abstract: All organisms need to be able to sense and respond to their environment. Much of this process takes place via proteins embedded in the cell membrane, the border between a living thing and the external world. Transient receptor potential (TRP) ion channels are a superfamily of membrane proteins that play diverse roles in physiology. Among the 27 TRP channels found in humans and other animals, TRP melastatin 8 (TRPM8) and TRP vanilloid 1 (TRPV1) are the primary sensors of cold and hot temperatures, respectively. They underlie the molecular basis of somatic temperature sensation, but beyond this are also known to be involved in body temperature and weight regulation, inflammation, migraine, nociception, and some types of cancer. Because of their broad physiological roles, these channels are an attractive target for potential therapeutic interventions. This dissertation presents experimental studies to elucidate the mechanisms underlying TRPM8 and TRPV1 function and regulation. Electrophysiology experiments show that modulation of TRPM8 activity by phosphoinositide interacting regulator of TRP (PIRT), a small membrane protein, is species dependent; human PIRT attenuates TRPM8 activity, whereas mouse PIRT potentiates the channel. Direct binding experiments and chimeric mouse-human TRPM8 channels reveal that this regulation takes place via the transmembrane domain of the channel. Ligand activation of TRPM8 is also investigated. A mutation in the linker between the S4 and S5 helices is found to generally decrease TRPM8 currents, and to specifically abrogate functional response to the potent agonist icilin without affecting icilin binding. The heat activation thermodynamics of TRPV1 are also probed using temperature-controlled electrophysiology. The magnitude of the gating enthalpy of human TRPV1 is found to be similar to other species reported in the literature. Human TRPV1 also features an apparent heat inactivation process that results in reduced heat sensitivity after exposure to elevated temperatures. The work presented in this dissertation sheds light on the varied mechanisms of thermosensitive TRP channel function and regulation. / Dissertation/Thesis / Doctoral Dissertation Biochemistry 2019

This study is intended to investigate and propose a systematic method for uncertainty quantification for the computer code validation application. Uncertainty quantification has gained more and more attentions in recent years. U.S. Nuclear Regulatory Commission (NRC) requires the use of realistic best estimate (BE) computer code to follow the rigorous Code Scaling, Application and Uncertainty (CSAU) methodology. In CSAU, the Phenomena Identification and Ranking Table (PIRT) was developed to identify important code uncertainty contributors. To support and examine the traditional PIRT with quantified judgments, this study proposes a novel approach, the Quantified PIRT (QPIRT), to identify important code models and parameters for uncertainty quantification. Dimensionless analysis to code field equations to generate dimensionless groups (�� groups) using code simulation results serves as the foundation for QPIRT. Uncertainty quantification using DAKOTA code is proposed in this study based on the sampling approach. Nonparametric statistical theory identifies the fixed number of code run to assure the 95 percent probability and 95 percent confidence in the code uncertainty intervals. / Graduation date: 2013 / Access restricted to the OSU Community, at author's request, from Dec. 5, 2012 - Dec. 5, 2013