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Appalachian Student Research Forum

Office of Research and Sponsored Programs

Division IV

Abstracts Submitted:Division IV - Medical students


THE REGULATION OF IL-1 b-INDUCED PRO-INFLAMMATORY CYTOKINE PRODUCTION IN MAST CELLS BY EPINEPHRINE AND IMMUNOSUPRRESANTS

Shannon Y. Pitts, Guha Krishnaswamy, Ellis King, Karen Cantor, David S. Chi, Department of Internal Medicine, East Tennessee State University, College of Medicine, Johnson City, TN 37614

Human mast cells are located throughout the body in association with mucosal surfaces and connective tissue. They function as essential elements in allergic reactions and inflammatory response. Previous studies show stress and mast cell mediated inflammatory response are linked through the expression of pro-inflammatory cytokines: Interleukin-6 (IL-6), Interleukin-8 (IL-8) and Interleukin-13 (IL-13). The purpose of this study was to examine the regulation of IL-1 b induced pro-inflammatory response by epinephrine and immunosuppressants. The human mast cell leukemia line (HMC-1) cells (0.75 x 106 cells/ml) were incubated for 24 hours in triplicate with various combinations of IL-1 b (10 ng/ml), epinephrine (1 x 10 -5 M), prednisolone (1 x 10 -6 M), dexamethosone (1 x 10 -7 M), and cyclosporine A (10 mg/ml). The cell free supernatants were analyzed for IL-6, IL-8, and IL-13 levels by enzyme linked immunosorbent assays. The cell viability was determined before and after each incubation period, and no significant difference was found (>90%). The data was analyzed by using the Students t-test, and a p-value <0.05 was considered significant. Epinephrine alone produced at most trace amounts of cytokines, while IL-1 b alone induced a significant amount of cytokines. The addition of epinephrine to IL-1 b significantly enhanced the production of IL-6, IL-8, and IL-13 by mast cells. However, the addition of the immunosuppressants decreased this enhancement. In order to study the involvement of two regulatory pathways, NF-kappa B and MAPK (p38) pathways, the HMC-1 cells were pre-incubated with the signal transduction inhibitors, Bay-11 (10 mM) and SB203580 (10 mM), for 2 hrs prior to addition of IL-1 b (10 ng/ml) and/or epinephrine (1 x 10 -5 M). Bay-11 and SB203580 significantly suppressed the enhancing effect of IL-1beta and epinephrine on cytokine production. The results suggest epinephrine enhances IL-1 b induction of HMC-1 pro-inflammatory cytokine production, which is inhibited by immunosuppressants. The mechanism of the enhancing effect of epinephrine on IL-1 b induction of HMC-1 pro-inflammatory cytokine production appears to involve both MAPK (p38) and NF-kappa B pathways. (Supported by NIH grant HL63070, the Ruth R. Harris Endowment, and RDC of ETSU.)


POTENTIAL FOR A RESIDENT WITH A DISABILITY IN THE FIELD OF OBSTETRICS/GYNECOLOGY

Matthew T. Standridge MS, Martin Olsen MD, John Kalbfleish PhD

Objective: To investigate whether a resident physician who has a congenital absence of the left hand at the wrist could succeed in an Obstetrics/Gynecology residency. Design and Participants: Surveys were mailed to program directors of obstetrics and gynecology residency programs. Interventions: Program directors were asked to fill out the survey and return it to our institution in the enclosed prepaid envelope. Main Outcome Measured: Surveys were scored based on the program directors responses, and chi-square tables were created to analyze the answers to questions compared to the comments offered. Results: Of the 252 surveys mailed, 146 were returned completed. A majority (58%) of program directors felt that a resident with one hand could not meet the standards of performance expected in an obstetrics and gynecology residency. Fifty-two percent felt that training a resident with one hand would be disruptive to the training of other residents. Program directors were unsure whether gynecologic surgery patients would accept a resident with one hand. There was a mixed response as to whether a resident with this disability should pursue a career in the field of obstetrics and gynecology, with 26% answering yes, 37% answering no, and 34% uncertain. Conclusions: Based on the data, the majority of program directors would discourage a resident with one hand from pursuing a career in the field of obstetrics and gynecology. However, there was a significant positive response that it would be possible for a resident physician to enter this field, particularly if they planned on office gynecology or a non-surgical practice. Whatever the case, we feel that this decision relies on many variables and must be decided on an individual basis.


FABRICATION OF 3D TISSUE ANALOGS: CELL VIABILITY, PLASMID STABILITY, AND DISTRIBUTION

Wan-Yin Chan and Benjamin M. Wu, University of California, Los Angeles, Department of Biomedical Engineering, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, Dept of Materials Science and Engineering, Los Angeles, CA 90025

One of the goals of tissue engineering is to repair or correct damaged tissue. One method is to implant scaffolds seeded with biological materials, including cells and signals at the site of need. A common problem is non-uniform seeding on scaffolds, which may require the control of spatial deposition of cells, signals, and matrix simultaneously. Objective: To determine the appropriate matter-deposition strategy of cells and DNA signals in terms of cell viability, plasmid stability, and partitioning effect of substrate material on plasmid distribution. Methods: NIH3t3 fibroblast cells were jetted through a piezoelectric-driven capillary microdroplet generator under a range of computer-driven waveform parameters (pulse amplitude, duty cycle, frequency). Cell viability was assessed with calceinAM and ethidium homodimer-1 stain and quantitative image analysis. 60ug of CMV-LUX, 1.6ug of 3xflag-CMV2, and 40ug of pDsred2-N1 were jetted and their structural integrity was analyzed on an agarose gel and compared to unjetted plasmids. In vitro transfection of pDsred1 and pDsred2 into NIH3t3 fibroblastic cells was performed to confirm plasmid functionality. Plasmid affinity was tested by exposing 1ug of pDsred diluted into 1X PBS to PLGA, PLGA/HA, PCL, and cationic gelatin scaffolding substratess at 37degrees Celsius. Results: Cells printed from pressure driven dispenser maintained 82+/-3% viability as compared to the positive controls (88+/-4% viability; p~0.17). Cells printed from piezoelectric generator only maintained a 23+/-7% viability compared to the controls (96+/-1% viability;p<0.005). Plasmid structural integrity was preserved after jetting. Cationic gelatin binds to DNA upon exposure to it, while PLGA and PLGA/HA appear to have a binding affinity to DNA. pDsred2-N1 transfected in the absence of serum yielded higher efficiencies. Conclusions: Cell viability and plasmid stability was maintained after printing from the pressure driven dispenser, which aids in the design of a printer to ultimately pr


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