In this project the PI will develop a cellular biophysics program at the predominantly-undergraduate Physics Department at Miami University in Ohio. By involving students in all stages of the research process, emphasis will be on discovery-oriented learning and undergraduate scholarship. The research activities focus on cellular biophysical studies at high hydrostatic pressures. The educational activities will be synergistic with research goals and will emphasize research mentoring and integration of discovery elements into biophysics courses for STEM and non-STEM majors. The PI will study physiological responses, in vivo, to hydrostatic pressure. Cellular energy metabolism related to respiration and mitochondrial function will be assessed by monitoring endogenous fluorescence from metabolic co-factors such as reduced nicotinamide adenine dinucleotide (NADH). The main goal of this project is to characterize free/bound NADH fluorescence from a model cellular system under metabolic conditions that are well defined under ambient pressure. The outcome will be the identification of pressure-sensitive metabolic states. The PI will continue to build a strong record for mentoring undergraduates who will be key participants in completing the research project.
. By involving students in all stages of the research process, emphasis is on discovery-oriented learning and student scholarship. Research activities focus on cellular biophysical studies at high hydrostatic pressures. Educational activities are synergistic with research goals, and include continued emphasis on research mentoring and integration of discovery elements into biophysics courses for STEM and non-STEM majors. i. Intellectual Merit Piezophysiology refers to physiological responses to hydrostatic pressure, typically in the 102 atm range. A relatively unexplored area of piezophysiology is the in vivo, in situ pressurized monitoring of cellular energy metabolism expected to exhibit pressure sensitivity because biological membranes are known to be amongst the most pressure-sensitive components of cells. Cellular energy metabolism related to respiration and mitochondrial function can be assessed by monitoring endogenous fluorescence from metabolic co-factors such as reduced nicotinamide adenine dinucleotide (NADH). Ambient-pressure fluorescence spectroscopy and imaging techniques for probing and monitoring NADH exist and are well developed. Here, we develop analogous approaches for metabolic monitoring at high pressure. The project has the following accomplishments to report: construction of a micro-perfusion system capable of monitoring fluorescence emission from biological samples while pressurized in the 102-atm range, development of rapid analysis approaches for the fluorescence-based monitoring of mitochondrial function, and demonstration that instrumentation and analysis approaches developed here are capable of metabolic monitoring under pressure. ii. Broader Impacts In the discovery learning paradigm, often called the student-as-scholar model, students are encouraged to inquire without boundaries and in the process produce knowledge. The most compelling way to engage students in discovery learning is to involve them in the actual process, i.e., in original research. To this end, this project directly involved 13 undergraduates and 6 master’s students on activities leading to the accomplishments described above. Undergraduate participation is substantial with an average of 4.8 semesters involvement per student. Students work in an interdisciplinary environment with undergraduate majors including physics, biological physics, engineering physics, zoology, biochemistry, and microbiology. Graduating students entered the work force or pursue graduate or professional education in physics, medical physics, or medicine. This project also informed the development of discovery elements into biophysical courses taught at the advanced and foundational level, and contributed to the development of a Biological Physics BS program. By establishing a program that seeks to involve students in discovery and scholarship, by developing and integrating discovery-elements into courses in biophysics, and by synergistically enhancing the inclusive, research-oriented education environment found in the Department, the project had a broad impact on society by training future professionals entering STEM fields, and by engaging non-STEM majors in the process of scientific discovery.
