Category Archives: Week 4

A Day in the Bilbo lab

I start the day by getting situated at my usual desk in Dr. Vaidyanathan’s office.

In the time I have before I dive into my lab work, I like to lay out my schedule and to-dos for the day in my bullet journal (anyone who knows me is all too aware of my obsession with good stationary). From there, I’ll go with Dr. Vaidyanathan to start our day of tasks for the project.

As someone who loves a structured routine, I’ve been really leaning into the weekly layout of the project. On Mondays, Dr. Vaidyanathan and I will typically start the day by collecting estrous smears from that week’s cohort of EEG/EMG mice. We’ll then start the EEG/EMG recordings for the mice–this marks the start of the experiment for the week!

Next, we have to image the estrous samples taken that morning. The estrous cycle is essentially the mouse equivalent of the menstrual cycle. It’s about 4 days long, and each day can be characterized into different stages. You can tell what stage in the estrous cycle a female mouse is in by examining the quantities of certain cell types in a sample taken from the vaginal epithelium–a process called vaginal cytology.

Our lab manager Dang Ngyuen (a former BSURFer himself!) trained me on using the brightfield microscope for this process. As I’ve been getting more acquainted with the lab, I’ve been able to perform some procedures with more independence. Lately, I’ve been imaging the estrous samples myself!

On Tuesdays, Dr. Vaidyanathan usually spends the day preparing next week’s mice for EEG/EMG. This part requires a lot of advanced animal handling, so I’ll shadow her for part of the process. Tuesdays typically give me a lot of free time, which I’ve been using to analyze data from the behavioral tests we conducted in the weeks prior. If there’s not much work to be done on the data, I’ll spend that time practicing MATLAB in preparation for the heavier data analysis that I’ll be doing down the line.

Wednesdays are a bit more unpredictable, but they usually end up being some of my favorite days in the lab. There’s not much to do with the cohort of mice on these days, aside from a quick check. Last Wednesday, I had the opportunity to shadow Lauren Green, another post-doc in the lab. She primarily works with zebrafish, another common model organism in neuroscience research. She taught me how to cross transgenic zebrafish and how to screen eggs for a specific genotype that would allow fluorescent tagging of microglia and serotonin-producing neurons. Then, she showed me how she uses a confocal microscopy to image these cell types in the brains of live zebrafish.

On previous Wednesdays, I’ve mostly been learning the procedure for astrocyte and microglia isolation and RNA extraction from Dang. Our plan is to run Q-PCR on RNA isolated from the cortical astrocytes of the mice to confirm that our isolation procedure did in fact isolate astrocytes. Then, although this step goes beyond the 8-week timespan of the BSURF project, we plan to perform RNA sequencing to analyze the astrocyte gene expression patterns of our treatment mice.

Thursdays tend to be another day of coding and data analysis. I’ve been working with Dr. Vaidyanathan to write a MATLAB script that will be able to efficiently compute statistics and  produce graphs from the data we’ve collected from the Forced Swim Test–which happens on Fridays.

We wrap up the experiment for that week’s cohort on Fridays. We stop the EEG/EMG recordings, and set up for the Forced Swim Test–a behavioral test meant to serve as a measure of depressive-like behavior. There’s many moving parts to this test, so Olivia, a new lab technician in training who I’ve been working closely with, helps me and Dr. Vaidyanathan with this part of the experiment.

Finishing up the Forced Swim Test is the conclusion of our week. I usually head out early, and begin with my weekend plans.

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

Episode 4-The Phenotype approach

For this past week, I have been listening to my peers present chalk talks on theirs works, providing me with insight into other areas of biological research. Of these projects, Sweta Kafle’s  phenotypic analysis of transgenerational seed stability in Arabidopsis thaliana could act as an indicator for the stability of transgenerational affects in these plants. These transgeneraiontal affects would be induced by  placing Arabidopsis thaliana  F0’s in hot or cold conditions, then either keeping these conditions constant throuighout the F2 generation, or alternting this patern of hot and cold until the F2 generation, Or altering the hot and cold conditions then keeping those conditions constant ( Hot cold cold, cold hot hot) until the F2 generation..The reason that her project was so interesting to me is while we are analyzing a similar phenomenon, we are taking 2 very different approaches. While S’weta is testing for phenotypic changes and then going on to test for epigenetic ones, I am doing the exact opposite. This talk helped provide me with insight into how the next steps for my own project should be to analyze the phenotypic affects( behavioral, physiological) that an epigentic insult has on my F1 rats. But  coming back to S’weta’s project, I really liked the way that Sweta provided a comphrehensive look into both the flaws and strengths of her study, and it in turn provided a wonderful experience.  I am looking forward to the results of Sweta’s study, and any further insight that future related projects have.

 

Until Next Time

Sea Urchins, Baboons and Mice, Oh My!

Over the past week, myself and other members of the BSURF program had to give chalk talks; 8-minute talks about our project with nothing but a whiteboard and a marker. Even though I was nervous about giving my talk, I was looking forward to the talks. I had previously talked to some of the other students about their projects, but I wanted to know more about everyone’s project. Hearing about the diverse research opportunities gave me the opportunity to expand my knowledge on topics including Alzheimer’s and sea urchin embryos.

One talk that I found fascinating was entitled “Neural Circuitry for Vocalization in Mice”. The chalk talk was given by Alina Xiao, who is working in the Mooney Lab. Her project involves looking at a specific area in the brain to see if it is activated when male mice vocalize, whether it be for defense purposes or courtship. The project got my attention because humans vocalize all the time, yet I rarely think about how and why other animals vocalize. Seeing how mice use this trait compared to humans is interesting. I also enjoy learning about how how the activation of different pathways illicit different responses in the body. Besides being fascinated by the project, I was also impressed with the quality Alina’s talk. It was apparent that she had thoroughly rehearsed her talk and she enjoyed what she was doing. I cannot wait to see the results for her project at the end program.

Overall, everyone gave an amazing chalk talk. Even though we have four more weeks until we present our results (we hit the halfway point!!), I am excited to see how everyone’s projects turn out.

Diverse cellular responses to oxidative stress

This past week, everyone had the opportunity to present their project to the other BSURFers through a chalk talk.

I found Julia’s talk particularly interesting, as her work on Oxidative Stress relates to what I am working on in the Pendergast Lab. Reactive Oxygen Species (ROS) cause cellular damage. By oxidizing amino acid residues, ROS can alter protein structure, causing protein unfolding, aggregation and plaque formation. There are two general mechanism by which the cell responds to such damage: damaged proteins are either degraded or repaired. Julia’s project focuses on a process called ubiquitination. It is the cell’s method of flagging damaged proteins for degradation. K48 ubiquitins are secondary ubiquitin molecules that are attached to the primary ubiquitin’s position 48 lysine (K48) residue. K48 ubiquitination is not understood as well as other forms of ubiquitination. Julia’s project focuses on identifying which enzymes flag proteins with specifically K48 ubiquitins. She is also interested in whether K48 accumulation on proteins following oxidative stress is the result of reduced proteasome activity, or reduced deubiquinating activity.

My project looks at ROS from a somewhat different angle. I focus on system xCT, which is a membrane antiporter responsible for importing cystine into the cytoplasm. Cystine is an important starting molecule in an oxidative stress response pathway. Through a number of intermediates, cystine is converted to glutathione, which quenches harmful reactive oxygen species through conversion to glutathione disulfide. While my project looks at how cells quench ROS before they can damage cellular components, Julia’s project focuses on how cells deal with proteins that have been damaged. It was nice to discover a connection between my project and that of another BSURFer, and I would be interested to see whether anyone in Julia’s lab studies the glutathione synthesis pathway.

Similar Research from a Different Perspective!

 

I enjoyed being able to here about the various research projects going on by my fellow B-Surfers. It was interesting hearing the different problems/questions posed and seeing the ways and which these research questions were being answered. I found it intriguing how some of us were looking at similar topics or ideas but the ways in which we tackled the problems and the methods we used were completely different. With that being said I found Ayana’s project particularly interesting.

Although me and Ayana are both focusing on Cryptococcus neofromans and the ways by which we can treat this fungal pathogen, her project project looks at this opportunistic human fungal pathogen from a different perspective. While my project focuses on controlling sexual reproduction of Cryptococcus by regulating its essential gene expression Ayana’s project is focusing on a transcription factor of Cryptococcus that could play a role in its virulence. Tests have shown that when you delete the BZP4 transcription factor in a host, the fungal pathogen is no longer able to kill that host which is a very interesting finding. She is current running more experiments to confirm this finding but if this is true then its a major breakthrough in finding an efficient method of treating illness caused by Cryptococcus neoformans. 

 

A Different Approach…

My week was highlighted by great chalk talks from every member of the BSURF program. I definitely learned a lot about various topics and projects. Further, these chalk talks spark my interest in new subjects and has gotten me talking about things I normally would have never even thought about! This week was a great lesson in the communication aspect of science and research: having to prepare a chalk talk, listen to presentations from others, and explore other topics through challenging question has certainly helped me to see the utmost importance communication and discussion has in research. Something to be noted is a question asked during my own chalk talk as given me a new idea in which to approach my project, something that might actually help move along my project objective.

That being said, a talk that particularly stood out was the one presentation by Simeon. Simeon gave a chalk talk on the study of essential genes involvement in cryptococcus neoform sexual reproduction expression.

He began with a bit of background information, presenting the case that cryptococcus is a fungi that can cause brain damage to those suffering immunodeficiencies. Cryptococcus is specifically hard to target and create successful drugs to combat it as through its sexual reproduction process different and various types of the cryptococcus fungi are formed and some of these will ultimately carry drug-tolerance. Therefore, going into his project, Simeon noted that he is targeting two essential genes involved in sexual reproduction. Using different constructs, the goal is to test three different approaches and see which conditions will ultimately repress sexual reproduction the most. Having more information on what successfully alters the reproduction expression within these fungi could build toward antifungal treatments in the future.

I found this presentation to be particularly interesting due to the way in which they approached this drug-tolerance issue. Approaching this issue from the perspective of altering sexual reproduction creates an opportunity to then create drugs to target specific types of cryptococcus to combat this fungus. Other studies focus on functional proteins within the fungi or creating drugs that will be undeterred by such tolerance, leaving this approach quite interesting. As this fungi issue is a big clinical problem around the world, this different approach could ultimately begin to lead toward a solution to this problem.

I am very much looking forward to see the information or conclusions gathered by this project and others at the end of the program!

 

 

Luke Sang

The Different Faces of Science

This week, I heard about the amazing research that my fellow BSURFers are conducting while listening to their riveting chalk talks. I learned about neurobiology, plant biology and all the different topics in between. Though all of the presentations were phenomenal, the one that stood out to me was Michelle’s talk. Her project this summer is on knocking out the gene Endo16 in Sea Urchins and whether or not that would affect gut development.

Endo16 is a gene in sea urchins that is fundamental in gut development. Her question is whether knocking out Endo16 will result in reduced gut development in sea urchins. To do this, she will design guide RNAs and inject them along with the Cas9 complex into sea urchin embryos with the goal of causing an indel in that particular gene. With a large deletion or insertion, a frameshift mutation may occur, causing a premature stop codon, and rendering the gene useless thus “knocking it out”. She will then monitor the embryos with a knocked out Endo16 gene to determine if gut development is able to carry on as it normally would or if the lack of the gene prevents the formation of a gut. Determining the function of this gene will aid in understanding how embryonic development works in sea urchins and could lead to information about its interactions with other genes. Her project will expand the Genomic Regulatory Network and she can continue to test other genes with unknown functions.

I enjoyed Michelle’s presentation because I saw the way it related to my own research project. The use of a CRISPR-Cas9 system to knockout a gene and test its function is a real-life application for the technique I am studying. While using CRISPR for medical treatment is what my lab focuses on, hearing about the many other ways it can be used for research makes me appreciate my project even more. I loved hearing about the different methods and aims for her project because I understood everything she discussed. I’ve had to design my own guides and I can relate to the various steps in her procedures. I am amazed by the various ways this genomic tool can be used, as determining the function of a gene is important for understanding its part in genetic diseases. By doing a gene knockout and finding its function, we grow one step closer to unraveling the mysteries of biology.

Episode 4- Chalk the Talk

As interesting as it was to hear about everyone’s projects and learn a thing or two from each one, I think we’re all pretty glad that’s over with. For now, anyway. Hopefully, the posters will be easier since we’ll have everything written down to be presented as is and we’ve already got our thoughts collected.

Out of all the intriguing projects from last week, I believe that the one that caught my interest most was Michael’s investigation on the epigenetic effects in the offspring of mice after the male parent’s usage of THC. In particular, it was curious though concerning that the F1 mice appeared to exhibit some symptoms of ADD and expression of genes associated with ASPD and schizophrenia, though from what I understood, those results are not conclusive.

However, if those current findings prove to be further supported, then that leads one to wonder if THC usage could have similar effects in the children of those who use it. It’s already well-known that women shouldn’t use a variety of drugs or consume fish with high mercury content while pregnant, but it could also be that men should avoid certain substances before planning on having a child, outside of certain drugs that are known to lower sperm count (unless the transgenerational effects of other drugs have been studied and shown to be detrimental?). That also makes me wonder how long the effects of THC on sperm lasts, though I would imagine it would be about however long it takes to produce new sperm.

But anyway, everybody’s projects were pretty cool, and I’m curious to see how the posters turn out at the end of this program!

Week 4 – Mating Galore

Thank you my fellow BSURFers for sharing your projects this past week during the Chalk Talks. It was interesting to see the wide range of research going on this summer and learn about the different topics and techniques used in your research. One of the Chalk Talks that caught my attention was Melissa’s project on the genes underlying neural circuits in male fruit fly courtship behavior.

In her project, she will be looking at how environmental factors affect the expression of the genes Fruitless (Fru), Doublesex (DSX), and Choline Acetyltransferase (ChAT), which have been tied to sexual behavior in male fruit flies. More specifically, ChAT is a gene involved in the production of the acetylcholine and neural circuits involved in courtship behavior. Fru is involved in all courtship behaviors in males, while DSX is expressed at varying levels depending on how much courtship experience male flies have. Male flies with either the wildtype or mutated versions of these genes will either be put into group houses or isolated. While observing how the males respond in the different environments, Melissa can determine what changes were made during gene expression using ChIP, an immunoprecipitation method that tracks interactions between DNA and proteins.

A big reason why I am fascinated by Melissa’s project this summer is because it looks at a behavior in a non-human species. When learning about DNA and genetics in high school, I looked forward to learning more about how environmental changes affect gene expression, which would in turn influence a human’s behavior. Since coming to Duke, I surprisingly became more and more interested in understanding behavior in non-human species, along with the various ways we can study it. Therefore, Melissa’s project appeals to my interest in behavior and genes, and I’m excited to see how her project culminates at the end of the program!

We Love Sea Urchins

This week, everyone presented their chalk talks about their projects. It was fun and interesting to see what everyone was working on! I liked that a lot of the same areas of interest and methods (e.g. epigenetics, use of gRNA, mice etc.) could be seen across projects. But even then, there were still key differences among projects and I could clearly see the contributions they would make to science.

With that being said, I was particularly interested in Michael’s chalk talk titled “To Transfate or not to Transfate: Recognition of Skeletal Cell Presence”, specifically in sea urchins. My lab and I would always walk past the sea urchin lab and think about how cool it would be to work with them, so I was excited when Michael started to present.

Michael’s project looks at PMCs (primary mesenchyme cells) in sea urchins, which become the skeleton of  the sea urchin later in development. However, a previous study found that the skeleton still forms without PMCs because of NSMs (non-skeletogenic mesoderm). This led Michael’s project to focus on how the sea urchin embryo knows that the PMCs are missing and what signals are given after this is determined. He will determine the PMC genes, clone these genes through PCR, and look at them using in situ hybridization (ISH). He will also determine the signaling molecules through ISH as well as using drug treatments. Michael’s project is important to science because his project results are important for just general knowledge about the skeletal system, and could potentially be used to help skeletal diseases in humans in the future.

Expo® Manifesto

Can we really call the presentations last week a chalk talk? I would say it’s more like a Expo® manifesto.

 

All (bad) jokes aside, I think the talks given were amazing. Everyone explained their projects extremely well, and I was surprised by the variety of biology topics presented. There wasn’t one project I wasn’t amazed by, and I am excited to in a cohort of such amazing individuals. One of the common themes was the world of epigenetic. As my project deals with these markers as well, I think a project that stood out to me was Michael’s “Differential Methylation of Shank 1 and DLG4 in THC exposed F0 -> F1 mice.” His project focuses on the transgenerational effects of THC on the methylation of the the Shank 1 and DLG4 genes.

THC is found in marijuana, and the doses that he uses on the mice are relevant to the doses that an average user of marijuana consumed. The two genes he is focusing on are important in the post synapsis, so this project can help us to see if THC can cause harm on the gametes that are exposed to THC. He studies male mice gamete, because it is extremely painstaking to see the effects on the female gametes. He will expose mice to chemically relevant doses of THC and measure the effects on the methylation of the gametes by using pyrosequencing and rt-PCR. With the movements to legalize marijuana in the US, it’s important to assess the risks of the THC in it.

Everyone gave an amazing chalk talk

This week, I had the opportunity to listen to everyone give a chalk talk about their project.  It was nice to understand details about their project,  such as Alzheimer’s, and DMD instead of thinking, okay he works in an x,y,z lab.  Additionally, I enjoyed how everyone was able to present such complex topics in a short amount of time in a way that was easy to follow.

As someone who attended the NC Zoo school, I had the opportunity to go into the zoo on a daily basis.  If time allowed me to, I enjoyed going to the African side of the NC Zoo to watch the chimpanzees and baboons.  When Christine presented the idea that both the highest ranking (alpha) and lowest ranking baboon experiences the same amount of stress, but it might be due to a different kind of stress I was all for it.  Over the summer she is planning to answer the question, “Do alpha baboons and low ranking baboons experience different stressors?”.  She is planning to look at energetic stressors, so she will analyze a thyroid hormone(1), T3, in a wide range of baboon fecal samples. Another thing that caught my attention about the presentation, was the fact that there was a significant decrease in stress from the alpha to the second rank baboon (beta).  Finally, it was great to learn the behind the scenes of this research project.  She talked about how they have an area in Kenya and scientists in Kenya watching the baboon’s behavior (looking for changes in rank), collecting the samples, and shipping them back to Duke for analysis.

This is an amazing project and I can not wait to see the outcome of this project and other projects on July 27, during the poster presentation.

Next week, I will provide a glimpse into my daily routine (A day in the life).

1-thyroid hormones allow you to indirectly measure energetic stress because it is based on changes in metabolism.

Alphas and Betas and Low-Ranked, Oh My!

I would like to start this post off by saying that all of the chalk talks given this past week were absolutely amazing! They all dived into complex topics like genetics and neurobiology in a simple way that made understanding them much easier. Seeing the research drawn as a visual tool made the talks more dynamic and really simple to follow. As someone with extremely limited biology experience, I really enjoyed that I could follow along and learn so much.

As someone who loves animals and grew up watching documentaries on their behavior, I found Christine’s talk extremely interesting. I knew that there were dominance hierarchies in baboons, but I had no idea there were distinctions between the types of stress that a baboon experienced depending on where was in the hierarchy. It was really interesting to learn about the distinction between energetic stress (more dominant in Alphas) and psychosocial stress (more dominant in low rankings males). I think the most fascinating thing for me was that stress levels could actually be quantified through measuring glucocorticoids in the feces. (Who knew poop could be so useful?) It is also really fascinating to think that you can tell what a dominant source of stress for a group of baboons is by measuring certain hormone levels and not just observing their interactions in the wild; I had no idea the level of T3 secretion indicated a certain type of stress (energetic) experienced. Additionally, it surprised me that just by moving down one rank– from an Alpha to a Beta– a baboon’s stress levels would decrease significantly! Overall, the talk gave a really interesting insight into the stressors of baboons in hierarchies from a hormonal standpoint. Christine did a great job in explaining her project in a clear, fascinating way. 

All the chalk talks this past week allowed me to dive deeper into fields with which I was unfamiliar. I really enjoyed learning more about each of the projects everyone else was working on and seeing common threads that linked them together. So far, BSURF has taught me so much and the talks really enriched my learning in a fun, simple, and visual way. Thanks everyone!

Social Hierarchy’s Affects on Male Baboon Stress Levels

The entirety of this past week’s BSURF meetings were dedicated to giving Chalk Talks, giving everyone in BSURF an opportunity to learn about and engage with each other’s research. My eyes were opened to the range of topics being investigated. Previously I had some concept of what my fellow BSURFers were studying using blanket labels like “neurobiology” or “plant biology” and what I had learned from their blog posts. But with time having passed and projects having progressed and evolved, Chalk Talks gave a new opportunity to understand the various research projects in a deeper way and have lingering questions answered. As someone who is very much a visual learner, the emphasize on drawings and schematics in these presentations really helped to develop my understanding.

A project I found really interesting was Christine’s. Her project was a little different from the other research being conducted in BSURF. Her research focuses on studying baboons and hierarchal stress amongst males. Her main question was, “Do alpha and low-ranking baboons experience different sources of stress?” I found the infrastructure involved in this research and the previous findings to be really interesting. Previous research has found that the alpha males experience high levels of stress due to energetic costs, while the beta males experience significantly lower levels of stress. Interestingly, as you move down the social hierarchy, relative levels of stress increase due to increased psychosocial stresses. I found this distinction between stress sources as they relate to social hierarchy surprising and fascinating.

Additionally, I was intrigued by the large scale, international logistics involved in this research operation. The population of baboons being studied is Kenya and monitored by a team of people that analyze social interactions to determine social hierarchy and collect fecal samples for analysis. These samples are sent to Duke and the relative stress levels of the baboons is determined by analyzing cortisol levels. The amount of coordination and cooperation between those in Durham and Kenya is truly remarkable.

I think Christine did a great job in her presentation. I learned a lot about an area of research I know very little about and I’m excited to learn about her findings  and their possible implications.

Life in the Perfect Lab

My work in the Perfect Lab varies from day to day. The day may start with me immediately working because at this point I know what I’m expected to do. Nevertheless, on days where I have no idea what I’m supposed to do, I sit and talk with my mentor, Jenny, about the next experiments I need to complete to finish my construct*. Most days consist of me performing PCR’s on particular segments of gDNA to make the constructs for biolistic transformation. However, every week I learn something new in order to move further in the project. Jenny primarly introduces different techniques and approches to solving problems in experiments. Besides performing PCR’s and gel electrophoresis, I have learned to cut bands out of gels, extract DNA from gels, innoculate microbes, isolate DNA from cells and a plethora of other tips and procedures. I’m usually doing 2-3 of these things in a single day.

In addition to this, I have also learned from my mistakes and through those mistakes learned more about myself. For example, I have noticed that around 4 or 5pm, I am more prone to making simple mistakes compared to the beginning of the work day. Nowadays, I avoid performing complicated procedures at that time of the day and simply prepare to do it the next day. In doing this, it forces me to be more congnicent of time and prioritize my responsibilities for the day.

Fortunately, I have been able to produce sufficient yeilds with my PCR’s and DNA extractions to be able to transform the C. neoformans fungi. The next couple of weeks will be a little more stressful because my mentor will be leaving and I have to prepare/start mice experiments.

*construct: DNA that is to be put into an organism’s genome

Big Is Small, and Small Is Big

Thank you to everyone for sharing their projects this week! It felt like looking through a kaleidoscope: so many different colors, angles, combinations in biology and in people’s attempts to understand life. One thing I learned from all of your talks is to remember to zoom in and out on this kaleidoscope.

By zooming in, big questions come down to acute focuses. Hsp70 is one of the many things that Bio201 hammered into my head this past spring. Nevertheless, I was still fascinated when Tamanna shared in her talk that the Hsp70 family can facilitate parasite survival by mitigating the effects of temperature change as the parasite enters from cold-blooded mosquito into the warm-blooded human host. Many of the malaria studies that I have read before relate to genetic modification in mosquitoes or massacre of mosquitoes, and Tamanna’s chalk talk directs me to look at malaria from a new perspective. Her zooming in from the big question that Plasmodium falciparum results in falciparum malaria, to the moment this parasite getting transmitted from mosquito to humans through a bite, to finally the C-terminal LID domain of PfHsp70-1 binding with PI3P turned this giant ball of problem into a sharp focus. With her clear delivery, it really became clear to me why her lab is doing what it’s doing (wow that’s some cool stuff) and how a big mystery can be teased apart bit by bit. By zooming back out and looking at the broader applications, one can also see clearly that the small dots on the ITC curves can link together something of great significance.

This applies to every single chalk talk that I had the great pleasure listening to this week. As we become more and more familiar with our lab schedules and more and more at ease carrying out each step after rounds and rounds of repetition, I believe it is important to keep thinking why we’re doing what we’re doing and how everything comes together (eventually). Big can be small, and small can be big.

Hot topic and cool project: Sweta looks into epistability of plants


This past week, I listened to multiple people talk about their amazing projects. From using CRISPR/Cas9 to alleviate symptoms of muscular dystrophy to stress between male baboons, I have thoroughly enjoyed hearing about the research my peers are doing.

The one project that really peaked my interest was Sweta’s project titled “Epistability: Transgenerational and Environmental Effects on Plant Development.”

Sweta is looking at early plant development in the model organism Arabidopsis (which she called Dobby) and how the epigenetic changes can be inherited (which is very much a Lamarckian idea). Sweta is looking at the effects temperature has on growth, knowing that Dobby grows better at 22°C compared to 10°C. She has two aims: looking at the stability of the transgenerational effects and looking at the stability of the genotype. She is mainly working on her first aim the summer in which she is looking at 3 generations of plants treated at both hot and cold temperatures (see figure 1). When I asked her what she expects from her data, she said that she expects the parental environment to have the most effect on the offspring compared to the grandparents.

I found this topic extremely interesting because it seems very much like Lamarckian evolution rather than Darwinian evolution. I entirely believe that the environment has a strong effect that can be passed on from parent to offspring, especially after doing a field work experiment with a graduate student in the Donohue lab (the lab Sweta works in) on niche construction as a method to cope with drought. The idea that a phenotype in addition to a genotype can be passed down is very exciting considering we all express genes differently, although we have many of the same genes.

I final thing I was very impressed by was Sweta’s use of the board and presentation because she clearly knew what she was talking about and was able to demonstrate it well using only a section of the already limited space.

Figure 1