Author Archives: Dang Nguyen

Final Stage: Thank You

During the poster presentation Friday, someone approached me and told me that he had read many of these blogs and liked my style, applauding my writing. Before I could really grasp that he was not simply after the 30-second rundown of my poster I had laboriously perfected the prior night, he continued through the parade of posters ephemerally as I squeaked a meek ‘thank you.’

I say it so dreamily because it certainly was one of the most touching experiences this summer, right with seeing my entire lab flood into the hall, looking around for me and Christine (who also presented under the Summer Neuroscience Program). It was somewhere in that moment that I realized they had become my family- a clutter of familar voices cutting through the mob and somehow I already know what they’re saying. An ineffable feeling that Dr. Colton is in front, like a flag-bearer in her blue blazar, and somewhere lagging behind is Hui Fang glancing around. Stuart, now unaddled by the broken A/C of the lab, is out of his lab coat and in a buttoned shirt- plaid -and shorts as things should be, and notices me first along with Angela. Joan wastes no time walking closer, steering the group in my direction, including Kendra undistracted by all the other flashy posters. Like the 5th grade science fair where your parents came despite having work, except with so, so many more questions about the science and not the closest bathroom. As BSURF concludes, I realized how many memories and experiences I have tied to this summer, and how many people I wish to thank beyond a 10′ by 5′ poster corner could.

Thank you Dr. Grunwald and Jason Long. I think you both are entirely aware of how impactful this program can be because you watch it happen every year, but for repeated emphasis, this summer has been LIFE-CHANGING. Perhaps not in a existential manner, but definitely in a navigational one. From working behind the scenes with food, finances, and fun to arranging informative experiences and talks, both of you have helped us tremendously to stay on track while still being so tolerant and flexible. I really enjoyed getting to know both of you these two months, especially in this environment where your mentorship skills really thrived. Interviewing with Dr. Grunwald had me in complete admiration from the start- both from his office’s many reptilian tanks and his scientific authority -and this has only continued to grow around your charisma towards students, and Jason’s welcoming and attentive chats. Although I had some trouble with my chalk talk, there was no dip in support, and in that atmosphere, how could I not learn to improve, to present, to communicate.

Thank you to Dr. Colton and my lab. Everyone has been incredibly friendly and helpful these two months: answering endless swarms of questions, guiding me towards what goals I should have, casual conversations and parties, and tireless jokes about my sleeping habits in and out of lab. Quite honestly, this has been the undergraduate lab experience I dreamed of, an appropriate balance of independence and mentorship, fascinating research topics with several directions that convene regularly, mellow lab environment without pressure to perform, and meetings to reinforce professionalism and ambitions while remaining friendly. I do mean it quite seriously when I claim everyone as like family, and that includes our now-gone, infant (or perhaps fetus would be more accurate?) Taylor (Good luck on college applications and the SAT/ACT!). I have picked up much from being around everyone, such as how to interact with lab members and the path to designing one’s own project about a topic. Even the finer nuances of the lab like project timelines, lab presentations, reagant costs, and technical tips and tricks to spitting out 5 Westerns in a week. I have so much more I’d like to attribute to everyone, but there will be plenty of time for me to pester the lab with them, since I will be sticking around for three more years to further look into UK114 🙂 See you in a month!

Thank you to my audience! This includes my peers, other researchers, and yes, those reading my blogs! I know how long these posts are- apologies -but I would like to ramble a bit more on how grateful I am for your support and future interactions. It is somewhat strange for me to process that my vehicle for transcribing my thoughts and project directions has actually been reviewed by a professional audience, and apparently enjoyed. Those small moments really are both moving and exciting, alerting me to just how interactive a community research can be. I hope to continue working hard to impress those looking on without deviating too far from my current style.

Weekly Highlights

“What is your name?”- a random PI grabbing a drink at the same time I was at the BioCore symposium whose name I unfortunately did not catch
“Oh, Dang”-Dang
“And where are you from, sir?”-PI
“South Carolina?”-Dang
“Oh are you from MUSC? (the Medical University of South Carolina)”-PI
“OH! Oh no. I am an undergrad.”-Dang *strangely explaining that he is not a medical student for the third time this year*

“IT’S FREEZING”-Stuart running around in lab coats for warmth trying to find out how to fix the thermostat

“Hey, Hui Fang, do you want to join a luncheon for my program?”-Dang
“What? Really?”-Hui Fang
“Uh, yeah”-Dang
“Really?!”-Hui Fang

“Good morning?”-Joan opening lab at 6 am
“Good morning Joan :)”-Dang cracking open his third Western gel
“…How long have you been here”-Joan
“I never left”-Dang

“You should sleep more Dang!”-Christine
“I feel so rested though!”-Dang
“You fell asleep three separate times during the talk”-Taylor

“What is that green ice cream?”-Marilyn
“I think it’s green tea?”-Angela
“It’s got some weird off green color. Oh Dang got some, how is it?”-Stuart
“…it has…like…no real flavor?? It is green tea I think”-Dang on his third bowl of it

“So like, no ulterior motive or anything, but could you hypothetically drink the 200 proof ethanol?”-Dang with serious but innocent inquiries
“Uh, I think? I think I asked that question when I was new too.”-Stuart

“Stuart what would weird black orbs in your culture mean?”-Dang
“Oh, is there an infection? What do they look like?”-Stuart
“Well I think they’re yeast, but they’re like orbish and black. Really round. Here let me show you, I caught it and cultured it in a well”-Dang
“You what”-Stuart
—-later—
“So, I accidentally treated the same well with hydrogen peroxide meant for one of my experiments, so I think I killed them all”-Dang
“What? Are you sure they weren’t debris or anything?-Stuart
“Yeah it definitely wasn’t! There were a ton of them and they were all different sizes!”-Dang
“Hmm, well show me if you find them again I guess?”-Stuart
—later—
“So, it turns out Stuart, that they were just bubbles”-Dang
“What the heck haha you had me worried. You were trying to grow bubbles?”-Stuart

SUMMER BLOOPERSHIGHLIGHTS

*in the midst of Lefkowitz’s talk about his career*
“Oh god I left the hot plate on”-Dang as he nervously begins staring at the clock for when he can run back to lab

“Oh thank god it didn’t explode”-Dang finding the hotplate still on with the bowl of water completely evaporated
*bowl shatters*

“zzz”-Dang asleep while waiting for chemiluminescent exposure time
*Hui Fang softly knocking outside the locked imaging room for 5 minutes*

“Now just carefully….”-Dang cracking open his first Western gel
*gel case shatters*
“…stab the gel with shrapnel…”-Dang
“How did the Western go?”-Hui Fang
“Did you learn how to solve jigsaw puzzles in your other lab?”-Dang turning around with 5 scraps of gel

“Wow we’re back at lab already?”-Dang
“You were snoring right after we left Dr. Colton’s house”-Joan

Are you a student here?”-a high schooler touring through the Bryan Research Building
“Yes, I am! A rising second-year”-Dang
“Oh wow, where did you go for undergrad?”-high schooler
“Oh, um. I actually am still an undergrad…Do I look that old?”-Dang

So you’re only a first-year?”-Dr. Colton
“Yup!”-Dang
“Well, our lab is always open to having medical students onboard!”-Dr. Colton
“Oh. Oh no. I’m an undergraduate.”-Dang’s first interaction with Dr. Colton ever (actually in October)

Funding provided by Duke University Trinity College of Arts and Sciences. THANK YOU BSURF!!!! 

Stage 7: I mean, as if PI wasn’t good enough but a Dean…married to another DEAN

After a troubling week in lab (5 inexplicably failed trials, time restraints, and deadlines deadlines deadlines!), it’s certainly nice to reflect on other research being done at Duke, especially when that research involves my favorite buggers: viruses! I introduce Dr. Mary Klotman, Dean and Professor of Duke’s School of Medicine, and HIV specialist who has fought at both the clinical frontlines and the bench operations. For years, I have assumed my interest in STEM and medicine to blossom directly as a physician of some manner, not really considering the entire prospect of researchers even existing, much less being one. Thus, choosing from research and clinical work whenever asked by peers and advisors alike seemed entirely too daunting, but Dr. Klotman is an amazing example of stringing the two fields together through her translational research.

Thoroughly Duke-bred since her undergraduate years, Dr. Klotman entered research in a slightly unconventional route to other scholars. Marked by the concurrent AIDS epidemic, her rotations and residency at Duke drove her to seek more ways to help her bedside patients out of compassion.

After pushing herself into the world of HIV research, she began investigating the mechanisms and properties of the virus as the research remained relatively fresh. Developing techniques at the time rushed to fill the dire gaps of the crisis, leading to major discoveries such as the limited function of failure drugs like AZT as potential treatments and the entire sequencing of the HIV genome. However, Dr. Klotman’s research focuses mostly on a fatal kidney disorder related to the virus targetting the young African-American male community. By studying the behavior of this disease, Dr. Klotman would be able to investigate the infection methods of the virus and potential treatment targets, as the virus is typically associated with infecting T cells, yet the kidney lacked many traditional immune cells. Combined with her continued bedside activity, Dr. Klotman labored tirelessly to resist the terminal diagnoses given to AIDS patients, even leaving Duke to Mount Sinai to work as the chief of the Infectious Diseases division after determining that HIV evolved and behaved distinctly differently in the kidney.

Since the end of the AIDS crisis, Dr. Klotman has persisted and furthered the field of HIV through her work, investigating specific factors of the virus heavily tied to the virus’ pathogenesis in immune cells. Despite the subsiding of the urgency for a cure, her projects have transitioned back to Duke’s Medical School to work with the Viral Vector Core on a potential vaccine for HIV using integrase-defective lentiviral vectors to stimulate the immune system. Since HIV itself is a lentivirus, using a vector with mimicking properties/factors that is unable to reproduce would allow for the development of a vaccine. Now, she juggles both her previous responsibilites to the clinic and laboratory, while adding the administrative workload of a Dean of the School of Medicine, which is simply incredible. Plainly put, Dr. Klotman’s story is inspirational, as it evinces that one doesn’t have to really decide strictly between M.D. or Ph D., as in the end, she never chose to be either but became both out of her passion and interest in her work. It is quite relieving, as I was worried about the practicality of pursuing research if I were to choose the M.D. path, but Dr. Klotman’s extraordinary work in both paths despite technically belonging to one shows how arbitrary those divisions can be in the face of hard work (and probably a whole lot of skill).

Weekly Highlights

“CRISIS CRISIS”-Dang as membranes dry up
“I’ll go get Huifang and take care of what she’s working on so she can help you”-Joan who leaves
*crisis immediately fixes itself*
“What’s wrong, I was feeding mice [an hour task]!”-Huifang
“…so, guess who doesn’t have to feed mice anymore?”-Dang

“Tell me when things are running low so I can buy them before we run out”-Joan
“Oh ok, um can you buy DMEM/F12 [$100], RSAD1 antibody from Novus [$400], West Femto ECL[$350], 6-well plates for cell culturing [$150], Stainless Protein Ladder [$100], anti-tag FLAG antibody [$300], 20x TBST [$100], 10x Tris/Glycine/SDS buffer [$40], BCA Reagant A [$60], and some precast Western gels.”-Dang
“Oof.”-Huifang

“So I put the marker in the well, and I was telling myself not to load the sample in that same well.”-Dang
“And then you did exactly that”-Joan

“Did you remember that you were supposed to present at the lab meeting today?”-Joan
“Yes, save meee.”-Dang
“Are you done preparing?”-Christine
“I mean, not completely…”-Dang
“Good, lab meeting cancelled!”-Joan

“Isn’t there a lab meeting today?”-Kendra who runs Mass Specs so you know these things are a journey
“Just the two of us have been sitting in the conference room for 20 minutes confused.”-Huifang
“Oh, uh, yeah we cancelled it, did nobody tell you?”-Dang
“Really? Oh I guess I didn’t, when?”-Kendra
“5 minutes ago”-Dang

 

Stage 6: I should probably have bookmarked those sources

Abnormal and excessive tau phosphorylation commonly characterizes Alzheimer’s disease (AD) neuropathology through neurofibrillary tangles (NFTs) causing axonal dysfunction and microtubular instability. Essential to DNA repair, protein modification, and gene expression, the methionine cycle generates a methyl group upon the conversion of S-adenosyl-methionine to S-adenosyl-homocysteine, which can activate protein phosphatase 2A (PP2A) and leucine carboxyl methyltransferase 1 interactions in neurons.1,2 These interactions are known to target serine/threonine phosphate groups prevalent in hyperphosphorylated tau, and prior studies have demonstrated increased aggregate formation in vivo upon PP2A inhibition.1,2 Furthermore, increased levels of homocysteine, a byproduct of the methionine cycle normally converted back into methionine or anti-oxidative glutathione, is strongly linked to the progression of AD.3 This suggests insufficient methionine resynthesis allowing the hyperphosphorylation of tau to form NFTs. Biochemical analysis of the different pathways homocysteine metabolism undertakes show many similarities to digestive reactions in the liver of the reactive intermediate imine deaminase A homolog (UK114) enzyme. We are studying the potential interactions and role of UK114 in transfected Chinese Hamster Ovary cells by recording protein expression levels via Western blot. The transfected cells are subjected to various conditions either translatable with symptoms of AD or saturated with components of the methionine cycle that are potentially catalyzed by UK114. The results of these studies will further expand our understanding of the methionine cycle in relation to AD pathology, as well as explore the antioxidant uses of UK114 in an immunological scope.

Weekly Highlights:
*LOUD THUD as Dang accidentally knocks over a giant bottle*
“Is Dang throwing things again? Bad boy.”-Joan

“Let’s go stuff some squirrels and hamsters in a fridge to induce torpor!”-Dr. Colton
“We already tried that. It didn’t work.”-Joan
“PARTY POOPER!”-Dr. Colton

“Wow, big pellets! This sample is good!
*BCA subsequently fails. negative protein concentrations returned. half the trial ruined*
“Oh no.”-Huifang

“What’s wrong with your milk? :/”-Huifang as she throws away milk and goes to her own stash

“And one day, you’ll be like these two amazing children. One graduating with distinction and the other just a lowly freshman!”-Dr. Colton

References

1.Qian, W., Shi, J., Yin, X., Iqbal, K., Grundke-Iqbal, I., Gong, C., Liu, F. (2010). PP2A Regulates Tau Phosphorylation Directly and also Indirectly via Activating GSK-3β. Journal of Alzheimer’s Disease, 19(4), 1221-1229.https://content.iospress.com/articles/journal-of-alzheimers-disease/jad01317

2.Stanevich, V., Jiang, L., Satyshur, K. A., Li, Y., Jeffrey, P. D., Li, Z., … Xing, Y. (2011). The structural basis for tight control of PP2A methylation and function by LCMT-1. Molecular Cell, 41(3), 331–342. http://doi.org/10.1016/j.molcel.2010.12.030

3.Seshadri, S., Beiser, A., Selhub, J., Jacques, P. F., Rosenberg, I. H., D’Agostino, R. B., Wilson, P. W.F., Wolf, P. A. (2002). Plasma Homocysteine as a Risk Factor for Dementia and Alzheimer’s Disease. The New England Journal of Medicine, 346, 476-483.
http://doi.org/10.1056/NEJMoa011613

Stage 5: So I can’t copy/paste my lab notebook here?

With a full day of benchwork planned, I try and wake up at 5 am, but the nauseating breach of sunlight through the blinds often send me back under the covers for another 5 minutes. Or maybe 50. Given the former, I head to the gym briefly, shower, and hustle it to lab to consolidate the day’s different goals into a precise schedule and starting it before the BSURF meeting. Given the latter, well, might as well round 5:50 am to 6 am, which is essentially too late for a whole workout routine, so might as well conserve sleep until 7 am, but I don’t actually need breakfast, so 7:45 am works, but construction is bothersome, and therefore, I hope my 8:40 am alarm catches me in time for the BSURF meeting…Maybe 8:41?

After BSURF, I drift back to the Bryan Research Building at around 10:20 am. Christine, the only other undergraduate in the Colton lab, is hard at work preparing and reviewing her PCR data on ground squirrel brain tissue but doesn’t hesitate to glance up and greet anyone who walks into the lab. Joan, a technician, is mysteriously working on the other side of the bench, somehow switching constantly between benchwork, lab management, and mice care. Three coffees, some slight joking, and one lively Dr. Colton visit later, the day officially starts. What I do depends on where I am on the experiment. Some days are laborious: passage the cells, treat the experimental lines, prepare buffers, read articles, prepare the next experiment. Other days are lazy, and I might sneak in a nap for my typically brisk lunch break, although 2 more hits of coffee will suffice before bugging Huifang and Stuart, another two techs in the lab, for guidance or just because. They’re really cool, and also somehow do the things that take me a day in an hour. And then some.

If I haven’t clumsily destroyed something (it’s only happened once or thrice), then the day is done! Which is usually around 5 pm because although precise, efficiency evades me on the best of days. *looks nervously over at all 8 BCAs I have done sans knowledge of the glorious multi-channel pipette* *furious glance at Western blot gel that blew up and had to be meticulously jigsawed back together* *desperate groans as water/bead bath takes an hour heating up a singular media for a one-step protocol* *flies into hysterics at all the reagents that have conveniently run out right before the last sample* Although sometimes, I just stay in lab a few extra hours just to peek at some IHC slides Angela, another tech specialized in churning out hundreds of these things (it’s really quite amazing), has prepared under a microscope for different stained proteins or cell types. Past 4 pm, most everyone has left besides Stuart or Huifang, and so, I pester them a bit more because they are awesome, and try and get the next day worked out. After 6 pm, everyone is most definitely gone, and if not, woe to them because their only remaining company would be me! I spend around 30 minutes to an hour completing my lab notebook and reviewing calculations or data from the day, and if I am still awake, I glance through some more articles to see what I want to do next. All this makes me feel productive, which is comforting because as soon as I get back to my room, I eat and sleep. Rinse. And repeat. I cannot imagine fitting this into a semester of classes and extracurriculars, but nights are long and I really enjoy it here so I’ll make it work. It better work. Usually, something works.

Weekly Highlights

“This is Dr. Colton’s place!”-Joan Wilson, proceeding to turn into a forest
“What”-Dang Nguyen as the next half mile is spent driving through straight gravel to get atop of a hill

“I prepared the cells!”-Dang Nguyen
“What? You prepare the cells the day before transfection.”-Huifang Dai
“Oh. Uh.”-Dang Nguyen ruining his weekend plans right before heading home on a Friday afternoon

“Now we’re finally done!”-Huifang Dai at 6:11 pm after a long day of Westerns
“Huzzah!”-Dang Nguyen as he turns around to head home just a tad too violently, merrily spilling the last drops of antibody dilution they had just finished

“We’re here!”-Joan Wilson breaking into Dr. Colton’s backdoor and walking straight in

“Happy birthday guys!”-Dr. Carol Colton with a birthday cake
“All [f]our birthdays were weeks ago”-Stuart Sundseth
“…It’s America’s birthday”-Dr. Carol Colton

Stage 4: A Quick Intermission

Although a life of research often leads to an extremely narrow field of expertise, reading and learning about new research and knowledge from other, sometimes completely unrelated, fields can be both exciting and intriguing. One such example is Ayana’s project on Cryptococcus neoformans, specifically on searching for links between the BZP4 gene and their virulence in humans.

C. neoformans is a very common yeast, often found clinging to the dirt and other plants and animals. While widespread, infections from the fungus is rare, as most people’s immune systems are capable of defending against the pathogen. However, occasionally, they opportunistically infect the lungs, which can then spread to the central nervous system where the fungi can cause meningitis or encephalitis- two very dangerous conditions. Therefore, Ayana’s lab is researching the pathology and potential factors that could help the development of some form of prevention or cure. Currently, her project revolves around the curious BZP4 gene within C. neoformans, as the gene has been previously known to fluctuate in gene expression levels in different conditions and upon knockout, the virulence of the fungi disappears.  Thus, she has decided to investigate its relationship to the virulence of the fungi by directly interfering with it function.

In Ayana’s first aim, she hopes to confirm the link between virulence and BZP4 in C. neoformans, by taking a BZP4-knockout strain and blasting it with the BZP4 gene to observe for a recovery in lethality. This is cool primarily because it involves the use of a gene gun, which is just a really awesome machine that exists apparently, transforming cells with DNA by blasting them with a gene-coated bullet- and it works! At some point in my life, I need to devise a transformation experiment that requires that machine just to see it in action. Furthermore, her work would further pinpoint a site of target for treatments of C. neoformans infections, which is a monumental success in the world of disease prevention and care. Additionally, it could demonstrate a stronger link between virulence and the BZP4 gene, as the gene’s interactions and expression post-transformation could be identical to pre-knockout levels which would indicate greater independence between the gene’s mere presence and virulence over the possibility of gene expression interactions and post-translational activity with nearby genes being the source of virulence.

In Ayana’s second aim, the goal is to determine if there a competitive advantage given by the BZP4 gene. To study this, she will be inoculating a BZP4+ strain of C. neoformans and a BZP4- strain in close proximity, and measuring their growth and interactions upon reaching one another. This will perhaps give some insight in the purpose of the BZP4 gene, as not much is known beyond the excitatory effects it has on melanin production, which at most can be suspected to boost tolerance to environmental oxidants such as UV radiation. This is also important because the results can be used to determine if virulent C. neoformans growth can be stymied by the introduction of nonvirulent BZP4- strains into important sites of infection. However, something I wondered in this component of the project but did not remember to ask was the activity of BZP4 in various environments. As antioxidants are so versatile and diverse, many function differently in different conditions, so I wonder if there are any environments where BZP4 activity is optimized, boosting the vitality of C. neoformans in that environment. Similarly, which environments is activity dampened in? This could perhaps explain what BZP4 is specialized for (or what the melanin is meant to do) and what pathway exactly that it manipulates in the body to cause so much damage. Studies like these fascinate me and remind me of just how essential research is to the field of medicine, even in straight biology settings. While not at all related to my research in Alzheimer’s disease, this project gave me some energy and exciting plans to bring to my work.

Weekly Updates

“A piece of my gel fell on the floor once and I didn’t notice, so renovations re-varnished the lab. Now it’s forever imprinted on the floor.”-Stuart Sundseth detailing one of many Western fails

“I accidentally threw my gel into the sink once.”-Stuart Sundseth

“I’ve torn a gel before”-Stuart Sundseth an hour before tearing another gel

“Have fun at the beach this weekend!”-Christine O’Connell
“Wow, you’re going to the beach again?”-Dang Nguyen
“Yeah, I left some potatoes in the cupboard”-Joan Wilson

“I feel…so dizzy…I think…I might fall over”-Hui Fang as she aggressively hoses the floor with liquid nitrogen

“And now I’ll just seal up this bad boy and throw him in the cold room for the-” *throws dead spider in box at Dang* “-night and let him blot since I can’t find the antibody.”-Stuart Sundseth
*graphic screaming*-Dang Nguyen

Stage Three: Fishy Business

Behind every publication lining the pages of Cell and Nature, there’s a scientist. A tinkering mad(wo)man with an insatiable lust for knowledge and also just being really cool. Sometimes, being cool just seems to run in the family, as it does for my PI, Dr. Carol Colton (or maybe its the self-professed madness slowly taking her). A perky yet wry neurobiologist, she is a pioneer in the immunological field of Alzheimer’s disease with an dignified, open attitude, and I fear the day she reads this.

Belonging to a family of scientists, Dr. Colton has always desired to do STEM research, particularly in Biology. Initially, she planned to enter paleontology at the University College London, but science is never a straight road, and that goes for the scientist too. While excelling in all her studies, there was one occasion that she could only describe as career-ruining. To this day, the trauma and shudders from the experience haunt her, and she still has a bone to pick with fish paleontology. On a fateful lab day for a class, the assignment happened to be to take rotting fish-market rejects, to boil their carcasses, to pick the meat apart, and to piece back together the skeleton and identify the fish. Nauseating.

After a prompt and well-deserved department shift, Dr. Colton found herself in a neurology lab. At the time, neuroscience focused heavily on studying the function of neurons, and so the most lending model to study using patch-clamps was of course the squid giant axon. After continuing her research and career at Rutgers University and the National Institute of Health, she began to discover the voice of dissent among scientists when she became interested in microglia.

Microglia, commonly summarized as brain’s immune cells, not too long ago were considered a myth. The brain was just a brain with many neurons, surrounded by a network of blood vessels. However, Dr. Colton found some research indicating a strange possibility of there being something more. After consulting with a fellow researcher who was centrifuging down brain cells to study, she discovered something strange about the heaviest layer which expressed traits common to those of macrophages. But others disagreed, there was opinion that those were just impurities of the sample from the nearby vascular system, pouring with immune cells. Nonetheless, Dr. Colton ignored symposium introductions “humoring” her microglia work, and studied these cells further, and later contributed to the slow recognition of the existence of microglia. An astonishing amount of work has been done since then on the cells, as it turns out, they are essential in many diseases and neural interactions, controlling many inflammatory and metabolic pathways in the brain.

Since then, Dr. Colton has worked at Georgetown University on one inflammatory factor released by microglia: Nitric Oxide. An important component in many neurological diseases, nitric oxide is used both as a neurotransmitter and a player in Alzheimer’s disease. However, the study of its levels in Alzheimer’s disease was scant because of Alzheimer’s posthumous diagnosis and the inability to collect brain samples from living humans. Therefore, for a long time, mice models were used to simulate human neural pathologies, but after a strange virus kept plaguing Dr. Colton’s mice colonies, she abandoned mice for hamsters. Oddly, her new data matched with none of her previous work and even conflicted with other research done on nitric oxide levels prior. After months of confusion and frustration, she discovered that hamsters actually have a different nitric oxide processing enzyme (known as NO synthase 2 as opposed to NO synthase 1) that actually mirror human immune systems much more accurately. This finding slammed her once again into the neuroscience spotlight, as she began working towards replicating the human neural system more and more accurately within rodent models.

More recently, Dr. Colton’s work has ranged from developing patented mice lines for research to studying various metabolic pathways related to Alzheimer’s disease, piecing together different mysteries found in more general brain tissue scans. Now residing at Duke and working with its Kathleen Price Bryan Brain Bank, she has investigated the various functions of arginine and other immuno-regulated chemicals in cell cultures, mice colonies, and even human samples. Her latest challenge to the field of science: viruses are the missing link to Alzheimer’s disease. Of course, the specifics are confidential, but I am sure she will be both excited and also apprehensive that her theory just recently got the Alzheimer’s field clamoring once more, as a heavily controversial study became published by Neuron last Wednesday supporting the possibility of viruses being a key factor to the onset of Alzheimer’s disease. I can already see her laughing at everyone’s face next week.

Weekly Highlights

“I’m smart and your minds are decaying. I’m going to tell you why.”-Joan Wilson imitating Dr. Colton at an Alzheimer’s Talk

“I told him about my idea on viruses, and he was like ‘naahhh.’ Well, when we have proof, I’m going to shove it up to him and be like ‘HA. Ha ha. Ha.'”-Dr. Carol Colton

“How was your vacation, Joan?”-Dr. Carol Colton
“Well on Monday a TV fell on me. There’s the bruise right here. I also got my ankle really swollen, that was my Tuesday.”-Joan Wilson, a survivor

“I do EVERYTHING?!”- Hui Fang finally realizing she doesn’t get paid enough for the 6 roles she plays across 2 labs

“The plasmids arrived!”-Dang Nguyen
“Good! We can start tomorrow.”-Hui Fang
“There’s two”-Dang Nguyen
“What.”-Hui Fang, as the next half hour is spent reading through both DNA sequences to find the right one

Stage 2: UK114 HTTP404

As universally known, science is a lot of waiting. From mice studies to monotonous PCR cycles, the Colton lab runs plenty of patience-intensive projects that can seem daunting. Therefore, I should begin as soon as possible to avoid an incomplete or hastily patch-worked project for the summer, so theoretically like this week. Theoretically. Who knew just getting the supplies would cost the most time, as the day turnaround estimate for the UK114 mRNA clone to arrive stretched into its second week. But I haven’t just been dawdling about the lab for a week, that requires getting out of bed. Instead, I have been spending the week reading further into the specifics of the lab’s projects and prized focus: the methionine cycle.

The methionine cycle, in short, is an essential and widely used metabolic pathway in the body where the namesake amino acid is converted into SAM, a prevalent substrate used for methylation in an extensive list of processes. These systems include gene regulation, neurotransmitter production, amino acid conversion, and various protein mechanisms; but the Colton lab is particularly interested in the methylation of PP2A which has been shown to downregulate the phosphorylation of tau. This is extraordinary to Alzheimer’s research, as hyperphosphorylated tau composes the infamous tau tangles in the brain that drive the neurodegeneration seen in Alzheimer’s disease. Thus, it is suspected that the methionine re-synthesis step of the methionine cycle is inhibited, preventing the production of SAM and consequently the accumulation of phosphorylated tau. Indeed, this hypothesis is supported by the elevated presence of homocysteine- a hazardous product of the methionine cycle that is normally recycled back into methionine -in many Alzheimer’s patients, suggesting some deficiency of the methionine synthesis step. While several factors are known to regulate this process, there are some intriguingly obscure proteins with unresearched functions that have a notably high presence in Alzheimer’s patients, such as RidA protein UK114.

Avoiding overly outlining the biochemistry and exact mechanisms of the methionine cycle and UK114’s potential interactions, my project involves the transfection of cells with the UK114 gene and subjugation of the cells to various conditions related to the methionine cycle to study UK114’s exact role in Alzheimer’s disease and why it is upregulated. I have spent the past week investigating much of UK114’s structural specifics and which enzymes it could potentially react with and when; however, I plan to test these theories for confirmation once I am able to produce a transient cell line expressing human UK114. Unfortunately, a delayed delivery that contained simply an empty vector without the sequence has put this plan at a standstill, but hopefully my project will commence at some point next week when the gene arrives. I feel like I have repeated this countlessly to everyone in my lab, but “until then, I’ll keep reading.”

Weekly Highlights:

“The methionine cycle is like a salmon wheel!” -Dr. Carol Colton
“A what”-Dang Nguyen as the lab meeting proceeds to derail into a half hour lecture on salmon wheels

“Did she just run outside and catch some wild ground squirrels for [brain] tissue samples?”-Christine O’Connell
“No. I don’t know. We should do that with hamsters.”-Dr. Carol Colton

“We should run into PetSmart and unlock all the hamster cages, so we can study them!”-Dr. Carol Colton

“Forget mice! We should go to New Zealand and study platypuses! And maybe hamsters.”-Dr. Carol Colton

“And here we see that- oh. That’s wrong. Oh well, it’s already in the grant!”-Dr. Carol Colton

Stage 1-Steady Steps

With a week past in the Colton Lab, I am absolutely confident that this will be a wonderful summer. Now, I say this recklessly of course, there will probably be plenty of moments where the urge to nap or to give up will be great, but nonetheless, I expect some of the greatest weeks of the year coming.

Like most, there are certain experiences I would be shocked not having: experimental failure, scheduling conflicts, basic protocol training, and tedious manual work. Although mingled with successful experiences- like discovering fascinating patterns, developing new skills, and exploring my interests-one might wonder, why even bother? I mean, personally, if I were to expect anything, why not purely good things like relaxing at home all summer and eating ice cream? Well, I certainly have this thought waking up early every morning, but something about arriving and just being in Dr. Carol Colton’s lab everyday brings forth some sense of contentment. And by contentment, I refer not to the complacency of finally being in an Alzheimer’s frontier-leading lab- though that thought by itself is very exciting – but instead, I speak of the eagerness that settles in every morning mapping out my plans for the day, for the summer, for the next 3 years. There is some ineffable emotion caught up in breath whenever I am researching or watching our amazing technicians, Joan Wilson and Stuart Sundseth, working.

But I digress, sometimes there is no romanticizing the day’s 6th BCA assay or getting entirely lost in the background literature for your own project. I could say, as a whole, this week has been a struggle; from relentless protocol practice to swamps of scientific articles, I have been playing catch up to hone my skills, knowledge, and the trust of others, so that I can work on an equal playing field as them. Slowly however, I am getting there. As I sift through my second window of 20-some article tabs, the words and techniques used in the lab is flowing more cohesively and efficiently. I am gaining ground in understanding the different roles the protein UK114 plays in the body, and why I will be transfecting it with Chinese Hamster Ovary cells containing human tau within the coming days. Furthermore, I hope to specialize in an area related to Alzheimer’s that will allow me to add my own input at lab meetings, and as suggested by Dr. Colton, I have begun diving deep into Herpes virus research and its links to Alzheimer’s for future ideas. While all of these experiences in the past week have been undeniably overwhelming, it has been and foreseeably will continue being incredibly rewarding. If there’s one thing to expect this summer from me, then of course, I expect not to hold back  to strengthen my lab in every way I can.

Highlights of the week:

“You will become THE VIRUS KING!” -Dr. Carol Colton
“Virus King just sounds like he’s carrying something”-Stuart Sundseth
“What? But I’d love to be known as the Virus Queen.” Dr. Carol Colton

“You just want to watch them have sex”-Joan Wilson with another tech on growing a mouse colony bigger

“You will probably reach 85 too, and then yes, you too will get Alzheimer’s”- Dr. David Bennett reassuring an elderly AD-symposium attendee of her future health

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