Art and science are often considered opposites, but Beatriz Praena Garcia sees overlap.
“I think in this job you need to be very artistic,” Praena said. “I have a basic methodology to do the essays … then I read a little bit online. You can search in another paper and do some research to see how you can apply it to your work. You can be more creative, so it’s not always the same.”
The postdoctoral researcher studies influenza vaccines in the Henry Wan lab, tackling them from three angles. She works to improve vaccine effectiveness by growing it in different types of cell lines and eggs. She also works to improve mouse model systems for her lab and studies the influenza virus receptors.
Beatriz Praena Garcia looks at epithelial cells through a microscope in the Henry Wan lab. Since Praena studies respiratory illnesses, she analyzes epithelial cells from lung lining. Photo by Cara Penquite | Bond LSC
Praena worked on antivirals for herpes before coming to Mizzou, and the switch to vaccines was a welcome change for her.
“I always wanted to study vaccines. . . there are a lot of antivirals already in the war against viruses. I want to give something [new] to the community,” Praena said.
Praena started in the lab right out of high school in a two-year technical training program where she worked in a biology lab.
“In high school, I was not a very good student, so when I finished high school I didn’t go to the university directly,” Praena said.
Once exposed to the lab, Praena knew that was where she wanted to be.
“I realized this is good for me. My score was very high in the class, and I said, ‘Oh, I will try to go to the university,’” Praena said.
Beatriz Praena Garcia shows boxes storing viruses and vaccines. They are stored at minus 80 degrees Celsius for long-term preservation. Photo by Cara Penquite | Bond LSC
Growing up in Spain, Garcia attended Autonomous University of Madrid for undergrad where she started research on herpes antivirals and its receptors. She stuck with that research for the next eight years, working in the same lab for her masters and Ph.D. studies.
Praena came to MU at the height of the pandemic in 2020. She had to gain a special visa, which she also worked on in the Wan lab during the pandemic. After accepting the position at MU, Praena remembers consulting a large map on her wall to find the landlocked state of Missouri.
“At the beginning it was complicated because when I accepted the position, I didn’t know where Columbia, Missouri was,” Praena said.
For Praena, finding passion for her work is vital to her success.
“You need to be a hard-worker, and you need to have a lot of resilience, because in academia you will never be rich, and you have to work a lot,” Praena said. “So, the first thing you need is to love your job [and] the science.”
Beatriz Praena Garcia shows a tube containing a flu vaccine stored in a freezer in the lab. It took Praena six months to produce the vaccine. Photo by Cara Penquite | Bond LSC
Her hard work translates to outside of the lab where she competes in triathlons and bikes trails in Columbia.
“I like to take my bike to the trails. I like the Katy Trail, the MKT Trail and I went to the Ozarks,” Praena said.
Praena and her husband also camp and explore different states. Over winter break, Praena took advantage of having two weeks off and traveled to Arkansas, Texas and Oklahoma.
Praena enjoys Columbia and traveling in the U.S., but she hopes to one day return to Spain and have her own lab. While in the Wan lab she works to improve her research skills and develops project ideas.
“You always have to ask ‘why’ and ‘how,’” Praena said.
Ajay Gupta learned biology basics as a first year undergraduate on the bumpy bus ride from his small hometown to Punjab Agricultural University. Just a few hours’ ride, he made the most of his time before he returned home to help his family’s agricultural goods business.
Working extra hours in the margins of his time has become a habit for Gupta. Now a plant science first year Ph.D. student in the Bing Yang lab and Department of Plant Science and Technology Millikan Endowment recipient, Gupta starts his day around 9 a.m. and works on three plant science projects until night.
“My advisor has to sometimes push me out of the lab to go home,” Gupta said. “I have that fascination with science.”
Gupta grows bacteria that carry recombinant DNA harboring CRISPR-Cas9 system. This DNA will be inserted into rice plant cells and the rice plant cells will duplicate with modified DNA. Photo by Cara Penquite | Bond LSC
For his Ph.D. thesis project, he studies how rice plants fight off bacterial leaf blight — a disease caused by the bacterium Xanthomonas oryzae. This disease can devastate rice fields in Asia and other areas that depend on the staple grain. His research focuses on genes that help plants resist the deadly disease.
“The second project, which is very close to my heart, … is the development of CRISPR genome editing toolkit,” Gupta said.
Gupta works to improve the gene-editing tool CRISPR-Cas9 in rice plants. Researchers currently modify chunks of DNA sequence with CRISPR, but Gupta’s goal is to modify the individual building blocks of DNA known as nucleotides. He simplifies the process to make gene editing easier for researchers with less CRISPR experience.
Gupta shows three stages of growing genetically modified plants. The plants start as cells called calluses, infected by agrobacteria that inserts the modified DNA into the cell. The cell duplicates the modified DNA as it multiplies into small groups of cells that are all genetically modified. Photo by Cara Penquite | Bond LSC
His third project started with self-taught skills rooted in a side passion. Gupta contributes to a global bioinformatics project investigating bacterial resistance genes in rice. By understanding the genetic make-up of plant resistance to disease, the researchers may find innovative solutions to prevent crop infections. Gupta’s input involves assembling the genome of a type of rice that is resistant to the pathogen that causes bacterial blight in rice.
“It’s a global project where everyone across the globe who uses that variety [of rice] can take advantage of that genome,” Gupta said.
Most of Gupta’s time is spent in the lab now, but Gupta started studying biology by accident. As an undergraduate he decided to major in agricultural biotechnology thinking there would be more math involved.
The genetically modified plants grow roots as the cluster of cells forms stalks of rice. Photo by Cara Penquite | Bond LSC
“It mentioned that it will have some math and some biology, and I knew nothing about biology,” Gupta said. “I said, ‘OK, it has math, I should go for it.’”
For the first year Gupta had six biology classes and one math class each semester.
“I said, ‘Oh my god, how am I going to survive,” Gupta said.
Gupta worked through basic biology courses in subjects like genetics, botany and zoology alongside his math courses. Then the math courses dropped off for the following years leaving Gupta’s schedule packed with biology classes.
“And then I started liking it,” Gupta said. “It seems more practical to me than math. I think if I would have gone to math I would have been in engineering, but I’m happy that I chose this path, and right now I’m working with plants which I love.”
To verify that the plants contain the modified DNA, Gupta mixes the plants’ roots with chemicals. The roots turn blue in plants with modified DNA but remain their natural color if they do not contain modified DNA. Photo by Cara Penquite | Bond LSC
Gupta’s small high school in his agricultural hometown left out many biology basics, but without the finances to attend a better school in another city Gupta was left to catch up at the university level while also helping his older brother at home.
“It was very challenging for the first year. Our financial situation was not great, so I had to support myself for my studies,” Gupta said.
Gupta returned every day to work as an accountant at his family’s small agricultural goods store to earn money for his education and to help.
“My father passed away when I was 15, and me and my brother who was 18 at that time, took care of our business without having much experience. So, I had to ride the bus four hours every day to go there and help my brother during my first year in bachelor because we could not afford at that time to hire somebody” Gupta said.
Back on their feet by his second year, Gupta continued working on weekends throughout the rest of his studies, and he only separated from the business when he left for his masters’ studies halfway around the world at South Dakota State University.
Gupta initially stayed with friends in South Dakota to soften the transition to the United States. Although difficult to live an ocean away from family, Gupta maintains relationships with his family.
“I talk with my mom almost every day. That’s a kind of an Indian or Asian thing. We are very close to our parents,” Gupta said.
Gupta started CRISPR research while working on his master’s degree at South Dakota State University. He focused on modifying wheat genes to increase yield and diversity, but on the side he also found a passion for bioinformatics.
“Most of this was self-learning, so I just found things on the internet and found out how they work and then made my own protocols and now have the pipeline,” Gupta said.
After this taste of research, Gupta was hooked. He applied to continue his studies as a Ph.D. student and asked Bing Yang, a collaborator with the lab’s wheat genetics research, for a letter of recommendation.
Instead, Yang asked Gupta to join his lab.
Gupta accepted his offer to continue working on CRISPR alongside expanding to more basic research — a change of pace from his previous work in applied agriculture.
“I want to develop a mindset where I can work on [basic science] well where there is little or no information and then I can think on my own,” Gupta said.
Gupta is the primary bioinformatics researcher in his lab and helps with data analysis for several projects alongside his bioinformatics project. While he spends most of his time hard at work on a research bench — and can often be seen walking across the Bond LSC atrium to the greenhouses late into the evening and over holidays — he also enjoys talking with lab mates.
In the Bond LSC rooftop greenhouses, Gupta grows the genetically modified rice plants at controlled conditions for optimum growth. Photo by Cara Penquite | Bond LSC
“Sometimes, post [research] talks, we sit together and then I discuss with them if they have some problem I can try to help, and I do the same as well. If I’m having some problem I talk to them,” Gupta said.
Gupta plans to stay in academia and continue research and teaching others. He enjoys mentoring other students and also works on outreach to high school students.
“I love to do those things and talk to students, and maybe in some ways help this university as well,” Gupta said.
Gupta holds on to the inquisitive spirit that drove him to spend hours studying basic biology on the bus despite now learning in advanced research facilities.
“I love to talk with people [to] find out what they’re doing and if I can learn something out of it,” Gupta said. “Because a lot of the time our knowledge is limited and when talking to people you can get something new.”
Cynthia Tang is an M.D.-Ph.D. student in the Wan lab. Photo by Cara Penquite | Bond LSC
Cynthia Tang’s academic career is marked by her propensity to multitask. From earning a major and three minors during her undergrad to making a documentary while getting lab and clinical experience, she makes the most of her time.
Recently Tang received the Excellence in Public Health Award from the United States Public Health Service, and a $181.734 National Institutes of Health grant to be used over four years . . . all while getting an M.D. and Ph.D. simultaneously.
The funding from the grant goes towards Tang’s research on SARS-CoV-2, the virus responsible for the COVID-19 pandemic, and its effect in rural areas. By comparing DNA sequences of the virus from patients in rural and urban areas, Tang looks for differences in variants. Her goal is to understand how the virus has been evolving and why patients in rural areas experience more severe symptoms.
“I’ve always found infectious diseases really interesting,” Tang said. “This pandemic came up and it was exciting to be in the position to help contribute to the knowledge base of it.”
Cynthia Tang creates protein structure models for the spike protein on the Sars-CoV-2 virus. Along with 3D models, Tang creates phylogenetic trees to compare the genetic makeup of variants of the virus. Photo by Cara Penquite | Bond LSC
In a family of immigrants, Tang saw the challenges in health literacy and cultural differences affecting patient care in the U.S first-hand, and she also caught a glimpse into a global view of healthcare.
“I had an opportunity to travel to Vietnam and actually see some of the health disparities of different health care structures in different countries,” Tang said. “We were visiting my grandfather who was in the hospital, and it was alarming to see the contrast between the quality of health care and availability of health care there compared to some of the hospitals that we have here.”
Tang recalls noticing lack of supplies, physicians and space for patients. She saw similar issues in the U.S., and shined light on public health disparities here by creating a documentary about the challenges immigrants face in the U.S. healthcare system. She also held competitions across Washington University’s campus to teach students more about public health issues while working as a clinical research coordinator.
“I wanted to help improve health equity,” Tang said.
A bachelor’s in chemistry and minors in philosophy, international development and pre-health professions at the College of Idaho may start to explain Tang’s public health focus and her path to bench and clinical research.
“My minor in international development was really useful for me because that gave me a lot more exposure to the different economies of different countries of the world, their health status and how politics, economics and health care all interrelate in different countries,” she said.
Her research at Washington University led Tang to an interest in its clinical applications.
“I really enjoyed the aspect of working with patients, actually getting to talk to patients and hearing their stories,” Tang said. “I felt like with clinical research, you can see … slightly quicker results from bench to bedside compared to bench research, and that was also when I decided I was interested in becoming a physician.”
Part of an eight-year joint M.D.-Ph.D. program at Mizzou, Tang started with two years of pre-clinical medical school before transitioning to Ph.D. research and then will return to finish the clinical years of medical training.
When she came to Mizzou, Tang continued fighting health disparities for immigrants.
“We organized a group of volunteers that were able to translate COVID-19 information to different non-English speaking communities in Boone County,” Tang said.
Always one to tackle several projects at once, Tang plans to pursue a career as a pediatric physician scientist interfacing with patients while continuing lab research.
“I like the idea of doing both. As a physician, you get that one-on-one contact and you get to make a very direct contribution to someone’s life,” Tang said. “And then with research, you don’t have as much contact with patients, but what you do can affect populations.”
When Samantha Yanders stepped to the front of Monsanto Auditorium, she followed presentations from two researchers with three degrees each.
Yanders only had three years of undergraduate research experience.
Nevertheless, she pinned the microphone to her tie, ran her fingers through her short curly hair, and explained her research with a calm certainty to her voice.
Having just finished her junior year as a plant science undergraduate, Yanders spent the first week of her summer sharing her passion for plants with fellow researchers during the 2022 Interdisciplinary Plant Group Symposium.
“I want to be inclusive in how I talk about my work to be able to educate people,” Yanders said. “Even if you’re talking to molecular biologists, they may have no idea about extracellular ATP.”
A clear communicator and advanced undergraduate researcher, Yanders was selected to present her research in Monsanto Auditorium during the symposium and often helps write and edit manuscripts for her lab.
Yanders began research in the Gary Stacey lab through Freshman Research in Plant Science, a program that places plant science freshmen in MU labs. Yanders’ research focuses on the signaling role of extracellular ATP when a plant experiences high-salt conditions.
Within a cell, ATP is a molecule used for energy. However, research shows that when a plant is in high stress or damaging conditions, ATP outside the cell signals for protective mechanisms.
“It’s really clever because instead of spending energy to make both a signaling molecule and a signal receptor, it already has something that’s really high concentrations in the cell and very low concentrations outside the cell,” Yanders said. “So it’s easy for the plant. It just has to make a signal receptor.”
Under normal conditions there is less ATP outside the cell and a high concentration of ATP inside the cell since it is used as energy. However, under high salt conditions there is an increase in ATP outside the cell, which binds to receptors on the cell surface that communicate with the cell to stunt plant growth. Yanders’ work explores the impact of high salt environments for plants — an increasingly relevant project as climate change raises sea levels and increases salt deposits in soil.
Although she now enthusiastically recounts her work to auditoriums of fellow scientists, biology was not originally Yanders’ first choice.
“In high school, I knew that I wanted to do something scientific because I’ve always liked science,” Yanders said, “But I didn’t think I wanted to go into biology, because I saw biology as mainly oriented towards animals.”
Sam Yanders shows the roots of arabidopsis plants. Yanders studies extracellular ATP in arabidopsis and tobacco plants. Photo by Cara Penquite | Bond LSC
With her geneticist grandfather piquing her interest from a young age and her enthusiasm for environmental science in high school, once she learned Mizzou had a plant science program she was ready to commit.
“So I was like, ‘Okay, I’ll go into biology and focus on plants.’ And then I’m looking on the Mizzou website and they [have] plant science. I didn’t even know that was a thing,” Yanders said. “So I switched to plant science.”
After enjoying high school chemistry labs, Yanders was ready to take the next step into research labs as soon as possible. Freshman Research in Plant Science allowed Yanders the opportunity to join a lab right away — fostering her interest in plant research.
“As humans, we move around. We can adapt to change by moving and changing what we do,” Yanders said. “But a plant has to do all of that management from a stationary position. . . It can’t change the circumstances that it’s in, so it has to adjust itself to be able to adapt.”
Tobacco plants grow under artificial light at Bond LSC. Photo by Cara Penquite | Bond LSC
Yanders’ love of plants extends beyond the walls of the lab, and she spends her free time in her herb garden where she learns more about plant and human interactions.
“The way that you interact with the plant fundamentally changes how the plant acts,” Yanders said. “So like with basil, if you just let a basil plant grow it just gets leggy and crazy, but if you do what you feel like is harming it by pinching the tops off, then it grows more compact and bushy, which is good for the plant.”
Yanders hopes to explore plant and human interactions in the future, potentially pursuing urban agriculture and ecology.
“I’m really interested in soil health, I feel like we’ve done a lot to deplete soils, and coming up with more renewable ways of doing things [and] producing things for human consumption,” Yanders said.
While uncertain which questions she will ask next, Yanders hopes to continue answering questions and explaining her research to others.
“[I like] being able to offer an explanation, and not just see it, but to create that explanation for other people,” Yanders said.
If the world can be taxing on a person as pressure mounts, just think about how stress must feel to plants.
Humans can add a layer of clothing when cold or get a glass of water when thirsty, but plants do not share this simple luxury and must endure whatever environment they sprout in.
As climate change, pollutants, and extreme weather patterns escalate, this poses a serious global threat to plants and our food supply.
Ron Mittler, a principal researcher at the Bond Life Sciences at the University of Missouri, recently looked at how this piling up of multiple stressors at once can significantly decrease plant survival.
“The principle is that you can have a combination of several different stressors, each by itself has no effect on the plant but when they come together, they’re causing severe effects,” Mittler said.
Studying plant response to stress isn’t a new thing, but Mittler’s focus on the compounded effects of stressors may give us a better idea of the threshold of stress plants can endure in our changing climate. Multifactorial stressors combine three or more stress factors simultaneously impacting plants. Four categories of stress— biotic, climate, anthropogenic, and soil threats — all become worse as climate change and environmental pollution progress, subsequently decreasing plant quality of life.
Biotic threats relate to enemies like pathogenic bacteria and insects. Similarly, soil threats are determined by poor nutrient soil and salinity. Climate threats include extreme temperatures and drought. Anthropogenic threats are man-made as humans use harmful pesticides and create microplastics.
Arabidopsis thaliana seedlings, a model plant used in experiments, were placed side by side on a plate and received a combination of stress conditions such as heat, salt, excess light, acidity, heavy metal, and oxidative stress. Researchers studied the growth, survival, and molecular responses of the seeds. Seedlings were grown on plates rather than in soil to isolate and study the impact of multifactorial stress.
Seedlings grew on separate plates and experienced different individual or combined stresses. Results showed that each individual stressor applied to the seedling had a minimal effect on the plants but with the increase in the complexity and number of stress factors affecting the plants, survival, root growth, and chlorophyll content declined. Similar results were also found for seedlings grown in soil.
Ecosystems are already seeing these impacts in Florida and Germany. Multifactorial stress of heat and pollution prompted algae blooms to grow exponentially. This toxic overgrowth led to thousands of manatees dying. Entire forests in Germany are experiencing massive storms followed by long periods of drought, insect attacks, and fires.
Mittler said things may not look dire now, but we will eventually reach a point of no return where plants die off in mass quantities or even go extinct.
“The harmful effects of stress on the nation can serve as a dire warning for society. We may not see the effects now but 10, 20, 40 years down the road we will be having severe problems with our food chain,” Mittler said.
Since there are multiple factors at stake, predicting negative impacts on agriculture and ecosystems is tricky as researchers are unsure how this domino effect may unravel. What they do know for certain is that there will be severe consequences and we are already seeing them today.
These sporadic weather extremes are weakening the plants, making them more vulnerable to insect predators and other stressors.
The research yielded alarming results, and Mittler highlights that this is a dire problem that people need to take seriously. Once people understand the severity of this problem, he hopes individuals and policymakers will take action before the consequences become irreversible.
The Mittler lab is working on several fronts to address this problem and is trying to find a solution to it. They are currently studying multifactorial stress combination in different crop species, such as soybean, rice, and tomato. In addition, also identifying key plant regulators that are activated during multifactorial stress combination. These will be used in future breeding efforts to increase the tolerance of different crops to multiple stresses.
This study is now an international collaboration between the Mittler and Zandalinas laboratories, as Dr. Sara Zandalinas took on a faculty position in Spain.
Investigators at Bond LSC take steps to apply basic research
By Cara Penquite | Bond LSC
Photo by Lauren Hines | Bond LSC
Scribbling in a lab notebook and planning experiments tucked between shelves of equipment, it’s easy to fixate on day-to-day lab operations. But scientists also face the challenge of finding how research can improve the world around us.
“The direction, the vision of the lab, ultimately comes from the principal investigator that bridges the research into applied directions,” said Jay Thelen, biochemistry professor and Bond LSC principal investigator
Despite the focus on basic research within the Bond LSC, many principal investigators choose to take their research to the next level with commercial partnerships.
Thelen’s lab researches ways to increase oil production in seeds and has three patents licensed to Yield 10 Bioscience, a sustainable crop innovation company who applies Thelen’s research to commercial crops. While seed oils like canola and soybean oil are known for their use in cooking, Thelen explains that increased production of these oils could play a larger role in sustainable fuel sources such as biodiesel and sustainable aviation fuel.
“We have to make more oil to balance out our need to eat it [and] our need to wean ourselves off of fossil fuels,” Thelen said. “To do that, we need to either plant more acres of oil seeds, or we have to raise the oil in existing oil seeds.”
Thelen researches enzymes with that potential application in mind. One is acetyl-CoA carboxylase, the enzyme which initializes the production of fatty acid chains found in plant oils.
“We’ve known this is an important enzyme, and we know that any tinkering you do with it has an impact on the oil production,” Thelen said. “In this case we’ve made new discoveries that permitted us to rationally engineer this enzyme to make it more active.”
Thelen suggests thinking of the enzyme as a “gatekeeper” to oil production which initializes the production of fatty acids and increases oil production. Thelen’s lab identified two different gene families that influence the activity of the enzyme in Arabidopsis and camelina plants. Yield 10 then applies these discoveries in other commercial plants.
While Thelen works closely with his commercial partner — having served on their scientific advisory board for three years and now stays in contact with Yield 10’s CEO to develop research projects — some labs stick with short-term arrangements.
Kamlendra Singh — assistant director of the Molecular Interactions Core at Bond LSC and Veterinary Pathobiology research assistant professor — studies HIV treatments. His lab identified a compound licensed by a commercial partner that targets the shell containing the virus’ genetic information.
Singh’s work in HIV started in 1994 with basic research investigating the enzyme that makes the viral DNA.
“I wasn’t into [studying] the drugs when I started working on HIV, I was mostly trying to understand how HIV enzymes works,” Singh said. “Once you know how the enzyme works, then you can target these enzymes for discovering the drugs.”
After years of studying how the enzyme works, Singh switched to HIV treatment. The first step to develop a treatment is to look for structures in the virus that the drug could potentially target to stop the viral replication. Singh targeted the shell around the virus’ genetic information known as the HIV capsid.
Building on previous research, Singh’s lab developed a compound able to bind the HIV capsid and prevent it from releasing the contained genetic information. Even with the licensing of his compound, Singh plans to continue researching ways to improve it.
“There are two reasons to keep working on it. One, well it’s my brainchild,” Singh said. “The second reason is as the company grows, we grow. We get more recognition and more funding. You can use it to [study] different viruses or use the same funding to improve upon it.”
While Singh plans to remain looking towards the applied side of his HIV research, he does not forget his roots in basic research.
“You have to put in time … [to] understand the system first, which is basic science, before you go to applied science,” Singh said.
Michael Roberts, a Chancellor’s Professor Emeritus of animal sciences and biochemistry who has had several patented projects, focuses on improving basic science projects and applies for patents if warranted.
“I don’t deliberately go into anything for commercial purposes,” Roberts said. “If I see something that I think does have commercial application, I’m happy to do it, but that is usually after you do [basic sciences].”
Whether starting a project with applications in mind or focusing on basic research, knowledge gained through research can be building blocks for the future.
“Science is simple. Even the most applied research project has its genesis in basic biology and basic research,” Thelen said.
One step into the Advanced Light Microscopy Core (ALMC) sounds an automated bell prompting Alexander Jurkevich, the core’s assistant director, to step out of his corner office into the open square room. With a friendly smile, Jurkevich coordinates biologists across MU’s campus to reveal the wonders of the microscopic world.
“Our mission is to provide researchers campus-wide with advanced microscopy instrumentation,” Jurkevich said. “We not only provide access to instrumentation, but we also train, advise users and support them during their early research at the core.”
The core hosts an annual image contest celebrates MU researchers’ microscopic imaging throughout the year. After being canceled for the past two years, this year contestants submitted their best images for consideration.
Christie Herd, a postdoctoral fellow in the Alexander Franz Veterinary Pathobiology lab won the Best of Show Award for her image of the La Crosse Virus in mosquito ovaries.
Image by Christie HerdDavid Porciani, assistant research professor under the supervision of Bond LSC’s Donald Burke, won Director’s Award for Best Technically Challenging Image with his image showing epidermal growth factor receptors in two types of resolution.
Image by David Porciani.
Janlo Robil, a graduate student in the Bond LSC’s Paula McSteen lab won Experts’ Choice Award with his image of hormones in a corn leaf.
Image by Janlo RobilA wonderful surprise
Herd was “blown away” when she got her colorful image that won the Best of Show Award. Not only had she tried imaging other viruses with less success, she also worked on different La Crosse samples with no luck.
“That image in particular, I was not expecting to see because the day before I was there for three hours and had to quit,” Herd said. “So, when I saw that image, I was surprised because I did not think I would see that much detail.”
Christie Herd is a postdoctoral fellow in Alexander Franz Veterinary Pathobiology lab. Herd won the Best of Show award for the 2022 ALMC Imaging Contest. Photo by Cara Penquite | Bond LSC
Herd’s image shows the La Crosse virus in developing eggs within mosquito ovaries. Herd uses La Crosse, a type of bunyavirus, as a model to study virus transmission from a female mosquito to her larvae to determine how viruses can remain transmitted within generations of mosquito populations. While La Crosse infects a small amount of humans a year, it transmits quickly, making it the perfect model to learn more about other bunyaviruses like Zika and Chikungunya.
“With bunyaviruses, they’re so multifaceted, and I like being able to research different aspects of them,” Herd said. “I just feel like they’re medically important.”
Herd dissects mosquitoes before imaging. She studies the transmission of bunyaviruses like the LaCrosse and Zika viruses from female mosquitoes to their progeny. Photo by Cara Penquite | Bond LSC
Herd’s image includes different colors to label different parts of the ovaries as well as the virus so she can see where in the ovaries the virus is traveling. Using a technique that allows her to see different focal planes, Herd can see where the virus is in every dimension, but it can be tricky to get high quality images.
“It does require a few hours of playing around, looking at the microscope,” Herd said with a chuckle. “It requires an investment of time, and sometimes there are days where it’s just not what you wanted and it doesn’t work … and you wasted days.”
Even with the challenges, the ability to see multiple dimensions of a sample at once is valuable for research projects like Herd’s.
“You just have to persevere and try again with a new set of samples,” Herd said.
From side project to passion
Although a deviation from his primary research, David Porciani’s project to image cell surface receptors slowly took over his focus.
“The biggest surprise was that I really had fun,” Porciani said. “This was not my primary project, but it became my primary project for a while.”
David Porciani is an assistant research professor under the supervision of Donald Burke. Porciani studies cell surface receptor interactions linked to lung cancer and his image won Director’s Award for Best Technically Challenging Image. Photo by Cara Penquite | Bond LSC
Porciani, an assistant research professor under the supervision of Donald Burke, studies molecules on the surface of cancer cells which are receptors for growth factors. These receptors act as a lock, with growth factors as a key. When the growth factors and receptors come together, the cells divide and create more cells. In lung cancer, there are more of these receptors, which can lead to uncontrolled tumor growth.
“This receptor, EGFR, has been widely studied,” Porciani said. “But for me there is definitely an interest because it’s one of the markers in lung cancer.”
Porciani tags those receptors with small molecules called fluorophores that glow under the light of the microscope, so he can see where the receptors are and how they move. However, the fluorophores cannot attach to the receptors alone, so he used aptamers — synthetic keys created by researchers that can bind receptor locks with specificity similar to the natural growth factors. Ultimately, the aptamers clip the fluorophores to the receptors.
“If you can follow the motion of the receptors, these receptors are kind of dancing,” Porciani said.
In his image, each dot is a different receptor made visible by the attached fluorophore.
However, fluorophores can become bleached, rendering them invisible after being exposed to the laser from the microscope for a while. If the synthetic keys, or aptamers, are still bound to those receptors, they cannot be imaged any longer. To address this, Porciani developed an aptamer which attaches to the receptor for a shorter time and then detaches so that even if the fluorophore bleaches, another aptamer can replace it and so receptors can be imaged for longer time.
“We engineered an aptamer with lower affinity that could work with this approach,” Porciani said. “By having lower affinity aptamers we can still determine localization of a high number of receptors and their motion.”
Porciani shows the process of imaging cells using super-resolution techniques. When a laser hits the cells with the fluorophore specific wavelength, the fluorophores glow while the rest of the cell remains dark, and Porciani can see where the fluorophores, and consequently the tagged receptors are in the cells. Photos by Cara Penquite | Bond LSC
For his winning image, Porciani split the image to show the difference between single molecule resolution on the bottom right and a lower resolution image on the top left.
“With [lower] resolution, you don’t have a single molecule solution. If there are two molecules close together you will see them as just one single dot,” Porciani said. “But with the image, after software analysis with the image on the bottom right, then you have single molecule resolution.”
With this technique — made possible with microscopes at ALMC in Bond LSC — Porciani saw his efforts come together.
“At the beginning, I was just focused on the aptamer engineering from a high affinity aptamer to low affinity aptamer, and making them was the fun part to play with the structure,” Porciani said. “But when we started doing the imaging experiments at the Bond Life Sciences Center I realized that it was not just fun, but it was actually meaningful and this approach could have lots of biomedical applications.”
The artist’s touch
Although passionate about biology and microscopy, Janlo Robil decided to submit his image based on aesthetics.
“I chose this one because I am also a graphic artist, and I appreciate the color and composition,” Robil said.
Janlo Robil is a Ph.D. Candidate in the Paula McSteen lab. Robil studies the hormones involved in corn leaf development. Photo by Cara Penquite | Bond LSC
Robil’s image shows a developing corn leaf with different colors labeling different plant hormone response proteins involved in stimulating growth. His unique image of an entire leaf, which is just under a millimeter in length, required piecing together images of sections of the leaf.
“About 0.75 millimeter, that’s still big in the microscope,” Robil said. “This is kind of difficult to make because it means that you need to tile several images [together] and sometimes it takes up to an hour just to [get] an image.”
The experiment requires planning ahead since the plants containing the fluorescent proteins must be crossed with plants with genetic mutations to determine the roles of the hormones in the leaf development.
To tag the plant proteins with fluorescent proteins requires planning ahead since the plants must be grown with genetic mutations.
“The beautiful image is actually a result of the expression of fluorescent proteins that are tagging the hormone response protein and also the hormone transport protein,” Robil said.
Robil looks at images from a confocal microscope. Using a laser, confocal microscopy brings clarity to Robil’s images. Photo by Cara Penquite | Bond LSC
Initially from the Philippines where the agricultural staple is rice, Robil came to Mizzou interested in genetic mechanisms to make rice plants more productive. One way to enhance the rice plants is to make the rice leaves more similar to corn leaves, so Robil found interest in the McSteen lab’s project understanding the role of hormones in corn leaves.
“This project is perfect for me because I am studying the leaf and also integrating genetics,” Robil said. “And I love microscopy so much. I had quite a good amount of training starting in 2021 on confocal microscopy, and that’s why I was able to image this.”
Mengran Yang sat perched on a stool too tall for the cart of lush green tobacco plants in front of her. Behind towering shelves of lab equipment, she hunched over the plants and steadily pricked each leaf with a syringe.
Yang works with Arabidopsis and tobacco plants to learn about plant immune systems as a postdoctoral fellow in the Gary Stacey lab. Her research focuses on signals plant cells send to coordinate a fight against pathogens.
“I think it’s very interesting to see how plants can defend against the pathogens,” Yang said.
Mengran Yang infiltrates the leaves of tobacco plants with a syringe. While her work mostly focuses on Arabidopsis plants, Yang also uses tobacco plants as a tool in her research since it is convenient to express the correct target proteins in their leaves. Photo by Cara Penquite | Bond LSC
Just like humans have immune cells that fight germs, plants have a network of cells and signals to deal with harmful microbes. This system is particularly important for plants since they are rooted in place and cannot avoid the attackers in their environment.
The plant immune system protects it both inside and outside its cells. Proteins inside the cell can recognize molecules from invaders and trigger an immune response, and the second — the part Yang’s research focuses on — uses receptors on the surface of the cells to recognize molecular patterns from microbes to signal for the plant to get rid of the pathogen.
“Our lab is focused on the extracellular ATP signaling pathway,” Yang said. “ATP, as maybe everyone knows, is an energy source, but we found it can also be a signal when it is released to the extracellular matrix, where it is referred to as extracellular ATP.”
Yang’s work with the lab started when she moved from China to Columbia for her postdoctoral research at Mizzou. Although originally drawn to the lab because of Stacey’s prominence in the field of plant immunity and signaling, Yang stays because of the community within the lab.
“Our lab is very international — some people are from China, Korea, Brazil, Vietnam and India,” Yang said. “A lot of the same-aged girls will go out for fun.”
This community stems from Stacey’s emphasis on the team’s collective success.
“You try to develop an esprit de corps in the lab where people care about each other’s success,” Stacey said. “They’re taking care of themselves, but at the same time, they care about other people’s success.”
Becoming a good researcher started with strong academics, and Yang remembers being the top of her class in biology, physics and math in middle school and high school. When she started her studies at the university level she followed her growing interest in biology.
Growing up with her twin sister in China, Yang recalls societal pressures to fill any spare time with more study. Her parents thought her teachers gave too much homework and pushed back on this expectation.
“My father thinks this kind of education is not good, he thinks you need to promote your efficiency and not just work hard,” Yang said. “You need to study smart, not just hard, and I think my dad influenced us a lot.”
Yang applies this mentality to her research and is supported by Stacey who encourages her to plan experiments and life goals.
“While I was a Ph.D. student, I spent the most time in the lab for fun,” Yang said. “But now I’m good at scheduling stuff, so I schedule my experiments at least a week ahead.”
Mengran Yang shows a dish with Arabidopsis plants. Yang’s work primarily focuses on studying extracellular ATP in Arabidopsis plants. Photo by Cara Penquite | Bond LSC
Stacey works with Yang to ensure her future impact as a researcher. When picking a project, Stacey guides his researchers towards projects with lots of potential.
“That’s the kind of discussion I have with people, what’s the best project for them where they have the chance to make an impact,” Stacey said.
With her lab mates and other research labs, Yang collaborates to strengthen each other’s weaknesses. If she does not have experience with an experiment, Yang asks other postdoc students with more experience for help with her project. When she publishes her research, she makes sure to give credit to the other students who helped her.
“In Chinese, ‘shuangying’ means ‘good to both sides,” Yang said. “That’s very common. You will read a paper, and you can also see most [papers] have a lot of authors there. That’s the cooperation.”
After finishing her research in the U.S., Yang plans to return to China to continue doing what she loves — research.
“I want to have my own lab in China,” Yang said. “It’s difficult, I know, but it’s my dream.”
Exploring new places and diving into the world of the unknown can be intimidating. At a young age, Leah Lepore was immersed in this world and grew to love it.
“The first time I left the country was to travel to Japan and it was an incredible culture shock,” said the current Chris Lorson lab member. “From my twelve-year-old perspective, I was learning about a different way of life but also drove my parents nuts because I would only eat McDonald’s while we were there.”
An avid traveler, Lepore loves immersing herself in new cultures and seeing the world from a different point of view. This has blended into other areas of life, especially when it comes to her passion for science.
“I think traveling is what formed this kind of wanderlust that I have, and that exploring unknown places gets you out of your day-to-day routine and puts things into perspective,” she said. “There’s so much in the world than what you see around you.”
Lepore is a sophomore majoring in biochemistry. She wasn’t new to working in a lab when she joined Bond LSC in summer of 2021. She worked at two other labs in high school, so she found it imperative to join a lab that would challenge and prepare her for the future. As a member of the Lorson lab and an employee of Shift Pharmaceuticals — Lorson’s business off-shoot of his research — she found that.
“It’s really interesting to see the industry science and the academic science be complimentary,” Lepore said.
At Shift Pharmaceutical, she works on Charcot-Marie-Tooth disease (CMT). CMT causes abnormalities in the nerves feet, legs, hands, and arms. This disease was unfamiliar to her before she joined, but that did not hinder her enthusiasm to learn more.
“What’s really interesting is that it doesn’t necessarily kill you, but it diminishes the quality of life because it is caused by this overexpression of protein,” she said. “It causes your limbs to contract but not able to relax while also impacting your balance and dexterity.”
Lepore uses gene silencing therapy with synthetic nucleotides that bind to mRNA in this research. mRNA codes for proteins and since there is an abundance of certain proteins in patients with CMT, she is trying to silence portions of mRNA that code for them.
Lepore aims to spread her desire to be involved in research as an ambassador for the Office of Undergraduate Research, and she encourages students to get involved as soon as possible. Many students are apprehensive to dive headfirst into working at a lab, but Lepore reassures that there is a place for everyone at Bond LSC.
“I find that everyone that I’ve met here is extremely intelligent, but also great at making anyone who hasn’t worked in a lab before feel extremely comfortable,” she said. “It’s a very welcoming and inviting atmosphere.”
When not in the lab or helping undergraduates, she focuses on fitness. At Orange Theory, Lepore lifts weights and runs on the treadmill, which helps her cope with stress and anxiety. No matter how hectic life might get, her positive attitude and ambitious personality shine through.
“Sometimes you have to realize that you’re not going to know everything even if you want to, but there are always going to be people that are willing to explain it to you,” she said. “However, not knowing everything is okay sometimes because it gives you room to learn and engage these critical skills that are not only going to help you in the lab but also in your day-to-day life.”
Hari Krishnan holds a handful of A. pavonina seeds. Known for their bright color, the seeds are known among many Asian and African communities as coming from the red bead tree. Photo by Cara Penquite | Bond LSC
By Cara Penquite | Bond LSC
An energetic and fulfilling day starts with a spread of healthy meals, and many people rely on nutrition labels to meet their daily quota of vitamins and nutrients. But how did scientists measure the Vitamin C in an orange or the protein content in peanuts for the label?
Finding out what is in food we eat starts with scientists like Hari Krishnan, a USDA-ARS molecular biologist and MU adjunct professor of plant science and technology. Krishnan calculated the type and degree of nutrients packed into seeds from red bead trees with the help of the Advanced Light Microscopy Core in Bond Life Sciences Center.
“We are highly dependent on the food that we eat,” Krishnan said. “Anything we can do in order to either improve nutrition or quality or finding alternative sources of food is very useful.”
Commonly known for its pods of seeds as bright as red M&M’s, the red bead tree is known as Adenanthera pavonina in research communities.Krishnan took an interest in A. pavonina seeds — turning away from his usual soybean research — to see if they could be a potential alternative protein source in developing countries. He quickly found a different, unusual characteristic.
“I’ve never seen a seed have such a high content of trypsin inhibitor,” Krishnan said.
Our bodies make trypsin to break down protein in our intestines, but trypsin inhibitors block the enzyme from its job of protein digestion. That makes it hard for us to benefit from nutritious proteins in the seeds. Although many Asian and African communities already incorporate A. Pavonina seeds in their diet, its unusual amount of trypsin inhibitors limits nutritional benefits.
“Soybeans have probably less than five percent of the entire seed protein made of trypsin,” Krishnan said. “This particular legume is 20 to 25 percent, which is fairly very high.”
Krishnan turned to Alexander Jurkevich, associate director of MU’s Advanced Light Microscopy Core, to find out where in the plant’s cells the trypsin inhibitors were located. Jurkevich and Krishnan used glowing molecules to mark the trypsin inhibitors and see where they are stored in the cells.
Hari Krishnan sits in his lab at MU Curtis Hall. A USDA-ARS molecular biologist and MU Adjunct Professor of plant science and technology, Krishnan’s research looks at the nutritional value of legumes. Photo by Cara Penquite | Bond LSC
Tagging protein here starts with an antibody. The first antibody acts like one side of a strip of velcro that can attach to a second antibody that also carries a molecule of a fluorescent dye. Through the double antibody system, the glowing, fluorescent flag attaches to show scientists which parts of the cell have trypsin inhibitors.
While Krishnan developed the antibody, Jurkevich used his expertise in light microscopy to take images of the glowing cells.
“This cooperation is very important, because modern research technologies are very complex and a single person cannot learn and excel at all techniques available in life sciences,” Jurkevich said.
Understanding the amount and locations of trypsin inhibitors, Krishnan looked to reduce inhibitors so the seeds would be more nutritious for humans.
Roasting or boiling the seeds breaks down the trypsin inhibitors, but Krishnan warns that it may break down other essential amino acids as well. In his work with soybeans, Krishnan looks for a way to grow plants without trypsin inhibitors at all.
“We were able to find some [soybean] mutants, which have lower levels of this trypsin inhibitor,” Krishnan said.
Natural mutants with lower levels of trypsin inhibitors could be cultivated on a larger scale to produce seeds that are easier for humans to digest.
Since trypsin inhibitors are not ideal for consumption, Krishnan decided to think outside of the box. He turned to USDA-ARS research scientist and entomologist Adriano Pereira to find another use for the seeds.
Pereira tested the seed proteins as an insecticide against corn rootworm larvae and found that it stunted larval growth but did not necessarily cause mortality. While it is still early in this research, there may be some insecticidal properties from the seed.
“It’s something that has to be investigated to make sure […], but we assume that since it is a trypsin inhibitor it could be inhibiting on some level the trypsin during the growth of the larvae,” Pereira said.
As Krishnan’s first look into A. pavonina seeds, the research opened many new questions yet to be answered, but he plans to continue investigating the role of trypsin inhibitors in soybeans and other plant protein sources.
“[Research] is a never-ending investigation, and one thing leads to another. It’s always interesting,” Krishnan said. “I come to the lab everyday thinking of what I’m going to end up doing today.”
The Advanced Light Microscopy Core provides technical imaging expertise to researchers who need state-of-the-art imaging for their experiments. That includes confocal, super-resolution, digital light-sheet and widefield microscopes, image analysis and processing and sample preparation.
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