Peddie EXP Summer 2018

The second week of my lab began with a small El Sistema group meeting. My PI Ellen, my undergrad student Elizabeth, and I met together to discuss details with data analysis. My new job was to compare the kindergarten (baseline) data to the first year data in SPSS. SPSS is a computer program that performs analysis of variance (ANOVA). ANOVA is a statistical measure that determines whether variance of group means is due to intervention or due to chance. Ellen gave me a quick lesson on using SPSS and analyzing the results that it gives. Then I started working with the data sheet that Elizabeth gave me. I first compared data in the dot-counting test. The within-group variable was age, and the between-group variable was orchestra intervention. My ANOVA analysis gave me results that there was a significant improvement of dot counting with the within-age variable, yet there was weak evidence that orchestral intervention, the between-group variable, actually improved children’s dot-counting s...

We started off our week with a congratulatory acai bowl trip to celebrate Preston’s acceptance into a training grant program. Acai bowls in California top Playa Bowls (no question about it). From what I can tell, its a pretty huge honor to be recognized by this grant, but he’s really humble about it. On Monday, Preston and I decided that testing antibodies that have never been tested on prostate epithelial cells before would be a good objective for my first Western blot on my own. We needed to probe for ASCT2, a glutamine transporter, and GLS in order to determine if their corresponding antibodies are functional. Antibodies are crucial for Western blots because they bind to the protein of interest (POI), allowing for us to qualify its expression after imaging. As such, Preston wanted to make sure they worked by probing for ASCT2 and GLS on three different cell lines. Cell lines are commercially purchased human cells that have been immortalized (modified to grow indefinitely) by telome...

The FitzGerald Lab is located directly across from the Children's Hospital of Pennsylvania (CHOP). The building is called the Perelman Advanced Research Center, and the reason for this title is due to the concept that there are medical doctors that meet with patients on the first four floors and the top 5-12 floors are devoted to research and many labs. The name of the institute that I am working in is known as the Institute for Translational Medicine and Therapeutics (ITMAT). Translational medicine is the concept of going from the preclinical trials (in model organisms, ie. mouse) to clinical trials. When I arrived at the lab, I was informed by the lab manager, Dropped . Tilo Grosser, that I will be working with nine other high school volunteers. Together, we are trying to answer the question of what is cyclooxyrgenase-2's (COX-2) role in inflammation. All ten of us were divided into teams of two, where each group is learning a lab technique and applying it to the research...

     During these past two weeks, I have been introduced to the various research projects my lab is working on. Luckily, the project is very similar to the primary literature I have read throughout the year, so I understood my postdoc's description of it. Our main project focuses on the research question: given that NF-κB signaling in muscle stem cells causes telomere shortening, what is the specific cause of the shortened telomeres? In the NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling pathway in muscle stem cells, spurred by growth factors and other stimuli, a phosphorylation cascade causes the transcription factor NF-κB to be released. Our lab has previously shown that NF-κB signaling causes telomere shortening, but we have repeated the experiment more meticulously to confirm these results and record clearer data. Elisia allowed me to prepare grids of telomere assays (muscle stem cells plated in wells) in which the telomeres were tag...

     I started work on June 4, just a week after spring term exams. Thankfully I didn't have much trouble finding my lab's building because I had visited it in fall term for an interview. For the duration of my lab, I start work at 9:30 AM and end at 5 PM.      "We're going to the mouse room to injure some mice," my postdoc voiced, walking me to said mouse room. It was only my first day in lab; talk about hitting the ground running. After selecting two boxes of mice in a room lined with shelves of them, my postdoc Dr. Elisia Tichy, or Elisia as we call her, explained to me that she would injure the muscles of the mice. To do so, she anesthetized them and injured their target muscles using injections of notexin, an enzyme found in snake venom. The notexin injuries would cause the muscle stem cells of the mice to activate and divide in abundance. Next week, we will collect their muscle stem cells and analyze them using telomere assays ...

My second week’s been a bit tedious. I finished scoring behavioral videos way before my postdoc, Giulia, thought I’d be done, which she said was impressive, but also left her with almost no work for me to do. On the bright side, I’ve gotten really good at reading literature and grasping main ideas, so I don’t feel like a complete dunce anymore at meetings and journal clubs. Giulia says that once my fingerprints get processed through their system, I’ll be authorized to work on the benches, where I can actually get my hands dirty, so hopefully that happens soon. On another note, I’ve been getting more time to explore center city, and had a really great cheese steak last night, but it kind of went downhill from there, and now I’m really regretting not getting the taco salad instead.

During my third week in the lab, I got the chance to do some work instead of just learning techniques. On Monday, I got to make the media for the HepG2 cells myself. This is media contains all the things the cells need in order to grow and proliferate for a few days. On Tuesday, both the grad students I follow were out sick. However, this was a problem because Danyelle, one of the students, needed to be in the lab to finish her western blot. This left Dara, a masters student, and I to finish it for her. Although it wasn’t a lot of work, Danyelle believed we were capable enough to finish her work for her. Later in the week, I started to do more and more things by myself. I got to run my own western blot (just with old samples), and I was excited to see that it worked (mostly). I also got to do a transfection with two plasmids, which also worked well. I’ve learned so much in my 3 weeks here, and I can’t wait for 3 more. Western Blot GFP transfection after 3 days