2012-2013 PROJECTS AVAILABLE FOR ANESTHESIA INTEREST GROUP MEMBERS & MEDICAL STUDENT SCHOLARLY PROJECTS: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Genetics of Post-Mastectomy Pain Inna Belfer, M.D., Ph.D., Associate Professor of Anesthesiology Molecular Epidemiology of Pain Program Persistent pain following breast cancer surgery is highly disabling and is a significant clinical problem due to limited treatments. Up to 47% of patients report pain after surgery, of which 13% is severe, and up to 40% moderate. Little is known about risk factors contributing to pain chronicity and their interaction that affects inter-individual variability in pain. This study will identify genetic & environmental (demographic, surgical, psychological) factors associated with persistent pain phenotypes. We collect comprehensive data on 2000 women who had standard therapies for breast cancer at Magee-Women’s Hospital. We perform Quantitative Sensory Testing in a subsample of these subjects. Genetic data will be collected using DNA extracted from frozen blood. Clinical: Interviewing patients for pain and pain-related traits assessment using webbased surveys; quantitative sensory testing using standard protocols for static and dynamic stimulation. Genetics of Labor Pain and Analgesia Inna Belfer, M.D., Ph.D., Associate Professor of Anesthesiology Molecular Epidemiology of Pain Program Significant pain is reported in up to 50% of pregnant women in the months prior to delivery, and in 30% of women months to years after delivery. Attempts to explain this variability with physical markers have thus far failed. This study seeks to understand the genetic and environmental (clinical, medical, demographic, psychosocial) factors associated with the incidence of persistent pain during and after childbirth. Prospective design, multiple time points for follow-up and comprehensive approach to data collection makes this project challenging. The ultimate aim of current pilot and future larger-scale studies is to predict which women are at risk for poor labor analgesia and chronic post-partum pain development. This will hopefully lead to more personalized peri-partum pain management. Student’s input will have tremendous value and will help to identify the unique individual “fingerprint” that may predispose women to chronic pain. Clinical: Interviewing patients for pain assessment and quantitative sensory testing; Laboratory: DNA extraction and genotyping of pain candidate genes. Student can participate in either or both components. Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: The North American Malignant Hyperthermia Registry Barbara W. Brandom, M.D., Professor of Anesthesiology and Jerome Parness, M.D., PhD., Professor of Anesthesiology Dr. Brandom’s group maintains the North American Malignant Hyperthermia (MH) Registry, which now contributes to the Global Rare Disease Registry. Research will focus on measures of muscular function that could document the sub-clinical chronic myopathy that is one of the manifestations of some mutations in the ryanodine receptor gene. Studies into the genetics of MH susceptibility and the phenotype of MH susceptibility are ongoing. Trainees will participate in research projects involving de-identified data already acquired by the North American Malignant Hyperthermia Registry (over 4,000 cases). They will develop and execute projects involving patients diagnosed as malignant hyperthermia (MH) susceptible or MH negative, and may prepare data for analysis, perform descriptive statistical analysis, and present the data at a research meeting. Preliminary Studies for Whole Genome Association Study (WGAS) in Acute Perioperative Pain Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Jacques E. Chelly, M.D., Ph.D., M.B.A., Professor of Anesthesiology and Orthopedic Surgery, Vice Chair of Clinical Research This preliminary, developmental, prospective genome-wide association study will explore genetic mechanisms responsible for variation in pain perception. The study will attempt to correlate markers on genes & single nucleotide polymorphisms w/ patient phenotypes (quantitative sensory/motor testing, psychometric questionnaires & other clinical variables). The aim is to devise a method for defining a pain phenotype sensitive enough to detect differences in genotype between subjects who undergo a standard surgical procedure. Subjects undergoing total knee arthroplasty will be genotyped for genes associated with pain & single nucleotide polymorphisms (SNPs), and phenotyped in the perioperative period. Fluids will be collected for biomarkers of inflammation analysis using multiplex immunoassay. Trainees will be responsible for all clinical research duties including but not limited to enrolling subjects, phenotyping using quantitive sensory/motor testing, administering psychometric questionnaires, processing of biological specimens, and data analysis. Laboratory interest can also be accommodated to include DNA extraction/genotyping and multiplex immunoassay. Inflammation-induced Changes in the Actions of Local Anesthetics Michael S. Gold, Ph.D., Professor of Anesthesiology Regional nerve blocks are now a standard component of post-operative pain management. However, we have recently obtained data indicating that there are a number of conditions in which there is not only a loss of local anesthetic potency, but where the use of local anesthetics may even have deleterious consequences. The goal of this project is to determine the mechanisms underlying the loss of local anesthetic potency as well as an increase in local anesthetic toxicity. Will vary depending on student interests but can involve anything between behavioral pharmacology using pre-clinical models, to biochemical and/or physiological analysis is isolated tissues. Research Topic: Mentor(s): Project Overview: Injury-induced Changes in the Regulation of Intracellular Ca2+ Michael S. Gold, Ph.D., Professor of Anesthesiology Ca2+ is plays a critical role in mediating an array of physiological as well as pathophysiological processes in peripheral neurons. We have evidence that changes in the regulation of intracellular Ca2+ contribute to the manifestation of persistent pain states. Interestingly, the type of tissue injury appears to influence the specific pattern of changes in intracellular Ca2+, suggesting that the pattern of changes may also contribute to the unique qualities of the pain state specific to particular types of injury. To tease out the basis for these differences as well as the underlying mechanisms, we have been studying changes in the regulation of intracellular Ca2+ in models of persistent inflammation and more recently, nerve injury. Student Role: Trainees will participate in research projects involving the function and biochemical analysis of Ca2+ regulation in sensory neurons from rat and human donors. Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Role of GABAa Receptors in Ethanol Action Gregg E. Homanics, Ph.D., Professor of Anesthesiology Research in Dr. Homanics laboratory applies genetic dissection of putative molecular targets of ethanol to explain the neurophysiologic basis of ethanol action. Knockin mice with alterations in specific GABAA-R and glycine receptor subunit genes will be created, characterized, and tested. These novel mice will be analyzed with tests spanning molecular, cellular, and behavioral levels. Such a multi-level approach allows a determination of the relevance of a specific drug target (receptor) as a mediator of a specific phenotype (e.g., motor ataxia). Techniques student may learn: Molecular biology: recombinant DNA procedures, PCR, Southern blotting, vector construction. Cell culture: embryonic stem cell culture, electroporation, drug selection of clonal cell lines. Animal analysis: whole animal drug responses, behavioral characterization. The Effect of Neuroinflammation on Neurologic Outcome after Deep Hypothermic Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Circulatory Arrest in Rats Patrick M. Kochanek, M.D. Professor of Critical Care Medicine & Director of the Safar Center for Resuscitation Research Tomas Drabek, M.D., Assistant Professor of Anesthesiology Cardiopulmonary bypass (CPB) & deep hypothermic circulatory arrest (DHCA) are used to provide a bloodless field during cardiac surgery while minimizing cerebral ischemia. However, neurological complications remain a significant concern. More subtle neurological deficits could be triggered by inflammatory reaction in the glia. We have developed a rat CPB model to elucidate mechanisms associated with ischemiareperfusion injury after DHCA, and to test novel therapies. We showed that deep hypothermia attenuates microglial proliferation independent of neuronal death after DHCA, and improves neurologic outcome. These findings underscore the role of neuroinflammation. In current studies, we are selectively targeting neuroinflammation to improve outcome. The role of the student would be assisting during rodent experiments involving cardiopulmonary bypass, assessment of neurobehavioral outcome after deep hypothermic circulatory arrest, evaluation of neurohistochemistry, and magnetic resonance imaging (in cooperation with Carnegie Mellon University). The Genetics of Pain in Rodents and Humans William R. Lariviere, Ph.D., Assistant Professor of Anesthesiology Dr. Lariviere’s lab is determining the genetics of variability in several pain traits in mice, including mechanical sensitivity, inflammatory nociception and hypersensitivity, and neuropathic hypersensitivity. Advanced mapping populations of mice, biostatistics and genome-wide and tissue-specific expression data are used to understand which genetic mechanisms are involved in the traits. Novel genetic targets are then tested with molecular and behavioral pharmacology methods. High priority targets are tested for genetic association in human cohorts. Dr. Lariviere’s lab has performed psychophysical testing of healthy human subjects’ sensitivity to capsaicin, the pungent substance of hot peppers, and continues to study genetic association with sensitivity and dynamic pain responses. Trainees will become familiar with commonly used behavioral pain tests that we use in the lab, online biostatistical archives and tools used to go from behavioral traits to the underlying genetic mechanisms, and common molecular and neuropharmacological techniques applied to the pain tests. Ankle Block in the Rat with Ropivacaine combined with Nanoparticles – Safety and Efficacy Studies. Venkat Mantha, FFARCSI, William Lariviere, Ph.D. Complications of peripheral nerve blocks from accidental intravascular injections of local anesthetic drugs (seizures, cardiopulmonary arrest), continue to occur in spite of taking precautions or using advanced techniques like ultrasound guidance. In our previous research with ropivacaine combined with magnetic nanoparticles, we found that rats tolerated a several -fold higher dose of the drug even with direct intravenous injection. In the proposed research, in collaboration with CMU, we will combine ropivacaine with non-magnetic nanoparticles and perform ankle block in the rat and test its efficacy and safety compared to plain drug. The hypothesis is that the complexes will be effective in producing the block, and safer than the plain drug. Thus, if proved safe and effective and used in humans, the ropivacaine/nanoparticle complexes could improve the safety of peripheral nerve blocks. The project is funded and is approved by the Institutional animal care committee. Students will be trained to perform ankle blocks in the rat with plain ropivacaine and different dilutions of ropivacaine/nanoparticle complexes, and to test for efficacy of the block. They will also perform intravenous injections of the complexes in the animals to test for toxicity, as evidenced by immediate cardio-pulmonary arrest. Using simulation in medical education William McIvor, M.D., Associate Professor of Anesthesiology By using simulation and other state of the art educational technology in the training and assessment of healthcare system professionals, we can create a safer environment for patients. Dr. McIvor has supervised many student SPs, in simulation Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Mentor(s): topics including developing mannequin- and screen (computer)-based simulations. Currently, Dr. McIvor is studying ways to automate the debriefing process after mannequin simulations, comparing the effectiveness of screen versus mannequin simulation, and ways utilizing simulation in medical education. Students meet with Dr. McIvor to determine their interests and explore on-going and potential simulation projects. They then perform literature searches to familiarize themselves with simulation education, and create an SP that meets the graduation requirement. Several students have presented their work at the American Society of Anesthesiology and International Meeting for Simulation in Healthcare annual meetings, as well as University of Pittsburgh student research forums. Cellular Ca2+ Homeostasis in Skeletal Muscle and its Relationship to Malignant Hyperthermia Jerome Parness, M.D., Professor of Anesthesiology We study the regulation of intracellular Ca2+ in skeletal muscle to excitationcontraction coupling and a rare, pharmacogenetic sensitivity to volatile anesthetics known as malignant hyperthermia (MH).. An affected individual exposed to volatile anesthetics responds with a massive rise in intracellular Ca 2+ in muscle which stimulates muscle contraction and various Ca2+-dependent metabolic processes resulting in elevated body temperatures and death. Dantrolene, a hydantoin derivative and the only specific therapy for MH, inhibits the rise of intracellular Ca 2+, presumably by affecting ryanodine receptor/Ca2+ channel function. The Parness lab uses dantrolene as a probe of the ryanodine receptor and its interacting proteins, and the molecular target(s) of dantrolene. Trainees in Dr. Parness’ laboratory will be introduced to the field of cellular Ca2+ homeostasis as it relates to the pharmacogenetic syndrome Malignant Hyperthermia (MH), an autosomal dominant syndrome of the hypermetabolic response of skeletal muscle to volatile anesthetics, and the mechanism of action of dantrolene, the drug that truncates this hypermetabolism. Trainees will learn the techniques related to understanding the process of excitation-contraction coupling and Ca2+ release in skeletal muscle. These include: skeletal muscle organelle isolation and gradient centrifugation, membrane protein purification, radioligand binding techniques and analysis, cell culture, cDNA purification, mutational analysis, PCR, transfection, and Ca fluorescence microscopy. Trainees are expected to learn hypothesis generation, experimental design, the importance of controls, and techniques of data analysis. Tetsuro Sakai, M.D., Ph.D., Associate Professor of Anesthesiology All projects are currently filled. Visceral Inflammation and Pain Erica S. Schwartz, Ph.D., Assistant Research Professor, Department of Anesthesiology To investigate the transition from acute to chronic visceral pain using a model of chronic pancreatitis. Studies will examine the basic mechanisms that underlie the transition from acute to chronic pain. In acute pain, tissue insult and pain generally resolve without development of persistent pain, suggesting that the process of nociceptor sensitization is normally reversible. In chronic pain states, nociceptor sensitization appears to be irreversible. In pancreatitis, pain and inflammation are predominant clinical symptoms. Experimental approaches include: whole cell patch clamp recording, calcium imaging, and single cell PCR from labeled sensory neurons. and procedures for the quantification of localization of ion channels, G protein-coupled receptors, and transcription factors. Will vary depending on the student interests but can involve anything between behavioral assays, pharmacology, and physiology of sensory neurons to histological analysis in isolated tissues. Pei Tang, Ph.D., Professor of Anesthesiology Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: The research is designed to determine high-resolution structures and dynamics of proteins in the central nervous system and to investigate the interactions between proteins and general anesthetic drugs. State-of-the-art experimental tools, particularly multidimensional NMR spectroscopy, will be used to provide protein structural and dynamical information as well as anesthetic binding properties. The ultimate goal is to understand molecular mechanisms of general anesthetic action. Students will gain hands-on experience with our state-of-the-art instruments, including high-field NMR spectrometers, to (1) determine high resolution structures of proteins that have been recognized as potential anesthetic targets; (2) characterize interaction sites of anesthetic drugs on proteins and the impact of anesthetic interaction to protein structure and dynamics; (3) evaluate potential factors that contribute to protein aggregations and the aggregation related diseases. Molecular Mechanisms of General Anesthesia: a Computational Approach Pei Tang, Ph.D., Professor of Anesthesiology This project focuses on computational investigations of the anesthetic action on ion channels with the long-term goal of determining the molecular mechanism of general anesthesia. Students will receive training in computational approaches to biomedical research. They will be exposed to the computational facility at the Pittsburgh Supercomputer Center and learn various computational methods, including (1) parameterizations of drug molecules for further use in molecular dynamics simulations; (2) all-atom molecular dynamics simulations of proteins in the absence and presence of anesthetic drugs; (3) simulations of collective protein motions using coarse-grained models. The Interplay of Oxidants with Nitric Oxide in the Pathogenesis and Pathophysiology of Vascular Dysfunction Margaret Tarpey, M.D., Professor of Anesthesiology This project focuses on the interplay of oxidants w/ nitric oxide in the pathogenesis & pathophysiology of vascular dysfunction, including hypertension & atherosclerosis. We are interested in involvement of the enzyme xanthine oxidoreductase in elevated steady state production of superoxide & hydrogen peroxide in diseased vascular tissue. Recent studies show that moderate hypoxemia enhances expression & activity of this enzyme. We are exploring the implication for enhanced vascular dysfunction in patients w/ hypoxemia secondary to cardiopulmonary disease & development of sitedirected antioxidants to ameliorate local production of oxidants. The lab also develops methods of accurate detection of these evanescent reactive species, particularly within a biologic or clinical milieu. Trainees will have the opportunity to study these questions from a variety of experimental approaches, such as kinetic modeling of enzyme inhibition, molecular and cellular biology studies of vascular cells in tissue culture, as well as organ physiology approaches and translational studies in patients with heart failure. Such investigations, aimed at better understanding of the chemical, biochemical and cellular genesis of vascular dysfunction will provide opportunities to develop targeted therapies for the increasing number of patients with cardiovascular disease. Efficay of Phenylephrine Infusion vs. Phenylephrine Bolus for Elective Cesarean Section Manuel C. Vallejo, M.D., Professor, Director of Obstetric Anesthesia at MageeWomen’s Hospital The primary aim of this study is to compare the efficacy of phenylephrine infusion vs bolus for elective cesarean section under spinal anesthesia. Trainees will help in the recruitment of patients, enter data and write an abstract for presentation at a national meeting. Inhaled Nitrous Oxide and Labor Analgesia Manuel C. Vallejo, M.D., Professor, Director of Obstetric Anesthesia at MageeWomen’s Hospital The primary aim is to determine if nitrous oxide can be an effective alternative technique for the provision of labor analgesia. Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Trainees will help in the recruitment of patients, enter data, and write an abstract for presentation at a national meeting. Modification of Shear Induced Hemolysis by Anesthetic Agents Jonathan Waters, M.D., Visiting Associate Professor & Chief of Anesthesiology at Magee Women’s Hospital The aim is to determine if red blood cell exposure to commonly used anesthetic agents will alter the cells’ ability to withstand mechanical stress & shear forces. Subjects are non-pregnant adults, w/ no known hemoglobinopathy, coagulopathy, or anemia, and scheduled to receive general anesthesia. A blood sample (control) is collected from the IV line prior to the operation. After patient induction with propofol, another blood sample (experimental) will be obtained by venipuncture. A control sample will be used to perform ABO typing & measure total hemoglobin. 4 samples from will be tested w/ standard protocol to determine the mechanical fragility index. Comparisons will be made using 1-way analysis of variance. Trainees will help with patient consenting, blood drawing, inducing hemolysis utilizing a validated shear model, blood centrifugation, measurement of hemolysis through spectrophotometry Inherited Coagulation Dysfunction in Women Who Have Had a Postpartum Hemorrhage Jonathan Waters, M.D., Visiting Associate Professor & Chief of Anesthesiology at Magee Women’s Hospital Postpartum hemorrhage (PPH) is the leading cause of death during childbirth & a source of significant postpartum morbidity. An estimated 2.5M American women have an inherited coagulation defect which could cause/ worsen PPH. This study aims to give an understanding of the relationship between these problems & to identify symptoms, risk factors, co-morbidities, & adverse pregnancy outcomes associated w/ a bleeding disorder. Our hypothesis is frequency of diagnosed bleeding disorders is undercounted among women of reproductive age. Study designs include: case-control study of women who had a transfusion or pharmacologic treatment for hemorrhage with carboprost tromethamine for PPH during the past 12 years at MWH & prospective study of women delivering during the two year study period. Trainees will help with patient consenting, blood drawing, and data collection activities Peripheral Nerve Blocks With Multimodal Analgesics Brian Williams, M.D., M.B.A., Professor of Anesthesiology Dr. Brian Williams aims to develop appropriate animal models (rat) addressing the multimodal analgesic nerve block, in order to further advance opioid-sparing analgesia, and aim to reduce patient dependence on opioid analgesics as the primary mechanism of analgesia in the first week(s) after orthopedic surgery. Continuous nerve blocks have helped to reduce opioid requirements, but are technically complicated and subspecialized, while local anesthetics can produce unwanted motor block that may lead to the injury of an insensate extremity. This current study is designed to develop multimodal single-injection nerve blocks, in order to provide (i) sustained pain relief after surgery, (ii) reduced motor block, and (iii) reduced opioid requirements. Trainee roles include rat habituation and training, nociceptive and locomotive behavioral testing, and data collation. Volatile Anesthetic Interactions with Membrane Proteins Yan Xu, Ph.D., Professor of Anesthesiology and Vice Chair of Basic Sciences Experimental and theoretical approaches are combined to study how low affinity neurological agents, particularly general anesthetics and alcohols, exert their effects on the central nervous system at the molecular level. The goal is to understand the molecular mechanisms of general anesthesia. Trainees will have the opportunity to learn a variety of modern techniques, including expression and purification of membrane proteins, immunohistochemistry, high-resolution nuclear magnetic resonance imaging and spectroscopy, imaging reconstruction, 3-D protein structure calculation, and molecular dynamics simulations Membrane Protein Structural and Dynamical Studies by NMR Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Research Topic: Mentor(s): Project Overview: Student Role: Yan Xu, Ph.D., Professor of Anesthesiology and Vice Chair of Basic Sciences NMR is used to determine the transmembrane domain structures of the human glycine receptor, which is the primary inhibitory receptor in the spinal cord and brainstem and responsible for a wide range of diseases. The long-term goal is to provide the structural basis for novel design of drugs that are disease specific and devoid of side effects. Trainees will have the opportunity to learn a variety of modern techniques, including expression and purification of membrane proteins, immunohistochemistry, high-resolution nuclear magnetic resonance imaging and spectroscopy, imaging reconstruction, 3-D protein structure calculation, and molecular dynamics simulations. Gene and Stem Cell Therapy for Brain Protection and Revitalization After Cardiac Arrest and Resuscitation Yan Xu, Ph.D., Professor of Anesthesiology and Vice Chair of Basic Sciences New therapy strategies are being developed to target reperfusion injury after cardiac arrest and resuscitation. Recently, Dr. Xu’s group used umbilical cord matrix stem cells in an effort to stop and reverse the neuronal loss after reperfusion from prolonged cardiac arrest or stroke. A novel mechanism of extracellular signaling between stem cells and host cells are being explored. Trainees will have the opportunity to learn stem cell transplantation, various stroke models, high-resolution magnetic resonance imaging (MRI), image reconstruction, and confocal microscopy. Glycine Receptor as a Target for Pain Medicine Yan Xu, Ph.D., Professor of Anesthesiology and Vice Chair of Basic Sciences From structural and functional analyses, we have identified a novel site within the glycine receptor alpha1 and alpha3 subunits that can mediate analgesia effects of cannabinoids without the psychoactive effects. We also developed novel channel proteins that can be potentially used to drugs to treat chronic pain. The goal of the project is to further develop drugs that have high affinity for the binding site identified in the glycine receptor, and to characterize the drugs’ in vivo efficacies. Students participating in this project will learn structure-based computational drug design, animal models of chronic pain and behavior testing for pain evaluation, and gene therapy and nanomedicine in treatment of chronic pain.