Students and Scientists 2015

Emily Shutman

This past summer I was fortunate enough to have been placed in a laboratory in the Stony Brook University School of Marine and Atmospheric Sciences (SOMAS) by the Huntington Breast Cancer Action Coalition (HBCAC).  This experience was so enlightening to me as I was able to see the inner workings of an actual laboratory to asses if research is a career that I would be interested in perusing in the future.  During my time at the lab, I worked with a graduate student named Abby Tyrell and another high school student named PJ to create an experimental design and conduct a subsequent experiment on the physiological mechanisms by which trace metals effect algae biology. 

            Initially, my first few days at the lab were daunting as I had never been in this kind of environment before, and I was still unsure of how to properly use the laboratory equipment and how to design a comprehensive research experiment.  Additionally, I was still worried about managing Stony Brook’s difficult transportation and bus schedule.  During these first few days I read many papers pertaining to the research topic, and while I was initially unsure of many of the terms the scientists used, after I continued to challenge myself with the literature I was able to understand it much better.  As the days passed I also began to become much more comfortable in the lab with all of the various equipment and laboratory procedures I was expected to perform.  During the first couple of days PJ, Abby, and I also mapped out the research plan for our trace metal experiment.  We decided to conduct testing on three metals: Chromium III, Copper, and Mercury.  These metals cause free radicals to arise, which can cause subsequent DNA and organelle damageThese metals were specifically selected because they are known to cause DNA and organelle damage via free radicals, but I sought to find which metal caused the most free radical production and subsequent damage.

            Over the course of the next three weeks, the three of us ran a variety of assays to quantify the cellular damage from the free radicals.  Not only did I attain an interesting data set on how the metals affected the algae at the end of this experiment, but I was also afforded the opportunity to really learn the scientific procedures in depth and see truly how much time and effort goes into conducting just one procedure.  We conducted four procedures during the time I was there, namely the SOD, TBARS, Protein quantification, and Lipid Extraction assays.  An “assay” is simply another word for a “test.”  Each assay was used to measure the chemical byproducts of trace metal induced cellular and DNA damage.  The SOD assay is used to determine the amount of an enzyme called superoxide dismutase (SOD), which is an enzyme that combats free radicals, a product of trace metal damage.  Therefore, if there is a higher concentration of SOD, then there are more free radicals present, so the metal had a more negative effect.  The TBARS assay measures lipid damage, an effect of free radicals through monitoring an end product, known as MDA.  Therefore, the more MDA present, the worse the effect of the trace metals was.

When I was first introduced to the methodology and underlying science of these assays I was a little overwhelmed because it seemed so complex, but after a little bit of diligent reading and discussion with Abby, I was able to feel confident in what I was doing and more importantly why I was doing it.  This was a wonderful feeling of accomplishment because this work was harder than anything that I had previously done in any science class in school, but once I understood it I felt like I had achieved something relatively profound. 

            Conducting the assays themselves was not that difficult of a process, except it could become a little tedious after a while and very time consuming.  Some days I would stay late at the lab to ensure that we had finished all the work we needed to achieve on that day.  For the most part, the experimentation was standard, but one of the assays did not work due to difficulties with culturing (growing) the algae, which was a little disappointing.  However, through this disappointment I learned that powerful lesson that in science, the procedure often does not go according to plan, but you need not to get discouraged, but just continuously tweak the experiment and strive for accuracy. 

            Over the course of my time at the lab I exponentially broadened my mind, learned complex laboratory techniques, and delved into the comprehensive research process.  As a high school student, I think that this experience is invaluable as it helped solidify my desire to have a career in the sciences when I am older.  The research I performed sparked my excitement in marine and environmental sciences specifically, which is an interest that I will certainly continue to explore in my studies when I get to college and as a possible career interest.  I am very excited for what still lies ahead pertaining to this research project, and continuing to increase my breadth of knowledge in the sciences.  


Abstract

            The presence of free radicals and Reactive Oxygen species (ROS) in biological systems was discovered almost five decades ago and was almost immediately linked to various diseases and aging due to their volatile redox states.  Various ROS including the Hydroxyl radical (*OH), Hydrogen peroxide (H2O2), and singlet O2, Superoxide anion (O2*-) are transiently present in aerobic organisms as normal byproducts of oxidative metabolism and cellular respiration when the oxygen is only partially reduced, resulting in the creation of oxygen compounds that have an unpaired valence electron. Some ROS are beneficial as they function as important signaling molecules to alter gene expression, regulate the activity of specific defense proteins, and aid in muscle relaxation, and free radicals and their derivatives also regulate multiple biological processes such as hormonal responses in plants and animals, but at high concentrations ROS oxidize proteins, lipids, and nucleic acids due to their high reactivity and reduction potential to fill their valence shells, causing mutagenesis and alteration of cell structure.  There is a natural antioxidant defense system in place to combat naturally occurring reactive species, but environmental pollution can elevate radical species levels past the point where the antioxidant defense system is effective, stimulating the production and accumulation of reduced oxygen intermediates due to the multiple oxidation states of metal ions, allowing them to be oxidized and reduced by other compounds and still have a stable valence shell.  This is a large issue in aquatic biomes due to increased incidence of metal pollutants, namely Copper, Mercury, and Chromium, due to urban runoff, industrial wastes, agricultural pesticide runoff, boating activities, heavy metal mining, and garbage dumps.