Pingry Community Research Journal - Winter 2017

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PINGRY COMMUNITY RESEARCH JOURNAL

WINTER 2017

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TABLE OF CONTENTS RESEARCHER ARTICLES

REPORTER ARTICLES SUMMER PROGRAM: RESEARCH IN THE BIOLOGICAL SCIENCES (RIBS) @ UNIVERSITY OF CHICAGO Felicia Ho

ANALYSIS OF APOPTOPIC PROTEIN BCL2L12’S ABILITY TO ACCELERATE MELANOMA GROWTH IN A ZEBRAFISH MODEL Raymond Chen, Annette Jones, Dr. Colleen Kirkhart

SUMMER PROGRAM: NEUROBIOLOGY OF BEHAVIOR Hannah Gruber

TESTING THE EFFICACY OF TUTORING BIOLOGY I Wesley Streicher, Charlotte Curnin, Amanda Celli

WAKSMAN SCHOLAR PROGRAM Ashana Makhija

THE EFFECTS OF CAFFEINE ON DAPHNIA MAGNA Allyson Bisgay, Jeffrey Zucker

SMART TEAM Grace Brown

THE EFFICACY OF HANDS-ON DISSECTIONS FOR LEARNING SNAKE ANATOMY Nell Beatty, Rose Beatty, Julia Dannenbaum

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MAXIMIZING THE GROWTH OF DUCKWEED Ellie Palmer, Rikki Borkowski

PINGRY COMMUNITY RESEARCH JOURNAL

DETERMINING SUCCESSFUL FOOD SOURCES FOR DAPHNIA MAGNA CULTURES Myla Stovall, James Parker OPTIMIZATION OF DAPHNIA MAGNA GROWTH Lucas Monserrat, Coby Weiss, Edward Johnson MEASURING THE TOXICITY OF AMYGDALIN ON MUS MUSCULUS LEUKEMIA CELLS Amy Kaplan, James Robertson, Heba Syed EFFECTS OF MEMORY-ENHANCING SUPPLEMENTS IN A DROSOPHILIA MELANOGASTERALZHEIMER’S MODEL Sofia Briones, Naiyah Atulomah, Shruti Sagar, Saxon Scott, Varun Seetamraju

Editor-in-Chief: Allie Riddell, Class of 2019 Head Copy Editor: Felicia Ho, Class of 2019

Copy Editors: Darlene Fung, Class of 2019 Brian Li, Class of 2020 Art Editors: Namita Davey, Class of 2018 Isabelle Sheyfer, Class of 2020 Layout Editor: Praesana Danner, Class of 2019 Faculty Advisor: Mr. David Maxwell

Winter 2017 Edition 2

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RESEARCHER ARTICLES WINTER 2017

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Analysis of Apoptopic Protein BCL2L12’S Ability to Accelerate Melanoma Growth in a Zebrafish Model

by Raymond Chen, Annette Jones, Dr. Colleen Kirkhart ABSTRACT Our project involved the analysis of the apoptotic protein Bcl2L12’s ability to accelerate melanoma development. To test Bcl2L12 for this ability, we used an assay in transgenic zebrafish in which transgenic BRAF(V600E) is overexpressed on a p53 mutant background.

Melanoma and melanocytes that develop in Tg(mitfa:BRAFV600E); p53(-/-) zebrafish are suppressed by a mitfa(-/-) mutation, Tg(mitfa:BRAFV600E);p53(-/-);mitfa(-/-). (3) We are engineering a Tol2 transposon-based “miniCoopR” vector that rescues mitfa, and therefore melanocytes and melanoma, and drives expression of Bcl2L12 in these rescued tissues.

We will then analyze and compare the development of melanoma to a zebrafish melanoma strain without the upregulation of Bcl2L12 to determine the effect of Bcl2L12.

INTRODUCTION Melanoma is the most lethal form of skin cancer that originates in the malignancy of melanocytes (skin pigment cells). In 2016, approximately 75,000 people in the US had melanoma and approximately 10,000 died from the disease. Melanoma accounts for less than one percent of skin cancer cases, but accounts for the vast majority of skin cancer deaths. (1) The race for treatments is on, and research discovering new pathways and proteins involved in melanoma malignancy is in progress.

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Many cancer studies involve analyzing the genomes of malignant tumors to isolate major oncogenes and tumor suppressor genes. In previous genomic studies of cutaneous melanoma tumors, Bcl2L12, a protein belonging to the Bcl2 apoptotic protein family, was found to be upregulated, implying its role as an anti-apoptotic oncogenic protein in melanoma. (2) The goal of this project is to determine Bcl2L12’s function as a possible oncogene to accelerate the melanoma growth using a zebrafish model.

Determining Bcl2L12’s ability to accelerate melanoma can lead to new treatments that inhibit melanoma growth. Due to Bcl2L12’s upregulation in genome analyses, we hypothesize Bcl2L12 to be an anti-apoptotic protein.

If this is proved to be valid through our experiments, drugs that inhibit Bcl2L12 or other related or interacting proteins can possibly retard melanoma development, leading the way for new treatments for melanoma.

MATERIALS AND METHODS To test Bcl2L12 for its ability to accelerate melanoma development, we will be using an assay in transgenic zebrafish in which BRAF(V600E) is overexpressed on a p53 mutant background. Melanoma and melanocytes that develop in Tg(mitfa:BRAFV600E); p53(/-) zebrafish are suppressed by a mitfa(-/-) mutation. (3) We are engineering a Tol2 transposon-based “miniCoopR” vector that rescues mitfa, and therefore melanocytes and melanoma, in a Tg(mitfa:BRAFV600E);p53(-/-);mitfa(-/-) strain and drives expression of Bcl2L12 in these rescued tissues.

Zebrafish Model The transgenic zebrafish model we are using relies on research previously done by Zon Lab at Harvard University. Transgenic BRAFV600E (Tg(mitfa:BRAFV600E)) zebrafish were found to develop benign nevi. (3) When p53 was also knocked out, the (Tg(mitfa:BRAFV600E); p53(-/-)) zebrafish developed nevi that became malignant. (3) In our model, Tg(mit-

WINTER 2017 fa:BRAFV600E);p53(-/-);mitfa(-/-), in addition to transgenic BRAFV600E and a p53 knockout, mitfa (microphthalmia-associated transcription factor a) is also knocked out. Mitfa is considered the “master” melanocyte transcription factor, and without mitfa, melanocytes and thus melanoma cannot develop. Thus, our zebrafish model has all the necessary ingredients to develop melanoma from melanocytes, but lacks the ingredients to develop melanocytes in the first place. miniCoopR Vector We want to see what effect our candidate oncogene, Bcl2L12, has on melanoma growth, so we want melanoma to develop. However, mitfa is knocked out, so we need to replace it using a Tol2 transposon-based “miniCoopR” vector, which we are constructing through a Invitrogen Gateway Cloning reaction. The final structure of the miniCoopR vector can be seen below in Fig. 1. The mitfa minigene is labeled the “mitfa ORF”, and is promoted by natural transcription factors existing within melanocytes, which allows it to be active in melanocytes when incorporated into the zebrafish genome. In the process, we can also include our candidate oncogene, Bcl2L12. Bcl2L12 is promoted by mitfa, and thus is active only when mitfa is present in melanocytes. To construct the final miniCoopR vector, we received 4 component vectors: Bcl2L12 ORF, 5’ Entry vector w/ mitfa promoter, 3’ Entry

vector w/ polyA sequence, and the miniCoopR backbone plasmid. With these components, a Gateway Cloning reaction can be performed to create the final miniCoopR plasmid in preparation for microinjection into the Tg(mitfa:BRAFV600E);p53(-/-);mitfa(-/-) zebrafish strain.

Figure 1 | miniCoopR Plasmid Vector Structure Simplified structure of final miniCoopR vector

Table 1 | miniCoopR Plasmid Components List of miniCoopR plasmid components and each component’s antibiotic selector and cell type used in bacterial transformation 5

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Bacterial Transformation Before constructing the final miniCoopR vector, we are first transforming the plasmid components into bacteria. The antibiotic selector and cell type used for the bacterial transformation process can be seen above in Table 1. To construct the final vector, we are using a technique called Gateway Cloning.

Gateway Cloning Gateway Cloning is a method that allows for an efficient transfer of sequences between plasmids using “Gateway att” sites and an enzyme mix, called “LR Clonase”. (4) Gateway Cloning Technique allows transfer of DNA fragments between

different cloning vectors while maintaining the reading frame. (4) Using Gateway, one can clone or subclone DNA segments for functional analysis. (4) Gateway Cloning can best be thought of as a puzzle. Flanking each sequence of the entry vectors sequences are att sites, in which one strand is cleaved by the Clonase enzyme mix.

Based on the sequence, the cleaved att sites will bind to another att site as seen in Fig. 2 by the numbering of the att sites. The entry vector sequences will combine to form the completed expression construct, which still contains att sites. These att sites are short and are unlikely to interfere with protein expression and structure.

Figure 2 | Gateway Cloning LR clonase reaction of three entry vectors and a backbone vector 6

WINTER 2017 DISCUSSION/CURRENT PROGRESS We have received all four plasmid components as seen in Table 1 and are in the process of transforming those plasmids into bacteria for long term storage. We have also received materials for the gateway cloning process. Our zebrafish facility has been set up and is fully operational. We are still working to breed zebrafish and care for zebrafish embryos. We have all the machinery necessary for the microinjection process, but we are currently working to find the optimal settings for pulling glass capillaries into microinjection tips that will allow for a safe microinjection of our miniCoopR plasmids into the zebrafish.

ACKNOWLEDGEMENTS Jacob Weiss Dr. Morgan D’Ausilio David Maxwell Dr. Leonard Zon Dr. Julien Ablain Christian Lawrence

Testing the Efficacy of Tutoring Biology I

Wesley Streicher (‘17), Charlotte Curnin (‘17), Amanda Celli (‘17)

REFERENCES 1. Skin Cancer Foundation. “Skin Cancer Facts & Statistics - SkinCancer.org.” The Skin Cancer Foundation - SkinCancer.org, www.skincancer. org/skin-cancer-information/skin-cancer-facts.

Accessed 27 Jan. 2017.

2. Gartner, J. J., Parker, S. C. J., Prickett, T. D., Dutton-Regester, K., Stitzel, M. L., Lin, J. C., …

Samuels, Y. (2013). Whole-genome sequencing identifies a recurrent functional synonymous mutation in melanoma. Proceedings of the National Academy of Sciences, 110(33), 13481– 13486. http://doi.org/10.1073/pnas.1304227110 3. E.Elizabeth Patton, Hans R. Widlund, Jeffery L. Kutok, Kamden R. Kopani, James F. Amatruda, Ryan D. Murphey, Stephane Berghmans, Elizabeth A. Mayhall, David Traver, Christopher D.M. Fletcher, Jon C. Aster, Scott R. Granter, A.Thomas Look, Charles Lee, David E. Fisher, Leonard I. Zon, BRAF Mutations Are Sufficient to Promote Nevi Formation and Cooperate with p53 in the Genesis of Melanoma, Current Biology, Volume 15, Issue 3, 8 February 2005, Pages 249-254, ISSN 0960-9822, http://dx.doi. org/10.1016/j.cub.2005.01.031. (//www.sciencedirect.com/science/article/pii/ S0960982205000916) 4. “Gateway Cloning.” Thermo Fisher Scientific.

N.p., n.d. Web. 16 Apr. 2017.

Abstract: Our hypothesis was that group tutoring would increase the biology knowledge of freshmen and therefore their test scores. We created a preliminary quiz based on the material they would be tested on later in the week. Next, each student attended a 45 minute tutoring session.

After the tutoring sessions, students took the quiz again. We compared the scores of those who had attended the tutoring sessions (experimental group) to those who had not (control group). We detected no significant differences.

Introduction: Tutoring is an established practice dating back to the times of Plato and Socrates in Ancient Greece and is thought to be the oldest method of teaching. In Greece, the children of the wealthy were provided tutors to supplement their edu7

PCR cation (1). We wanted to test the efficacy of group tutoring. In 2001, the government released the report “Evidence That Tutoring Works”, emphasizing the importance scheduling frequent tutoring sessions and assisting students with learning disabilities (3). This 2011 study concluded that students tutored in math and language performed better overall on standardized testing (2). Similarly, tutoring for standardized tests, such as the SAT and ACT, has become increasingly popular to increase the chances of success on these tests. We decided to specifically test the efficacy of AP Biology students tutoring a group of freshman biology students to see how this would affect their success on a quiz that we designed. With group tutoring in mind, our foundational paper was “The Role of the Lecturer as Tutor: Doing what Effective Tutors Do in a Large Lecture Class” (3).

Materials and Methods: After our proposal, we began working with Ms. Torres to gain her support for our project. She accepted our proposal and granted us total access to her Schoology and class roster. We began finalizing timelines according to her class schedule. Additionally, we formulated lesson plans according to Ms. Torres’s first unit’s curriculum. Furthermore, we used their textbook and class materials to create an assessment that required students to respond to questions about various subunits within the unit. We read through the textbook and important terms and divided the four chapters into three equal parts.

Each of us individually studied our units to generate questions for the quiz, created a 20 minute review of the material. We collected the data and performed a chi-squared analysis. This chi-squared analysis, which looked at whether or not scores improved after students had been tutored, proved that our tutoring was ineffective and led to inconclusive data.

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Figure 4a-f) The questions asked on the preliminary and follow-up quiz are pictured in figure 4.

During class we introduced our project and incentivized their participation with extra credit on their upcoming test. Next, we administered our preliminary quiz to all of the participants via Google Forms. After collecting baseline information, we conducted two tutoring sessions containing identical content. Each tutoring session lasted 45 minutes and reviewed previously learned material that would be on their upcoming test. During the session, we reviewed information from the first three chapters of their textbook, class PowerPoints, and homework worksheets all found on Ms. Torres’ Schoology page. Each tutor (Wesley, Amanda, Charlotte) discussed separate subtopics, dividing the four chapters into three equal sections. We then worked collaboratively to answer questions and summarize the unit. Following both sessions, we administered a follow-up quiz to all the participants. This follow-up quiz included the same questions as the preliminary quiz. We then compared the difference in baseline and follow-up scores of the students who were tutored to the untutored group. The untutored group were those who were unable to attend a tutoring session and were therefore self-selected.

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PCR Results and Discussion: Our data showed no statistically significant differences. The average scores for the preliminary test were 72%. The average scores for the follow up test were 75%, indicating an increase, but not a significant one. Figure 1b shows the percentage of people that got each question correct, comparing the preliminary results with the final results.

Figure 2 demonstrates the specific results of the percentages correct on each individual question.

Eleven of the fifteen questions had an increase in the percentage correct; the largest increase was 25%, regarding the question on unsaturated fats. While there was marked improvement for specific questions and the class’s performance overall increased, our experiment proved mostly ineffective.

After conducting a chi-squared analysis, we had to reject the null hypothesis that the scores between the preliminary and follow-up quiz would increase significantly as the scores were not significantly different. One question, “Which of the following statements is not true� regarding the characteristics of water, had a lower score (75% originally and 45% follow up).

The decrease in performance on this question skewed our data; we are currently examining what misled the students the second time. Our investigation has mostly pointed towards a miscommunication/misinterpretation during the

Figure 1: The graph shows the difference between the control and experiment groups performance on the follow-up evaluation.

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Figure 2: The percent of students who correctly answered each individual question. The average scores overall show an increase from the preliminary quiz to the follow up.

WINTER 2017 tutoring session. The second biggest decrease in overall class performance was of 5% for one question. Additional variables that negatively affected our data was the small sample size, given our total was 20 students when we planned for around 30. Although our results did not show as strong of an increase in before and after, our results still yielded some improvement.

We hope to conduct further research to eliminate variables and outliers to receive a more considerable increase in performance. Further research could support the implementation of group tutoring as a supplemental aid to the Biology I course at Pingry. Continued investigation would then research the possible implementation of group tutoring for other subjects, specific tutor to student ratios, and male versus female performance.

Literature Cited: (1) Lasiewicz, Bob. “From Socrates to the SAT: A Brief History of Tutoring.” From Socrates to the SAT: A Brief History of Tutoring. Larnin100, n.d.

Web. 08 Feb. 2017. <http://www.learning100. com/public/316.cfm>.

(2) Rothman, Terri, and Mary Henderson. “Do School-Based Tutoring Programs Significantly Improve Student Performance on Standardized Tests?” RMLE Online 34.6 (2011): 1-10. Research in Middle Level Education, 2011. Web. <http:// files.eric.ed.gov/fulltext/EJ925246.pdf>.

(3) Usgpo. Evidence That Tutoring Works.

(2001): n. pag. Evidence That Tutoring Works.

Department of Education, Washington, DC. Web. <https://www.gpo.gov/fdsys/pkg/ERIC-ED464343/pdf/ERIC-ED464343.pdf>.

(4) Wood, William B., and Kimberly D. Tanner.

“The Role of the Lecturer as Tutor: Doing What Effective Tutors Do in a Large Lecture Class.”

CBE Life Sciences Education. American Society for Cell Biology, 2012. Web. 08 Feb. 2017. <https://www.ncbi.nlm.nih.gov/pmc/articles/

Cell Biology, 2012. Web. 08 Feb. 2017. <https://www.ncbi.nlm.nih.gov/pmc/articles/ PMC3292071/>.

The Effects of Caffeine on Daphnia magna Allyson Bisgay (‘17), Jeffrey Zucker (‘17)

Abstract We explored whether the small crustacean Daphnia magna can build up a tolerance to caffeine. The first part of our experiment cultivated a flourish-ing population. The second part of the experiment administered caffeine to the D. magna and observed their heart rates. After conducting the experiments, we concluded that the D. magna were able to build a tolerance, up until one cup of coffee’s worth of caffeine (8.02 mg/20 mL). After that, their bodies start to shut down.

Introduction Many people criticize coffee, arguing that it has negative effects on health, growth, and development. Nevertheless, 83% of United States adults consume the beverage (4). People are often accused of being “coffee addicts”; however, this is a false accusation considering that most people are actually addicted to the the drink’s most crucial component, caffeine (5). Caffeine is a stimulant that affects the body by increasing heart rate (6).

D. magna are small, transparent crustaceans with a heart that is visible under a light microscope (1). Males are 2 mm, and females are 5 mm. Their transparent shell is extremely sensitive to environmental changes such as light, temperature, etc. As a result, they have proven useful for experiments such as bioassays (7). Our goal was to successfully cultivate a population. We hypothesized that the more caffeine the D. magna are exposed to initially, the greater tolerance they would have to additional caffeine later.

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PCR Materials and Methods The D. magna was grown in plastic containers filled with 1.2 liters of water from Round Valley Reservoir in Clinton Township, NJ. The initial populations were approximately 30 D. magna per container. We gave them 16 hours of light and eight hours of darkness each day using LED lights connected to a timer. We monitored the pH, temperature and water hardness. Ideal values for pH are between 7.5 and 8.2, and ideal values for temperature are between 20oC and 25oC. When the water in the containers started to dissolve due to the light and develop a film over the top, we refilled the containers with more reservoir water.

We used Carolina Biological Daphnia Food, which comes in pellet form, to feed the D. magna. We administered more food once all of the food in the pellet had dissolved in the water. We fed them approximately every two weeks.

We then tested three caffeine concentrations of 0mg/20mL, 4.01mg/20 mL, and 8.02mg/20 mL on the D. magna. The highest concentration is equivalent to the typical concentration of caffeine in one cup of coffee. We crushed a No-Doze caffeine pill and added the powder to the 20 mL of reservoir water in the petri dishes.

We measured heart rate under a light microscope by putting the D. magna on a slide with water and cotton. We then recorded video in slow motion using an iPhone. We used five D. magna from each treatment. We watched the video in slow motion and counted how many times their heart beat in ten seconds and then multiplied by six to get beats per minute. We initially checked the heart rate to get a baseline average. Then, we administered caffeine to two of the containers and checked their heart rates 10 minutes after being exposed to caffeine. Next, we checked their heart rates after 1.5 hours after we administered an additional 16.04mg/20mL of caffeine to each container.

Results and Discussion Environment: We recorded the initial environments for each of the containers. Container 1’s pH was 7.41.

Its temperature was 20.8ºC, and its water hardness was 7.47 mg/L. Container 2’s pH was 7.17, the temperature was 20.5ºC, and the water hardness was at 6.86 mg/L. Container 3’s pH was 7.55, the temperature was 20.6ºC, and the water hardness was at 7.32 mg/L. Container 4’s pH was 7.54, the temperature was 20.6ºC, and the water hardness was 6.59 ml/L. After ten minutes of exposure to caffeine the heart rates of the experimental groups of D. magna differed from the control group. The average heart rate of the control group was 280 bpm. The average heart rate of the group with 4.01mg of caffeine was 318bpm. The average heart rate of the group with 8.02mg of caffeine was 324bpm (Figure 4). After one and a half hours, the average heart rate of the control group remained constant. The average heart rate of the group exposed to 4.01mg of caffeine went up to 368 bpm, and the average rate of the group exposed to 8.02mg dropped to 296 bpm (Figure 4).

Figure 4: Heart Rate of D. magna When Exposed to Caffeine Heart rate after administering additional caffeine: After administering a solution containing 16.04mg of caffeine (2 cups worth of caffeine) per 20 mL of water to each of the groups, the heart rates of the D. magna spiked. The control group’s heart rate went up to 350 bpm (25% increase). The second group, which had been originally administered 4.01mg of caffeine, had an increased average heart rate of 362 bpm (2% decrease), and the third group, which had been originally administered 8.02mg of caffeine, had an increased average heart rate of 374 bpm (26% increase) (Figure 4).

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WINTER 2017 When we first administered the caffeine, the control group’s heart rate was the lowest, the 4.01mg/20mL group’s heart rate was slightly faster, and the 8.02 mg/20mL group’s heart rate was the highest. This showed that caffeine initially increases the heart rate of the D. magna.

After leaving the D. magna exposed to the caffeine for 1.5 hours, the 4.01 mg/20mL group’s heart rate increased even more, but the 8.02 mg/20mL group’s heart rate decreased. This shift in trend shows that D. magna cannot tolerate 1 cup’s worth of caffeine for an extended period of time before their bodies start to shut down.

Later, when given 2 cups worth of caffeine, the control group’s heart rates spiked, but the 4.01mg group’s heart rate changed minimally. The full cup group’s heart rate also spiked, showing that with a higher initial dose of caffeine the D. magna did not build a tolerance because the initial amount of 1 cup was above their tolerance level.

Literature Cited (1)Corotto, Frank, et al. Making the Most of the Daphnia Heart Rate Lab: Optimizing the Use of Ethanol, Nicotine & Caffeine. North Georgia College & State University, web.as.uky.edu/Biology/faculty/cooper/ABLE/Daphnia_Concentration_Paper.pdf. Accessed 8 Feb. 2017.

(2) “Culturing Daphnia” Environmental Inquiry - Bioassays Using Daphnia. Cornell University, n.d. Web. 08 Feb. 2017. http://ei.cornell.edu/toxicology/bioassays/daphnia/culture.html.

(3) Heckmann, Lars-Henrik, and Richard Connon. “Culturing of Daphnia magna - Standard Operating Procedure.” (n.d.): n. pag. Daphnia Research Group. University of Reading, July 2007. Web. 8 Feb. 2017. http://www.reading. ac.uk/web/files/biosci/Culturing_Daphnia_201KB.pdf.

(4) Republic, Karen Fernau The Arizona. “Coffee Grinds Fuel for the Nation.” USA Today. Gannett Satellite Information Network, 09 Apr. 2013.

Web. 08 Feb. 2017. http://www.usatoday.com/ story/money/business/2013/04/09/coffee-mania/2069335/.

(5) Stromberg, Joseph. “This Is How Your Brain

Becomes Addicted to Caffeine.”Smithsonian. com. Smithsonian Institution, n.d. Web. 08 Feb. 2017. http://www.smithsonianmag.com/science-nature/this-is-how-your-brain-becomesaddicted-to-caffeine-26861037/.

(6) “University Health Service.” Caffeine | University Health Service. University of Michigan, n.d.

Web. 08 Feb. 2017. https://www.uhs.umich.edu/ caffeine.

(7) “Why Daphnia?.” Environmental Inquiry Bioassays Using Daphnia. Cornell University, n.d. Web. 08 Feb. 2017. http://ei.cornell.edu/toxicology/bioassays/daphnia/.

Acknowledgements David Maxwell The Pingry School

The Efficacy of Hands-on Dissections for Learning Snake Anatomy.

Nell Beatty (‘17), Rose Beatty (‘17), Julia Dannenbaum (‘17)

Abstract:

We tested the efficacy of learning snake anatomy through dissections compared to learning from a PowerPoint presentation. We created a multiple choice test about the snake anatomy that the control and experimental groups took before and after dissection, and compared the percent increase in score from pretest to posttest of the control group to the experimental group.

Our results showed a higher percent increase in the experimental group than the control group, suggesting that dissections are a more effective means of learning than oral powerpoint presentations.

Introduction In a time of rapid advancement in science and technology, private and public schools should make an effort to keep developing their science programs. Though students may not wish to focus their career in science, knowledge of science is 13

PCR important to everyday life (1). Beyond research in a lab, science dictates our society, impacting human health and public policy (1). A deeper knowledge and understanding in science can help people make informed decisions, with the ability to differentiate facts from opinion (1).

When increasing the efficacy of science programs, schools have to look into methods for teaching material effectively. In some cases, young students struggle with understanding information and interpreting diagrams in their textbook (2). Hands-on learning is a more effective teaching method (2). Using physical objects, students can explore different topics for themselves, and students are kept ontask and interested (2). Most students agree that projects, experiments, and labs are most interesting to them in science class (2). Within science programs, Anatomy class is “gateway course for health majors” (4). But, across the U.S., Anatomy classes have high dropout rates (4). One study that looked at the relationship between motor systems and memory suggests that hands-on learning plays a role in longterm memory (3). A class that examines the body and organs of different species, Anatomy is greatly focused on “manipulable objects”, and can thus benefit from adopting hands-on learning.

We examined the impact of dissections on high school students’ understanding of anatomy. Students were able to explore the body systems of real animal specimens. In a classroom of about 20 people per dissection, we walked around to the groups of students and asked them if they could locate a specific organ of the specimen. We helped them find the organ if they struggled. To test the efficacy of these dissections, we gave them a pretest and posttest made up of multiple-choice questions, asking where certain organs were located in an animal. As a control, one group of students took the pretest, listened to a PowerPoint presentation of an anatomy lesson (in place of a dissection), and then took the posttest. Our hope was to find that hands-on dissections were more effective in teaching anatomy than inactive, lecture-based lessons.

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Materials and Methods SIGN UPS We created a Google document with the emails of the 81 people who signed up for Anatomy Club at the club fair in September. For each dissection, we sent out an initial form telling the members what they would be dissecting, and which days we decided to hold them. We gave the members a week to sign up, sending a reminder the day before we need to order the specimen. When we had to order the specimen, we determined the number that we had to order based on the amount of people who signed up, knowing that they would be working in groups of two or three. The members signed up for the days that they were available, whether it was both days, one day, or neither. Ultimately, we assigned people to each day, giving priority to members who could only attend one day and filling in the remaining spots so that both days had an equal number of dissectors.

DETERMINING DISSECTIONS: To determine the specimen for dissection as a club, we reviewed the specimens from Home Science Tools, a website that carries a variety of specimens for learning purposes. We ordered five different specimens to dissect ourselves, to see which ones we would dissect as a club.

These dissections helped us decide which specimen would be best to teach anatomy, most interesting for the members, and how we should execute the meetings. We ultimately chose to use a snake as our specimen for dissection.

PRETESTS AND POSTTESTS Before each dissection, we created a multiple choice test on Google Forms, which served as the pre and post tests for both the experimental and control groups. This test consisted of 10 questions asking the locations of organs in the snake. Each question included a snake diagram labeled with numbers corresponding to different organs. Students were asked which number corresponded to which organ. The organs we asked students to identify were the heart, lung, liver, stomach, gallbladder, pancreas,

WINTER 2017 small intestine, fatty tissue, large intestine, and kidneys. We had 2 groups of students who received this test: a control and experimental group. The control group consisted of members who came to a pre-dissection meeting to learn about the anatomy of the specimen from a PowerPoint slide. This control group took the test before coming to the pre-dissection meeting and then after listening to the lesson using a PowerPoint slide. The experimental group took the test before performing the dissection, and then took it again after the dissection.

PERFORMING DISSECTIONS

We used a tray for each specimen, a scalpel, forceps, and scissors for every dissection. Every participant wore goggles and nitrile gloves. After every dissections, all utensils were washed with soap and water. In January, we ran two meetings where we dissected water snakes. Each student worked with a few other students to dissect their specimen. After working for about 30 minutes, we disposed of the specimens and cleaned up.

Results and Discussion After the students finished the pretests, posttests, and dissections, we calculated percent increases in the scores. For each student, we found the percent increase from their pretest to their post-test. Then, we found the average percent increase for the control group and the experimental group. The percent increase for the experimental group was 36%, while the increase for the control group was 17% (see Fig. 1).

Figure 1: The graph above compares the percent increase in the non-dissection group (red) to the dissection group (blue). dissection group (blue).

We can conclude that performing the snake dissection was more effective in teaching snake anatomy than the PowerPoint and oral presentation. While this sample size is small, this research highlights the importance and benefits of hands-on learning. If we were to continue this research, we would obtain a larger sample size. We would also explore hands-on learning in other areas of science.

Literature Cited 1. Dutriaux, Léo, and Valérie Gyselinck. “Learning Is Better With The Hands Free: The Role Of Posture In The Memory Of Manipulable Objects.” Plos ONE 11.7 (2016): 1-11. Academic Search Premier. Web. 27 Jan. 2017.

2. Entezari, Maria, and Mohammad Javdan.

“Active Learning And Flipped Classroom, Hand In `Hand Approach To Improve Students Learning In Human Anatomy And Physiology.” International Journal Of Higher Education 5.4 (2016): 222-231. ERIC. Web. 27 Jan. 2017.

3. Marincola, Elizabeth. “Why Is Public Science Education Important?” Journal of Translational Medicine 4 (2006): 7. PMC. Web. 27 Jan. 2017.

4. Martin, Jennifer S. “The Impact Hands-on Experiences Have on Interest and Attitudes of Middle School Science Learners.” Ohio.edu, Ohio University, 28 June 2011, www.ohio.edu/ education/academic-programs/teacher-preparation/department-of-teacher-education/masters-programs/loader.cfm?csModule=security/ getfile&PageID=2184610. Accessed 27 Jan. 2017.

Acknowledgements David Maxwell Lilliana Torres Colleen Kirkhart 15

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Maximizing the Growth of Duckweed

By Ellie Palmer (‘17) and Rikki Borkowski (‘17)

Abstract The species Lemna minor is commonly found floating on the surface of still or slow-moving freshwater habitats. L. minor has recently gained attention as a nutrient rich crop that has the potential to assist in sustainable agriculture. L.minor, commonly known as Duckweed, has begun to be used in the process of bioremediation (the process of purposefully introducing microorganisms to break down environmental pollutants) in aquaculture by small farmers to support crop and animal systems, and as a method of low-cost remediation for wastewater. The central goal of this study was to determine the optimal growth of L. minor in order to maximize its growth and assist later studies relating to the uses of duckweed in such fields. Our experiment tested the productivity of two different growing methods to grow duckweed. We found that Sherman and Hildebrandt salts were not productive in producing duckweed, while Hoagland’s solution was productive in producing duckweed.

Introduction As researchers search for sustainable agriculture and methods for bioremediation, the duckweed species L. minor has gained attention for its potential as a nutrient-rich crop (1).

Bioremediation is the deliberate introduction of microorganisms to break down environmental pollutants to purify a polluted site (2). Despite the inevitability of global warming and the decline of oil resources, nonrenewable fuels remain a major part of industrialized agriculture (1). The use of duckweed as a fast growing crop 16

can reduce this reliance. It can contribute to the evolution of sustainable agriculture as an alternative food source for organisms that does not rely on the use of fuel in its farming (1). Recently, duckweed has been used in aquaculture by small farmers to support crop and animal systems. With its high protein content and low lignin content, duckweed is a low-cost source for animal feed that can be easily processed (3).

Additionally, duckweed is a low-cost method for remediation of wastewater because it is easily harvested in municipal sewage and agricultural runoffs, and it is present on water’s surface, where it collects nutrients. Ultimately, duckweed has potential in securing continuous food production, particularly by small farmers, as it can provide fertilizer, livestock feed, and human food, in addition to decreasing water pollution and increasing the potential for water reuse (4).

L. minor is a tiny, aquatic, angiosperm (flowering plant). It is commonly found floating on the surface of still or slow moving, freshwater habitats (3). L. minor is typically made up of 1-3 small green disks that are each connected to a single root. These plants are found in dense colonies (4), often covering the surface area of the water (5). The thallus (leaf-like body) has a circular shape 2-5mm in diameter (5). Under ideal growth conditions, such as nutrient rich waters, a single L. minor plant can reproduce every three days by growing a bud from the parent thallus that later breaks off to become a new plant, making it a fast growing crop (5).

In certain environments L. minor is an invasive species and can kill other aquatic plants by blocking sunlight from their leaves under water, or by causing oxygen depletion (4).

The central question of this study is to determine the optimal growth of L. minor. By varying light, water, temperature and pH through multiple trials, the optimal environment of duckweed will be determined. By testing different nutrients, specifically Schenk and Hildebrandt Basal salts and Hoagland’s Solution, the favorable source of nutrients for the plant is discovered.

Materials and Methods Over a liter of water was obtained from the Cold Brook reservoir in Oldwick, NJ. L. minor was

WINTER 2017 collected from a pond on Long Rd., Martinsville, NJ. Two experiments were performed to compare and determine which nutrients better support the growth of L. minor. The first experiment was completed using Schenk and Hildebrandt (SH) salts as the source of nutrients. In this primary experiment, duckweed was grown in three 1L beakers. Following protocol from the Rutgers Waksman Institute of Microbiology (3), each beaker contained a varied measure of SH salts. The first beaker contained 1.006g of SH salts. The second beaker contained .506g of SH salts. The third beaker contained 1.506g of SH salts. The salts were weighed and placed into the bottom of each beaker. Then each beaker was then filled with 333mL of water from the reservoir. The pH was then balanced to 5.8 using KOH and HCl and 3.3g of sucrose was added to each beaker. Then, the greenest leaves from the collected L. minor were evenly placed into the beakers and photos were taken of the contents. The beakers were left uncovered in an incubator set to 23 degrees C and received 14 hours of light and 10 hours of dark for 17 days.

Photos were taken of the results.

The second experiment used protocol for Hoagland’s solution. We used 0.815 g/500mL of Hoagland’s solution (Sigma Aldrich). We did not vary the concentration in this experiment in order to focus on growth rather than optimization. The greenest leaves of L. minor obtained from the same pond on Long Rd., Martinsville NJ were again evenly placed into containers.

The beakers were covered with parafilm and incubated at 23o C with 14 hours of light and 10 hours of darkness for 17 days. Photos were taken of the results.

Results Using the SH salts was unsuccessful due to the fungal growth that occurred. Figures 1 and 2 show the first and last days of the experiment, respectively. There is a clear difference in the material inside of the beakers between the two sets of images. The duckweed from the first day died and by the 17th day, and fungi had grown throughout the beakers instead.

Fig 1. SH BEFORE 12/16/16 Healthiest duckweed in Sherman Hildebrandt solution

Fig. 2 SH AFTER 01/03/17 Fungal growth in all three beakers is evident after 17 days.

Hoagland’s solution proved to be more effective in growing duckweed. This is evident from the growth depicted in Figures 3 and 4. Based on the particle count, there are 611 more particles in the images from the final photographs of duckweed after exposure Hoagland’s Solution, as compared to the first photographs of duckweed prior to exposure to Hoagland’s Solution. This indicates about a 111.7% increase in the total duckweed originally collected in the beakers.

Fig. 3 01/18/17 Hoagland’s Solution BEFORE ImageJ Particle Count: 547

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Fig. 4 02/07/17 Hoagland’s Solution after 17 days of growth in an incubator kept at room temperature. ImageJ Particle Count: 1158 Discussion and Conclusion From these results, it is evident that Hoagland’s solution optimized the growth of duckweed while the Sherman and Hildebrandt salts did not. It is unclear whether the results from the Sherman and Hildebrandt salts experiment were due to the SH salts or if they were caused by a pH imbalance. It is possible that since these beakers were not covered, the moisture evaporated, causing the duckweed to dry out to allow fungal growth on the added sucrose. In the Hoagland’s experiment, the beakers were covered in parafilm, suggesting the possibility that ensuring that all beakers containing L. minor are covered will lead to successful growth. To test this, we could complete another experiment wherein we use SH salts to grow duckweed. We would assess the necessity of adding sucrose by adding sucrose to a control group and not adding sucrose to another. After a period of 17 days we would assess the success of using sucrose with the SH salts. We would also need to assess the pH balance of the solution before and after 17 days to determine if it changed throughout the growth period. Overall, optimization of growth is shown in the Hoagland’s beakers, in which particle counts increased from 547 to 1158 after seventeen days, just about doubling the original amount of duckweed in the beakers.

Duckweed has many possible applications in the sustainability sphere. With the ability to filter metals from water in an eco-friendly way or even act as a dietary supplement for other animals, students at Pingry can test a variety of uses of duckweed. Our research can help future projects attain an understanding of how to 18

grow duckweed faster. Our project can thus form a basis for future research on various duckweed proteins. Overall, this project can help future phases of the environmental agenda at Pingry.

Literature Cited (1) Leng, R. A. “DUCKWEED.” DUCKWEED. Food and Agriculture Organization of the United Nations, 1999. Web. 27 Jan. 2017.

(2) “Environmental Inquiry - Bioremediation.”

Environmental Inquiry - Bioremediation. Environmental Inquiry, Cornell University and Penn State University, 2009. Web. 08 Feb. 2017.

(3) “Rutgers Duckweed Stock Cooperative.”

Rutgers Duckweed Stock Cooperative. N.p., n.d.

Web. 27 Jan. 2017.

(4) “Common Duckweed « AQUAPLANT.” Common Duckweed « AQUAPLANT. N.p., n.d. Web. 27 Jan. 2017.

(5) “Lemna Minor (lesser Duckweed) and Lemna Trisulca (star Duckweed).” Lemna Minor (lesser Duckweed) and Lemna Trisulca (star Duckweed). Washington State Department of Ecology, 1994. Web. 27 Jan. 2017.

Acknowledgments David Maxwell The Pingry School

Determining Successful Food Sources for Daphnia magna Cultures Myla Stovall, James Parker

Abstract This project measures the success rates of algae and yeast in Daphnia magna cultures by measuring the rate of procreation and mortality over a span of four weeks. At the end of the four weeks, yeast proved to be the more successful food source, but due to our small sample size, our results were not statistically significant.

Introduction Daphnia magna are common crustaceans found in most ponds, lakes and streams (3). They are about 3mm or less in size and are related to crabs and shrimp (3). D. magna belong to the Phyllopoda subclass, which have flattened leaf-

WINTER 2017 like legs which are used to produce a water current for their filtering apparatus (2). D. magna are essential to the food chains of ponds and lakes as they are responsible for harvesting tiny algae cells that convert sun energy into food (3). To establish healthy D. magna cultures, it is essential to provide optimal oxygen, pH and food so that they will be able to reproduce (1).

Bacteria, yeast, and algae have been reported to be successful food sources for culturing D. magna (1). In our experiment, we fed D. magna cultures of algae and yeast to determine which food source would be most successful. We hypothesized that the culture fed algae would be the most successful.

Materials and Methods

In order to properly culture D. magna, containers with a wide surface area were required. Four bins that fit this criteria were used in this project. D. magna were placed into water from Round Valley Reservoir in Clinton Township, New Jersey, with a pellet of D. magna feed to allow for an easy transition and a better initial survival rate. Approximately two to four hours later, we placed 20 D.magna into each bin. Another pellet of D. magna feed was then placed in the bins to further acclimate the D. magna to the new environment. The D. magna were then fed every other day. Rate of mortality as well as rate of procreation were then continually recorded.

Results & Discussion Although there are many other factors that determine the survival of a culture of D. magna, it was determined that yeast are the most successful food for growth and reproduction.

Figure 1: Death and Reproduction Data with Algae

Figure 2: Death & Reproduction Data with Yeast

Figure 3: Averaged Daphnia Count by Bin, Weeks 1-4 At the culmination of the data evaluation, we performed a paired t-test to determine whether our results were statistically significant. Due to our sample size of only 2 bins per food type, our results came out to be .2577, which is not statistically significant. If our sample sizes were doubled or tripled, the results would be statistically significant. This is a minor experimental error and can be easily corrected with the collection of a new set of data and larger sample sizes.

We believe some factors that contributed to the rate of mortality in D. magna with algae included lack of light or decreased water temperature. As we were recording our data, a thin layer of algae was discovered growing over our D. magna cultures with the algae sustenance.

This is a variable that did not appear in our yeast trials and could account for the differences in mortality and reproduction rates. The legacy of this project provides a baseline for any future Pingry projects involving D. magna.

By using yeast as the primary sustenance for 19

PCR D. magna, Pingry students can now further research D. magna.

Works Cited “Daphnia.” Daphnia. N.p., n.d. Web. 09 Mar. 2017.

Ebert, Dieter. “Introduction to Daphnia Biology.”

Ecology, Epidemiology, and Evolution of Parasitism in Daphnia [Internet]. U.S. National Library of Medicine, 01 Jan. 1970. Web. 09 Mar. 2017.

Gerber, Gwen. “The Biology Classics: About Daphnia - The Water Flea.” Home Page. N.p., n.d. Web. 09 Mar. 2017.

Optimization of Daphnia magna Growth

Lucas Monserrat ‘17, Coby Weiss ‘17, Edward Johnson ‘17 Abstract:

We found the optimal conditions for culturing Daphnia magna, a small crustacean that is extremely sensitive to water conditions.

After our first attempt failed, we changed our water source and food supplement to see if these factors were the causes of death. We found that using the water from the Round Valley Reservoir proved to be a success given the ample growth that occurred following the water source change. For our final test, we put baker’s yeast in two of the four tanks and put D. magna food pellets in the other two tanks to test which food source would be better suited for their cultivation. In conclusion, we found that water from the Round Valley Reservoir combined with D. magna food pellets are the most favorable combination for culturing D. magna.

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Introduction: A wide variety of human influences has jeopardized the ecological integrity of many different water systems around the world. Not only have these factors affected marine ecosystems, but they have also led to a shortage of uncontaminated freshwater for both human consumption and agriculture needs(1). Chemical contaminant runoffs from synthetic organic compounds, such as fertilizers, have increased the amount of nutrients in water systems. This has led to algae blooms, which subsequently lower dissolved oxygen levels. (3) The effect of less dissolved oxygen in water has led to the decreased water quality of many streams, destroying habitats, and lowering biodiversity. (1) Daphnia magna, a small crustacean, is extremely responsive to toxic substances, can reproduce at a rapid rate, and has a short generation time, making them ideal organisms to use when testing toxicity. (2) The use of D. magna as a medium to test water quality is accepted in many countries around the world and by the Environmental Protection Agency. (4) The organisms are specifically used to determine permissible concentrations of pollutants and the success rate of sanitation. (4) In this study, our primary goal was to successfully culture D. magna. We planned on culturing four separate tanks of D. magna that would all receive different feeding schedules with different supplements for each nutritional regimen.

Our secondary goal was to test water toxicity in two streams located near housing development sites. Lastly, we hoped to collaborate with some of the other groups workng with D. magna in order to compare the efficacy of certain food supplements.

Materials and Methods: We followed a protocol that we used was by the University of Reading entitled “Culturing of

WINTER 2017 Daphnia magna - Standard Operating Procedure.” (5) D. magna requires 20 oC water, 16 hours of light and eight hours of darkness each day which we supplied with LED lights on a timer. We used 1.2L of water as described by ISO 6341. We fed marinure stock extract supplement and a baker’s yeast solution to the samples.

Figure 1. Table of Recipe for Making ISO 6341 Water We poured 1.2L of the water into the four separate containers that the D. magna would be cultivated in. We placed our four containers on a shelf in complete darkness inside of a closed cupboard. We then applied 16 hours of light and eight hours of darkness to optimize the culturing environment. We also ensured that the water was at room temperature, between 20 and 21 degrees Celsius. Once we had the optimal conditions, we placed 15 full grown D. magna into each of the four containers.

For the first nutritional supplement, we used 400mL of the marinure extract solution each day, Monday through Friday. For the second nutritional supplement, we used 0.5mL of a 0.1mg/mL baker’s yeast to water concentrated stock for each container daily, Monday through Friday.

After the above protocol failed, we decided to use a much simpler one. Instead of using water prepared to ISO 6341 standards, we used water from the Round Valley Reservoir in Clinton Township, New Jersey. According to our biology teacher, Mr. Maxwell, the water from Round Valley has been known to have the correct and natural amount of salts and other matter that are essential for organism growth. Also, instead of using the marinure and baker’s yeast food supplement, we used D. magna food pellets.

We changed the water every two weeks to ensure that the amount of dissolved oxygen would never be too low. Also, we pipetted out the remnants of the pellets every week and added in one fresh one.

Results and Discussion: The first attempted cultivation of D. magna resulted in failure. After starting with fifteen D. magna per tank, all of them died after five days.

During this attempt, we used water prepared to ISO 6341 standards, which we obtained from the protocol of the University of Reading, and fed them using the marinure and baker’s yeast supplements. Although we can not point to any one factor that caused the deaths of the D. magna, we can speculate as to say that it was either the homemade lake water, prepared water, and/or the prescribed food supplements of baker’s yeast and marinure extract. We could determine the principal cause of death by treating groups of D. magna with each one of these factors independently and seeing the effects of this singular change. Using the model of organism viability over time on the y and x-axis, respectively, one can see how each tank fared during our first cultivation attempt (2).

Figure 2. Graph of the average amount of D. magna per tank using ISO water and marinure extract and baker’s yeast.

The graph below (3) shows that the combination of reservoir water and food pellets proved to be successful. Over the course of sixteen days, tank 4 had over 30 D. magna and tanks 1 and 2 both had over 19 specimens. The outlier of the group was tank 3, which from our water tests 21

PCR had the lowest or most acidic pH level on average.

Figure 3. Graph of the average amount of D. magna per tank using Round Valley Reservoir water and D. magna food pellets.

Next, we tested the effects of baker’s yeast and marinure extracts on growth. We used food pellets as a control. The results can be seen in the following figures (4). The supplements proved ineffective as all test specimens died by day 10.

Figure 4. Graph of the average amount of D. magna per tank. Tank’s 1 and 3 had marinure extract and baker’s yeast as their food supplements. Tank’s 2 and 4 had D. magna food pellets as their food supplement.

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Literature Cited Kennish, Michael J. “Environmental threats and environmental future of estuaries.”

Environmental Conservation 29.01 (2002): n. pag. Cambridge University Press. Web. 22 Jan. 2017. <https://www.cambridge.org/core/ journals/environmental-conservation/article/ div-classtitleenvironmental-threats-and-environmental-future-of-estuariesdiv/830CE2DF3F482B87AA60ADAB773906D6>.

Persoone, G., R. Baudo, M. Cotman, C. Blaise, K. Cl. Thompson, M. Moreira-Santos, B. Vollat, A. Törökne, and T. Han. “Review on the acute Daphnia magna toxicity test – Evaluation of the sensitivity and the precision of assays performed with organisms from laboratory cultures or hatched from dormant eggs.”

Knowledge and Management of Aquatic Ecosystems 393 (2009): 1-29. Web. 22 Jan. 2017. <http://www.kmae-journal.org/articles/kmae/ pdf/2009/02/kmae09009.pdf> Ryther, J. H., and W. M. Dunstan. “Nitrogen, Phosphorus, and Eutrophication in the Coastal Marine Environment.” Science 171.3975 (1971): 1008-013. Science . Web. 22 Jan. 2017. <http://science.sciencemag.org/content/171/3975/1008>.

Tyagi, Vk, Ak Chopra, Nc Durgapal, and A. Kumar. “Evaluation of Daphnia magna as an indicator of Toxicity and Treatment efficacy of Municipal Sewage Treatment Plant.” Journal of Applied Sciences and Environmental Management 11.1 (2007): 61-67. Bioline International.

Web. 22 Jan. 2017. <http://www.bioline.org.br/ info?id=bioline&doc=about>.

University of Reading. Culturing of Daphnia magna - Standard Operating Procedure. 2007. www.reading.ac.uk/web/files/biosci/Culturing_ Daphnia_201KB.pdf. Accessed 27 Apr. 2017.

Acknowledgements A special thank you to: David Maxwell Luke De Nathaniel Conard The Pingry School

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Effects of Memory-Enhancing Supplements in a Drosophila Melanogaster Alzheimer’s Model Sofia Briones, Naiyah Atulomah, Shruti Sagar,

Abstract Alzheimer’s disease (AD), the most common cause of dementia internationally and the sixth leading cause of death in the United States (1), is an irreversible neurodegenerative disorder that causes significant loss of memory and cognitive skills. AD is associated with the buildup of amyloid plaques in the brain, formed by a protein called amyloid beta 42 or Aβ42. Previous research has indicated that the selective expression of Aβ42 in the eye of Drosophila melanogaster creates a detectable rough eye phenotype (Fig. 1), making D. melanogaster a suitable model for our project (4). We aim to determine whether or not commercially available memory-enhancing supplements can lessen or exacerbate the neurodegenerative effects of AD. In this paper we report results from the control portion of our experiment, in which controlled dosages of select memory-enhancing supplements were administered to wild type CantonS flies and from which survival data was collected. Our data indicated that D. melanogaster have the ability to survive and reproduce in the presence of acetyl-L-Carnitine, Rhodiola rosea and Gingko biloba.

Introduction Alzheimer’s disease (AD), the most prevalent form of senile dementia in humans, is diagnosed by the presence of neuritic plaques, composed mainly of amyloid-beta peptides and neurofibrillary tangles made of of tau protein.

ifestation is age-dependent, with the incidence of Alzheimer’s rising from 6% in those over 65 years to 30% in those over 85 years in the general population (1). The amyloid precursor protein (APP) is a transmembrane protein most commonly found in neurons. APP is initially cleaved by α-secretase and β-secretase, with the β-secretase cleavage further cut by γ-secretase (8). The resulting peptides are known as amyloid-beta40 (Aβ40) or amyloid-beta42 (Aβ42).

Aβ42 contains two extra hydrophobic amino acids that resist movement out of the lipid bilayer, and are the basis of the toxic plaques that are a hallmark of AD (8). We are interested in researching the varying effects of different herbal memory-enhancing supplements in a Drosophila melanogaster model of AD. Materials and Methods

Figure 2. Flow chart of experimental design.

Fly Strains. UAS-Abeta42 and Gal4-ninaE.GMR were obtained from the Bloomington Stock Center in Bloomington, Indiana. CantonS and Cyo/ Bl;TM2/TM6B strains were obtained from the Scott Lab at UC Berkeley. All strains were raised on standard fly food.

Crosses - GMR-Abeta42. As shown in Figure 3, four virgin female Cyo/Bl;TM2/TM6B flies were crossed with four male UAS-Abeta42 flies. From the resulting F1 generation, four male Cyo/+;UAS-Abeta42/TM2 flies were crossed with four virgin female Cyo/Bl;TM2/TM6B flies. From the resulting generation, male Cyo/Bl;UAS-Abeta42/TM2 flies were selected and set aside.

Four virgin female Cyo/Bl;TM2/TM6B flies were 23

PCR crossed with four male Gal4-ninaE.GMR flies.

From the resulting F1 generation, four male Gal4-ninaE.GMR/Cyo;TM2/+ flies were crossed with four virgin female Cyo/Bl;TM2/TM6B flies.

From the resulting generation, female Gal4ninaE.GMR/Cyo;TM2/TM6B flies were selected and set aside. Four male Cyo/Bl;UAS-Abeta42/ TM2 flies were crossed with four virgin female Gal4-ninaE.GMR/Cyo;TM2/TM6B. Resulting Gal4-ninaE.GMR/Cyo;UAS-Abeta42/TM6B (GMR-Abeta42) flies were selected for later experimentation.

Figure 3. Chart depicting the process of the genetic cross in order to create the flies needed for later experimentation.

Supplement Trials.We added 0.5g, 0.25g, 0.1g, and 0.05g of Gingko biloba, Rhodiola rosea, aceytl-L-Carnitine and omega 3 (BulkSupplements) to 10g of standard fly food. We placed 10 female and 10 male CantonS in each of the supplement vials. Fly death and reproduction were scored over the next two weeks as a baseline in multiple populations and the F1 generation was examined for any defects.

Baseline results indicated needed changes in the concentrations of supplements and a second trial was repeated with the necessary adjustments: Gingko biloba was re-administered in dosages of 0.75g, 0.5g, 0.25g, 0.10g, 0.05g, Rhodiola rosea was re-administered in dosages of 0.35g, 0.25g, 0.10g, 0.05g, and aceytl-L-Carnitine was re-administered in dosages of 0.4g, 0.25g, 0.10g, 0.05g, while omega 3 was removed from the trial. The second supplement trial was repeated with GMR-Abeta42 flies expressing the rough-eye phenotype. A control trial with no supplements is currently in progress.

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Results and Discussion As trial and error was used for determining optimal concentrations concentration of each supplement, multiple populations of control trials using CantonS flies were set up with the varying concentrations described previously, with fly death and reproduction rates being calculated over a span of two to three weeks.

Figure 4 depicts the survival rates of wild type flies in 0.10g of acetyl-L-Carnitine in 10g of food, with the black line indicating the average life span. The longest life span Figure 5 depicts the survival rates of wild type flies in 0.25 grams of Rhodiola rosea in 10g of food, with the black line indicating the average life span. Figure 6 depicts the survival rates of wild type flies in 0.50g of Gingko biloba in 10g of food, with the black line indicating the average life span. These results have helped us determine that D. melanogaster have the ability to survive long enough to reproduce in a supplement-enhanced environment. Therefore, the GMR-Abeta42 D. melanogaster are expected to survive in the supplement food in later testing.

Figure 4. Survival rates of wild type Drosophila in 0.10g of acetyl-L-Carnitine and 10g of food.

The black line represents the average, and the vertical line represents the standard deviation.

WINTER 2017 Currently, we have restarted the mating scheme with the intention of finishing it during May 2017 and hope to set up a control baseline test of CantonS flies in regular fly food to collect data to compare other supplemental control tests with.

Once we complete our genetic cross and have viable stocks of our eye-specific Aβ42 mutant fly, we will be performing the same supplement experiments that we completed with the CantonS flies.

We will be using the concentrations Figure 5. Survival rates of wild type Drosophila in 0.25 grams of of supplements that we deterRhodiola rosea and 10g of food. The black line represents the mined from our control experiment average, and the vertical line represents the standard deviation. and sectioning large populations of Aβ42 into different vials containing concentrations of these supplements mixed with 10g of fly food.

After the flies ingest the supplements and successfully reproduce, we will examine their progeny. We will be using a genetic screening method to do so, which will allow us to compare the eye of control Aβ42-mutant fly that was put in regular food to a Aβ42-mutant fly that was born in the supplement.

This screening method will allow us to determine whether or not the administration of the supplements Figure 6. Survival rates of wild type Drosophila in 0.50g of Gingko lessens or exacerbates the severity biloba and 10g of food. The black line represents the average, and of the rough eye phenotype in futhe vertical line represents the standard deviation. ture generations. If the eye shows signs of recovery, we will proceed to use the GAL4 method with a The genetic mating cross proved to be difficult in achieving. As mushroom body-specific driver shown in Figure 3, the setup involves two separate systems of line to express Aβ42 in the brain. crossings running parallel to each other, which eventually meet up We will then follow up by testing in the final cross. In the initial first crossings between Cyo/Bl;TM2/ memory recovery through behavTM6B x Gal4-ninaE.GMR and Cyo/Bl;TM2/TM6B x UAS-Abeta42 ioral assays. were successful in producing the next generation. However, it was noted that the UAS-Abeta42 strain had relatively low progeny rates Literature Cited compared to the Gal4-ninaE.GMR strain. This proved to be a huge “What Is Dementia?” Alzheimer’s problem later on, when we were at the final cross of the mating Association. ALZ.org. 12 Apr. 2016. scheme with no viable Cyo/Bl;UAS-Abeta42/TM6B flies from the Bahadorani, Sepehr, et al. “The previous Cyo/Bl;TM2/TM6B x Cyo/+;UAS-Abeta42/TM6B cross.

Effects of Vitamin Supplementation 25

PCR on Drosophila Life Span Under Normoxia and Under Oxidative Stress.” J Gerontol A Biol Sci Med Sci, https://doi.org/10.1093/gerona/63.1.35.

Cao, Weihuan, et al. “Identification of Novel Genes That Modify Phenotypes Induced by Alzheimer’s β-Amyloid Overexpression in Drosophila.” Genetics, doi:10.1534/genetics.107.078394.

Iyer, Janani, et al. “Quantitative assessment of eye phenotypes for functional genetic studies using Drosophila melanogaster.” NCBI, doi:http://dx.doi.org/10.1101/036368.

Moloney, Aileen, et al. “Alzheimer’s disease: insights from Drosophila melanogaster modelsTr.” Trends Biochem Sci, doi:10.1016/j.tibs.2009.11.004.

Prüßing, Katja, et al. “Drosophila melanogaster as a model organism for Alzheimer’s disease.” BioMed Central, DOI:10.1186/1750-1326-8-35.

Prokop A. 2013. A rough guide to Drosophila mating schemes” G3 (Bethesda). 3(2): 353-358.

Shulman, Joshua M., et al. “Functional screening in Drosophila identifies Alzheimer’s disease susceptibility genes and implicates Tau-mediated mechanisms.” Human Molecular Genetics, doi:10.1093/ hmg/ddt478.

Acknowledgements Dr. Kirkhart Mr. Maxwell The Pingry School Ethan Blum, Joshua Garrett Metzger, and Matthew Ludwig

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REPORTER ARTICLES

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Summer Program: Research in the Biological Sciences (RIBS) @University of Chicago

By Felicia Ho (V) This past summer, from June 16 to July 14, I attended the Research in the Biological Sciences Program (RIBS) at the University of Chicago. I was in the lab every day from 8AM to 5PM, which gave me an eye-opening experience to research. Throughout the program, my classmates and I had guidance from professors and lecturers at UChicago, as well as from recent graduates or rising seniors (TAs, or Teacher’s Assistants) at UChicago. After class and on the weekends, my friends and I explored the the Navy Pier, went to Taste of Chicago, and tried deep dish pizza. In addition, the RAs (Room Advisors) were an amazing group who always supported us, especially when we had a heavy course load, bringing us out to eat sushi in the surrounding neighborhood of Hyde Park.

During the first two weeks, I learned many basic lab techniques, including working with cell cultures, using fluo28

rescent microscopy, and even using CRISPR. CRISPR, which stands for Clustered Regularly Interspaced Short Palindromic Repeats, is a cutting-edge gene technology that selectively removes or introduces a specific gene into an organism’s genome. In other words, the CRISPR-Cas9 system can be used as a “scissor” for the genome. Thus, CRISPR holds great potential in removing undesirable genes, such as genes that increase the risk of cancer, from the human genome in the future.

As an introduction to this technique at camp, we knocked out a gene involved in yeast metabolism. Although CRISPR has been controversial as it could lead to the rise of “designer” babies, it was exciting to learn a technique currently at the forefront of gene research.

Lab experience with CRISPR at RIBS also inspired me to propose an iRT project with Luke Ittycheria (V). After project

approval, we hope to investigate the newest developments in antibiotics research in developing drugs that use CRISPR to kill bacteria. Current antibiotics do not specifically target invasive bacteria but instead affect the entire bacterial genome, even harming beneficial bacteria living in our digestive tracts. With CRISPR, we hope to build upon current research in selectively targeting and killing only the invasive bacteria.

In the final two weeks, we developed our own independent research projects with a group, and my group chose to focus on using cell culture techniques to compare certain treatments of model leukemia cells. Specifically, we compared the effects of treating BaF3/p210 mouse cells, a common cell model for chronic myelogenous leukemia (CML), with telomerase inhibitor drugs against the radiation treatment of the cells. Although Gleevec, a drug developed in

WINTER 2017 2001, helped increase the remission rates of CML patients, Gleevec is losing efficacy as CML cells are mutating over time. Thus, new treatments for CML need to be developed.

We specifically focused on telomerase inhibitor drug treatments and radiation treatments. An overactive telomerase is linked to an increase in the number of cell divisions a cancer cell can have by adding on telomeres to the ends of chromosomes after each division. Wee hypothesized that its inhibition would reduce cancer cell proliferation. We also used radiation, a traditional form of treatment, to see its efficacy on CML cells. We used cell culture techniques, and performed cell viability assays and TRAP (telomeric repeat amplification protocol) assays in our experiment to quantify the effects of the two treatments (telomerase inhibitors and radiation) on the cancer cells. Ultimately, however, we were inconclusive as to which treatment would be more effective, and hope to perform later research to look into this issue further. The final group project culminated in a poster presentation on the last day of the program, as well as a formal scientific paper.

Applications are not yet available online, but the following website, https://bscd.uchicago.edu/content/ ribs, should give additional information on the program.

Overall, I highly recommend this program to anyone who is considering pursuing biomedical research in their future and wants to gain some experience in the lab by learning standard techniques. Although spending eight hours in the lab on a daily basis may sound daunting at first, it is a rewarding experience that will give you a taste of lab-based undergraduate biomedical research.

Summer Program: Neurobiology of Behavior

By Hannah Gruber (V)

This summer, I attended the Pre-College Program at Harvard University for two weeks and took a class called, “Neurobiology of Behavior”. In this class, students from across the world came together to learn about what researchers in neurobiology face in their studies. We performed labs to produce stimuli in crayfish and observe their reactions.

My class focused on the neural patterns of an “escape route”. An escape route is triggered by an animal when they notice a sign from their environment that tells them a predator is near. A signal is sent to the brain, and a split-second decision must be made to determine whether the situation is life-threatening or not. In my class, we had the opportunity to visit Harvard University and learn about research that is currently being done by scientists there. My two teachers happened to be graduate students at Harvard who study bees and crayfish and their habits. I found this experience to be insightful to my possible future career as a researcher, as I learned what the daily life of a researcher was and how to get involved in potential research projects.

When a crayfish senses a predator, it performs a tail-flip to turn in the opposite direction and swim away as an escape route. The process begins when a sensory neuron detects a change in electricity, which is triggered by changes in the environment. The brain receives the signal from senso29

PCR ry neurons and then must quickly decide if this change in environment is life-threatening, like when a predator is present, and decide if a tailflip needs to be performed. However, if the neurons that are used for this detection are inhibited or not present at all, the crayfish will either tail-flip constantly when a signal is perceived or not flip at all, causing it to die at the hands of its predator.

There are two different ways in which the brain decides if the animal needs to perform its escape route: neuronal dictatorships and neuronal democracies. A neuronal dictatorship is when a command neuron, the first neuron to receive the signal from the sensory neuron, is the only neuron that decides whether or not to engage an escape response. Its decision triggers a chain reaction of neurons around it that then cause the movement of the crayfish. Conversely, a neuronal democracy is when one neuron uses the other neurons near it to decide whether or not using an escape response is necessary. Seeing as the brain uses either neuronal dictatorships or neuronal democracies when they are in immediate danger, the accuracy of the choice is essential. Thus, the animal requires a lot of energy in order to reach the correct decision before their predator reaches them.

Many researchers have found that neuronal democracies are more common than neuronal dictatorships. Scientists can confirm this by studying animals that do not have many neural paths, such as crayfish, zebrafish, fruit flies, and bees. These animals are ideal as neurons that are used are easily identified and can be traced directly to the source. In humans, there are trillions of neural paths, making it impossible to track every single one; however, some researchers have come close to mapping the entire brain, an astronomical feat that has taken many generations of scientists.

For our final project, we were split into groups and had to choose an animal that we had not discussed before. Our goal was to find something unique about the animal’s neuronal paths that we could present in video form.

My group and I decided to study hammerhead 30

sharks and their use of electroreception, which is the ability to recognize electrical stimuli.

Hammerhead sharks have eyes on the sides of their heads, making it difficult to see prey beyond their peripheral vision. To solve this dilemma, hammerhead sharks have adapted sensors around their body that can detect when the electrical waves around them change.

These sharks use a neuronal democracy to decide if what they are detecting in their environment is prey. Once prey is detected, the shark begins to move towards the prey’s location, continuously tracking the prey’s movements.

To catch the prey, signals are sent to open the shark’s mouth, and close it once the prey is inside. In this situation, a neuronal dictatorship is used. The action of opening and closing the mouth of a hammerhead shark requires a great deal of energy. So, if the shark has not located its prey in its precise spot, the shark has to wait until they can regain the energy and locate their next prey. This is crucial because if the shark is consistently inaccurate, it can die from starvation. There are many different neural paths that can be traced and, as a result, researchers have been able to apply their found knowledge and apply it to the human brain. How? Explain

In conclusion, this summer I spent two weeks researching neural paths in the brains of animals and finding ways to apply that research to the human brain. In doing so, I met so many new friends from around the world, and learned from their experiences and knowledge. As for the application process, I applied in the winter to the Harvard Pre-College Program and once I was accepted, I chose from a variety of classes and between three different sessions that they offered. I highly recommend this program for rising juniors and seniors who want to experience college classes in a new environment, as applying to the Harvard Pre-College Program allows you to discover a topic that you may have never been interested in before, or to continue to pursue your dream career.

Application Due: May 7 https://www.summer.harvard.edu/high-schoolprograms/pre-college-program

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Waksman Scholar Program

By Ashana Makhija (IV)

As the end of my freshman year at Pingry came to a close, I found myself more and more interested in science, specifically biology. The summer provided a 3 month gap during which I could explore my interests without the added pressure of schoolwork or extracurriculars.

After looking at the different biology programs linked to Pingry that were offered in the summer, one in particular caught my eye: Waksman Student Scholars Program or WSSP. WSSP is a 2 week intensive program in the last two weeks of June which allows students to conduct real research, as well as publish their findings. In particular, I took part in WISE, or the Waksman Institute Summer Experience.

The WISE program is a 10 day full day course for students particularly interested in biology.

The program focuses on a plant called Duckweed, specifically Landoltia Punctata. Students in the WISE program isolate, sequence, and compare expressed RNA sequences from the Duckweed plant, and have the opportunity to publish these findings on national databases maintained by the National Center for Biotechnology Information (NCBI). In other words, following along the central dogma, students first isolate RNA fragments from Duckweed cells,

or the segments of the DNA that have been expressed in the cell. Once the RNA fragments are isolated, students run gels. Gels help tell students the number of base pairs the RNA fragments are, indicative of their length. Each student gets six RNA fragments, all of which differ in function and length. Once the length of the RNA sequences are known, they are sequenced and returned to the students in the form of a waveform graph. This graph enables students to identify the specific nucleotide order in the RNA fragments that they have isolated.

This nucleotide code is then searched and translated into its corresponding amino acid chain in other NCBI databases, where other RNA or DNA fragments have been isolated and shared.

If a match comes up, then depending upon how closely a studentâ&#x20AC;&#x2122;s sample matches up with the best result, the student can then identify the function of the fragment he/she has isolated.

Oftentimes, these fragments can be proteins with particular functions in the cell. For example, one of my fragments was a very common motor protein. However, many other times the RNA fragments could also be regulatory fragments, whose functions are still unknown. Once the nucleotides match up in the nucleotide database search, a student then searches their sequence in two more databases which search common proteins. They then study the protein their RNA codes for by looking at their structure, function, and even charge. Finally, at the end of the two weeks, the students present one of the proteins they have found at a poster exhibition where 31

PCR WISE faculty and parents are free to browse and ask questions, culminating a student’s experience at Waksman.

There are two main sections to any particular day at WISE: lecture, and lab. Every morning, I would first go to lecture, spending 1-2 hours reviewing and studying the concepts behind the projects we were doing in the lab. In lecture, there would be a lot of emphasis on the specific science behind the experiments we were conducting, as well as the basis or thought process behind the program in general.

Once our basic concepts were clear, we would go to the lab, and would isolate the cell data (RNA sequences), and run a variety of tests to gage new data on our individual RNA excerpts.

Each student was able to isolate 6 sequences over the course of the 2 weeks. After morning lab, we would go to lunch, then back to lecture, and then finally lab once more. In these labs, we would perform a variety of experiments from cell digests to running labs, to enable us to learn more about our RNA fragments. In lecture, we would study how the experiments we were going to perform in lab worked, basically the science behind them and why we were performing them. In the lab, we implemented these experiments, learning how to pipette, run digests and perform PCR amplifications. At the end of the program, lab groups were expected to create posters on their most interesting set of information, or protein. These posters were then presented to all the 32

all the parents as well as the Rutgers staff and faculty within the Waksman Institute.

Taking part in this program is a great way for high school students to explore their interests within biology, as well as get initial exposure to research, and how to work in a lab. Through this program, I began to value how much I really enjoyed biology, and I learned a lot of the basic essentials for working in a lab.

There are two different sections of the WISE program, one in June, and another in August. Pingry’s Waksman club works to do the same things as the summer program over the entire school year. The club meets Wednesdays and Thursdays during CP, and is led by Ally Hosler (‘19). The Waksman club at Pingry goes through the same format: first explaining the science behind the experiments being conducted, and then actually conducting the experiments. Every student in the club will have the opportunity to isolate an RNA sequence, as well as publish their end results.

If interested, please reach out to Mr. Maxwell, or any of the club’s leaders!

SMART Team

by Grace Brown (V)

During my sophomore year at Pingry (2016-2017) I was a member of the Pingry SMART Team. Led by Dr. D’Ausilio, the faculty advisor, the 2016-2017 SMART Team included Drew Beckmen, Jeffrey Xiao, Felicia Ho, Ethan Malzberg, Brandon Spellman, Udochi Emeghara, Ketaki Tavan, and myself.

SMART Team, or Students Modelling a Research Topic, is a national program which gives high school students the opportunity to work with teachers, researchers, and other students nationwide to understand and model a molecular biology research topic. As a part of a SMART Team, I essentially worked to understand the functional “story” of a protein and design a 3D printed model of this protein which could be used to learn about the protein.

The culmination of SMART Team is the opportunity for teams to attend the Experimental Biology conference, presenting their research posters and models alongside scientists and college students.

On the Pingry SMART Team, we worked with Dr. Ryan Jensen of Yale University to To get involved in Walksman, you learn about the protein BRCA2. can contact Ashana Makhija at BRCA2 is a protein involved in amakhija2020@pingry.org, the DNA repair, and mutations in leader of the club, Ally Hosler, at the BRCA2 gene are linked to ahosler2019@pingry.org or Mr. breast and ovarian cancers. In Maxwell at dmaxwell@pingry.org. response to a double strand break (DSB) in DNA, BRCA2 interacts with another protein, RAD51, to repair DNA through homologous recombination

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(HR), a process which repairs DNA using an identical strand of DNA from a sister chromatid. Dr. Jensenâ&#x20AC;&#x2122;s lab is currently working to further understand the role both BRCA2 and RAD51 play in HR, as well as to identify specific mutations which contribute to tumorigenesis.

By discovering which mutations within BRCA2 contribute to development of cancer, Dr. Jensen and other researchers hope to not only apply new treatment methods directed at these mutations, but also to give patients with a specific BRCA2 mutation a better understanding of their diagnosis and whether or not the mutation will definitively lead to cancer in the future.

After gaining an in depth understanding of BRCA2 through interpreting a number of papers and studies on the protein, many of which were written by Dr. Jensen himself, we began the design of our 3D model, one of the many unique experiences given by SMART Team. We started with a basic protein structure model, and used an editing software to design the structure by highlighting specific amino acids, regions, and interactions. We decided to created two models, which, when combined together, would effectively highlight the role both RAD51 and BRCA2 play in DNA repair. First, we created a model showing the interaction between RAD51 and BRCA2, particularly demonstrating the BRC4 motif of BRCA2 which binds to RAD51.

For this model, we utilized magnets to show the binding capability of their interacting region, allowing the proteins to be detached from each

each other. Our second model consisted of two RAD51 units bound together, with the region in RAD51 which structurally mimics the BRC4 region emphasized to portray the parallel mechanism BRCA2 uses to bind RAD51.

Creating our model was a particularly memorable experience on SMART Team; I can still remember the excitement in the room as we opened the boxes containing our 3D printed models for the first time. Even better was at the end of the year, when we gifted one copy of our models to Dr. Jensen. He was enthusiastic to have a tangible model which could be used to teach others, including his college and graduate students, about his research on BRCA2 and RAD51.

As a part of SMART Team, we also took two trips throughout the year. The first trip was to Yale University in January; on this trip, we met with Dr. Jensen to talk about our project and his research. Not only did we get to discuss his research on BRCA2 and learn about the protein on a deeper level, but we also toured his lab and gained insight into the world of research.

In April, we attended the Experimental Biology conference in Chicago. While there, we had the opportunity to visit Argonne National Laboratory along with other SMART Teams from across the country. Along with learning about some of the research projects conducted at Argonne and even talking with researchers currently working there, we also got to learn about the Advanced Photon Light Source (APS) at the facility.

The Advanced Photon Light Source provides ultra-bright, high energy X-Ray beams which can be utilized in research across many scientific fields. During our day at Argonne, we had a chance to learn about current, cutting edge scientific research as well as see technology such as the APS. Additionally, while in Chicago, we attended the Experimental Biology conference, the annual meeting of the American Association of Anatomists, the American Physiological Society, the American Society for Biochemistry and Molecular Biology, the American Society for Investigative Pathology, and the American Society for Pharmacology and Experimental Therapeutics.

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PCR At the conference, we were able to attend talks given by researchers in various fields, giving us the opportunity to learn about many scientific topics, from breast cancer to nutrition. We were also able to hear presentations from other researchers, including many college students, at our own poster session; during this session, we were able to talk and ask the presenters about their research, truly giving us an opportunity to delve deeper into any research areas in which we were interested.

Of course, the highlight of the conference was the ability to present our own poster and models at the poster session. While at first presenting alongside researchers as a high schooler seemed scary, we all quickly gained confidence and enthusiasm in teaching others about our project. It was really incredible to realize that, after spending nearly the entire school year learning about a protein, we could easily talk about complex concepts regarding the protein and explain our work to anyone at the session.

Aside from the actual research and design we did on SMART Team, gaining experience in presenting research and learning about the broader scientific world was certainly an unforgettable opportunity.

In reflection, I am extremely thankful that I had the opportunity to be a part of SMART Team. I applied at the beginning of my sophomore year, inspired by the topics we had covered in Honors Biology II. Looking back, my greatest takeaway from SMART Team is not only the specific information we learned about BRCA2, but also the ability to become directly involved with the scientific community through working with Dr. Jensen and attending the Experimental Biology Conference.

IMAGE CITATIONS https://www.tradelineinc.com/reports/2007-8/gordon-center-integrative-science https://vaw.msu.edu/project/preventing-secondary-victimization-by-educating-systems-about-neurobiology-of-trauma/ https://www.andrews.edu/cas/biology/ https://en.wikipedia.org/wiki/BRCA2

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