Archive for the 'Uncategorized' Category

Back for More!

Monday, September 10th, 2007

Although initially it was a little depressing to return to school having been there less than a month before, now that classes have gotten underway and I have returned to the lab I am once again thoroughly enjoying my time at Duke.  My non-science classes this semester are extremely interesting - from a class concerning the leadership and institutional capacity in Africa to an English class analyzing bioethics and narrative.  As for genetics and molecular biology and organic chemistry, despite their bad reputations, I am really enjoying them as well.  The material is quite interesting and the concepts are relevant to the branches of biology that interest me.  Of course, we’ll see how my feelings change after exams begin!

This semester I am also continuing my research in Dr. Wang’s lab as an independent study.  While I am not working on the exact project that I did this summer, I will be assisting my graduate student mentor with his project, which is rather similar to mine.  I really enjoy being able to spend some time in the lab each week, as it is a nice alternative to sitting in lecture and I learn equally as much, if not more.  Well, that’s all for now but I will try to give updates as much as possible throughout the semester!

“Time Flies When You’re Having Fun…”

Monday, July 23rd, 2007

It’s hard to believe that this research experience is coming to a close.  Although it may sometimes seem like I’ve been passaging cells my entire life, it often feels like I just began my fellowship.  While these eight weeks were far from a typical summer, I would not have spent them any other way.  As is apparent from my previous posts, I absolutely loved my time in the lab and I am excited about continuing my work there during the school year.

I’m not sure I have ever learned so much in eight weeks, as this experience not only taught me about a particular subject in science, but also how to do science.  I learned how frame a scientific question, how to plan an experiment, how to control for variables, and how to make appropriate conclusions from data.  I also learned the importance of never losing sight of the big picture.  It is easy to become wrapped up in one small experiment, and easily frustrated if things are not working out as expected.  However, taking a step back and looking at the big picture can help you to troubleshoot and simply to keep you sane! 

Now that I have begun this project, it seems as though I have opened up a huge can of worms – there are so many more questions that have arisen and I cannot help but want to try and answer them.  While a large part of me would love to just drop everything and fully immerse myself in laboratory research, I know that the undergraduate experience is equally as enjoyable and educational.  I’m sure one day in graduate school I’ll probably wish I were an undergraduate again!  So for now, I am going to rest my brain in preparation for next semester and look forward to supplementing my undergraduate experience with some exciting laboratory work on the side!

Career Considerations

Saturday, July 14th, 2007

As a child, my parents succeeded in occupying me for hours by buying me my own toy microscope.  I have always been interested in science, particularly biology and its medical applications.  Growing up I thought that a career as a doctor would be an exciting way to combine an interest in biology and a desire to help people.  I volunteered at a local hospital in high school and really enjoyed the environment.  However, as I have learned more about the variety of science careers available I have realized that a career in research would also satisfy my passion for biology.  I have always been one to constantly ask questions, and as a scientist I would have the ability to ask, and then attempt to answer, the infinite number of questions that science offers. 

I have thoroughly enjoyed my experience in the lab this summer and I am confident that I would love a career as a scientist.  However, I still feel like I would enjoy the medical environment, especially the direct interaction with those you are assisting.  A combined MD/PhD degree would allow me to combine both the research and clinical aspects of medicine.  More specifically, I feel that the fields of cancer biology and infectious disease are exciting, dynamic fields that could be pursued as a physician scientist.  I hope that in the next few years I can take advantage of additional opportunities to gain experience in both fields and really understand what I am best suited for.

Update

Monday, July 9th, 2007

It’s about time that I update everyone on my recent work!  These past few weeks, I have been repeating several experiments in which I study the effects of TGF-b on specific cells through various assays.  These assays have the ability to quantitate TGF-b’s effects on cellular proliferation and migration.  I am working with lentivirus-infected 4T1 cells, which means that the cells have been infected with a retrovirus carrying a vector plasmid of genetic material.  In this case, when the plasmid is transcribed in the host 4T1 cell it yields a strand of RNA that subsequently folds into short hairpin RNA (shRNA).  This shRNA, through a process known as RNA interference (RNAi) has the ability to knock down a designated protein in the cell by binding and degrading the mRNA segment coding for the protein and therefore preventing its translation.  By knocking down specific proteins in the cell through this method, one is able to understand the roles of certain proteins in a cell.  If, for example, a cell knocked down for a specific protein ceases to migrate compared to a control, it can be proposed that the knocked down protein is required for migration.  For my experiments, I am attempting to elucidate the roles of certain proteins involved in the TGF-b signaling pathway and more specifically the pathway that is involved in promoting cellular migration, a key precursor to metastasis in cancer cells.

Thus far, despite about six weeks of nearly fifty hour work weeks, I find myself just as enthusiastic about science as when I started.  I enjoy the infinite amount of unanswered questions that encapsulate science and the freedom, as a scientist, to attempt to answer them.  Each experiment, even when it answers its intended question, opens up an entirely novel area for exploration.  Interested in pursuing similar work as a career, these facts are encouraging – not only does it seem that I will always love what I am doing, but I should also always have something to do! 

 

Responsible Conduct in Research

Sunday, June 24th, 2007

Today concluded a weeks worth of seminars focused on responsible conduct in research.  The movies, speeches, and articles that I viewed in the past week convinced me that all forms of research, not just those involving human subjects, significantly affect society and for that reason ethics in the laboratory is a requisite.

The first seminar began by detailing appropriate protocol for research involving human subjects.  At first, I was having a difficult time understanding how such suggestions could apply to my work with a mouse cell line.  However, as the week unfolded, I quickly began to understand how negligence in my work could negatively affect individuals.  This first became apparent when I read the article about an investigator who lied about his research in order to secure funding.  Such an act places money that could be better used to advance human knowledge, and therefore better society, in hands where this will not occur.  Furthermore, to turn science into a competition among investigators in search of notoriety often hinders the release of knowledge in a timely manner, as evidenced in the movie “And the Band Played On.”  Additionally, negligence in research could lead to incorrect conclusions that, once applied to society, could be detrimental.  Improved knowledge of the world allows us to better interact with it, and obviously incorrect conclusions could be very detrimental.

This set of seminars really emphasized to me the importance of ethical research.  Even though it may seem that the slightest negligence or alteration of data could do no harm, this small change could adversely affect individuals in various ways.  As a scientist, it is important to always be aware of this and to not succumb to negligence in a rush to publish. 

“A Picture is Worth a Thousand Words…”

Tuesday, June 19th, 2007

Sorry I have not updated in a while, my computer was being repaired making it difficult to blog.  Anyway, I spent the majority of last week conducting three simultaneous assays that were a part of one general experiment in which I disrupted the TGFb signaling pathway by inhibiting the ligand receptor.  Through a Western blot, transwell migration, and thymidine incorporation assay, I was able to visualize the several different, somewhat contradictory, effects that TGFb exerts on 4T1 cells.

What I have found most interesting about working in this lab is the ability to actually observe the effects of experiments through a microscope.  Often times, molecular biology requires one to appreciate the science behind the daily pipetting of clear liquids, which can still become tedious.  However, working with actual tissues that can be seen under a microscope allows one to visualize the effects of various perturbations.  For example, after treating the 4T1 cells with TGFb, I was able to notice changes in the cellular morphology.  The cells appeared more stretched and the colonies less compact. Such phenotypic changes were suggestive of TGFb-induced cellular migration, which could lead to metastasis in vivo.  The pictures below illustrate these changes – scroll your mouse over them to view the captions.

With two weeks of experience under my belt, I feel a lot more confident and independent about my work in the lab.  Tomorrow I will begin the actual experiments that will hopefully answer my research question – I’m pretty excited!

This is a picture of cells that have not been treated with TGFb.  The cells are rather round and the colonies densely packed.   These cells have been treated with TGFb.  If you look closely, the cells appear more spread out.                    

       

   

 

First Week

Friday, June 8th, 2007

This past week has been exciting, intense, and extremely educational.  My mentor worked closely with me the past few days to teach me various protocols that will be useful when I begin conducting independent experiments in the next few weeks.  After discussing some basic questions to be answered during the course of my fellowship, my mentor taught me basic techniques involved in cell culture.  Because the cells grow rapidly, the cultures must be passaged often so that they do not reach complete confluence on the culture dish as this would inhibit further division of the cell line.  It was very exciting to be able to see actual cancer cells under the microscope and subsequently work with them.  I also conducted two brief experiments involving TGFB signaling to better understand the signaling pathway and further practice basic scientific techniques.  I analyzed the results of my experiment by conducting polyacrylomide gel electrophoresis to separate the Smad proteins that had been activated by TGFB and used immunoblotting, or Western blotting, to detect the proteins. 

 This is a picture of my first western blot.  In western blotting, fluorescent antibodies bind to proteins allowing them to be detected on film.

Welcome!

Tuesday, June 5th, 2007

Hello, my name is Kristin and I am will be conducting research in the Department of Pharmacology and Cancer Biology at Duke as a part of the Howard Hughes Research Fellows Program.  More specifically, I will be working with a graduate student who is studying transforming growth factor beta (TGFB) and its effect on cancer cells.  In normal cells, the TGFB signaling pathway acts as a tumor suppressor by inducing growth inhibition and/or apoptosis.  Interestingly, TGFB has also been found to act as a tumor promoter in cancer cells by initiating angiogenesis, metastasis, and immunosuppression.  As a part of my fellowship, I will be working with my mentor to study how the various Smad proteins, proteins that are activated by TGFB and exert its effects by acting as transcription factors, encourage cellular migration and metastasis in 4T1 cells, an especially invasive breast cancer cell line derived from mice.  I am very excited about participating in this fellowship, as I am interested in pursuing a career in cancer research.  I see this program as a wonderful opportunity to better understand what such a career would be like as well as to prepare for future research opportunities by gaining significant laboratory experience.

 I will be working with the 4T1 cell line, a particulary invasive murine mammary carcinoma line.