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James Dieffenderfer has had a varied work experience, beginning in 2008 as a Personal Assistant at North Carolina State University. In 2009, they worked as a Systems Engineer at C Speed, followed by a Mask Layout Designer role at Qualcomm in 2010. From 2012 to 2019, James worked at North Carolina State University in various roles, including Research Assistant and Teaching Assistant. In 2016, they Co-Founded VieMetrics Inc, and in 2017, they Co-Founded VitalFlo Inc.

James Dieffenderfer graduated from North Carolina State University in 2010 with a Bachelor's Degree in Electrical Engineering. James then returned to North Carolina State University and obtained a Doctor of Philosophy - PhD in Biomedical/Medical Engineering in 2017.