Vascularization of Tissue Engineered Bone Grafts

What?

From May-August of 2014 I spent my time as an undergraduate research assistant in the Center for Regenerative Medicine and Cell-Based Therapies in Dr. David Dean’s lab.  The laboratory’s mission focuses on the creation of tissue engineered bone grafts through 3D printing. My work aims to test new methods of pre-vascularizing these bone grafts using a fibrin hydrogel seeded with stem cells, thereby inducing angiogenesis of new vasculature and anastomosis with host vasculature and ideally creating a more functional bone graft. What I really love about the Dean lab is the level of involvement that undergraduates have in their projects. In my projected, I have an active role from start to finish. I search the literature and determine methods, discuss with collaborators across the globe,  plan my experiments, execute my experiments, analyze the data, and even work to publish the results.

So What?

Research has taught me resilience and patience above all. The ability to independently work to create novel solutions to difficult problems has been invaluable to my personal growth. Class is really good at teaching you to find “the right answer”, but research teaches you to be an independent thinker, to go out in the world and use every skill at your disposal to hammer out a solution to life’s unsolved problems. In a world overflowing with college grads and never enough spots to put them, I think this is the quality that distinguishes you from the masses. After graduation everyone can hold up a degree and a GPA, but I can say that I contributed something to this world through my work.

Now What?

I hope to continue my education in veterinary school following the completion of my undergrad. Beyond the animal experience my research has allowed me, I believe my research makes me a unique candidate for future veterinary work. One of the biggest challenges I faced in previous veterinary internships was the inability of families to pay for their animal’s medical care. A human receives medical care regardless of whether or not they can pay, but many animals are euthanized simply for lack of funds. I believe that my experiences in research and engineering help me to not just shrug it off, but to continually ask, “what can I do to find a solution?”. My research has encouraged me to consider completing a senior thesis and even to pursue a PhD program in graduate school.

Throughout my research project I was able to perform surgeries on mice with the future possibility of working on larger animal models. These surgeries have allowed numerous hours of both animal and veterinary experience required for the common vet school application. Additionally, in my research I spend the vast majority of my time in meetings, discussing strategy and new ideas and making sure to go through all the proper approval channels. If there were such a thing as logging “communication hours”, I could set a new record. If I chose not to enter the veterinary world, I now have significant experience in experimental design, execution, the fundamental principles of tissue engineering, and plenty of essay writing for funding. In order to complete my time in research for which I used my STEP funding, I also left behind my family and friends and gave up my summer break to work 2000 miles away in a lab. Surely this displays a certain resiliency in character. All in all, a research laboratory is not where I hope to end up. However, I believe I have taken many valuable skills away from this experience.

 

 

Megan Posukonis- Biomedical Engineering 2016

 

Music and Tech in the City of London

What?

I never imagined I’d travel across an ocean and to another country for my STEP experience, but that’s exactly what I did.  In July 2014, I attended NIME – that is, New Interfaces for Musical Expression – in London, England.  NIME is an annual conference that unites academics, students, inventors, artists, musicians, engineers – an incredible variety of people from all different disciplines  – to explore how people express and interact with music.

I first heard about the conference after attending a lecture at OSU by Dr. Rebecca Fiebrink, an OSU alumna-now-professor who earned dual degrees in Computer Science and Engineering (CSE) and Music.  As a CSE major, I’m interested in combining music and technology for assistive projects in areas such as music therapy, healthcare, and education.  Dr. Fiebrink’s background and led me to sit in on her lecture and to later explore the research she has done – and the conferences she had attended. Consequentially I discovered NIME. Midway through the year, I was excited to hear that Dr. Fiebrink had moved to the very university that was hosting NIME in 2014. I soon found myself contacting her and developing my STEP proposal with the intention of attending NIME, and by May of 2014, I was booked for a flight to London.

Goldsmiths Music Studios

NIME was held at Goldsmiths College, University of London, which is well known for its music, arts, and media programs. The conference consisted of paper, poster, and demonstration sessions during the day, interspersed with performances in the afternoons and evenings.  Unlike most of the attendees, I was simply an observer, rather than a presenter or a well-established academic.  However, I do not say this with disappointment.  Attending just as I was, a curious undergraduate who happened to find her way to NIME, allowed more freedom to explore and reflect.  Wherever I went, my small black notebook was at my side, ready for my overzealous writing hand to jot down some important observation or fleeting amusement.

The conference had everything.  Topics included, collaborative music, motion and sensing, instrument design, and many others. The paper sessions were more academic in nature, whereas the demos and displays were often highly interactive.  In the same day, I could listen to a speaker from Stanford present on manipulating sounds with movement, then try my hand at something similar in nature by playing a modified version of Guitar Hero. And in the process of floating from room to room, my observer eyes wide with excitement, I met people from all different academic backgrounds and countries, each of whom had unique perspectives on music, technology, and life.

City of London Baglione

Aside from the conference, I got the chance to explore an incredible city.  Finding my wandering traveler’s feet to be the best mode of transportation (after the London rail system, of course), I walked myself from London Bridge to St. Paul’s cathedral, along the Thames, to the Globe Theater, and through the Tate Museum of Modern Art. I ventured into coffee shops and through the charming streets of SoHo.  I journeyed to Big Ben, Buckingham Palace, Westminster Cathedral, and Covent Garden Market.  I even got to spend a day in the English countryside with the kindest relatives of a family friend.  I had a taste of London culture, and it was truly an experience.  By the time I returned to the U.S. just a week later, my shoes were marked with travelers’ miles and my notebook and head were bursting with ideas.

So What?

I went to NIME with the impression that I already knew what music was.  But from day one, the conference challenged me to rethink the way I heard (and saw and felt) music.  As I wove my way through countless displays, demonstrations, and paper and poster sessions, I became a part of a world where sound, movement, and creativity were one entity.  And no, I didn’t think all of it was music.  Yes, I found some sounds and performances harsh, sporadic, or simply “ugly” to my ears.  Others I found to be absolutely beautiful.  I also discovered that I tend to like the sound of music played on more “traditional”- sounding instruments (ones made of wood or brass or that have strings) rather than music generated on a computer (quite ironic, yes, for a student of computer science).

But music wasn’t the half of what I learned.  Through interacting with so many different people, I gained insight into how the rest of the world conducts itself in academics, politics, and many other realms.  One day I ate dinner with a student from New Zealand who had done his graduate studies in Japan and had experience with robotic musical instruments (as well as a keen knowledge of American politics). Another day I had a conversation with a man from the Finland about his music therapy research.

Why does all of this matter?  Because in one short week, I got to experience the world of academia in the context of subjects about which I am passionate – music, technology, and engineering.  And now I have a “game plan” for how to achieve my own research and personal goals in those fields.

Now What?

NIME has caused me to reflect deeply on what I want to do in my graduate studies and where I want to pursue them.  The experiences of that week in London that affected me the most were those that bridged the gap between music and people – specifically, how music and people’s thoughts and emotions link together. Having participated a previous summer research experience in Human-Computer Interaction, I feel like NIME reaffirmed that yes, I really do want to focus on the “human element” of technology in my graduate studies and in my life.

I want to contribute to the growth of music and technology in music therapy, healthcare, and educational settings. NIME really helped me to expand my network so that I can move toward this goal.  During the week I interacted with several professors from U.S. institutions who seemed very receptive of my reasons for attending NIME and who may, in the future, offer the opportunity to pursue one of their graduate studies programs.  If not for the STEP program, I may never have met these professors, nor seen all the ways in which music and technology can work to enrich the human experience.

I remember sitting in on a paper discussion about music therapy and dementia patients and thinking, “I can’t wait until I can contribute something to this!”  Thanks to STEP and NIME, I’m well on my way to doing so.

For more on music and tech and what I did at NIME, follow me on my blog: http://techiegonemusical.wordpress.com

and on Twitter: @TechGoneMusical

Undergraduate Research in Mitotic Bookmarking

What?

I participated in undergraduate research with the Parvin lab during summer 2014 for my STEP experience. Before my STEP experience, I had hoped that participating in research over the summer would give me new opportunities that I don’t get when working in a lab during the school year when I can only work part time and can only perform limited experiments. In addition to being able to participate in more research than I normally do, I was hoping that I would be able to experience what it would be like to be a grad student or full time lab employee and determine whether or not I could see myself doing that as a career later in my life. Because of my STEP experience, I have discovered many things about myself and the direction I want to take my life and career.

My summer research project was to finish up the work of a grad student who had just graduated from our lab. She did most of the work for a paper and it was my job to validate her results. Throughout her time in our lab, she characterized a protein called RING1A. RING1A is an essential protein in a process called mitotic bookmarking. When cells undergo mitosis, or cell division, DNA condenses and almost all active genes are turned off. However, it is essential that a new cell turns on the same genes that were on in the parent cell. The cell’s solution to this problem is to bookmark the genes that were active in the parent cell with a biological marker so that the daughter cell knows which genes to turn back on after the cell divides.

The RING1A protein affects a couple different bookmarks but the bookmark that I focused on is an unknown protein that sits on the start site of active genes during cell division. We don’t know much about the unknown protein but we do know that it is attached to another small protein called, ubiquitin, when it sits on DNA. When the unknown protein is not attached to ubiquitin, it probably doesn’t act as a mitotic bookmark. The graduate student before me found that RING1A was responsible for attaching ubiquitin to the unknown protein so that it could act as a mitotic bookmark. Without RING1A, the protein does not get attached to ubiquitin and thus cannot act as a mitotic bookmark. If RING1A is not present, the cell cannot divide.

The way that we determined that RING1A is necessary for mitotic bookmarking to take place was by “depleting” RING1A. The way we do this is by using something called small interfering RNA, which are small double stranded molecules of RNA that are specific to a specific gene. They interfere with the process of how a specific gene makes a specific protein and thus make it so that none of the specific protein is present in a cell. From this, we can infer the function of a protein by observing what happens when it’s not there. When we deplete RING1A in cells, we observed that the unknown protein is not attached to ubiquitin and does not act as a mitotic bookmark and thus can infer that RING1A is the protein that attaches ubiquitin to the unknown protein.

My job this summer was to confirm the results of the grad student before me and provide more evidence that RING1A is an essential protein in mitotic bookmarking. As is the case with a lot of scientific research, things didn’t go as well as I had hoped. Although I did eventually confirm her results, it took several months longer than we had hoped and I experienced just about every problem that could have happened. But the important thing is that I did eventually get it done and we have submitted our results for publication. In addition to the publication, I learned several important things about myself that will guide me to become the person that I want to become.

You can read more about the experiments and methods that I used and the many challenges I faced in my blog: http://stepsummerreseach.blogspot.com/

So What?

I am very grateful for my STEP experience because it has taught me many things. By spending the summer working full time in a research lab I got to experience what it is like to a grad student or lab technician. I got to talk to many different people in many different parts of their lives, from first year grad students to post doctorate students to lab technicians, who gave me a lot of good advice. Most of the people I talked to said to not go into grad school unless I really want to. The funding is pretty bad at the moment and a lot of people are having trouble finding labs that can support them. Plus, a lot of the people I’ve talked to don’t seem very happy with what they’re doing and want to get out of research as soon as they can.

I tend to agree with a lot of the people I’ve talked to. I actually really like research; conducting experiments to discover things that no one else has ever known before is really exciting to me. But the other aspects of working in research aren’t very appealing to me. First, I’ve noticed that I don’t get to interact with many different people. It’s mainly the same people in my lab that I see every day. I think I’d rather work in a place where I meet a lot of different kinds of people. Second, there’s a lot of down time in research that bores me. It’s difficult to conduct experiments all day because most of the experiments we do involve waiting for cells to grow or waiting for slow biological reactions to happen. It’s gets pretty boring sometimes.

I think that instead of doing research, I want to go to med school and become a doctor. I don’t think I want to see patients all day every day and I think I’d like to participate in some research. Med school probably gives me the best option to get a career that I’ll enjoy while meeting a lot of different people and achieving my personal goals.

Now What?

From a research perspective, we are attempting to identify the unknown protein that is attached to ubiquitin. I think this will be relatively difficult but we’re hopeful that we can do it. If we do identify it, we’ll characterize it and hopefully show a similar effect when we deplete this protein as we saw from RING1A. This could potentially be a big paper if we can do it, which is pretty exciting to me.

From a personal perspective, I’m continuing to do research with the Parvin lab because I really like it there but I don’t think I want to go to grad school and do research. I’m pretty set on med school at this point because I think it gives me the best opportunity to accomplish my goals. In the meantime, hopefully I’ll be able to publish another paper or two and continue working towards my academic and personal goals.

 

Hepatocellular Carcinoma Research

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For my STEP experience, I conducted undergraduate research on the study of Hepatocellular Carcinoma. I am a part of Dr. Jacob and Dr. Ghoshal’s Laboratory in the Comprehensive Cancer Research Center at The Ohio State University under the instruction of Dr. Samson Jacob and Dr. Kalpana Ghoshal.

The purpose of our laboratory is to identify microRNAs highly expressed in the liver, and to determine the cancerous consequences if those microRNAs are deregulated. Primary hepatocytes, cell culture, diseased liver tissues, and mouse models are utilized to better understand the biological roles of microRNAs, and to aid in development of therapies for hepatocellular carcinoma (HCC).

MicroRNA 122 (miR-122) is the most abundant liver- specific microRNA. After undergoing cellular processing, it is expressed during late stages of embryonic development. In the laboratory of Dr. Jacob and Dr. Ghoshal, miR-122 was found to be reduced in both rodent and human HCCs. Furthermore, loss of miR-122 in the liver causes HCC to spontaneously develop in liver-specific knockout (LKO) mice. Overexpression of miR-122  in HCC  cell lines reduces tumerogenesis, which suggests that it functions as a tumor suppressor. MiR-122 also targets small molecules in signaling pathways associated with apoptosis and metastasis.

My contribution to these goals included researching four miR-122 targets. Those gene targets are TIMP2, Prakaca, SNAI2, and LIMK1. The reason that these particular four genes are important is because of their involvement with apoptotic pathways, Epithelial Mesenchymal Transition, and are tumor suppressors.

Data obtained from real-time PCR (qPCR) showed significant upregulation of SNAI2 and LIMK1 in miR-122 KO mice. Since gene expression of SNAI2 and LIMK1 is downregulated in normal, healthy mice, this upregulation shown in miR-122 KO mice is suggestive that miR-122 functions to suppress these genes. Based on HITSCLIP data, microarray analysis, and qPCR data, SNAI2 and LIMK1 are both potential miR-122 targets. As targets, this indicates that these two genes are involved in the HCC pathway, and contribute to the development of liver cancer in cells.

To complete my own experiments and to aid in the experiments of others in our laboratory, I learned and mastered the following techniques:

  • Tail Extraction
  • Gel Electrophoresis
  • Polymerase Chain Reaction (PCR)
  • Real-Time Polymerase Chain Reaction (qPCR)
  • Primer Design
  • Cell Culture
  • Vector Cloning & Luciferase Reporter Assay

Because of STEP, I was able to utilize the $2000.00 funding to cover living expenses May through August. By living in Columbus, I was able to effectively commute to the Tzagournis building on Ohio State’s campus. I was also able to successfully complete courses Physics 1201, History 3642, and Classics 2210 during this time period. If not for the STEP program, I would not have had an enriched academic experience during the summer, nor would I have been able to develop new skills through research.

One of the most incredible parts of the STEP program included being able to interact one-on-one with faculty. I felt that my interactions with not only my cohort faculty member, but also the faculty members of my entire house really enhanced my experience. 

I learned that my personal strengths include verbal and written communication. I am proud of the poster I was able to present at the STEP expo, and that I was able to convey the meaning and relevance of my project. Being awarded the Dean’s Best In Show Award was an incredible honor, and was personally inspiring to me to continue forward in my research with even greater enthusiasm than before.

Moving forward, I am currently continuing my research in Ohio State’s Department of Pathology. I will continue to research and identify other gene targets of mir122. My future directions include:

  • Determine if TIMP2 and Prakaca are also potential targets using qPCR
  • Complete TIMP2 Luciferase Reporter Assay  to confirm as target
  • Conduct Luciferase Reporter Assays for Prakaca, SNAI2, LIMK1
  • Continue to contribute to overall laboratory purpose through continued genotyping of mice

Following the STEP Expo, I have since completed the TIMP2 Luciferase Reporter Assay, and have confirmed with statistically significant evidence that TIMP2 is a miR-122 target. We are continuing to investigate the TIMP2 target gene region since it appears that the region has several properties unique to itself, including a “bulge” binding region. I will then conduct a final Luciferase Assay with a mutant miR-122 TIMP2 binding site.

 

Functional Connectivity in the Brain – Summer Research

In my STEP experience, I spent May-August 2014 doing undergraduate research in a neurological stroke rehabilitation lab at the Wexner Medical Center under the oversight of Dr. Alexandra Borstad. Working full time hours with the help of STEP covering my living costs, I spent my time in the lab independently learning the track-weighted functional connectivity analysis procedure, a technique combining both functional data and diffusion-track data into one comprehensive image.  We hypothesize that track-weighted functional connectivity is sensitive to sensory-motor changes caused by stroke. Through the TWFC method, we performed a whole brain analysis in order to assess frontoparietal white matter network connections.

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Essentially, the objective of my STEP undergraduate experience was to learn the TWFC analysis procedure, and to be able to perform the data processing with a high degree of understanding. This will allow me to utilize TWFC analysis in my future undergraduate research on diminished white matter microstructure in persons with stroke. Previous studies have suggested white matter connections are critically important for hand and arm function, and that these connections are diminished after stroke. These studies also suggest that through motor rehabilitation efforts, reorganization of white matter connections occurs. TWFC may be a potential resource for insight into efficacy of rehabilitation, overall damage to the brain network, reorganization of networks, and even predicting the success of rehabilitation methods.

In regards to my thoughts on my STEP experience, I overall felt like this research experience was a substantial challenge that I was happy to face head on. I had not previously had the opportunity to spend a significant amount of hours in a lab due to financial limitations, and STEP made this possible for the first time in my academic career.  I feel that I am much more confident in my ability as a student researcher, and am capable of performing such research in the future. Most of all, through my interactions with stroke patients attending therapy sessions in the Borstad lab, I also realized that I greatly enjoy and respect the clinical atmosphere of medical recovery, and that it is very rewarding improving the lives of patients that you interact with face to face. With this in mind, I will undoubtedly be pursuing a clinical position in my later career, so that I may continue to improve the livelihoods of persons suffering from ailments such as stroke.

I plan on continuing the research I began through my STEP project throughout the remainder of my undergraduate career, as this research is still far from completed. With the framework and understanding of the methodology, it is now a matter of applying it to have a positive impact on our understanding of stroke. Nevertheless, I gained an important understanding of what I want to pursue as a career through this research, as I realized I greatly enjoyed interacting with stroke patients and found it to be immensely rewarding. While I am not currently set on a clinical specialization, I do know now that I want to work with patients in the future, a huge leap forward from before my experience.  Previously, I had little to no idea of what I truly wanted to do with my field or science background, and was greatly confused about which path to take moving forward. Now, I have a solid general direction towards clinical graduate programs, and past research experience to back it up. I am truly thankful to have been provided the opportunity to learn, improve myself, and develop future goals through my STEP experience.

Luke Lundy – Neuroscience 2016

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Akron General Summer Research Fellowship

What? – A detailed description of what you did during your STEP experience.

Over this past summer I conducted research through the Akron General Medical Center (AGMC) Emergency Department Research Fellowship. The fellowship consisted of writing up case reports for unique presenting patients in the ER, revising AGMC’s emergency medicine milestone rubrics, attending various lectures and expos, shadowing various doctors, and taking part in simulation labs (i.e. lumbar puncture, intubations, etc.).

So What? – A personal response to your STEP experience, including feelings, thoughts, judgments, and what you have learned about yourself and your assumptions from what you did and how you reacted.

Through this experience I was able to gain valuable experience working within a hospital, and mre specifically, in the field I would like to be in, Emergency Medicine. I met many doctors, and doctors-to-be that told me about their path towards medicine, the hardships they encountered, and how to overcome them. I enjoyed working with the other fellows as well, who came from programs like NEOMED and other prestigious pre-med programs. One of the case reports I wrote, Cervical Artery Dissection, has been published in the American Journal of Emergency Medicine (AJEM). And another case report, Mesenteric Ischemia, is being considered for publication in the New England Journal of Medicine (NEJM). The part that impacted me most was the interactions I was able to have with the patients. While shadowing doctors I was able to talk to the patients about what they were going through and learn how the doctors helped with both the physical and emotional aspects of their respective trauma.

Now What? – Discuss how the things you experienced and learned during your STEP experience will affect your academic, personal, and life goals moving forward.

Most importantly, the experiences I had at AGMC encouraged and reinforced my path in life to pursue a career in medicine. My interactions with the doctors and hearing about their own path toward medicine taught me how to better organize my life so that I may work to my full potential in order to continue to pursue this rigorous course. I am excited to be involved in more research fellowships like the one at AGMC. This summer I hope to take part in Ohio University’s Summer Undergraduate Research Fellowship (SURF).

The following link is the final presentation I gave on my work:

Research Presentation

Undergraduate Research Molecular Genetics

What?

During the summer I worked with Dr. Helen Chamberlin in the Department of Molecular Genetics. I continued my research project: C.elegans vs. C.briggsae lin-3/ EGF gene comparison. During the summer I identified one gene that gon-14 and that the gu102 mutant allele causes a splicing defect in this gene.

The mutation in gu102 mutant is changing how the splicing is occurring causing introduction of stop codon. The most common outcome of the mutation that we have is the failure to splice because of the stop codon. After identifying this gene through a series of experiments, I started to sequence the entire gene gon-14 gene of the C.briggsae mutants. I am currently doing experiments to determine how the gon-14 gene affects the EGF/lin-3 pathway. Previous experiments in C. elegans show that this gene has only a minor role in altering EGF signaling, arguing that the signaling network in C. briggsae is more sensitive to disruption of this gene compared to C. elegans.

So what?

The STEP experience helped me because the results of the research I did this summer will be included in my undergraduate research. I will be writing my undergraduate thesis during my senior year. Completing an undergraduate thesis will be a great accomplishment in my undergraduate career. I want to graduate with undergraduate research honors. Also, I want to continue presenting in research forums and symposiums (NMS Research Forum, Denman Research Forum, SACNAS National Conference, among others) to gain more experience in the research field. . I learned that I love research and I want to pursue a PhD in Neuroscience with a focus on molecular genetics and systems. Thus, helping shape my plans for the future in terms of Graduate School and the research topic for my PhD.

Now What?

As I explained before, the STEP Undergraduate experience helped me realize that I want to pursue a PhD. During this experience, I realized that I love research and I want to purse a career as a researcher. In terms of academics, I will target my classes to fit the PhD Program requirements. I want to take several Graduate School courses on my last semester at OSU to be more prepared for Graduate School. As of my personal and life goals moving forward, the STEP experience helped me narrow my future plans. Now I know what I want to pursue a PhD degree and I also have a better idea in which area- Neuroscience with a Molecular/Cellular and System specialization. Moving forward, I will continue with my studies and will be applying to Graduate Schools next summer. I’m really grateful to have participated in the STEP Experience this past summer, this experience gave me an idea of my future plans and for that I am very thankful.

Molecular Genetics Lab

Molecular Genetics Lab

 

Presenting my research at the SACNAS National Conference in Los Angeles, California.

Presenting my research at the SACNAS National Conference in Los Angeles, California.