Kub, Ethan
Summary of MIDD research assistantship
The goals for the Fall semester 2021:
- Quantification of ceramide and sphingosine-1-phosphate levels in vitro in U87MG human glioblastoma cells after administration of ASAH1 inhibitors (carmofur, ARN14988) using multiple-reaction monitoring (MRM) mass spectrometry (MS).
- Quantification of ceramide and sphingosine-1-phosphate levels in vivo in mice after administration of ASAH1 inhibitors (carmofur, ARN14988) using MRM MS.
- Isolation of potential active metabolite fragment of ARN14988 using high performance liquid chromatography (HPLC).
- Synthesize ARN14988 fragment that will be used for comprehensive characterization in vitro.
- Establish the metabolite activity of ARN14988 fragment using MTT assay in U87MG cells.
- MALDI-Imaging MS for the quantification of ceramides in mouse brain.
Undergraduate student: Jermarcus Lewis
As a member of Dr. Shama Mirza’s research group, I had the opportunity to gain experience with mass spectrometry and culturing the U87MG glioblastoma cell line to contribute to the lab’s goal of developing a drug for glioblastoma. This occurred under the mentorship of Ethan Kub, I learned the protocol for lipid extraction for the preparation of samples for analysis and quantification of ceramides and sphingosine-1-phosphate levels using high- performance liquid chromatography mass spectrometry (HPLC-MS). I also learned to use the cryostat for mice liver tissue preparation for H&E staining and experienced the rigor of developing a protocol for staining that is cost effective and produce optimal results. The experience as a member of Dr. Mirza’s lab has given me a renewed interest in the field of chemistry so much so that I conducted literature review on mass spectrometry and presented on the topic in BSC 490 Molecular Genetics.
Undergraduate Student: Ranjak Joshi
My name is Ranjak Joshi, and I'm currently a sophomore majoring in Biology on the pre-med track. I believe research is an essential part of the advancement of the medical field, and before going into the medical field, I wanted to gain some hands-on bench lab experience to really understand what it means to be a scientist. After seeing many of my family members suffer from various CNS neurological conditions, I've always had an interest in understanding what the normal vs. disease state mechanism looks like. I researched a couple of faculties that were doing chemistry/neuro-related research, and after reading through Dr. Mirza’s work on protein characterization by using mass spec to reveal Glioblastoma-related therapeutics, I couldn't hold back my interest and curiosity. With my long-term personal connection with neurological conditions through my family, and with a high level of interest in getting to know protein structure and function in normal vs. disease state conditions, I felt as if I would be able to perfectly advance as a scientist in the Mirza lab. I have been in touch with Ethan since the Spring of 2020. We had weekly online meetings since meeting in person at the lab was not allowed due to COVID restrictions. This fall semester, I was able to come to the lab and perform hands-on experiments alongside Ethan. Prior to starting this lab, I had very limited knowledge about mass spec and research in general. Ethan and I spent a significant amount of our time focusing on cells. I learned how to culture cells, make media, count cells using a hemocytometer, freeze cells, and many other procedures. I also got to improve my lipid extraction technique throughout the semester. These are just some of the main procedures I worked on throughout the fall semester. I had such a wonderful time and learned so much information throughout this semester and I am beyond excited to expand my knowledge throughout the next couple of years!