwallowingQuestionnaire(SSQ)wasusedasameasureofselfreporteddysphagia
symptoms.Swallowprecision(variability)wasmeasuredbycurvefittinganalyses,andswallowingcoordinationwas
measuredbypressurepeaklatency.Weusedlogisticregressiontotestpredictivepoweroftheaboveparametersin
determininghealthstate.
Results:LogisticregressionincorporatingSSQscores,variabilityinpressuregenerationacrossthepharynx,and
coordinationofpharyngealpressuregenerationfromthevelopharynxtoupperesophagealsphincterclosurepredicted
healthstatusofthesubjectswith<99%accuracy(p<0.001).Pharyngealpressures&durations,MBSImPparameters,
andairwayprotectiondidnotpredictgroupdifferences(p>0.05).
Conclusions:ResultssuggestthatHRMandtheSSQarevaluablediagnostictoolstodetectsubtlechangesinswallowing
functionnotseenwithVF.Improvingearlydetectionhasthepotentialtoincreaselongtermswallowingoutcomes.

120
iPS-MSCs reduce incidence of Delayed Graft Function in the Non-human Primate
Brain Death and Renal Transplant Model
Laura Zitur, Juan Danobeitia, Saritha Dsouza, Xiaobo Ma, Anthony DAlessandro, Igor Slukvin, Luis Fernandez

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

ZITUR LAURA
(608) 628-2151
ZITUR@SURGERY.WISC.EDU
Scientist
Transplantation
FERNANDEZ
Translational/Clinical
KIDNEY TRANSPLANT, BRAIN DEATH (BD), NON-HUMAN PRIMATE (NHP),
MULTIPOTENT STROMAL CELLS (MSC, DELAYED GRAFT FUNCTION (DGF)

Introduction:
Brain death (BD) kicks off an inflammatory response and cytokine storm known to increase the immunogenicity of
transplantable organs, and impair organ function and quality. All eyes have turned to Multipotent Stromal Cells (MSC),
with their local immunosuppressive properties and capacity to repair injured tissue, as a potential player in the
transplant environment. Here we have attempted to harness their immuno-modulatory effects to address delayed graft
function (DGF) using our unique model of non-human-primate (NHP) brain death and renal transplant. These particular
MSCs represent a new paradigm since they have been differentiated from episomally-induced pluripotent stem cells
(iPSC), are a pure homogenous population with no MHC Class I/II expression, and have 10^22 expansion potential, far
greater than their bone-marrow derived counterparts.
Methods:
MSC derivation: Cynomolgus iPSCs were differentiated on OP9 for 4 days, then sorted for APLNR+ cells and plated in
semisolid colony forming medium. MSC colonies were then picked from culture on Day12, expanded on fibronectincollagen in serum-free MSC media, and tested for CD31,CD45,CD73,CD90,CD105,CD145,CD13 mCD29, and CD44
expression.
In-vitro studies: Monocytes/macrophages from Cynomolgus peripheral blood were examined for MSC induction of
cytokine secretion. Natural Killer (NK) cytotoxicity was tested in the presence of MSCs.
NHP Renal Transplants: Our NHP donor model attempts to mimic the clinical setting of BD; 8-hours of clinical braindeath followed by renal nephrectomy and 40-hours of cold ischemia. Recipients underwent bilateral nephrectomy and
heterotopic kidney transplantation. Animals were allocated to two groups: Group1 (G1) -untreated recipients or Group2
(G2) -MSC treated recipients. Treated recipients received 107MSCs/kg delivered intravenously 1-hour prior to
reperfusion. Criteria for euthanasia were 1) progression to DGF and severe azotemia incompatible with life within 1week of transplantation or 2) survival for 28-days.
Results:
In-vitro: MSCs induced IL-10 secretion from monocytes/macrophages in the presence of LPS and reduced cytotoxicity of
NK cells.
In-vivo: 5/7 G1 animals progressed to DGF within 1-week of transplant and were euthanized. In contrast, 2/2 G2 animals
survived 28-days in good health. The peak serum creatinines were 144.0mg/dL in G1 animals and 8.00.4mg/dL G2
animals. The average urinary output for post-transplant days 1 through 5 was 72.5107mL/day for G1 and
29619mL/day for G2. At Day-5, G1 kidneys showed significant infiltration of neutrophils and macrophages, and
extensive tubular cell vacuolization and cortical necrosis consistent with severe ATN (Acute Tubular Necrosis). In
contrast, G2 kidney biopsies showed a dramatic decrease in neutrophil and macrophage infiltration and preserved cytoarchitecture.
Conclusion:
Our in-vitro data suggest that iPS-MSCs dampen innate immune responses through monocyte expression of the potent
anti-inflammatory cytokine IL-10 and through reduction of NK cell cytotoxicity; this confirms previous publications using
bone-marrow derived MSCs, demonstrating that these iPS-MSCs maintain an immuno-regulatory capacity. Our in-vivo
data suggest that recipient treatment using iPS derived MSCs may reduce incidence of DGF and increase overall organ
quality and function. We are currently exploring the reproducibility of our findings and evaluating long term safety. The
potential for these cells, their human counterparts, and future iterations in targeted therapy for transplant recipients is
exponential.

121
Inofficebiopsyforsuspectedlaryngealandpharyngealmalignancy:safety,
tolerance,subsiteanalysis,andtimetotreatment.

DylanLippert,M.D.;MatthewR.Hoffman,Ph.D.;PhatDang,M.D.;GregHartig,M.D.;TimMcCullochM.D.;SethH.Dailey,M.D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DylanLippert
(720)9333328
dlippert@uwhealth.org
Resident
OtolaryngologyHead&NeckSurgery
Dr.SethDailey
Translational/Clinical
Laryngealcancer,OfficeProcedures,Endoscopy,Pharyngealcancer,Biopsy

INTRODUCTION:Definitivetissuediagnosisforsuspectedthroatcancerisshiftingfromtheoperatingroomtotheoffice.
Safety,tolerance,factorsrelatedtosuccessfulbiopsy,andtimetotreatmentarenotwelldefined.
METHODS:116patientsundergoinginofficebiopsyoforopharynx,larynx,orhypopharynxwereincluded.Logistic
regressiondeterminedifdemographics,subsite,Tstage,orapproach(transoral/transnasal)wererelatedtosuccess.
RESULTS:92transnasaland24transoralbiopsieswereperformedon73laryngeal,35oropharyngeal,and8
hypopharyngeallesions.97of116diagnosesweremadeinoffice.Therewerenocomplications.Twopatientsdidnt
toleratetheprocedure.Successwasntrelatedtoage(p=0.374),sex(p=0.566),subsite(p=0.527),Tstage(p=0.587),or
approach(p=0.7599).Timetotreatmentwas24.213.9dayswithsuccessfulofficebiopsyand48.849.4dayswithout
(p<0.001).
CONCLUSIONS:Highsuccesswasobservedacrosspatients,subsites,andapproaches.Allpatientsarecandidatesforin
officebiopsy,amoretimeandcosteffectiveoptionleadingtoearliertreatment.

122
FunctionalevaluationofhumanDCDdonorheartsusingacontinuousisolated
myocardialperfusiontechnique;potentialforexpansionofthecardiacdonor
population.

SatoruOsaki,MD,PhD,MatthewR.Locher,PhD,EntelaB.Lushaj,PhD,ShahabA.Akhter,MD,andTakushiKohmoto,MD,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MattLocher
(608)2179195
mrlocher@wisc.edu
PostdoctoralFellow
CardiothoracicSurgery
TakushiKohmoto
Translational/Clinical
DCD,myocardium,exvivoperfusion,pressure,contractility

Objective:Toinvestigateresuscitationpotentialandcontractilefunctioninadulthumandonationaftercardiacdeath
(DCD)heartsbyexvivoperfusion.
Methods:UnderourinstitutionalreviewboardapprovalandDCDprotocolintheUniversityofWisconsin(UW)Organ
ProcurementOrganization,5braindead(BD)and5DCDdonorheartswereevaluated.AllBDheartsweredeclinedfor
transplantduetocoronaryarterydiseaseorage,althoughcardiacfunctionwasnearlynormal.Followingfull
heparinizationandharvestingsolidorgansfortransplantation,allheartswerepreservedbyflushingwith2LofUW
solutionimmediatelyafteraorticcrossclampandstorageinUWsolutionat4 C.Usingourperfusionsystem,theaortic
rootwascannulatedandheartswerereperfusedwithbloodcardioplegiasolution(20C)for20minfollowedby
oxygenatedbloodwithgradualrewarmingfrom20to37C.Afterinitialcontrolledreperfusion,leftventricular(LV)ex
vivofunctionwasassessedevery30minfor2hrs.
Results:AllBD(n=5)and4DCDheartsweresuccessfullyresuscitated.OneDCDheartwasunabletoberesuscitated,
duetoprolongedwarmischemictime(174min).MeanwarmischemictimeforresuscitatedDCDhearts(from
extubationtoflushingwithcoldUWsolution)was34 3min(range,26to40);meancoldischemictimeforBDdonors
was211 31mincomparedto215 67minforDCDdonors.InitialmeanLVendsystolicpressure(LVESP)forBDhearts
was113 25mmHgvs.77 26mmHgforDCDdonors(p=0.35),whileafter1hrofcontinuousperfusionmeanLVESPforBD
heartswas76 17mmHgcomparedto67 40mmHgforDCDhearts(p=0.84).ThecalculatedLVendsystolicpressure
volumerelationship(ESPVR)forBDheartsinitiallyandafter1hrofreperfusionwas7.4 0.9and6.1 1.4mmHg/mL,
respectively,whileLVESPVRforDCDheartsinitiallyandafter1hrreperfusionwas5.3 1.4(p=0.28)and
6.0 1.3mmHg/mL(p=0.63),respectively.HistologicalanalysisofLVsectionsbeforereperfusionrevealednoappreciable
differencesingrossmorphologybetweenthegroups.
Conclusions:Usingcontinuousisolatedmyocardialperfusion,wesuccessfullyresuscitatedandmeasuredexvivo
cardiacfunctioninhumanDCDandBDdonorhearts.ResuscitationpotentialinDCDheartswasachievedwhenwarm
ischemictimewaslessthan40min.ExvivocontractileperformanceinresuscitatedDCDheartswascomparableto
thatofBDheartsinacutephaseofreperfusion.OurresultssuggestthathumanDCDheartsprocuredwithinanoptimal
warmischemicperiodhasthepotentialtoexpandthedonorpopulation.Furtherinvestigationofmyocardialviability
andventricularfunctionwithlongerreperfusionperiodsappearswarranted.

123
Sensor-Based Assessment of Cast Placement and Removal
Anne-Lise D. Maag, MD, Shlomi Laufer, PhD, Calvin Kwan, BS, Elaine R. Cohen, MEd, Rachel L. Lenhart, MS, Natalie C. Stork, MD,
Matthew A. Halanski, MD, Carla M. Pugh, MD, PhD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Anne-Lise Maag
(650) 400-0850
annelise.maag@gmail.com
Resident
General Surgery
Dr. Carla Pugh
Translational/Clinical
Force, Sensor, Casting pressure, Extremity, Simulation

Introduction: Appropriate pressure during cast application is critical to adequately stabilize fractures and prevent castrelated complications. Changes in fracture management practice and work hour restrictions have decreased orthopedic
surgery residents exposure to cast application. Our prior work showed sensor enabled medical simulation allows for
quantitative evaluation of hands-on clinical skills. Additional research has demonstrated the feasibility of measuring
casting pressures. The goal of this project is to develop a casting simulation model that provides immediate, reliable
feedback regarding skin pressure during cast placement and removal.
Methods: A plastinated arm model created from PlatSil Gel-10 was utilized and a single persons right forearm served as
a comparison. Data were collected from seven force-sensing resistors (FSR): six on the dorsal and ventral surface of the
forearm and one on the palm. The same experienced provider performed cast placement and removal with pressure
recordings taken throughout these seven steps: 1)cotton undercast padding placement; 2)fiberglass casting tape
placement; 3)cast sawing down the dorsal side; 4)cast spreading through the dorsal cut; 5)cast sawing down the ventral
side; 6)cast spreading through the ventral cut; 7)cast removal. Pressure data were collected and plotted over time for
each of the sensors. The data from FSR #2 were representative of the sample, and therefore, was used for further
analysis.
Results: After application of the cotton undercast padding, the average skin pressure of the plastinated arm
(20.46mmHg (SD=0.76)) was similar to that of the human arm (20.60mmHg (SD=0.37)). Following application of the
fiberglass cast, the average skin pressure of the plastinated arm (42.20mmHg (SD=2.52)) was similar to the human arm
(45.58mmHg (SD=2.71)). During the cast application process, the maximum and minimum skin pressures tended to be
higher for the human arm (59.49mmHg and 39.68mmHg) than the plastinated arm (49.07mmHg and 34.63mmHg). With
the human model, recordings were taken during hand movement, which showed increased variability of pressures with
a maximum pressure of 71.51mmHg and minimum skin pressures of 33.57mmHg. The average skin pressure in the
human arm model after hand movement was much lower (26.67mmHg (SD=4.06)) than prior to hand movement
(45.58mmHg (SD=2.71)).
Conclusion: Application and removal of the cast on both the plastinated and human forearm demonstrated a signature
pattern of changes in skin pressure giving validity to monitoring pressure changes during the task with FSRs. The
plastinated and human arm had similar pressures following placement of the cotton undercast padding and fiberglass
cast indicating the materials used to create the plastinated model are a good simulation for human tissue. There were
some differences between the human and plastinated skin pressures likely related to muscle contraction and skin
movement intrinsic to the human model. These factors will be important to consider when developing an effective
teaching simulation model for cast placement.
This project demonstrated an innovative strategy for monitoring skin pressure during cast application in the process of
developing a simulation model that provides immediate, reliable feedback during cast placement. Further research is
needed to establish an expert standard of cast application pressures for our simulation model.

124
Use of Advanced Sensor and Motion Tracking Technology to Classify Clinical
Breast Examination Techniques
Anne-Lise D. Maag, MD, Shlomi Laufer, PhD, Elaine R. Cohen, MEd, Calvin Kwan, BS, Carla M. Pugh, MD, PhD

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Anne-Lise Maag
(650) 400-0850
annelise.maag@gmail.com
Resident
General Surgery
Dr. Carla Pugh
Translational/Clinical
Force sensor, Clinical Breast Examination, Pressure, Simulation, Physical Examination
Technique

Introduction
The clinical breast examination (CBE) is a vital component of breast care; however, objective assessment of CBE skills are
lacking. Current recommendations for performing the CBE include using one hand to palpate varying levels of pressure
with the fingertips of the middle three fingers in a vertical strip-like fashion. Our prior work has shown that clinician and
patient factors significantly affect CBE technique. In addition, we have successfully used pressure sensor data to
delineate physical examination techniques using different simulation models. This study explored the feasibility of using
a newly developed sensor-based algorithm to quantify breast exam technique.
Methods
Data were collected from clinicians (n=32) attending the American Society of Breast Surgeons meeting. Participants
performed CBEs on a task trainer while sensor mapping and video-based motion tracking systems captured performance
data. Sensor data were analyzed for exam time and average force applied. Sensor outputs were also utilized to develop
a mathematical algorithm to automatically classify the number of hands used during the CBE. Algorithm classification
was verified in a blinded fashion using video recordings. Our hypothesis was that the newly developed mathematical
algorithm would properly classify one and two-handed users. In addition, we hypothesized that exam characteristics
(amount of pressure used) will have a higher correlation with accuracy than number of hands used during the exam.
Results
Participants who correctly identified the mass, applied more average pressure (12.53 N (SD=5.23)) throughout the exam
than those who did not find the mass (6.67 N (SD=0.21)) [p=.001]. Blinded video analysis classified 8 users as onehanded and 24 as two-handed. The mathematical algorithm correctly identified 6 of one-handed CBEs (75%) and 21 of
the two handed CBEs (87.5%) (kappa=.82). There was no correlation between number of hands used during the exam
and diagnostic accuracy.
Conclusions
The combination of video recordings and pressure mapping served as a feasible and reliable method to collect new data
on CBE techniques. Our results show that examiners applying more pressure during the CBE were more likely to find the
breast mass. Accuracy was independent of the number of hands used. This finding is important because published
recommendations for performing the CBE promote the use of one hand.
This project is a first step in objective classification of CBE techniques. Further study is needed to evaluate additional CBE
characteristics to gain a better understanding of the complex relationship between CBE technique and accuracy.

125
AKTExpressionisAssociatedwithDegreeofPathologicResponsein
AdenocarcinomaoftheEsophagusTreatedwithNeoadjuvantTherapy

NadiaSaeed,MakiYamamoto,JillWeber,RaviShridhar,SarahHoffe,KhaldounAlmhanna,andKennethL.Meredith

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

KenenthMeredith
(608)2622025
MeredithK@surgery.wisc.edu
Faculty
GeneralSurgery
SurgicalOncology
Translational/Clinical
EsophagealCancer,AKT,PathologicResponse,NeoadjuvantTherapy,Microarray

Background.Neoadjuvantchemoradiation(NCRT)hasbecomestandardinthetreatmentoflocallyadvancedesophageal
cancer(LAEC)withsurvivalcorrelatedtodegreeofpathologicresponse.TheactivationofthePI3K/Akt/mTORpathway
playsanimportantroleintumorigenesisandresistancetoanticancerdrugs.Theaimofthisstudywastoelucidatethe
roleoftheAkt/mTORpathwayinchemoresistanceandtheprognosisofpatientswithesophagealadenocarcinoma(AC)
whoreceivedNCRT.
Methods.AfterIRBapproval,aprospectivetrialwasinstitutedinwhichpatientswithLAECrequiringNCRTwere
consentedforendoscopicbiopsiesofnormalandtumortissuepriortoinstitutingtherapy.AftercompletionofNCRT
patientsthenproceededtoesophagectomy.Specimenswerethencategorizedbyresponsetotherapy.Pathologic
completeresponse(pCR)wasdefinedasnoresidualtumor,partialpathologicresponse(pPR)asa50%reductionin
tumorsizeornodaldownstaging,andnonresponse(pNR)asnodifferencebetweenpreoperativeandpostoperative
stagebaseduponendoscopicultrasound.Thetissuesobtainedpreoperativelyunderwentgeneexpressionprofilingusing
theAffymetrix133Plus2.0Genechip.SAMmethodwasusedtoanalyzesignificantdifferentiallyexpressionofAKT
withinnormalandtumortissue.Expressionwasthencorrelatedtodegreeofpathologicresponse.
Results.Onehundredpatientswereconsentedforthestudy,ofwhich67metfinaleligibility.Nineteenpatients
ultimatelyunderwentgeneexpressionprofilingviamicroarray.ThedifferentialexpressionofallAKTprobesetsintumor
tissuewasmarkedlyoverexpressedcomparedtonormaltissue(p=6x105).Therewere3patientsdesignatedaspNR,5
aspPR,and11aspCR.InvestigatingtherelationshipbetweenthedegreeofexpressionofAKTandpathologicresponse
demonstratedalinearcorrelationbetweenoverexpressionofAKTanddegreeofpathologicresponse.Partialandnon
respondershadhigherexpressionsofAKTcomparedtopCRwiththenonrespondersconsistentlyillustratedthehighest
expressionofAKT(p=0.02).TherewasasignificantcorrelationbetweenindividualprobesetsAKT1,AKT2,andAKT3and
degreeofpathologicresponse(p=0.002,0.04,and0.04respectively).
Conclusions.AKTisoverexpressedinpatientswithACoftheesophagus.Moreover,pathologicresponsetoNCRTmaybe
correlatedwithdegreeofAKTexpression.Additionaldataisneededtoclarifythisrelationshipfurtherandpotentially
addtargetedtherapiestotheneoadjuvantregimen.

126
TheStateofGlobalHealthTraininginPlasticSurgeryResidency:Pragmatic
ConsiderationsandFutureDirections

HarryS.Nayar,MD,MBE;DeloraL.Mount,MD;MichaelL.Bentz,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

HarryNayar
(412)6565454
hnayar@uwhealth.org
Resident
Plastic&ReconstructiveSurgery
MichaelL.Bentz,MD
Translational/Clinical
Education,GlobalHealth,Residency,PlasticSurgery,Surgery

Introduction:Thegrowingproportionofglobaldiseaseanddisabilitythatisamenabletosurgicalcarehascreateda
needforpractitionersthatarecapablytrainedinmulticulturalsettings.Wesoughttodescribethecurrentstateof
globalhealthtrainingandexposureinU.S.plasticsurgeryresidencyprograms.

Methods:AnelectronicsurveywasdistributedtoRRC/AAMCcertifiedplasticsurgeryresidencyprogramsintheUnited
States.Demographicdata(programtype,size,andlocation)werecollected.Programshavingaformalglobalhealth
curriculumforresidentswereaskedtodescribetheexperiencewithregardstoclassification,collaborationdetails,
regionstravelled,conditions/proceduresencountered,expensesupport,accreditation,andpersonalsentiment.
Programsnotofferingglobalhealthtrainingwereaskedtoselectbarrierstoimplementation.
Results:Sixtyfourquestionnaireswerereturnedcompletedfromeightyoneresidencyprograms(responserate:79%).
Programsidentifiedasindependent(n=16;25%),integrated(n=25;39%),orboth(n=23;36%).Geographically,these
werelocatedinthesouthern(n=21),northeastern(n=15),central(n=14),andwestern(n=14)regions.Twentysix
programs(40%)reportedaformalglobalhealthcurriculum;thirtysevendidnot(60%).Whenaskedtobestclassifythis
curriculum,mostselectedaclinicalcareexperience(n=24;92%)followedbyaneducationalexperience(n=19;73%).
Personalreferencewasthemostcommonmeanstoestablishtheinternationalcollaboration(n=19,73%).Thethree
mostcommonlyencounteredconditionswere:cleftlip/palate(n=26;100%),thermalinjury(n=17;65%),andpost
traumaticreconstruction(n=15;57%).Regardingdominantfundingsources,53%(n=14)selectednonprofit
organizations,46%(n=12)werefundedbyfacultypersonalexpenditures,and38%(n=10)report
section/division/departmentsupport.AlthoughthemajorityofprogramshadnotappliedforPSOLaccreditationand
RRCrecognition(n=23;88%),manywereconsideringapplying(n=16;62%).Overall,96%ofprograms(n=25)reported
beingsupportiveofglobalhealthtraininginresidency,withmostchoosingexposuretodifferenthealthsystems(n=22,
84%)andsurgicaleducation(n=17;65%)asreasons.Forprogramsnotofferingaglobalhealthexperience,themost
commonlyreportedbarrierswerealackofRRC/PSOLrecognitionofcasesperformedabroad(n=27;73%),lackof
fundingfortripexpenses(n=25;67%),andlackofsalarysupport(n=24;65%).
Conclusion:Programsendorsingglobalhealthtrainingdescribetheexperiencepositively,emphasizingclinicalvariety,
exposuretodifferenthealthsystems,anditsimportanceinsurgicaleducation.Issuesrelatedtofundingandcase
accreditationaremajorobstaclestoincorporatingaglobalhealthcurriculum.

127
ElevatedPTHAfterCurativeParathyroidectomyDelaysSymptomImprovement

PriyaR.Pathak,SaraE.Murray,MD,SarahC.Schaefer,NP,GlenLeverson,PhD,HerbertChen,MD,RebeccaS.Sippel,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

PriyaPathak
(608)6982538
prpathak@wisc.edu
UndergraduateStudent
GeneralSurgery
Dr.RebeccaSippelandDr.HerbertChen
Translational/Clinical
EndocrineSurgery,PrimaryHyperparathyroidism,PrognosticFactors,Symptom
Improvement,Timing

Introduction:Curativeparathyroidectomyforprimaryhyperparathyroidism(PHPT)hasbeenshowntoresolvevarious
nonspecificsymptomsrelatedtothedisease.Between8to40%ofpatientswithnormocalcemiaaftercurativeresection
haveelevatedparathyroidhormonelevels(ePTH)atfollowup.WeinvestigatedwhetherePTHintheearlypost
operativeperiodwasassociatedwiththetimingofsymptomimprovement.
Methods:ThisprospectivestudyincludedadultpatientswithPHPTwhounderwentcurativeparathyroidectomyfrom
November2011toSeptember2012.Biochemicaltesting2weekspostoperativelyidentifiedePTHversusnormalPTH
(nPTH).Aquestionnaireadministeredpreandpostoperativelyat6weeksand6monthsaskedpatientstoratethe
frequencyof18symptomsofPHPT,whichincludedsymptomsrelatedtothemusculoskeletal,urinary,gastrointestinal,
andneuropsychiatricsystems.Symptomfrequencywasreportedona5pointLikertscale,rangingfromnevertovery
frequently.Incomparingdifferenttimepoints,statisticalanalysiswasperformedonthechangeinscoresforeach
individualsymptom.
Results:Of194patientsthatunderwentparathyroidectomyforPHPT,129(66%)participatedinthestudy.Pre
operatively,allpatientsendorsedhavingatleastoneormoresymptom(s),withameanof134symptoms.Twoweeks
postoperatively,20patients(16%)hadePTH.Thepercentageofpatientswhoshowedpostoperativeimprovementfor
individualsymptomswasthencomparedbetweengroups.Attheearlytimepoint(6weeks),theePTHgroupshowedless
improvementin14outof18symptoms.Thisdifferencereachedstatisticalsignificanceforfoursymptoms:anxiety,
constipation,thirst,andpolyuria.Bythe6monthstimepoint,thesedifferenceshadresolvedandsymptom
improvementwassimilarintheePTHandthenPTHgroups.
Conclusions:ElevatedPTHaftercurativeparathyroidectomymayresultinadelayinsymptomimprovementat6weeks
fornonspecificsymptomsofPHPT,butthisdifferenceresolvesby6months.

128
UseoftheAnkleBrachialIndex(ABI)asapossiblediagnostictoolforperipheral
arterydiseases(PAD)inDiabeticpatientsatTikurAnbessaSpecializedHospital,
Ethiopia.

CharlesPenn

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

CharlesPenn
(240)6205784
capenn@wisc.edu
MedicalStudent
VascularSurgery
DrGirmaTefera
Translational/Clinical
ABI,Prevalence,PAD,Diabetic,Patients

Background:Inlowandmiddleincomecountries,muchfocushasbeenoninfectiousdiseaseshowevernon
communicablediseasessuchascardiovasculardiseasearegettingtoepidemicproportions.Theobjectiveofthisstudy
wastouseAnkleBrachialIndex(ABI)testtoestimatetheprevalenceofPADinDiabeticpatientsatTikurAnbessa
SpecializedHospital(TASH)inAddisAbaba,Ethiopia.
Method:ThisstudywasconductedattheTASHdiabeticclinic.Wecollectedinformationpertainingtopatient
demographic,physicalexamination,measuredandcalculatedtheirABI.TheABIvaluewasusedtodefinethepresence
orabsenceofperipheralvasculardiseaseamongstallpatients.Normalvaluewas09to1.3.ABIvaluesoflessthan0.9
wereconsideredabnormallylawandthose>1.3suggestedseverevascularcalcificationandnonecompressibility.Other
vascularriskfactorssuchassmocking,lipidprofile,andotherevidenceofconcomitantvasculardiseasewerealso
documented.

Results:DuringthemonthsofMayandJune2013,atotalof140patientswereevaluated.Thisincluded73females,and
67maleswithameanageof51(range1785).9.5%ofthepatientshadABIlessthan0.9while10.1%hadABI>1.3.
Mostpatientssufferedfromtype2diabetes(65%).2.6%hadapreviousamputation,4.5%hadfootulcers,3.6%had
historyofpreviousmyocardialinfarction.Only1.4%ofthepatientswerecurrentsmokers,however,16%ofthepatients
admittedtohavensmokedinthepast.Wedidobserveastatisticallysignificantcorrelationbetweenthepresenceof
previousamputationsandthepresenceofPAD.
Conclusion:ThisisthefirstattempttoscreedforvascularpatientsatTASHdiabeticclinic.Ourresultssuggestedabout
20%prevalenceofPADamongstthesediabetespatients.Thiscrosssectionalstudyisprobablyanunderestimationof
theactualprevalenceduetothesmallsamplesizeandselectionbiasfpatientswhocanaffordtobeseenatthis
specializedhospital.

129
ValidationoffinalioPTHlevelandriskofdiseasepersistenceandrecurrence

MohammadH.Rajaei,DavidF.Schneider,RebeccaS.Sippel,HerbertChen,SarahC.Oltmann

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MohammadRajaei
(608)4463869
rajaei@surgery.wisc.edu
PostdoctoralFellow
GeneralSurgery
Chen'sLab
Translational/Clinical
PrimaryHyperparathyroidism,MinimallyInvasiveParathyroidectomy,intraoperative
ParathyroidHormone,SurgicalOutcomes,Followup

Introduction:Previousstudiessuggestedacorrelationbetweenafinalintraoperativeparathyroidhormone(ioPTH)level
of40pg/mLorhigherandanincreasedriskofdiseasepersistenceand/orrecurrenceinpatientswithprimary
hyperparathyroidism(PHPT)followingparathyroidectomy.Thisstudywasdesignedtotestthisobservationinalarger
populationofPHPTpatientswithextendedfollowuptobetterevaluatepersistenceandrecurrencerisk.
Method:AretrospectivereviewofpatientswithPHPTwhounderwentparathyroidectomywithioPTHmonitoringatan
academicendocrinesurgerypracticewasperformed.Caseswith<50%declineinioPTHattheendoftheoperation
(operativefailures),thosewithfamilialparathyroiddisease,previousparathyroidsurgery,lessthan6monthsfollowup
and/ormissingdatawereexcluded.Thestudypopulationwascategorizedinto5groupsbasedonfinalioPTHlevel:<10
pg/ml(group1),1019pg/ml(group2),2029pg/ml(group3),3039pg/ml(group4),and40pg/ml(group5).Analyses
wereperformedusingchisquareandANOVAasappropriate.DataareexpressedasmeanSEM.
Results:Between2001and2012,1371patientsmetinclusioncriteria.Meanagewas610.4years,with1064(78%)
femalepatients.Patientgenderwasnotdifferentamonggroups.CaseswithfinalioPTHlevels40pg/mLwerenotedto
beolder,withhigherpreoperativeserumcalcium,PTHandcreatininelevels(allp<0.01).Glandweightwassignificantly
greaterwithingroup5(p<0.01).Diseaseetiologyandsurgicalmanagementdidnotdifferbetweengroups.Patientswith
finalioPTH40pg/mLhadsignificantlyhigherratesofdiseasepersistenceandrecurrenceandmadeup17of19(89%)
persistentcases,and26of39(67%)recurrences(p<0.01).Meanfollowupwas210.6months.
Conclusion:ElevatedfinalioPTHlevels40pg/mlattheconclusionofparathyroidectomyisassociatedwithhigherrate
ofdiseasepersistenceandrecurrence.PatientswithfinalioPTHlevel40pg/mLwerealsonotedtohaveadditional
factorspresentwhichmayalsoinfluencerateorspeedofhormonedeclineresultinginhigherhormonelevelsatcase
conclusion.PatientsidentifiedtohavefinalioPTHlevels40pg/mLshouldbemonitoredclosely,andmoreoften,for
promptdiseaserecognitionandintervention.

130
Magneticresonanceimagingevaluationoflateralpterygoidcontributionsto
Eustachiantubeopening

MichaelH.McDonald,M.D.;ThomasR.Richards;JohnPark,M.D.,MatthewR.Hoffman,Ph.D.;JackJ.Jiang,M.D.,Ph.D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ThomasRichards
(608)2288204
trichards@wisc.edu
UndergraduateStudent
OtolaryngologyHead&NeckSurgery
JackJiang
Translational/Clinical
Eustachiantube,pterygoidmuscles,magneticresonanceimaging,Eustachiantube
dysfunction,patulousEustachiantube

INTRODUCTION:ThephysiologicalmechanismofEustachiantube(ET)openingisnotentirelyunderstood;specifically,
whichmusclescontributetoopeningispoorlydefined.Weperformedmagneticresonanceimaging(MRI)duringtasks
elicitingETopening.Ourobjectiveswereto1)determinewhichMRIsequenceswereconducivetoETopening
evaluation,and2)determineifthemedialandlateralpterygoidmusclesplayaroleinopening.
METHODS:Brainvolumeimagingpulseswereusedtoobtainstaticimagesforanatomicreference.Steadystatefree
precession,fastspoiledgradient,andsingleshotfastspinechopulsesweretovisualizedynamicopeninginseven
normalsubjectsandsevensubjectswithETdisorders(3withETdysfunctionand4withpatulousET).Imageswere
obtainedatrest,duringelicitationofETopeningbyswallowing,andduringjawmovement.Contractionofthe
pterygoidswasevaluatedbasedonchangeincrosssectionalarea.ETopeningwasmeasuredbydisplacementof
referencepointsonthewallsofthepharyngealorifice.
RESULTS:SSFSEandFIESTAcinesequencesshowedmusclemovementandopeningoftheETpharyngealorifice.The
crosssectionalareaofthepterygoidsdecreasedduringcontraction.ThismeasuredmovementduringreportedET
openingsuggestsapossiblerelationshipbetweenpterygoidactivityandETopening.
CONCLUSIONS:MRImaybeausefulmodalityforevaluationofETphysiology.Additionally,thisstudyprovidesfurther
evidencethatsuggeststhepterygoidsmayplayanamplificatoryroleinETopening.Furtherstudiesarewarrantedto
evaluatemorerefinedimagingsequencesanddevelopmorerigorousquantitativemethodsofevaluatingmuscle
contributionstoETopening.Greaterunderstandingofthisrolewillbebeneficialtosurgeonswhoareincreasingly
performingEustachiantubeinterventions.

131
CostAnalysisofColonCancerComparingColorectalSurgeonswithintheSame
Institution

DavidRivedalBS,SarahTevisMD,EugeneFoleyMD,BruceHarmsMD,CharlesHeiseMD,GregoryKennedyMD,PhD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

DavidRivedal
(414)8011582
rivedal@wisc.edu
MedicalStudent
GeneralSurgery
Dr.GregoryKennedy
Translational/Clinical
coloncancer,cost,surgery,variability,laparoscopy

Introduction:Costsassociatedwithcolectomyforcoloncancervarywidely.Laparoscopicresectionhasbeenshownto
beassociatedwithhigherintraoperativecostsascomparedwithopenprocedures.Complicationsandlongerlengthof
stayhavebeenshowntoleadtohigherpostoperativecosts.Specializedsurgeoncarehasbeenshowntodecreasecosts
inotherspecialties,howevercostvariationinsurgeryforcoloncancerhasnotbeenwelldescribed.Ouraimswereto
(1)identifyperioperativevariablesassociatedwithoperativeandpostoperativecostsincolectomypatientsand(2)
evaluatecostvariationbetweensurgeonsatasingleinstitution.
Methods:WeperformedaretrospectiveanalysisofourinstitutionalAmericanCollegeofSurgeonsNationalSurgical
QualityImprovementProgram(ACSNSQIP)databasefrom20062012.Thisdatabasewaspairedwithourinstitutional
billingdataandinvestigationsintothecombineddatabasewereapprovedbyourinstitutionalIRB.Billingdataincluded
thedirectcostofeachpatientencounterandthedirectORcosts,neithercontainedoverhead.Adultpatientswho
underwentcolonresectionforcoloncancer(ICD9153.*)byacolorectalsurgeonatUniversityofWisconsinHospital
wereincludedinthestudypopulation.PatientswithNSQIPdefinedpostoperativecomplicationswereexcluded.
StatisticalanalyseswereperformedusingSPSSv20.Chisquaredanalyseswereusedtoevaluateforassociations
betweenexplanatoryvariablesandcosts.Variableswithapvalueof0.10onunivariateanalysiswereincludedin
multivariateanalysis.Wecontrolledforcollinearitybycombininghighlycorrelatedvariablesinthefinalanalysis.
Results:Weidentified170patientswhounderwentcolectomyforcoloncancer.Theoverallcomplicationratewas34%
(N=57)andthe30dayreadmissionratewas8%(N=14).Patientsofnoncolorectalsurgeonsandthosewith
complicationswereexcludedyieldingastudypopulationof103patients.Weidentifiedgreatercostvariationinthe
postoperativeperiodascomparedwithoperativecosts.Laparoscopicproceduresandtotalandleftcolectomy
(comparedwithrightcolectomy)wereassociatedwithhighercostsintheoperatingroom.Also,beingmaleledto
highertocosts.Patientswithmorecomorbiditiesandbeingdischargedtoahigherlevelofcareweremorelikelyto
havehighercostsduringthepostoperativehospitalstay.
Conclusion:Costvariationincoloncancerpatientswhohaveundergonecolectomyishighlyvariable,evenin
uncomplicatedcasesoverseenbyacolorectalspecialist.Variationsinoperativecostsaremainlyafunctionofoperative
complexityandtechnique,whilepreexistingpatientcomorbiditydrivespostoperativecosts.Amoredetailedanalysisof
variablecostsincolectomypatientsmayidentifyspecificpracticeswhichcontributetocostvariability.Changesin
postoperativemanagementtargetingthesourcesofcostvariabilitymayleadtomoreefficientpatientcare.

132
TheCrossTrainingandDetrainingEffectofTongueExerciseintheCranial
SensorimotorSystem

AllisonJ.Schaser&NadineP.Connor

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

AllisonSchaser
(608)2659160
schaser@surgery.wisc.edu
GraduateStudent
OtolaryngologyHead&NeckSurgery
NadineP.Connor
Translational/Clinical
Exercise,Tongue,Detraining,CrossTraining,Neuroplasticity

Exercisebasedtherapiesarecurrentlyusedtotreatvoiceandswallowingdisorderswithoutaclearunderstandingofthe
mechanismsthatalterthecranialneuromuscularsystem.Forinstance,itisnotknowniftongueexercisesthattarget
improvedswallowingalterbehavioralparametersintargetedstructures(tongue)aloneoriftheseexercisesalsoaffect
otherswallowrelated(laryngeal)structures(crosstraining).Itisalsounknownifbehavioralchangesendureafter
therapyisdiscontinued.Therecentapplicationofprinciplesofneuroplasticitytorehabilitationhasrevolutionizedhow
wethinkabouttreatment,highlightingtheneedforchangeinbothbehaviorandneuralsubstratesforcreatinglasting
benefits.Itisdifficult,however,tostudyneuralsubstratesinhumanpatientswhilecontrollingforfactorsthatmay
influenceplasticitysuchasgeneticandenvironmentaldifferences.Theuseofaratmodelallowsthesecontrols.The
proposedresearchaimstofurtherourunderstandingoftheneuroplasticpotentialofexerciseinthecranial
sensorimotorsystemwiththeultimatelongtermgoalofguidingcareofindividualswithvoiceandswallowingproblems.

Researchfromourlaboratoryhasshownthat8weeksoftongueexerciseinadultratsleadstoincreasedtongue
forces.However,itisunknowniftongueexercisecreateslastingandgeneralizabletherapeuticbenefitsthroughoutthe
cranialsensorimotorsystemandacrossthelifespan.Itishypothesizedthat:1)tongueexercisewillresultinimproved
behavioraloutcomesinthecranialsensorimotorsystemacrossthelifespanwithgreatereffectsinlingualversus
laryngealstructures,and2)thatexerciseeffectswilldiminishwhentheexerciseprogramisdiscontinued(detraining).
Wewilltestthesehypothesesinaratmodelbycomparingbehavioralparametersinratsofdifferentagesthathave
undergoneatongueexerciseprogram,adetrainingprogram,oracontrolcondition.

Preliminaryresultsconfirmpreviousresearchinourlaboratory,showinganincreaseintongueforcesafter8
weeksoftargetedtongueexercise.Detrainingandcrosstrainingresultsareforthcoming,asthecurrentstudyisstill
ongoing.

Theproposedresearchissignificantbecauseitwillexaminethecrosstrainingpotentialofexerciseinthecranial
sensorimotorsystem,alongwiththeenduringeffectsofexercise(detraining),withthelongtermgoalofusingour
resultstoguidecurrenttherapytimelinesandprotocolsusedinclinicalpopulationswithvoiceandswallowingproblems.

133
PRECLINICALANDCLINICALEVALUATIONOFAHUMANSKINSUBSTITUTE
ENGINEEREDTOSECRETEELEVATEDLEVELSOFAHOSTDEFENSEPEPTIDE

ChristinaThomasVirnig,CathyRasmussen,AllenComer,MaryLokuta,LeeShaughnessy,SandySchlosser,ColetteJohnston,
RebeccaBauer,ThomasCleven,NathanWieczorek,B.LynnAllenHoffmann

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ChristinaThomasVirnig
(608)2630832
clthoma1@wisc.edu
Scientist
GeneralSurgery
LynnAllenHoffmann
Translational/Clinical
Antiinfective,HumanSkinSubstitute,DiabeticFootUlcer,ChronicWound,Tissue
Engineering

Introduction:Bacterialcontaminationisoneofthemostcommoncausesofimpairedwoundhealing.Duringthehealing
response,keratinocytesproducehostdefensepeptides(HDPs)thathaveantimicrobialactivityagainstadiversesetof
pathogens.
Objective:Thegoalofthisstudyistodeterminewhetherahumanskinsubstitutegeneticallymodifiedtoexpress
enhancedlevelsofthehumancathelicidinHDPisabletoreducemicrobialinfectioninananimalmodel.Thisproductwill
betestedinaphaseIclinicaltrialtoenhancethehealingofdiabeticfootulcers(DFU).
Methods:Anonviralvectorencodinghumancathelicidinwasstablyintroducedintoauniquekeratinocyteprogenitor
cellline.Theabilityofskintissueexpressingcathelicidintopreventbiofilmformationbydrugresistantmicrobial
pathogenswasevaluatedinafullthicknessexcisionalrodentmodel.Woundswereinoculatedwith~100CFUofa
disaggregatedbiofilmofA.baumanniiandgraftedwithantiinfectivetissue.Graftedtissueandwoundbedfromeach
animalwereharvestedforbacterialquantification5daysafterinoculation.

Results:ApplicationofantiinfectivetissuetoexcisionalwoundsinoculatedwithA.baumanniibiofilmresultedina
statisticallysignificantdecreaseinbacterialburdencomparedtounmodifiedtissue(99%,p<0.05).AnINDisbeing
preparedforsubmissiontotheFDAinordertoconductaphaseIstudyclinicallyevaluatingthisantiinfectivetissue.The
multicenter,unblindedstudywillevaluatethesafetyoftheantiinfectiveskinsubstituteinthetreatmentofrecently
occurringDFU.Subjectswillbeenrolledinthreecohortsevaluatingthenumberandfrequencyofapplicationsofthe
antiinfectivetissue.
Conclusions:TheapplicationofthisHDPexpressingskinsubstitutetoDFUisanticipatedtonotonlyprovidetemporary
epithelialcoveragewhileactingasametabolicallyactivesourceofgrowthfactorstopromotewoundhealing,butour
studiesprovidecompellingevidencethatthetissuemayalsoreducethelikelihoodofinfectionbynosocomialstrains
thatareofgreatconcerntothemedicalcommunity.Clinicalevaluationofthistechnologyisanticipatedtobeginin
2014.

134
PharyngealPressureandTimingDuringBolusTransit

ChelseaWalczak[1],TimothyMcCulloch[2],CorinneJones[3],WilliamBleifuss[4]

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

ChelseaWalczak
(715)2228754
walczak@surgery.wisc.edu
Scientist
OtolaryngologyHead&NeckSurgery
Dr.McCulloch
Translational/Clinical
BolusTransport,HighResolutionManometry,Videofluoroscopy,Deglutition,
PharyngealCompression

Purpose:Manometricpressureduringpharyngealcompressionhasbeenwelldescribed,butitisnotwellknownhow
pressureschangeduringbolustransport.Thepurposeofthisstudywastodescribethepressures,velocities,andtiming
surroundingbolustransportduringa10mLliquidswallow.
Method(s):Simultaneoushighresolutionmanometryandvideofluoroscopywasperformedon10healthysubjects(21
52years).Bolusheadandtailcoordinateswereplottedagainstsensorpositiontoobtainthecorrespondingpressures
andvideoframeinformation.SwallowswerescoredaccordingtotheModifiedBariumSwallowImpairmentProfile
(MBSImP).Comparisonsweremadeusingnonparametrictests.
Result(s):Throughoutthepharynx,theaveragepressureatbolustailpassagewasatleast2timesgreaterthanthe
averagepressureatbolusheadpassage(p=0.012).WhentheMBSImPTongueBase(TB)retractionscorewas1
signifyingatracecolumnofcontrastbetweenthetonguebaseandposteriorpharyngealwallthebolustailvelocitywas
significantlyslowerthanifthescorewas0,signifyingcompletecontraction(p=0.016).Inaddition,whentheTB
retractionscorewas1,thetimingofthebolustailrelativetothepressurerise(p=.0001)andpressuremaximum
(p=.0014)atthesensor(s)wasdelayedcomparedtoswallowswhichhadTBretractionscoresof0.
Conclusions(includingclinicalrelevance):Resultssupporttheideathatabolusmovesstrategicallyfromaplaceofhigh
pressuretolowpressure.BolusvelocityandpressuretimingvariationsarereflectedinMBSImPscores.Thus,tracking
theboluscouldbeausefultoolfordetectingswallowingabnormalities.

135
HighthroughputScreeningIdentifiesIdarubicinasaPreferentialInhibitorof
SmoothMuscleversusEndothelialCellProliferation

ShaktiA.Goel,LianWangGuo,BowenWang,SongGuo,DrewRoenneburg,GeneAnaniev,F.MichaelHoffmann,K.CraigKent

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

BowenWang
(608)6097933
wangbowen.glia@gmail.com
GraduateStudent
VascularSurgery
Dr.CraigKent
Translational/Clinical
IntimalHyperplasia,HighthroughputDrugScreening,SmoothMuscleCell,Endothelial
Cell,DifferentialEffect

Aims:Intimalhyperplasia(IH)isthecauseoftherecurrentocclusivevasculardisease(restenosis).Drugscurrentlyused
totreatrestenosiseffectivelyinhibitSMCproliferation,butalsoinhibitthegrowthoftheprotectiveluminalendothelial
cell(EC)lining,leadingtothrombosis.ToidentifycompoundsthatselectivelyinhibitSMCversusECproliferation,we
havedevelopedahighthroughputscreening(HTS)formatusinghumancellsandhaveemployedthistoscreena
multiplecompoundcollection(NIHClinicalCollection)
Methodsandresults:Wedevelopedanautomated,accurateproliferationassayin96wellplatesusinghumanaortic
SMCsandECs.UsingthisHTSformatwescreeneda447drugNIHClinicalLibrary.TheassayZvaluerangedfrom0.71
0.89withanoverallZof0.73.Weidentified11compoundsthatinhibitedSMCproliferationgreaterthan50%,among
whichidarubicinexhibitedauniquefeatureofpreferentiallyinhibitingSMCversusECproliferation.Concentration
responseanalysisrevealedthisdifferentialeffectmostevidentoveran~10nM5Mwindow.Invivotestingofidarubicin
inaratcarotidinjurymodelat14daysrevealedan80%reductionofintimalhyperplasiaanda45%increaseofresidual
lumenwithnosignificanteffectonreendothelialization.
Conclusion:WehaveproducedarobustHTSassayofhumanvascularcellproliferation,andidentifiedidarubicinasa
selectiveinhibitorofSMCversusECproliferationbothinvitroandinvivo.Screeningoflargerandmorediverse
compoundlibrariesmayleadtothediscoveryofthenextgenerationoftherapeuticsthatcaninhibitIHwithout
impairingreendothelialization.

136
DeterminationofanAccurateandConsistentMethodtoMeasuretheTakeoff
AngleofSupraaorticBranchVessels

BradleyWendorff,TaylorSmits,HillaryAlberta,WendyMeadows,JonMatsumura,M.D.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

BradleyWendorff
(262)2249818
wendorff@surgery.wisc.edu
UndergraduateStudent
VascularSurgery
JonMatsumura,M.D.
Translational/Clinical
EndovascularSurgery,RadiographicImaging,AorticMorphology,StentDesign,
MeasurementTechnique

Objective:Thepurposeofthisstudywastodevelopatechniqueprocessthataccuratelyandconsistentlymapsthe
morphologyoftheaorticarchtofostermoredatadrivendevicedesignandassuranceofdeliverabilitywhilemaintaining
patencyinthetortuousanatomyofthearch.
Methods:Accuraterepresentationofthevesselstakeoffangle(TOA)isdescribedastheanglewhosevectorintothe
branchvesselstartsinthecenteroftheostium,andterminatesdistallyinthecenter,whilethepairedvectormimicsthe
lineofflowintheaortaatthediscretepointcorrespondingwiththecenterofthebranchvesselsostium.Aortic
morphologycandifferlargelyfromsubjecttosubject,thustheadvantagesanddisadvantagesofusingtheDouble
ObliqueandCenterlineTOAmeasurementtechniqueswereexposedthroughdatacollectionon40subjects.The
DoubleObliquemethodisbasedonthereadersabilitytocreateaframeworkofgeometricrelationsintheaxial,
coronal,andsagittalviews,whiletheCenterlinemethodutilizesacenterlinecreatedbythereaderinthe3D
reconstructedviewtodeterminetheangle.Tobetterdescribetheaorticmorphology,thecircumferentialarrangement
ofthebranchvesselsrelativetothebrachiocephalicartery(BCA)wascombinedwiththeTOAtoyielda3Dmappingof
thearch.TeraRecons3DimagingworkstationwasusedcompletemeasurementsonsubjectsXrayComputed
Tomographyscans.TheTOAmeasurementtechniqueswerecomparedusingcriteriaofaccuracyandprecisionas
dictatedbythelimitsofagreementintheBlandAltmanPlot.
Results:InitialresultssuggestthattheDoubleObliquemethodlacksprecisionduetocompoundingerrorthrougha
seriesofmanipulations,eachwithaitsowndegreeofreadersubjectivity.TheCenterlinemethodposestobemore
preciseandpotentiallymoreaccurate,becauseitlimitssubjectivityandcompounderrorwithfewermanipulations
required.Additionally,theCenterlinemethodperformsbetterwithcomplicatedmorphologiesthatmaketheaorticline
offlowdifficulttodetermineusingtheDoubleObliquemethod.InterpersonalagreementusingtheClockfacemethod
wascalculatedwithCohenskappastatisticonabinarybasisofwhetherthereadershadthesamemeasurementresults.
Usingtheresultsofthemostaccurateandreproduciblemethod,anaveragemorphologywascalculated.
Conclusion:Withthecontinualprogressionofendovascularsurgerytechniques,accuratemethodsformeasuringthe
TOAofbranchvesselswillbecomeincreasinglyimportantasthenumberofproceduresinvolvingbranchvesselsrises.
ThisTOAmeasurementofthesupraaorticvesselscouldplayanimportantroleinthesurgeonscaseplanning,orin
decidingwhethertheintroductionofastentgraftwillhaveahighsuccessrate.Patientswitharchpathologiesoftenwait
monthsforacustomdevicespecifictotheirarchmorphology.Bydeterminingareliablemeasurementtechnique,
populationdataonaorticarchmorphologycanbeusedtoestablishanarrayofstandardOfftheshelfdevicesthatfit
amajorityofthepatientpopulation,thusshorteningthetimebetweenconsultationandoperation.

137
Measuringlisteningeffortusingpupildilation:implicationsformanagementof
hearingloss

MatthewWinn,JanEdwards,RuthLitovsky

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

MatthewWinn
(302)7407449
mwinn2@wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
RuthLitovsky
Translational/Clinical
Hearingloss,cochlearimplants,listeningeffort,pupillometry,outcomemeasures

Listeningeffortiselevatedforpeoplewhohavehearingloss,withconsequencesonhealth,occupationallife,andsocial
life.Conventionalclinicalmeasuresofspeechrecognition(e.g.accuracyscoresforwordandsentenceidentification)
adequatelydescribesomeaspectsofhearingloss,butfailtoquantifylisteningeffort.Previousliteraturehasshownthat
pupildilationisareliable,objectiveindexofcognitiveloadduringlisteningandisanextremelyfinegrainedmeasure.
Largerpupildilationisindicativeofgreatereffortexerted,andinalisteningtask,canbeusedtogaugehowdifficultitis
foralistenertounderstandspeechinvariouschallengingconditions.Inthisstudy,weaimedtoestablishpupillometryas
ameanstoevaluatetreatmentforpeoplewithbilateralcochlearimplants(BiCIs).Itwaspredictedthatpupillometric
measureswouldrevealdifferencesthataredifficulttodiscernusingconventionalbehavioralmeasures.
Resolutioninthespectral(frequency)domainremainsamajorproblemforCIs.Inthisstudy,wemodifiedthe
frequencyelectrodeallocationintheirspeechprocessorsusingamethodpurportedtoimprovespectralresolutionby
limitingelectricalcurrentfieldinteractionswithintheimplantedelectrodearray.Patientsusedboththeireveryday
clinicalCIsettingsandtheaforementionedexperimentalstrategywhenparticipatinginthreeexperimentaltasks:
conventionalwordrecognition,modifiedwordrecognition(tailoredtospecificallygaugespectralresolution),and
sentencerecognition,wherepupillaryresponsesweremeasuredduringlistening&response.Todate,11patientshave
completedallthreetests.
Inthispresentation,wedemonstratehowpupillarymeasuresduringsentencerecognitioncleanlydividedparticipants
intogroupsof:1)individualswhobenefitedfromthenewstrategy(n=4),2)individualswhoexperienceddeclinewith
thenewstrategy(n=4),and3)individualsforwhomnodifferencewasobserved(n=3).Thesemeasurescorrelated
wellwithperformanceonthemodifiedwordrecognitiontest(r2=0.644;p<0.01),However,wordrecognitiondidnot
predictimprovementsinspectralresolution(r2=0.001;p=0.91),orlisteningeffort(r2=0.01;p=0.75).Theseresults
underscorethelimitationsofsomeconventionalmeasurements,andhighlighttheimportanceofdirectsensitive
measureslikepupillometryinoutcomesassessment.
Insummary,weprovideamodelbywhichindividualizedtreatmentforpatientscanbeevaluatedonthebasisof
listeningeffort,beyondtheconventionaltechniquesofwordandsentencerecognition.Byusingstimuliand
measurementtechniquesthataretailoredtospecificaspectsofhearingandtheexperienceofincreasedeffort,wewere
abletorecognizethebenefitordetrimentofaspecificclinicaltreatmentthatwasnotcapturedbyconventional
outcomemeasures.Inthispresentation,wediscussthepotentialforthistechniquetoanswerotherimportant
questionswithinotolaryngologyandaudiology.
ResearchsupportedbyNIDCDR01DC002932(Edwards),NIDCD5R01DC03083(Litovsky),andP30HD03352(Waisman
core)

138
ResolutionofThyroglobulinAntibodiesafterTotalThyroidectomyforCancer

JimmyXu,B.S.,RyanBergren,B.S.,DavidSchneider,M.D.,HerbertChen,M.D.,F.A.C.S.,RebeccaS.Sippel,M.D.,F.A.C.S.

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

JimmyXu
(917)3718912
jlxu@wisc.edu
MedicalStudent
GeneralSurgery
RebeccaS.Sippel,MD,FACS
Translational/Clinical
thyroid,cancer,thyroidectomy,thyroglobulin,antibody

Background:
Thyroglobulin(Tg)levelscanbeusedpostoperativelytoidentifyresidualorrecurrentthyroidcanceraftersurgical
treatment.Thyroglobulinantibodies(TgAb)areproducedby1025%ofthyroidcancerpatientsandcaninterferewith
thyroglobulinmeasurement.ThepurposeofthestudywastodescribethetimecourseofTgAbresolutionandthe
significanceofapersistentlyelevatedlevelinantibodypositivepatientsafterthyroidectomy.
Methods:Adatabaseof247consecutivepatientswithpreoperativeTgAbmeasuredwhounderwentsurgicaltreatment
ofdifferentiatedthyroidcancer(DTC)betweenJanuary2007andMay2013wasreviewed.Patientswerestratified
basedonTgAbstatus(positiveornegative)anddiseaserecurrence(definedbybiopsyprovendiseaseorunplanned
secondsurgery).SurvivalandregressionanalysiswasusedtodeterminethetimecourseofTgAbresolution.LogRank
(MantelCox)wasusedtodetermineanassociationbetweenpersistentlyelevatedantibodylevelsandcancer
recurrence.SPSSstatisticalsoftwarewasusedforanalysis.
Results:
Of247patients(77%women,23%men;meanageSE,45.71.0years)withTgAbmeasuredpreoperatively,34(14%)
hadpositivepreoperativeTgAblevels(greaterthanorequalto20IU/ml)(mean298.199.2IU/ml).Themediantime
toTgAbresolutionwas11.02.3months,andthemajorityresolvedby32.4months.Regressionanalysisofthepatients
withcompletelyresolvedantibodiesyieldedanaveragedeclineof11%IU/mlpermonth2.2%.Diseasefreesurvival
wasequivalentbetweenTgAbpositiveandTgAbnegativegroups(p=0.4).In9of34patients,antibodieshadnot
resolvedatlastfollowupbutimagingcouldnotidentifyrecurrentdisease.
Conclusion:
TgAbarecommoninpatientswiththyroidcancerbuttheyresolveaftersuccessfultreatmentatapproximately11%
IU/mlpermonthfromtheirpreoperativelevelswithmedianresolutionat11.0months.Persistentlyelevatedlevels
afterthyroidectomywerenotassociatedwithdiseaserecurrenceinourseries.

139
Yellowcoronalvocalfoldscarauniquepattern

WilliamGYoung,MD;SethHDailey,MD

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

WilliamYoung
(206)8195019
young@surgery.wisc.edu
PostdoctoralFellow
OtolaryngologyHead&NeckSurgery
Dailey
Translational/Clinical
Vocalfoldscar,Coronalscar,sulcusvocalis,hoarseness,vocaleffort

Introduction:
Vocalfold(VF)scarringistypicallydescribedinalongitudinalorientationalongtheVF;however,wehaveidentifiedan
intriguinganduniquecoronalscarringpatterninagroupofpatientspresentingwithdysphoniaandincreasedvocal
effort.
Methods:
Retrospectivecaseseriesdescribingacousticandstroboscopicfindings,aswellasoperativetechniquesforcoronalVF
scarmanagement.
Results:
4womenpresentedwithvisiblereductionofmucosalwavepropagationfrombilateral(n=3)orunilateral(n=1)
yellowishsubepithelialprominencesorientedperpendiculartotheVF.Tissuereorientationviamicroflapexposure
yieldedpathologyconsistentwithfibroconnectivetissue(n=3).Operativeinterventionwasassociatedwithimproved
closure(n=4)andanimprovedVoiceHandicapIndex(VHI)(p=0.05).Mucosalwavereturn,phonationthreshold
pressures,maximumphonationtime,andjitterchangeswereinconsistent.

Conclusions:
YellowcoronalVFscarpresentsanintriguingclinicalscenarioofunknownetiology.Tissuereorientationcanbe
associatedwithanimprovementinVFclosureandVHI.

140
OverexpressionofDysadherininThyroidCancerisAssociatedwithTumor
AggressivePhenotypes

XiaoMinYu,MD,PhD,CelinaMontemayorGarcia,MD,PhD,EricWeinlander,BA,RicardoV.Lloyd,MD,PhD,HerbertChen,MD,
FACS

Submitter
Phone
EmailAddress
Classification
Division
Lab
ScienceType
Keywords:

XiaoMinYu
(608)2653749
yux@surgery.wisc.edu
Scientist
GeneralSurgery
HerbertChen
Translational/Clinical
thyroidcancer,dysadherin,prognosis,lymphnodemetastasis,aggressivephenotype

Background:Dysadherin,anewlydiscoveredcellmembraneglycoprotein,hasbeencharacterizedtobeactivelyinvolved
intheregulationofcellcelladhesionandcancermetastases.However,littleisknownaboutDysadherininthyroid
diseases.ThepurposeofthisstudywastodeterminetheexpressionpatternandclinicalrelevanceofDysadherinin
bothbenignthyroiddiseasesandthyroidcancers.
Methods:Dysadherinexpressionwasexaminedbyimmunohistochemistryonaconstructedtissuemicroarraywith
triplicatesamplesofeachcase.Theimmunohistochemistryscoringwasperformedbytwoindependentobservers
basedonthestainingpatternandintensity.Thedatawerecomparedamongthepatientswithdifferentthyroid
pathologyandcorrelatedwithpatients'clinicopathologicfeatures.
Results:Atotalof158caseswereevaluatedincluding10normalthyroids,10nodulargoiters,11Hashimotosthyroiditis,
32follicularadenomas,57papillarycarcinomas(PTCs),28follicularcarcinomas(FTCs)and10anaplasticcarcinomas
(ATCs).NoneofthenormalthyroidsexpressedDysadherin.Inall53benignthyroidtissues,onlyonewithnodulargoiter
showedweakpositivestaining.Mostofthethyroidcancers(59/95)demonstratedmoderatetostrongcellmembrane
immunoreactivityforDysadherin(62%vs.2%inbenigngroup,p<0.001).DysadherinwasstronglyexpressedinallATCs
(100%)whilemoderatepositivitywasseenin42PTCs(74%)and7FTCs(25%).Aslightexpressiondifferenceof
DysadherinwasseenbetweenclassicalandfollicularvariantofPTC(86%vs.62%positivity,p=0.043).InPTCs,the
overexpressionofDysadherinwassignificantlyassociatedwiththepresenceofextrathyroidalextensionandlymphnode
metastases(p=0.045and0.017,respectively),butnotwiththetumorsizeorolderage.
Conclusions:Dysadherinisoverexpressedinabovehalfofthethyroidcancercases.Thepositiveexpressionof
Dysadherinisassociatedwithundifferentiatedandmoreaggressivethyroidcancerphenotypes,whichmayserveasa
prognosticmarkerforadverseclinicaloutcome.

141
Protective Role of Complement Blockade in the Context of Renal Ischemiareperfusion Injury and Kidney Transplantation.
Martynas Ziemelis, Juan S. Danobeitia, Xiaobo Ma, Gengwen Huang, Drew A. Roenneburg and Luis A. Fernandez

Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Martynas Ziemelis
(608) 262-8459
ziemelis@surgery.wisc.edu
Scientist
Transplantation
Luis Fernandez
Translational/Clinical
transplantation, ischemia-reperfusion, complement, fibrosis, kidney

Introduction:
Currently, organ donation following brain death (BD) is the most common source of transplantable kidneys. BD induces
ischemia/reperfusion injury (IRI) and initiates an inflammatory state detrimental to graft survival. Specifically, activation
of the complement cascade is known to play a role in the local inflammatory response to tissue injury and has been
associated with an increased risk of rejection, allograft failure and progression to fibrosis. However, the link between
complement activation and progression to fibrosis remains unclear. We hypothesized that complement blockade using a
complement inhibitor (rhC1INH) should decrease inflammation following IRI, decreasing inflammatory damage thus
limiting progression to fibrosis.
Methods:
We used a mouse (C57/B6) model of unilateral renal IRI followed by contralateral nephrectomy. Animals were assigned
to one of four groups. G1: Sham no IRI (n=6), G2: IRI + vehicle (n=6), G3: IRI + rhC1inh (750U/kg) treatment (n=6), and
G4: IRI + vehicle + no contralateral nephrectomy (n=6) to assess fibrotic changes at 90 days post-injury. We evaluated
structural damage, renal function, inflammatory response and overall survival at 1, 3, 7, 30 and 90 days post-IRI.
Results:
Animals in G3 showed a significant increase in survival with 83.3% of animals reaching the 90 day endpoint compared to
14.2% in G2 (p=0.019). Serum creatinine was significantly decreased in G3 animals at days 1 and 3 post-IRI compared to
G2 animals receiving vehicle prior to injury (p<0.05) and not different from controls in G1. H&E stains of recovered
kidneys revealed preserved renal tubular architecture in G1 and G3 at all-time points in marked contrast to G2 and G4
kidneys which evidenced extensive necrotic damage. A significant reduction was also observed in IL-6 and MCP-1 mRNA
expression in kidney tissue recovered from rhC1INH treated mice in G3 at days 1 and 3 when compared to G2
counterparts (p < 0.05). Subsequent staining showed a significant reduction in tissue infiltration by Macrophages-(F4/80)
and Neutrophils-(Ly6G) in kidneys from G1 and G3 animals along with decreased renal C3d deposition and a reduction in
C5a/C5aR signaling when compared to G2. To assess progression to fibrosis we stained kidneys at 30 and 90 days post
reperfusion targeting markers of fibrosis (SMA and Desmin). Kidneys from G3 mice had significantly lower levels of
both markers at 30 and 90 days post-IRI and lower renal levels of TGF-1 and p-SMAD2 when compared to untreated
animals in G4. Next, we used an in-vitro system to test leukocytic and renal epithelial cell (RTEC) response to
complement stimulation. Interestingly, our preliminary data suggests that RTEC but not leukocytes respond to C5a by
increasing TGF-1 and SMAD2 phosphorylation after C5a treatment. This suggests a novel paradigm in which
complement induces direct tubular fibrosis in a C5aR/TGF-1/SMAD2 dependent manner.
Conclusion:
Our findings indicate that intravenous delivery of rhC1INH protects kidneys from inflammatory injury, prevents immunecell infiltration, limits progression to fibrosis and results in an overall improvement in renal function and survival after
ischemic injury. Complement inhibition constitutes a promising strategy in the prevention of ischemic injury in the
context of organ transplantation.

142
The Evolving Integrated Vascular Surgery Residency Curriculum
Brigitte K. Smith, MD,1 Jacob A. Greenberg, MD, EdM,1 Erica L. Mitchell, MD,2 Madison, WI; and Portland, OR

Submitter
Phone
Email Address
Classification
Division

BRIGITTE SMITH
(608) 265-4420
bsmith5@uwhealth.org
Resident
Vascular

Background and Objectives:


Since their introduction several years ago, Integrated (0+5) Vascular Surgery Residency Programs are being
increasing developed across the country. To date, however there is no defined universal curriculum for these
programs and each program is responsible for creating its own curriculum. The aim of this study was to review
the experiences of current 0+5 Program Directors (PD) to determine what factors contributed to the curricular
development within their institution.
Methods:
Semi-structured interviews were conducted with 0+5 PDs to explore their experiences with program
development, factors influencing the latter, and rationale for current curricula. The interview script was loosely
structured to explore several factors including: time of incoming residents first exposure to the vascular surgical
service, timing and rationale behind the timing of core surgical rotations throughout the 5 year program,
educational value of non-surgical rotations, opportunities for leadership and scholarly activity, and influence the
general surgery program and institutional climate had on curricular structure. All interviews were conducted by
a single interviewer. All interviews were qualitatively analyzed using emergent theme analysis.
Results:
26 0+5 PDs participated in the study. 69% believed establishing professional identity early reduces resident
attrition and recommend starting incoming trainees on vascular surgical services. 62% spread core surgical
rotations over the 1st three years to optimize general surgical exposure and the majority of programs have
eliminated specific rotations as they were not considered valuable to the goals of training. Factors considered
most important by PDs in curricular development include: building upon existing institutional opportunities
(96%), avoiding rotations considered unsuccessful by experienced programs (92%), and maintaining a good
working relationship with general surgery (77 %). 58% of PDs voiced concern over the lack of standardization
amongst the differing programs and the majority of PDs agree that some degree of programmatic
standardization is critical for the continued success of the 0+5 training paradigm.
Conclusions:
Qualitative evaluation of PD experiences with the development of 0+5 vascular surgery residency programs
reveals the key factors that commonly influence program design. Programs continue to evolve in both
structure and content as PDs respond to these influences. Learning from the collective experience of PDs and
some standardization of the curricula may help current and future programs avoid common pitfalls in curricular
development.

143
Learning from developing countries in strengthening health systems:
An evaluation of personal, professional, and institutional impact of global health
volunteers
Busse H, Aboneh E, Azazh A, Wubben R, Tupesis J, Tefera G

Submitter
Phone
Email Address
Classification
Division

HEIDI BUSSE
(608) 263-0327
hbusse@wisc.edu
Assistant Researcher
Vascular

BACKGROUND: The positive impact of global health activities by volunteers from the United States in low-and middleincome countries has been well recognized and reported. However, what has failed to be captured in traditional
monitoring and evaluation plans is what kinds of change happens to US-based partners due to engagement in global
health partnerships, both at the individual and institutional levels. This type of impact is referred to as reverse
learning. Current evaluation indicators for global health partnerships tend to focus on what knowledge, ideas, and
activities the institution from the developed country transferred to the developing country. Even if a partnership was
developed with expectations of reciprocal relationships, shared accountability, and equity, implicit in this traditional
global health paradigm is the assumption that change and knowledge are solely transferred from developed to
developing countries. There is a need for more evidence about how global health partnerships can transform US-based
individuals and institutions to know what kinds of impact could be measured.
METHODS: To assess potential indicators for measuring impact, 80 US-based health care professionals who participated
in a global health partnership between Addis Ababa University and the University of Wisconsin were surveyed to
understand the personal and professional change they experienced as a result of their global health service. Surveys
were web-based and included multiple choice and open-ended questions.
RESULTS: Of the eighty surveys distributed, 62 responded (77.5% response rate). Eighty-three percent reported they
accomplished their trip objectives, 95% would participate in future activities and 96% would recommend participation to
other colleagues. Eighty-nine percent reported personal impact and 73% reported impact on their professional
development. Previous global health experience, multiple prior trips, and the desire for career advancement was
associated with positive impact on professional development. Statistically significant changes were seen in seven World
Health Organization health systems competencies.
DISCUSSION: Asking what kinds of impact can be attributed to participation in global health activities for US partners is
rarely assessed; however, it has important policy, economic, and programmatic implications. Further, it represents a
change in how partnerships are structured and recognizes the important lessons developing countries have to teach.

144
Re-differentiation of human T cell-derived induced pluripotent stem cells into
the T cell lineage
Matthew E. Brown and William J. Burlingham
Submitter
Phone
Email Address
Classification
Division
Lab
Science Type
Keywords:

Matthew Brown
608.263.2485
brownm@surgery.wisc.edu
Research Assistant
Transplant
William Burmingham
Basic
iPSC, T-Cell, Tolerance

Induced pluripotent stem cells (iPSCs) hold great potential for regenerative medicine therapies and for
the study of lymphoid development. Derived by reprogramming autologous somatic cells via transfection
with pluripotency related transcription factors, iPSCs provide a scalable cell source from which to
differentiate clinically relevant quantities of syngeneic replacement cells. T cells are an attractive starting
cell source for iPSC derivation because of their abundance in peripheral blood as well as the fact that the
original T cell receptor gene rearrangement of the source cell is preserved throughout the conversion to
iPSCs and into differentiated terminal cell types, akin to a bar code. In the work presented here, we
explore the isolation of T cell clones relevant to transplant tolerance, including regulatory T cells and
Th17 cells, reprogramming them via episomal scar-less plasmids, differentiating the iPSCs in vitro and
in vivo into CD34+ hematopoietic progenitor cells, and further differentiating the cells in vitro and in vivo
into the T lineage to examine their phenotype and functionality. Utilizing iPSCs for transplant research
will increase understanding of T cell biology, tolerance mechanisms and facilitate development of future
regenerative medicine therapies.