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"Engineering Platforms for Cellular and Molecular Processes" Seminar Series


High School Macrofluidics Lab 2

Elizabeth Mansfield, one of the 2010 High School Teacher Fellows, presents part two of her work on how to demonstrate microfluidic concepts to high school students.

High School Macrofluidics Lab 1

Elizabeth Mansfield, one of the 2010 High School Teacher Fellows, presents part one of her work on how to demonstrate microfluidic concepts to high school students.

Dr. Jeffrey C. Petruska - Nov 4, 2009

June 7, 2010 - "Neural Plasticity in the Adult Nervous System: New Tools and Approaches"

Abstract:  Laboratory of Neural Physiology and Plasticity seeks to understand the basic principles and the molecular and cellular mechanisms that regulate growth of axons in the adult mammalian nervous system with the goal of advancing clinical therapeutic interventions to address chronic pain and central nervous system trauma and stroke.

This seminar will present the concepts and characteristics of neural plasticity in the adult mammalian nervous system and some of the work done to identify and manipulate the extracellular and intracellular molecular mechanisms influencing neuronal function and connectivity. We have identified genetic changes and molecular signalling pathways associated with the native neuronal plasticity processes called “Collateral Sprouting”, where an existing intact axon extends a new branch in order to generate a new connection. We are now validating these observations and attempting to determine the mechanistic role of the highlighted factors and pathways in collateral sprouting. As a result, we have broken new ground by determining that the molecular profile of collateral sprouting is dramatically different from that of axonal regeneration – another major form of neuronal plasticity in the adult nervous system.

The seminar will also present an overview of the challenges inherent to these efforts, and the novel means being employed and developed to overcome these challenges, including novel surgical, genetic, electrophysiological, and bioinformatic/analytical approaches. Specifically, we seek to enable efficient systems- and organism-level testing of those hypotheses derived from molecular-, biochemical-, and cellular-level experimentation.

Biography:  Dr. Petruska received a B.A. in Psychology from Boston College where he was a scholarship athlete and examined the neural circuits regulating maternal behaviors with Dr. Michael Numan. He received a Ph.D. in Neuroscience from the University of Florida, the first Department of Neuroscience chartered in the U.S., where he worked with Dr. Richard Johnson examining the neural plasticity process of collateral sprouting, with Dr. Brian Cooper detailing a system for translating sensory neuron identities between in vivo and in vitro contexts, and co-authored a textbook for Medical Neuroscience. He received post- doctoral training at SUNY Stony Brook with Dr. Lorne M. Mendell, former President of the international Society for Neuroscience, where he examined the mechanisms of neural plasticity as an Associate of the Christopher and Dana Reeve Foundation’s International Consortium on Spinal Cord Injury Research. Dr. Petruska joined UofL in May, 2008 to establish the Laboratory of Neural Physiology and Plasticity. He has filed technology disclosures at UF, SUNY-SB, and UofL, with a patent issued to UF for a neural- interface device, and a patent application in-process at SUNY-SB for a sensory-selective anesthetic. He has won the BF Goodrich “Collegiate Inventors Award” and the International Campaign to Cure Spinal Cord Injury Paralysis “Outstanding Young Investigator Award”, and been awarded research support from the Paralysis Project of America, and the International Institute for Research in Paraplegia.


Michael D. Martin - Oct 28, 2009

May 27, 2010

Abstract: Lipid bilayers offer significant promise as elements in microfabricated devices with applications ranging from highly sensitive biochemical sensors, platforms for studying structure function of membrane proteins to metabolic machines such as biobatteries. These fragile membranes, only 5nm thick, must be stabilized before they can be practically applied. The majority of current stabilization techniques for these fragile structures require depositing the membranes on a solid or semisolid support to extend their functional lifetimes. This approach limits their applicability due to the non-physiological environment in this configuration and to lack of access to one side of the membrane.

In order to explore techniques for the stabilization of freely suspended planar bilayer lipid membranes (BLM), devices were developed composed of a 3.5μm thick polyimide films with a variety of diameters and numbers of perforations through them. Initial studies were performed using devices with arrays of 30μm, 75μm and 100μm diameter holes and egg phosphatidylcholine (PC). Formation and evolution of lipid membranes on the arrays was examined as a function of time via white light microscopy. Then the tinned membranes were coated with 200nm of vapor deposited polymer (parylene) essentially fixing them for further study. The resulting parylene coated lipid membranes were characterized using SEM, EDAX, AFM, interferometry and confocal microscopy. For electrical characterization, devices with a single 75μm or 100μm diameter hole were captured in a Teflon housing with reservoirs on either side for perfusion of buffer. Lipid membranes were formed with 1,2-Dioleoyl-sn-Glycero-3-Phosphocholine (DOPC) solvated in decane at a concentration of 5mg/ml. The lipids were applied to the microfabricated aperture by “folding” together solvated DOPC after deposition on an air-water interface. Electrical properties of the resulting lipid membranes were measured in 1M KCl using a commercial patch clamp amplifier (Axopatch 200B), oscilloscope and waveform generator. The formation of a true bilayer was demonstrated by measuring gigaohm resistances and specific capacitance of 1.13±0.21μF/cm2. To further demonstrate the efficacy of the apparatus and techniques, gramicidin, a small ion channel protein, was incorporated into bilayer membranes. Electrical measurements demonstrated functional reconstitution of the channel protein.

Biography: Michael Martin has 10 years experience in various microfabrication techniques ranging from conventional photolithography to micromilling and excimer laser microfabrication. He has extensive experience developing laser microfabrication techniques for projects such as radiation detectors for a NASA gamma ray telescope, plasma display production, and field emission display prototyping. Mr. Martin’s current position is Research Engineer in the Dept of Electrical and Computer Engineering at the University of Louisville. He is presently working on DHS and DoD grants funded to develop microfabricated preconcentrators for use in trace chemical detection. In his previous position as a Research Engineer for Plasmion Displays LLC in Hoboken, NJ, he was responsible for development of prototype micro machined plasma display panels and refinement of a Monte Carlo simulation of plasma discharges. Mr. Martin holds a B.S. in physics from Austin Peay State University and a M.S. in physics from the University of Louisville.



Dr. Venkatakrishna Rao Jala - Oct. 14, 2010

Abstract:  Seven transmembrane G-protein coupled receptors (GPCRs) play a critical role in several biological and pathological processes and have been favorite molecules for drug targeting. Leukotriene B4 (LTB4), a pro-inflammatory lipid molecule, mediates its actions via a high affinity GPCR known as BLT1. The LTB4/BLT1 axis was shown to be an important mediator of inflammatory disorders including atherosclerosis, arthritis and asthma. Molecular insights into the structure-function of LTB4 receptors and LTB4-mediated signal transduction pathways will be discussed. Briefly, we have identified the critical amino acid residues involved in the ligand binding, activation mechanism, phosphorylation of the receptors. We have also demonstrated a critical role for BLT1 in the development of atherosclerosis and identified mechanisms involved in the disease progression. Currently, in collaboration with bio-engineering department we are developing new systems for understanding the GPCR functions in cells under stress. Specifically, we are investigating how dynamic shear stress alters growth and expression patterns of BLT1 and other atherogenic genes in human umbilical vascular endothelial cells.

Biography:  Dr. Jala is an Assistant Professor and Chairman of Curriculum Committee of the Dept of Microbiology and Immunology at the University of Louisville since 2004. He received his Ph.D. in Biochemistry from the Indian Institute of Science, Bangalore, India in 2001. He joined the James Graham Brown Cancer Center, Dept of Microbiology and Immunology at UofL as a Postdoctoral Fellow in 2001 and joined the Faculty in 2004. Dr. Jala’s research has been recognized by the “best Poster Award (1st prize)” at Research Louisville in 2003 and he received “Prof. A. Krishnamurthy Award” for the best paper published in Indian journals from Society for Biological Chemists in 2004.

Dr. Jala’s research focuses on structure-function relationships of G-protein coupled receptors (GPCRs) and their role in the development of inflammatory diseases. His main focus is in investigating the causative effects and mechanisms in the development of inflammatory diseases in mouse models, especially in atherosclerosis, arthritis and colon cancer. GPCRs are the largest known class of molecular targets with proven therapeutic value. GPCRs are instrumental in the transmission of a wide range of chemical messages from the extracellular environment to the interior of the cell. GPCRs have been implicated in a wide range of disorders including allergies, cardiovascular dysfunction, depression, obesity, cancer, pain, arthritis, diabetes, AIDS and various central nervous system disorders.