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Regular updates from students in TRIP

We’re Actually Doing This! by Madison Shelton

8/31/2019

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Hello again! I can’t believe how fast TRIP has been progressing. The past three weeks have been great and I am so excited to carry out my independent project. I am continuously shocked by how much I am learning, and I love that interacting with flies is beginning to feel more natural. I have learned numerous lab techniques and different ways of communicating my results. I didn’t think that I would get along with everyone so seamlessly.  I love the way we can just talk to each other and give ideas on projects and different approaches to a topic. Everyone is able to express their ideas and opinions in a casual way which I don’t experience much at school. 
As for my independent project, I have chosen a substance that we all know well - caffeine. Caffeine seems to make its way into everyone’s day whether it be in coffee, Diet Coke (Dr. Purdy’s favorite!), tea, or even in pills. Everybody is looking for their next cup to stay awake or for that little burst of energy. I will be testing the impact of increasing concentrations of caffeine on adult activity when given in a short term setting through the Negative Geotaxis assay. ​
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In addition to adult activity, I will be testing larval activity using the larval locomotion assay with methylene blue in larvae whose parents were continuously fed caffeine. The larval locomotion assay with methylene blue uses methylene blue and the computer application “Fiji” to compare the distance traveled by larvae. I wanted to test the effects of caffeine on activity because I have read how excessive caffeine consumption is banned in some sports due to its effect on energy and activity levels.
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Capillary tubes
I also wanted to test how higher concentrations of caffeine can create a dependency on caffeine. To test the dependency, I will be using the Capillary Feeding assay on adult flies which measures caffeine flies drink in comparison to a yeast sugar solution, a liquid they are naturally inclined to drink. I was drawn to test this because it seems as though my peers don’t always want more caffeine, but they need more caffeine. There is no legal limit on how much caffeine one can consume, just a recommended amount for daily consumption. ​Therefore, I’m intrigued to research how pushing the limits of caffeine consumption can impact activity, dependency, and development.
​I’m excited to see what the results show and for the last few weeks of TRIP! ​
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Zantac and the Nervous System by Jacqueline Munis

8/25/2019

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Hi again! 
So much has happened in the few weeks since TRIP started. So far, we have conducted assays to test the effects of different stressors and drugs, and we presented the findings from our screen projects. I tested the effects of melatonin and constant darkness on locomotion. While working on my screen project, I learned a multitude of important lab skills, such as how to sort flies, collect developmental data, and perform different assays. 
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Recently, we started to work on our independent projects. I knew I wanted my project to relate to the nervous system in some way. After a lot of researching and brainstorming, I decided I wanted to focus on drugs that are linked to cognitive impairment. 
So, for my independent project I am studying the effects of Zantac on the fly nervous system. Zantac is a drug used to treat heartburn and other gastrointestinal issues. Zantac is a drug that blocks acetylcholine (a neurotransmitter) which in turn can block neuron signaling over time. I wanted to see how long and how much Zantac it takes to have adverse effects on the nervous system. To test the effect of Zantac, I am using locomotion assays and the proboscis extension response (PER) assay. These assays will allow me to see the drug’s effect on both sensory and motor function. I can’t wait to see how my independent project turns out and for what's in store for the rest of TRIP! ​
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Collecting developmental data
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Instructor, Dr. Amanda Purdy, teaching the Negative Geotaxis assay
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Imaginary Math Becomes Real! by Sebelah Sheriff

8/24/2019

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For this blog I will focus on the crazy amount of math involved in my independent project. Math and Science are both my favorite subjects. I will be in advanced geometry, advanced algebra 2, advanced chemistry, and advanced physics in the 9th grade, so you can tell that I am very passionate about science and math. I assumed that going into TRIP would not be awfully challenging since I have very good math skills, but I was proven wrong the second I opened the How to Make Solutions Packet. Making solutions and drug stocks has been hands down the hardest thing for me this summer. The session on 7/25 was surprisingly comfortable for me even though I had to make four drug stocks for pomegranate juice and coffee. 
I think that making drug stocks is so hard for me because I am used to a different type of math. By this I mean that I am used to working with numbers and numbers that will result in a useless number in life. I am not used to needing to know the concentration for a drug stock, or the size conical tube needed for a drug stock, or even converting micrograms, nanograms, and milligrams to gram instantaneously.  Though this type of math is challenging, I am glad that I get to expand my horizon of math, so that when I start my fiscal year of 9th grade, I will be ahead of the game.
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I can transition from theoretical math to real life math problems. Coincidentally, real life math problems have been the area I struggled with the most in my advanced math class in 8th grade, meaning that I have trouble comparing equations and variables to their definitions in real life. For example, slope can be expressed as 5x, but in real life that means you are going 5 miles per 1 hour. Those types of questions mess me up. 
In conclusion, if it were not for TRIP, I would struggle a lot in high school when relating real life math problems to numbers.
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Flies, Starvation, and More! by Annabelle Jin

8/23/2019

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​Hey everyone! It’s Annabelle. 
The last few weeks of TRIP have been super informative and exciting, with new things to learn every day. Before I started this program, I would swat away fruit flies if I saw them. Now that I’ve learned about all their applications to science and medicine, I see them differently— they’ve been the catalyst for so many groundbreaking discoveries, from understanding genes to finding the molecular mechanism for circadian rhythms. And, with the help of TRIP, I can use them for my own independent project:
​How does the length of recovery from starvation and the length of starvation affect female fertility and male activity levels?
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You may be wondering why this matters…
Well, with the rise of social media portraying what the “ideal” body is and more pressure to fit in and feel accepted, there has been an upsurge in eating disorders. A significant portion of them are restrictive, meaning that sufferers actively seek to reduce their calorie intake and/or eliminate entire food groups. This mental illness happens more often among adolescents. In fact, more than half of female teenagers and nearly a third of male teenagers have engaged in unhealthy weight control behaviors such as fasting and skipping meals.  
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It’s clear that this is an issue that needs to be addressed.

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Many long-term effects of eating disorders, such as osteoporosis and amenorrhea, have been well documented. However, there is conflicting information about whether or not women with a history of eating disorders are at a higher risk for infertility. Some studies have found that fertility rates among women with anorexia nervosa do not differ from that of the general population, while others have found that many women seeking treatment at infertility clinics have a history of eating disorders. No study has investigated how long recovery must be in order for fertility to return to normal, despite the fact that the mean age for the onset of anorexia nervosa is 18 years old, the duration of anorexia nervosa is on average 8 years, and the average age for American women to have children is 28 years old. With only a short time for recovery, it is important to know the length of recovery needed in order for fertility to return to normal, if it ever will. 

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Collecting developmental data
In addition, no study has investigated whether the duration of the restrictive eating disorder has an effect on fertility. Some eating disorders last for years, even decades, while others last only for a few months. Therefore, it is important to know if the severity of the eating disorder has any impact on fertility. It is also critical to know if recovery from starvation can allow activity levels to return to normal, since eating disorders can result in a drastic loss in energy. 
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Dr. Amanda Purdy teaching the Female Fertility assay
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I have just started my experiment this week, and I performed a female fertility and negative geotaxis assay (which measures activity levels). Already my preliminary results have surprised me. I’m excited to explore them further, so stay tuned for more details!
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How Will Starvation Affect My Flies? by Zoe Moreland

8/22/2019

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Planning my project was centered around what I wanted to test and what interested me the most, which is something I rarely encountered in my high school science labs.
For my independent project, I chose to look at how starvation and intermittent fasting affects female fertility. Intermittent fasting is becoming increasingly popular among people looking to lose weight and find alternative diets that fit a flexible lifestyle, however, restricting food intake can have negative effects on overall health. For example, anorexia is a common eating disorder where an individual restricts the amount of food they eat and it can lead to malnutrition, mental health issues, and even decreased fertility when a woman stops menstruating as a result of being unhealthy.
My goal for this project is to see the short-term effects
restrictive eating can have on a woman’s fertility and overall health.
So far, the first few weeks of TRIP have been informative, independent, and supportive. Planning my project was centered around what I wanted to test and what interested me the most, which is something I rarely encountered in my high school science labs. So far for my independent project, I have been able to test two seperate groups of flies by fasting them for 8 hour periods, and I am looking forward to seeing the results from the assays in the coming week.
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Fun with Flies and Fertility by Georgia Flego

8/21/2019

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As stoked as I was to begin my own study, I encountered a couple bumps in the road that made things just a bit more complicated (or exciting-- it depends how you want to look at it).
Hello everyone! I hope you can tell that I have a love for alliteration as seen in my title. I’m back to give a little update on my experience in TRIP thus far, and on the beginnings of my independent research project (YAY!). It is now coming up on four weeks since we have all started this program, and it has been an absolute whirlwind. 
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Soon after I wrote my last post, we began getting acclimated in the lab environment with our assigned preliminary screens in which I was tasked to determine how Vitamin D3 affects flies in both light and dark conditions. As many know, Vitamin D3 is a nutrient created in the skin when people are exposed to sunlight, or can be consumed through dairy, vegetables, and fortified products. With this information, it was fascinating to see if this Vitamin could supplement flies deprived of light, or if the flies were deprived by the trauma of darkness in the first place. Once the flies were exposed to their drugged food and/or deprivation of light, I utilized the social space assay, in which I placed flies into a two dimensional glass chamber to quantify their sociability by measuring the distance between each animal. Through this screen-- my first ever lab screen at that-- I was introduced to a variety of techniques and programs that I thought I would never be able to use until later on in my career, affirming my love for research. This made me even more excited to begin my own independent project!

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Social Space chambers
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Setting up the Social Space Assay
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Dr. Amanda Purdy teaching how to sort flies by gender
As stoked as I was to begin my own study, I encountered a couple bumps in the road that made things just a bit more complicated (or exciting-- it depends how you want to look at it). I initially thought that I wanted to test how soy milk in comparison to other milks affects female fertility in flies. Soy contains a compound called phytoestrogen, a fancy term for a substance that makes the body believe it is estrogen. If consumed in large amounts, soy isoflavones, the phytoestrogens in soy, can detriment female fertility in humans, so as a lactose-intolerant individual who is often exposed to soy, I wanted to explore this idea more. However, it was soon brought to my attention that flies do not use estrogen in their reproductive systems (an oversight by yours truly), so I had to slightly alter my course of action. 
Now, instead of soy, I am testing three drugs known to impact female fertility in one way or another: Fenugreek, Black Cohosh, and Folic Acid. By combining these drugs in a variety of manners, I am hoping to provide insight as to how people should treat these herbal supplements in regards to their fertility, and the possible dangers or benefits that could accompany consuming these supplements separately, or in reaction with each other. Using an assay-- an experiment-- testing female fertility, I will begin to record my first rounds of data. So far, I have had to do a little more math than most (because I am a bit ambitious and decided to test three drugs) and copious amounts food vial preparation.
​Long story short: I owe most of the TRIP initiative staff Diet Coke or other compensation for their much appreciated help.
I may not get the results I desire or expect by the end of this study, but what I want most is to acquire an experience that will further equip me for problem solving and solution developing that I can use in science or elsewhere; so far, my time in TRIP has been doing just that. Wish me luck!
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Cancer and the Diet! by Urmi Choudhary

8/20/2019

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The process of learning about the fruit fly life cycle to communicating information through graphical abstracts to hashing out our own experimental designs has been an incredible experience. 
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So far, the first few weeks of TRIP have been going really great! It is a completely different kind of environment than what I’m used to, but it’s a fantastic challenge that has pushed me to be more creative and thoughtful. The process of learning about the fruit fly life cycle to communicating information through graphical abstracts to hashing out our own experimental designs has been an incredible experience. I am super excited to continue with the independent projects and present my results!
For my project, I wanted to focus on ways we can simulate cancer for the flies. So, after doing some background research, I found information on how our diet affects our chances of getting certain types of cancer in our lifetime. Based on this, I decided to focus on salt (which Americans consume at almost double the recommended daily dosage) and Vitamin C, which is an antioxidant with several known benefits throughout our body. 
My plan is to give my flies large doses of both drugs and then expose them to UV radiation. After some flies are exposed to the radiation, I am going to perform the negative geotaxis assay to assess activity and a microbiome assay to assess overall health. I am then going to compare the results for all the fly groups. I am really excited to see what happens to my flies and the overall effect that these copious amounts of drug have on their systems!
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Getting Into the Swing of Things! by Tara-Al-Saleem

8/19/2019

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Hey, everybody! These past few weeks have been so amazing and full of new friends, routines, and most importantly, flies. When I was asked to hone in on one question to explore as an independent experiment, my mind ran at a hundred miles an hour in all directions. I’m so excited about many different aspects of behavior/anatomy; how can I possibly focus on just one thing?
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After some soul searching, I finally decided upon a topic that would not only pique my interest, but help answer a question I’ve been asking for years. 
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Do artificial food colorings cause hyperactivity in children?
After searching online, I found that this connection between colors and chaotic behaviors has never been concretely proven. A study by the University of Southampton in the UK tested the hypothesis on 300 children and found a connection between artificial colors and adverse behavior shown at school and home. Many countries in the European Union have taken this study seriously and pushed for colors to be removed from popular children’s foods. The United States, however, has not taken any strong action against food dyes. For example, popular candy “Starburst” is dyed artificially when sold in the US, but in Europe the candy contains only naturally derived dyes. 
This stark contrast has inspired me to delve deeper into the controversy of artificial dyes and further study its effects on children’s behavior.
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I cannot subject children to my scientific question, but I do have larvae at my disposal! I have set up some vials with a control versus a vial with Red #40 mixed in with the food. The level of dye was adjusted to match that of a 2-5 year old child’s maximum daily intake in the United States. When larvae are produced, I will test their memory and activity with a larval memory assay. This assay will allow me to see whether or not larvae can remember associations between scents and rewards. I can also set up a camera to quantify the larval activity levels and test for hyperactivity.
​Check back later for an update on my results!
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In Which I Cram Flies Into Tiny Spaces and See If They Get Nervous by Cailean Cavanaugh

8/16/2019

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Displaying anxiety in response to stressful environments is a trait that flies share with humans, so remember: if you’re ever feeling anxious, just know you aren’t alone, because a lot of flies are currently feeling that way as well.
Hello again!
​It’s Cailean, and now that you know who I am, I can skip all personal details and move on to the actual experiments.
A quick update on the effects of senna and sugar on the flies’ microbiome: there weren’t many. The flies that were fed sugar had roughly the same microbiome quantity and diversity as the control flies, while the flies fed senna had lower microbiome diversity. The differences were small, however, as were the total number of colonies. Really, this is nothing to write home about: not only because the results aren’t all that novel, but also because your family will probably think you’re weird if you choose to make fruit fly bacteria the subject of your long-awaited letter home.
Also, those flies are dead now. They outlived their usefulness, but most of them had already died anyway. 
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​In another topic, have you ever been trapped in a densely-packed room, nervously waiting for a chance to leave? As tensions rise, claustrophobia sets in, and the space begins to feel smaller and smaller and… well, hopefully you haven’t been, but my flies are right now. 
The subject of my independent experiment is how living in densely populated environments affects fly mood, with a focus on anxiety and social interactions. Displaying anxiety in response to stressful environments is a trait that flies share with humans, so remember: if you’re ever feeling anxious, just know you aren’t alone, because a lot of flies are currently feeling that way as well. ​


The actual process for keeping flies in densely populated environments was fairly simple: I used more or less fly food than usual and adjusted the vial’s cap to control the height of the flies’ living space. The only problem came when I had to repeat my volume calculations three separate times because of various errors that I could have avoided if I was paying more attention. 
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What proved harder was figuring out how much space to give the flies, and how much food I needed to make that space. I used a “standard” vial of flies (60 flies in 19 cubic centimeters) to calculate that the flies’ normal density is about 3 flies per cubic centimeter. This is probably one of the strangest units I’ve ever worked with, but I was able to use this average to make vials that gave the flies twice as much space, standard space, half the space, one third of the space, and one quarter of the space. After they’ve spent several days in these vials, I will test their response to the social space and centrophobism assays. 
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​In the social space assay, 5 flies are placed in a small environment as tall as they are, preventing them from flying but leaving them free to walk around. Every 5 minutes, I take a picture of the flies and measure the distances in between them. The closer together they are, the happier and more sociable they are and the further away they are, the more irritable and moody they are. 
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The centrophobism assay is based on the fact that fruit flies are a bunch of adorable little agoraphobes, and the more anxious they are, the more they stay away from open spaces. In this way, how often the flies cross the center of a small petri dish can act as a measure of their anxiety. 
Stay tuned to hear the amazing scientific results of putting flies in small spaces! It’s a very dense subject, but nothing to be anxious about.
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My Independent Project by Julia Morris

8/15/2019

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For my setup, I first made 4 vials. One was my control group, so there was no coffee in their food. Then I had 3 other vials with caffeine in the food at an increasing amount. I then sorted and placed 60 flies in each vial: 30 females and 30 males. They were next  placed into the incubator, where I will continue with them on Tuesday. ​
Hi TRIP community!!!
This past week I started my independent project, where I am trying to answer the question: How does the withdrawal of coffee affect the mood and activity of flies and their offspring? I derived inspiration for this question from my school peers, as many drink so much coffee. Most consider stopping but they don’t know how it will affect them… so, I was curious about this and decided to use it as my experiment. For my experiment, I will be placing coffee into the food of my flies, then after some time, I will place the flies in different vials with no caffeine so as to simulate withdrawal. 
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During the first few weeks of this program, I learned so much about being in a lab and how to manage fruit flies.
Did you know that you could use CO2 to anesthetize flies and sort them?
You can also use ice to knock them out for assays.
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It’s also pretty easy to distinguish the males from the females, as males have a prominent black rear and females don’t. I was able to figure out how much caffeine I would need to put in each vial after performing some drug calculations. I had to create two stock solutions because the weight of the coffee that I calculated was too small to weigh. So, I dissolved the coffee into 10 ml of water and 50 ml of water. I used a micro-pipette (pictured right) to dissolve the coffee. 
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I’ve learned so much in the past few weeks and I can’t wait to continue my experiment and collect some data!
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