Develop organoid cultures (colonic and hepatic) and assay systems to evaluate pathologic parameters of the cells following exposure to chemical compounds. My primary research focus is to characterize the molecular signatures of chemical-exposed organoids and better understand the transition of normal cells to cancer cells.

Implemented primary cell isolation for clinical use process at the GMP Cell Isolation Facility at Harvard's MGH. Duties included writing stepwise work instructions, creating and implementing IQ, OQ, and PQ documentation, and training staff. The other side of my job involved the opening of a research laboratory for next-generation product development. In addition to creating a quality system that covered all the equipment and test methods used in the laboratory, I designed and executed experiments using 2d and 3d (hydrogel) model systems. Techniques employed and evaluated included, cryopreservation, real-time cell analysis (impedance-based), metabolite analysis (adenine-related molecules), enzyme assays (colorimetric-, Glo- fluorescent-based), and microscopy.

My research is broadly focused on the role of Glycine-N-Methyl-Transferase (GNMT), folate binding proteins (ALDH1L1/2), and folate metabolism on parenchymal cells of the liver (2d and 3d culture systems) and at the system level. I used a wide variety of molecular and biochemical techniques to accomplish the task including the analysis of the metabolome by Metabolon, NMR-based methods, and various -omic-based methods including 2-d gels with MALDI-based protein identification as well as more traditional immunoprecipitation, HPLC, and western blot methods. Additionally, I employ light and confocal microscopy to monitor live cell cultures, stained tissue, or cells. More recently I have employed CRISPR technology to create models for the loss of function phenotype of specific genes using normal murine hepatocytes. One of the more interesting technologies I use ( and continuing from work at MUSC) is the use of a Seahorse to analyze energetics.

Taught Human Anatomy and Physiology I and II to college and nursing program students. I very much enjoyed interacting with students and guiding them through biology and the human body.

At Overhills High School, I taught Biology(2) and Anatomy and Physiology (2), Chemistry(1) and Physical Science(1). It was very challenging to create lesson plans for 4 different classes but I enjoyed covering all of the various curriculums throughout the day. Nothing was repetitive and some of the students were really impressive. They always kept one on their toes.

Taught PLTW Robotics and Automation curriculum to middle school students (grades 6-8). We used Vex and Lego MindStorm robotic systems. In addition to teaching classes, I ran robotic exploration and competition clubs at my school and the nearby elementary school. Without a doubt the most insane year of my professional life and also one of the most enjoyable.

As the manager of the research and development laboratory at the biotech company, Cytonet LLC, I isolated and cultured primary human hepatocytes and liver cells. In addition to metabolite analysis by HPLC, ELISAs and colorimetric assays, I performed molecular and genetic analysis to confirm phenotypes and differentiation states of cells in combination with FACS. Other duties included the creation and conduct of investigational studies to analyze and improve the GMP-production process, obtain data for patent applications, and the creation and validation of QC methods regarding the cryopreserved liver cell therapy developed by the company. I had direct oversight of one associate scientist and was a co inventor on one patent (Encapsulated Liver Cell Composition: 9517247).

Within the laboratory of John Lemasters, I continued our collaboration exploring kinase-signaling pathways regulating mitochondrial function. I isolated primary rat hepatocytes, performed primary and transformed cell culture, mitochondria isolation, 1D- and 2D- PAGE, Seahorse Extracellular Flux Analyzer and Clark oxygen electrode to measure oxygen consumption (OCR) and mass spectroscopy-based proteomics. Also used high content imaging (confocal and fluorescent microscopy) and propidium iodide fluorescence (plate reader) to screen for transition pore activators and inhibitors. Other duties included training graduate students and technicians in specialized techniques and procedures performed in the lab. In addition to presenting findings at scientific meetings, I wrote manuscripts for scientific journals.

In conjunction with Jeff MacDonald (UNC) and Jan Wooten (PMUSA), I set up a biological laboratory at UNC with the aim to develop an NMR-based fluxomic model to describe in real-time and nondestructively, metabolic changes of hepatocytes and BEAS-2B cells exposed to water soluble agents and total particulate matter (TPM) arising from cigarettes or extracts from chewed tobacco products. In addition to operating the wet laboratory and cell culture facility, I managed two technicians, used a high field NMR, trained personnel in aseptic cell isolation and culture techniques and authored all laboratory procedures for all cell culture, histological, molecular assays assays. Routine techniques included western blot, PCR and metabolite extraction. I gained valuable experience in validation strategies of analytical methods and an introduction to project management. Other duties included preparing and giving progress reports to Philip Morris scientists on a periodic basis, writing and managing animal use protocols including all IACUC applications and reviewing manuscripts and presentations by other members of the laboratory.

Funded by an individual NIH Ruth L.Kirschstein National Research Service Award, I researched the mitochondrial permeability transition-associated cell death in ischemia reperfusion injury. In addition to performing cell-killing assays with primary hepatocytes, I isolated mitochondria for energetics assays. Other techniques used included fluorescent and confocal microscopy, cell fraction by centrifugation, FPLC chromatography, 1-D/2-D SDS-PAGE, western blot analysis, proteins labeling, ELISAs, immunoprecipitation and PCR analysis (quantitative and nonquantitative). Additional duties included training new graduate students and directing research rotations, peer reviewing manuscripts (Gastroentorology and Hepatology), presenting oral and posters presentations at national meetings and writing research papers. For one semester , I taught the undergraduate class, Cell and Developmental Biology which included a laboratory course. I successfully wrote and received a pilot grant from the Division of Gastroenterology at UNC where I became an associate member of the Center for Gastrointestinal Biology and Disease.For the final year of my NIH Ruth L.Kirschstein National Research Service Award, I worked a in the laboratory of Lee Graves in the Department of Pharmacology and gained experience with mass spectrometry (Maldi-TOF-TOF)-based methods in protein and phosphorylation site identification, which tend to yield high informational content files. I performed thin layer chromatographic and electrophoretic separation of proteins and tryptic peptides. Also, I performed western blot and PCR analysis on cells cultured to evaluate differentiation state. Other duties included acting as a peer reviewer for research manuscripts submitted to the Molecular Pharmacology and the Journal of Biochemistry.