Dong Xu, Bond LSC principal investigator and Shumaker Endowed Professor in the University of Missouri’s College of Engineering. | photo by Roger Meissen, Bond LSC
A Bond Life Sciences Center researcher has been inducted into an elite organization comprised of two percent of all medical and biological engineers.
The American Institution for Medical and Biological Engineering (AIMBE) this week announced the induction of Dong Xu, a Bond LSC principal investigator and Shumaker Endowed Professor in the University of Missouri’s College of Engineering.
“Election to the AIMBE College of Fellows is among the highest professional distinctions accorded to a medical and biological engineer,” said Kamrul Islam, chair of the college’s Electrical Engineering and Computer Science department.
Xu was selected for his “distinguished contributions to bioinformatics and computational biology, and extensive services to University of Missouri and his research community.”
In addition to his endowed faculty position, Xu serves as director of the Information Technology program, whose core facility is housed in Bond LSC.
Membership to AIMBE’s College of Fellows recognizes those who have made outstanding contributions to engineering and medicine research, practice or education, and to those pioneering new and developing fields.
Because of health concerns, AIMBE’s annual meeting and induction ceremony scheduled for this spring was canceled. Under special procedures, the induction was held remotely.
The Sears Greenhouse Complex at the University of Missouri. | photo by Becca Wolf, Bond LSC
By Becca Wolf | Bond LSC
Picture this. It’s 25 degrees Fahrenheit outside and snow is falling in Columbia. The weathermen have projected 4 inches of snow in the next 24 hours. As wind whips the snow around, students hope the schools call a snow day the next day. Snow starts to accumulate as the sun sets and people all throughout town are staying inside, some eating soup with their families, others curled up with a book near a fire. Looking out their windows, they see their lawns covered in a blanket of snow.
All the plants outside are dormant or dead, but plant research does not stop when the seasons change. In fact, greenhouses make winter a highly productive season, despite Mizzou being located in the not-so-balmy Midwest. Complex systems help balance the temperature and lighting plants need to survive when it is not naturally provided.
You only have to go as far as the roof greenhouses on Bond Life Sciences Center to see an example of the total 91,250 square feet of plant growth facility space at Mizzou, part of more than 70 greenhouse rooms that are used year-round. Locations include the Sears Greenhouse Complex, the Ashland Road Complex and the new East Campus Growth Facility, and house everything from corn and soybeans to tomatoes, broccoli and model plants like Arabidopsis.
“Probably 95% of greenhouse space is research,” said Michelle Brooks, MU’s greenhouse coordinator, “And then about 5% of the space is used for teaching undergraduate plant science classes that have a hands-on lab in the greenhouse.”
To optimize growth, the heating and cooling temperature is kept within an 8-10-degree Fahrenheit range. Brooks explains, “you have to have that distance between it so the systems don’t battle each other, because there’s much more fluctuation in temperature in a greenhouse.”
These systems help keep the temperature balanced, especially once it gets cold outside. Greenhouses have to be kept as close to outside summer conditions as possible.
“You’re working to keep them warm and light enough to grow the crops year round because most of these plants, like corn and soybeans, they’re really high light plants,” said Brooks.
To maintain favorable lighting, greenhouses at MU have high intensity (HID) lights that provide both light needed for plants to grow. These lights mimic the sunlight plants would get outside during the summer. The lights have a nice side effect, said Brooks, “turning the lights on raises the temperature in the rooms by 10 degrees,” further promoting plant growth.
Another way greenhouses are heated is through hot water heaters. This system works by pumping hot water through thin pipes that are around the perimeter of the greenhouse. These pipes then radiate heat from the water into the room. The pipes allow heat to get closer to the plants than the HID lights because there is no concern of singeing or burning. MU also uses steam pipes in some greenhouses, which use the same process except with steam instead of water.
To prevent this heat from escaping, shades are used on the roof of the greenhouse. These shades act as insulation to save energy and keep heat in. Greenhouse shades are often made out of polypropylene, saran, polyethylene, and polyester and prevent direct sunlight from getting in, along with insulating the greenhouse. Shades are used year-round because in the summer, they help keep the greenhouses cool and keeps the temperature balanced, even if it is 90 degrees Fahrenheit outside.
Shades drawn over the ceiling of a greenhouse at Bond LSC. | photo by Becca Wolf, Bond LSC
Wet walls are also used in the summer to cool the greenhouses. These are comprised of cooling pads in an aluminum wall that circulates water through it. There is a fan on the opposite side of the wall that blows the cooled, evaporated water into the greenhouse, thus lowering the temperature. These are typically the length of the wall and are about 4-5 feet in height.
A wet wall in a greenhouse at Bond LSC. | photo by Becca Wolf, Bond LSC
How to Prepare for Snow
When heavy snowfall occurs, MU wants to prevent snow from accumulating because it blocks out the sun and cools the greenhouse.
Brooks explains, “we would disable the heat retention function of the shade cloth, because we want the heat to go up into the peak to melt the snow off the greenhouse and out of the gutters.” Luckily, this can be done automatically as most of the greenhouse system is computerized. “It’s pretty automated,” said Brooks, which is helpful because that means no one has to be at the greenhouse to make these adjustments. For example, if there is a lot of snowfall in the middle of the night, caretakers like Brooks do not have to wake up and drive to MU to pull the shades.
To make sure the greenhouses maintain their heat in cold temperatures, Brooks lists preventative measures taken, “you have to make sure that people are not propping doors open or turning on their exhaust fan because they’re working hard and you know, get hot for a minute, and then forget to turn it off.” Doing this saves energy and keeps the plants in a stable environment.
New MU Facility
Last fall, the new East Campus Plant Growth facility opened. Being a total of 22,880 square feet featuring 24 greenhouses and 27 growth chambers, there is a lot of space to keep warm in the winter.
Fortunately, thanks to new technology, there have not been many problems this winter there. “We had some heat valves that were wired wrong,” Brooks says. “Luckily, it was the hallway that was getting too cold, it wasn’t a room that was occupied.” These heat valve issues were caught early and have been fixed, and there have been no issues since.
The East Campus Plant Growth facility is full of advanced technology, including exhaust fans with variable speeds, instead of just ‘low,’ ‘high,’ or ‘off.’ This allows coordinators to change the speed by increments so they, “can have the fan come on slowly to where it doesn’t drop the temperature too fast because you don’t want the cooling and the heat in wintertime to battle each other.” The gradual increase and decrease in speed balance temperatures and gives scientists more control in their plant growth.
Another system that is new at the East Campus facility is the reverse osmosis (RO) water system. This system takes out minerals in water and purifies it, which plants like. Purifying the water also gives scientists more control in their plant growth because it gives plants clean water, eliminating detrimental factors. The new facility also has higher walls and ceilings, creating the potential to do research on trees and other tall plants, like biofuel grasses.
Aside from new technology, the East Campus facility has several other benefits as well.
“We have the potential now for new hires that could potentially come in and bring in research dollars,” Brooks said. MU does not have to turn away any researchers due to the lack of facilities anymore. Space at the new facility also offers the opportunity to expand in the future, and there are plans to add greenhouse ranges as the need arises.
Now as the temperature gets warmer and the days get longer, the greenhouses will have to be managed accordingly. Water pipe heating will be turned off and the wet walls will be turned on in order to combat the heat of Missouri summers.
But soon enough, it will be winter again and the process will start all over.
Beverly Agtuca graduated with her Ph.D. in Plant, Insect and Microbial Sciences in May of 2019. She is currently teaching at Adams State University in Alamosa, Colorado. | Photo by MJ Rogers, Bond LSC
By Mariah Cox | Bond LSC
At the age of eight, Beverly Agtuca held lofty aspirations for her future. While many of her classmates wanted to be a veterinarian or a doctor, she dreamed of finding a way to increase plant productivity on her relatives’ farm in the Philippines.
While many children’s career goals change — sometimes more than once — Agtuca held on to her interest in plant science, wrapping up five years of work on her dissertation with a Ph.D. in Plant, Insect and Microbial Sciences last May. Months later, the fruits of her labor made it into the American Phytopathological Society Journal in December and was chosen as the journal’s Editor’s Pick in February.
Agtuca’s research found a potential novel alternative to fertilizing grass species, such as corn — a global staple for human and livestock consumption.
A large-scale problem
Much of the agricultural industry relies on nitrogen fertilizers to support farming. However, when plants can’t absorb nutrients from nitrogen fertilizer quickly enough, soil bacteria convert it to nitrate. It is then flushed out of soils in runoff, polluting groundwater, streams, estuaries and oceans.
“Nitrogen fertilizers are being used worldwide to increase plant productivity. Unfortunately, nitrogen fertilizers have a lot of environmental impacts like pollution and use a lot of energy to make it,” Agtuca said.
Nitrogen runoff from farm fields causes harmful algal blooms — such as red tides, blue-green algae and cyanobacteria — and can have severe impacts on human health, aquatic ecosystems and the economy. In 2018, a red tide event off the coast of Florida killed at least a hundred manatees, a dozen dolphins, thousands of fish and 300 sea turtles to wash ashore in masses. A giant dead zone where the Mississippi River meets the Gulf of Mexico is also a side effect of these algae blooms removing oxygen from the water and rendering it uninhabitable by animals.
While it may seem like we’re only experiencing the effects of nitrogen fertilizer now, it has been around for over a century. More than 175 years ago, scientists in Europe were looking for ways to feed their growing nations and were on the cusp of discovering nitrogen’s role in plant growth. In 1909, German scientist Franz Haber developed a high-temperature, energy-intensive process to synthesize plant-available nitrate from the air, thus creating nitrogen fertilizer.
For over a century, nitrogen fertilizers have helped feed global populations. But its effects are becoming harder to ignore. To keep up with the constant demand for food while considering nitrogen fertilizer impacts, scientists are looking for alternative means to support global agriculture.
Finding an alternative
Recognizing the global impacts of traditional fertilizers, Agtuca set out to learn more about bacteria’s role in promoting plant growth in grass species.
The endophytic bacteria, Hebaspirillum seropedicae, was known to provide hormones to grass species, but not much else was understood about its interaction in plants. To see the interaction between her model species of grass, Setaria, and the bacteria, Agtuca’s collaborator, Sylwia Stopka built a laser ablation electrospray ionization mass spectrometry machine that performed in situ metabolic analysis.
Beverly Agtuca and Sylwia Stopka pose next to the laser ablation electrospray ionization mass spectrometry machine. | Photo contributed by Beverly Agtuca
The LAESI-MS setup for Setaria analysis. The top right image shows the long-distance working microscope to visualize the root samples. A motorized xy-stage was used to move the sample easily. Two computers were used – 1) to visualize the samples and 2) to move the sample during analyses. Below the long-distance working microscope is the LAESI-MS including the laser, electrospray ionization source, sample holder, and mass spectrometer. | Photo contributed by Beverly Agtuca
“We found a lot of metabolic pathways that were significant promoters of plant health,” Agtuca said. “Those metabolic pathways were similar to a known nitrogen-fixing bacterium called rhizobium in soybeans, which people previously thought wasn’t similar. And then we found out that it’s more complex than other types of bacteria.”
Agtuca and her colleagues found purine, zeatin and riboflavin pathways in the plant-bacteria interaction — chemical pathways that support plant growth.
“The purine pathway helps to give nitrogen to the plant, whereas zeatin and riboflavin help for plant growth,” Agtuca said. “The bacteria that was found to be beneficial to soybeans formed external nodules on the plant, but this new bacterium does not create nodules. The bacteria live inside the roots intercellularly and only benefit grass species like corn. Corn is an important crop for farming and for our productivity to feed the world.”
While Agtuca’s research won’t be debuting in farm fields anytime soon, it laid the groundwork for other scientists to continue researching new natural fertilizers in hopes of someday replacing chemical fertilizers. The next step is finding a particular molecule to add to the plant, rather than adding bacteria.
After graduation, Agtuca moved to Colorado to teach at Adams State University, a small undergraduate institution in the San Luis Valley. She’s spent her time teaching classes such as microbiology, cell biology, bioinformatics and cell physiology. Although she enjoys teaching, she misses being in the lab.
“It’s great to teach the overall broad concepts to the students and then I can relate it to research, which is fascinating to the students. I try to talk about why we’re learning specific material and why It’s important to our everyday lives,” Agtuca said. “Since I’m freshly graduated, I can express how research works, despite not having research here on campus.”
Beverly Agtuca reflects on her collaboration with Sylwia Stopka:
Sylwia and I shared the same “Ph.D. experience,” where we were both graduate students and we struggled when experiments don’t work. Together, we both planned how to make the experiment work. We supported and helped each other out. For instance, I was the plant biologist, while Sylwia was the chemist and we both have the same goal in this project. I thank Sylwia because without her we won’t get our findings rapidly and obtained numerous publications including this one.
Some scientists go into research for basic science, such as finding an enzyme and figuring out its functions and properties. Others like Hsin-Yeh Hsieh, gravitate toward applied sciences where they use what they know to develop technology.
“Because I can use everything I learned in science as building blocks to create a technology that actually can do something and make impacts in real life, that’s what I like about science,” said Hsieh, a research scientist in the George Stewart lab at Bond LSC.
Hsieh began her science journey in Taiwan, where she went to school for several years, studying molecular and microbiology as well as medical technology. Her love of science began in high school, she explained, “I was very curious about what happens in human health.”
After completing her Bachelor of Science at Kaohsiung Medical University in Taiwan, Hsieh worked in the Cancer Research group of Institute of Biomedical Sciences in Acdemia Sinica in Taiwan, before she came to the U.S. for her graduate study in the Pathology and Anatomical Department of MU Medical School. For her Ph.D. study, she had the opportunity to work on a research in blood banking and transfusion, which focused on enzymatically converting type A and type B blood into universal blood type O. This is where she really found her niche, “I am in the field I enjoy,” she said. However, her biggest challenge for the blood type conversion is how to remove the enzyme from the converted red blood cells before transfusion.
After her doctorate, Hsieh joined David Setzer’s lab in Biological Sciences of MU and later, George Stewart offered her a postdoctoral position in his Molecular microbiology lab.
Collaborating with a multidisciplinary team on MU campus including faculty in Veterinary Pathobiology and in the School of Natural Resources, and students in biochemistry and in bioengineering, Hsieh’s team came up with a design for a cartridge that can immobilize enzyme for blood type conversion, and filter enzyme out of the converted red blood cells (RBCs).
“Let’s say, after the blood goes through this cartridge, it gets converted. The converted RBCs flow through the cartridge, but the enzyme is locked behind so it won’t go along with the product,” she explained.
Hsieh and her team actually filed a patent on this process in December 2019 and are currently seeking more funding to continue testing the process in other industrial applications, such as biofuel and special chemical production.
“I can see its potential,” Hsieh said. “We think it can work in biofuel and it can work in pollutant degradation, but we need to prove it with scientific evidence.”
Said Hsieh, “The technology could turn cellulose to sugar, into glucose, that becomes an energy source,”
It can be expanded beyond these applications as well, with potential to help clean the environment.
She explains, “We actually can break down the herbicide pollutants, like atrazine, in the environment using this enzymatic process.”
It took four years to create this cartridge, overcoming several challenges along the way. That did not intimidate Hsieh.
“I like to see if my idea is right on the track or way off, so that’s quite challenging but also fun,” said Hsieh with a chuckle. “You should always explore something new and challenge yourself.”
Day to day, Hsieh is busy working with undergrads in the lab. Their willingness to learn and try anything intrigues Hsieh’s curiosity and expands her horizons in research.
“The students are so open minded. I say, ‘well, let’s take a new approach and do it,’ and they just go ahead and do it. If we get a good result that proves the concept, that’s great. If not, we learn how to troubleshoot and make it work. In the process, students are trained for critical thinking and research strategy” Hsieh explained. “I think that’s a win-win situation, for them to advance in the career path and also for me to move the project forward.”
Expanding her horizons is important not only in the lab, but as well as in her community. When talking about the friends she has made here in Columbia, Hsieh says, “I met most of my friends while playing tennis, and at a point in time, I counted there were people from 46 countries among us because students come to this campus from all over the world.”
Creating a community with different cultures keeps the life interesting and helps her learn of the world, while staying in Columbia all these years. With unfinished tasks in hands, Hsieh is not planning on leaving anytime soon. Describing Columbia, she “enjoys the atmosphere.”
Anand Chandrasekhar is a Bond LSC principal investigator studying the mechanisms involved in the development of the nervous system. He is also a biological sciences professor. | Photo by Justin Stewart, Bond LSC
It’s an asset to be able to visualize and think about the nervous system from the perspective of an electrical engineer.
Cell biologist Anand Chandrasekhar — whose work focuses on the movement of neurons within the brainstem of mice and zebrafish, as well as on the consequences of that movement or lack of movement for the animal’s behavior— brings that angle to his work all the way from his undergraduate degree in electrical engineering he received in his native country of India. Although much of the nitty gritty details of his engineering knowledge have been lost to time, the legacy of those concepts has stayed with him in his neurological research at MU’s Bond Life Sciences Center.
“I’ve always been interested in seeing how things work and how processes are connected, and I think that’s because of my engineering background,” he said. “The nervous system was a natural thing to study for me because I’ve always been curious as to how circuits work within an organism’s brain and how those circuits allow or disallow certain biological processes to happen.”
That circuit-related curiosity led to Chandrasekhar’s two-decades-long investigation of the movement of neurons within the brain.
“Neurons are not just in a spot in the brain from the beginning,” he said. “They are told to move there at some point in the embryonic stage. If they do not move to the proper spot within the brain, the consequences can be quite severe.”
He uses mice and zebrafish for his experiments because they are both vertebrates, meaning they have a lot in common with the human brain in structure and function. Previous scientific research shows when neurons don’t move properly within a human brain the consequences are devastating, resulting in a spectrum of cognitive and motor disabilities. In severe cases, the individual does not survive to birth. So, the consequences to the animal of neurons that don’t move to their rightful place during development can be quite dire.
Two projects related to neuron movement take up the bulk of the Chandrasekhar lab’s time, with graduate students Emilia Asante and Devynn Hummel diligently studying each. Asante’s project focuses on zebrafish, and looks specifically at the consequences for the fish when neurons in the brainstem do not migrate properly. “We have some understanding what the actual consequences of defective neurons are in humans, but it’s not so clear with fish,” she said.
Emilia Asante is pursuing a Ph.D. in biological sciences and works in the lab of Anand Chandrasekhar at Bond LSC. | photo by MJ Rogers
A snail toxin, MVIIA, is illuminated in a zebrafish embryo. The toxin blocks calcium channels needed for neurotransmitter release. Chandrasekhar’s lab studies motor neurons using zebrafish. | Photo by Devynn Hummel
Zebrafish embryos and larvae are transparent, and Asante takes advantage of this fact with her work. For one particular series of tests, Asante feeds the larval fish tiny fluorescent particles so that she can tell how much food the fish eat when she puts the live fish under a microscope. Compared to wild type (normal) fish, the mutant fish (those with defective neuron movement) ate less food than their normal counterparts.
Armed with this conclusion, Asante took her work a step further, and she asked why the mutant fish didn’t eat as much. So, she designed a test to look at the jaw movement of the zebrafish to investigate whether that had something to do with their reduced food intake. Asante devised a method such that fish could move their head but were not able to swim away. This allowed her to record the jaw movement of the fish. Asante found that the fish who did not eat as much as others were, by and large, the same fish whose jaw movement was less frequent. At the same time, Asante observed that the fish whose jaw movement was reduced were normal in every other way, including their ability to swim and capture food. Thus, Asante identified slowed jaw movement as the definite effect of defective neuron movement.
Now, Asante is investigating what upstream or downstream issues could cause this reduced jaw movement.
“In science, processes often don’t work in isolation,” she said. “So, there could be any number of things that are causing the fish to have slower jaw movement. They may not be getting a signal from their brain that tells them they’re hungry. Even if they do get that signal, they may not be able to move it downstream and tell their jaw to move so that they can chew on, swallow and eventually digest as much food as they need.”
Hummel is taking his investigations in a different direction. He works with mice and is currently studying how a set of genes functioning within the brainstem interact with each other in order to allow for proper neuron migration. Hummel’s work is focused on how cells move in general, in contrast to Asante’s. However, Hummel’s work still uses the neurons that control the jaw as a model cell type. As is the case with zebrafish, neurons that don’t migrate properly can cause severe behavioral defects in mice.
Several years ago, the lab discovered a gene that controls the direction in which neurons migrate, as compared to other genes required for the ability to migrate.
“This is exciting, because such a gene had not been previously identified,” he said.
Unlike the zebrafish that Asante works with, Hummel must dissect pregnant female mice in order to determine the extent of neuron migration within a developing embryo. This means tracking pregnant females over several days prior to ever beginning an experiment. Managing a colony of over 200 mice is no easy task, yet Hummel stays motivated since his work can yield such meaningful results.
“One of the coolest things about doing this kind of work is that sometimes, no one in the world has ever investigated specifically what you are looking at,” he said. “By that criteria, once you make a discovery, you know more about that subject than anyone else in the world.”
Chandrasekhar agrees that seeking answers to these big questions is what makes his work so fulfilling.
“Neuron movement within the brain is similar across the board – in humans, mice and zebrafish,” he said. “If we can understand how these cells move — as our work with mice tries to figure out — or why they move the way they do — as our work with zebrafish tries to figure out — in simple systems, the hope is that we can then transfer that knowledge to improve our understanding of the human brain.”
Protein is important in balancing iron and reactive oxygen in plant and cancer cells
Arabidopsis growing in Ron Mittler’s lab. | photo by Becca Wolf, Bond LSC
By Becca Wolf | Bond LSC
You might tend to think durability is more of an issue in building a car or engineering a building, but environmental stress makes resilience vital for plants, too.
In the world of understanding and engineering more durable crops, scientists recently identified a protein that’s key to some of that resilience in a neat way. Actually, it’s literally a NEET protein.
Ron Mittler, a Bond Life Sciences Center principal investigator and professor of plant sciences and surgery, is probing the use of the NEET protein to balance iron and reactive oxygen species (ROS) in the chloroplast to make plants stronger and healthier, and it plays a similar role in cancer cells. These proteins belong to a unique family of iron-sulfur (Fe-S) proteins ideally maintaining the proper balance of the heavy metal in and outside of plant cells. Along with several other scientists, Mittler published his most recent plant findings in The Plant Journal last October.
Since NEET proteins regulate both iron and ROS, Mittler teamed up with heavy metal expert David Mendoza, fellow Bond LSC primary investigator and associate professor of plant sciences. Through a Bond LSC Grant for Innovative Collaborative Research, the two started work at the intersection of two labs.
“The main philosophy of Bond LSC is to establish collaborations between researchers to advance the field of whatever sciences that you do,” Mendoza said.
Mittler and Mendoza looked at how NEET impacts iron and ROS, working together to figure out how to make plants more durable. Doing so, they have made a lot of progress in the field and believe that their collaboration has been a good one. Says Mendoza, “It is a very productive collaboration.”
Trays of Arabidopsis in Ron Mittler’s lab. | photo by Becca Wolf, Bond LSC
A Balancing Act
But the work doesn’t stop with plants. Mittler found NEET proteins also play a similar role in cancer cells, regulating the level of iron and potentially toxic ROS compounds. Humans and plants need iron and ROS, but too much of either can be destructive. NEET proteins allow cells to grow and spread quickly which is beneficial for plants, but detrimental for humans when cancers grow out of control. In humans, NEET proteins also play a significant role metabolism related to diabetes and progression of neurodegenerative diseases.
Despite needing both iron and ROS to function, there can be too much of a good thing. To keep iron levels balanced, oxygen is needed. While oxygen on its own is a fairly stable molecule, adding electrons or exciting the present electrons can make the molecule want to react with things around it creating oxidants. Electrons are the negatively charged particles of an atom that keep balance within the cell. When balanced, iron and ROS helps cells grow, but when placed together, the iron turns ROS into hydroxyl radical, an ion that reacts with anything and destroys everything. If not controlled, this desire to bond with things nearby can damage cells and lead to cancer.
Cancer cells like ROS and iron because it enables them to spread quickly and take over more of the body. To stay balanced, NEET proteins are needed.
“These proteins apparently enable the cells to keep high levels of both iron and ROS and to survive,” says Mittler, “So it looks like cancer needs these proteins because if you suppress these proteins or if you target them with drugs, the cancer cells die.”
Sara Zandalinas holding a dying Arabidopsis seedling and a living seedling in Ron Mittler’s lab. | photo by Becca Wolf, Bond LSC
Poisoning the cell
Knowing NEET proteins make cancer and plant cells stronger, Mittler wanted to look at “what happens if they’re not functioning well,” in hopes to understand their function. To test this, Mittler and his collaborators found a way to make the NEET proteins not function. Mittler called this “poisoning the cell.”
“If there is no protein, there’s no life,” he states. “We were able to control when we introduce this protein, and being able to introduce this protein enabled us to actually find out what happened to the cell if this protein goes bad.”
It was found that if the function of this protein is disrupted, the chloroplast is destroyed. Mittler described it as “completely blown to pieces.” The destruction of the chloroplast is caused by too much iron due to a broken NEET protein. Similarly in cancer cells, if there are no NEET proteins, the cells do not spread as quickly and die.
Mittler elaborated, “What we found is that we can target those proteins in cancer, so then we basically destroy the cancer cells.” Currently, NEET proteins are used as markers in the cancer field. Doctors and scientists look for them in order to help diagnose patients and to figure out life expectancy rates. But in the future, doctors hope to use targets these proteins to kill off cancer cells, a treatment that would be less toxic than chemotherapy.
In plants, Mittler has concluded that the NEET protein would make them grow bigger and faster, leading plants to be “more tolerant to stresses because it has the ability to modulate the levels of iron and reactive oxygen.” Findings like these may one day help plants deal with stress like drought and temperature extremes. With humans continuing to contaminate the environment and climate change escalating conditions, Mittler hopes his findings on iron, ROS, and NEET proteins will make plants able to withstand changes better.
“If I can improve the way plants balance reactive oxygen and iron, I can make plants tolerate much more of these bad combinations of stresses even, and maybe even make bigger, stronger, healthier plants.”
Read more about this research in the October 2019 edition of The Plant Journal article “Expression of a dominant‐negative AtNEET‐H89C protein disrupts iron–sulfur metabolism and iron homeostasis in Arabidopsis.”
The Bond LSC Grant for Innovative Collaborative Research program aims to foster inter-laboratory collaboration to make pilot projects that have potential for later federal funding.
Kamal Singh was in the town of Allahabad in his native India, preparing for competitive exams to become a government official. As he craned his head to the left, he saw a highly respected official getting berated by an arrogant and disrespectful political leader. While Singh always knew that, in these positions, one was under the government’s thumb to an extent, that incident was what sealed the deal for him and made him realize he did not want to spend his life simply taking orders and being subject to verbal abuse by corrupt politicians.
It was at that moment — roughly 30 years ago — he decided he wanted to dedicate his life to something more personally fulfilling: research.
Singh was raised in a small village in rural India. While his primary and high school education came from a military school, he obtained a Master’s degree in physics from Agra University, in Agra, the city where Taj Mahal is located. Still, Singh thought he wanted to work as a public official until that incident served as the last straw. After that, he decided to complete his graduate degree in quantum physics, and committed fully to a life in academia.
That led him to cross the Atlantic in 1994 when he moved to the U.S. and joined Rutgers New Jersey Medical School. He realized that now was his chance to focus on a different area of science that he had always been fascinated with: biochemistry.
“I taught myself everything from scratch in New Jersey,” he said. “I had always wanted to work with DNA polymerase, and I felt like now was my chance to really get involved with that.”
Over the span of 15 years in New Jersey, he occupied a number of positions including postdoctoral fellow, instructor and assistant professor. His work transitioned from research related to DNA polymerase to his current focus on Human Immunodeficiency Virus (HIV) and other viruses including Foot-and-Mouth Disease Virus, SARS and MERS coronaviruses.
That’s where Singh’s story came to Mizzou where he arrived in Jan. 2009. While he has always been highly motivated and driven by collaborations, the bread and butter of his work for more than a decade has come at the Bond LSC focused on HIV.
The goal of Singh’s HIV-focused research, in a nutshell, is to find drugs that bind to the active site of the enzymes that replicate HIV, thereby blocking the spread of the disease. According to Kyle James Hill, a postdoctoral student working in Dr. Singh’s lab, the function of his research is twofold: to better inform human understanding of the viral life cycle and to help identify novel targets of drug identification to potential treatments.
Kyle James Hill works on a bacterial transformation in Dr. Singh’s lab.
More precisely, Singh’s work is focused on enzymes/proteins that serve particular functions for the virus after the virus enters human body. Most enzymes have a suffix of -ase, and what comes before the suffix generally states the function of a given enzyme. For example, protease has to do with proteins, specifically the breakdown of proteins into smaller proteins or other subunits. Enzymes can have a positive function such as promoting growth and generation of muscle tissue, or a negative function, such as what happens when HIV highjacks their function for its own benefits. The HIV infection, in turn, weakens a person’s immune system and lowers his/her her ability to fight disease, in this case AIDS. Each enzyme has an active site, or a region where its specific function is carried out. All research that deals with enzymes, therefore, must center on what goes on at the active site.
Singh’s HIV-focused research is centered specifically on three HIV enzymes: reverse transcriptase, integrase and protease. Reverse transcriptase catalyzes reverse transcription: that is, the formation of DNA from an RNA genome of HIV. Reverse transcriptase converts RNA genome of HIV into DNA.
HIV then uses another enzyme called integrase to insert viral DNA into the genetic framework of the host cell. This allows the virus to further embed itself in what was once a healthy cell. A third enzyme called protease breaks down long proteins into smaller functional proteins, and these smaller proteins then combine with the genetic material of HIV to infect another cell.
The practical application of Singh’s work is that it is focused on moving research toward tangible results that physicians, in turn, can use to better diagnose and treatment plans for patients. Singh’s current lab work focuses most of all on finding drugs that are able to work to treat HIV patients from different parts of the world.
Singh became interested in investigating why certain drugs designed to treat HIV in the developed world, have less effect on the particular strains of HIV that exist in low- and middle-income countries. To help him with this process, Singh developed a collaboration with scientists at the renowned Karolinska Institutet in Stockholm, Sweden. Singh is also an Associate Faculty member at Karolinska.
While HIV infects people all over the world, there are actually many different strains of the virus, and some strains are more abundant in particular regions of the world. According to Singh, there are 10 subtypes of the virus, and more than 100 hybrid or combined types of it. Some drugs treat a particular subtype better than others. If a given drug does not work to treat a particular subtype, then it is useless in terms of treating patients that are infected with that strain of the virus. According to Singh, most drugs produced in the U.S. treat HIV subtype B, which is most prevalent in the developed world. However, patients in many other parts of the world, such as India, South Africa and Brazil are infected with subtype C of the virus. Singh says that 95% of patients in India are infected with subtype C.
“Because these patients are infected with a different strain of the virus, they, in many cases, do not respond to the type of drugs that are traditionally given to patients in the developed world,” he said. “Our job is figuring out why they don’t respond to that particular drug, and then coming up with something that can more effectively treat them.”
This practical application of his work is what’s most rewarding to Singh. His lab is able to provide concrete results that physicians then apply when making decisions about how to best treat their patients. Based on Singh’s work, physicians can better determine which type of drug would most likely effectively treat patients infected with a specific subtype of the virus, also considering the specific set of genetic variations and mutations that the patient may have. It is in this way that Singh’s work makes a very real difference in the world, and that is ultimately what keeps him so passionate about his work.
“To be able to apply what I work on in the lab and see it at work in real patients is very rewarding for me,” he said. “I am able to see the entire process unfold, starting with the very basic biochemistry all the way to direct application to the patients. Seeing that I’m making a real difference means everything to me.”
Christian Lorson poses next to a microscope in his lab. | photo by Lauren Hines, Bond LSC.
By Lauren Hines | Bond LSC
From developing a question to discovering a potential solution and putting it into practice, the journey from research to practical application is a long one. Nevertheless, each step brings that solution one step closer to reality.
Shift Pharmaceuticals — the brainchild of Bond LSC’s Chris Lorson — has taken another of those steps with a new patent issued last month and meetings with the Food and Drug Administration in October 2019.
Lorson — a Bond LSC primary investigator, professor of veterinary pathobiology and associate dean for research and graduate studies for the College of Veterinary Medicine — co-founded Shift three years ago in March 2017 after Lorson’s lab had a breakthrough in his research on the genetic disorder, Spinal Muscle Atrophy (SMA). He worked with Mizzou’s Technology Advancement Office and entrepreneur-in-residence Steve O’Connor to dive into this business endeavor.
“Mizzou does a great job of supporting entrepreneurial activity, and working with Sam Bish has been astounding,” Lorson said. “He really lowers the hurdles and helps take care of the business stuff on the MU side.”
After developing the new technology, Lorson went to Sam Bish — the senior licensing & business development associate in the Technology Advancement Office (TAO) within the Office of Research — to help protect the discovery. As a faculty member, Lorson shares the rights to his discoveries with the university since he used its resources and funds. The initial funding for this project came from Cure SMA, a non-profit patient advocacy group focused upon SMA. Through licensing and patenting the technology, Shift is allowed to commercialize it even though the university owns it.
With the university’s help, this product moves closer to the clinic and those with SMA are a step closer to healing.
SMA is a genetic disorder that develops in children missing the survival motor neuron-one gene. The disorder is the leading genetic cause of infantile death worldwide and causes muscle weakness, respiratory failure and premature death.
SMA is caused by the loss of a gene called SMN1. Humans have a “back-up” copy called SMN2. SMN2 does not make enough SMN protein to prevent disease but has been the focus of considerable interest for drug development within the SMA field. The compound that the Lorson lab developed targets SMN2, causing SMN2 to be “turned up” so it makes high levels of SMN protein, like SMN1. In pre-clinical models, a single dose can extend survival ~800%.
The compound is an antisense oligonucleotide (ASO), which is a synthetic string of nucleic acids that bind to a sequence of the SMN2 gene. Normally SMN2 produces a protein that has an important region that is missing. This compound forces SMN2 to make the “normal” full-length protein which would prevent disease development.
Highlighting the promise of personalized health care and the impact of large-scale interdisciplinary collaboration, this type of research is part of the University of Missouri System’s bold NextGen Precision Health Initiative. The NextGen Initiative unites government and industry leaders with innovators from across the system’s four research universities in pursuit of life-changing precision health advancements.
“The university is not in the business of selling products. That’s just not what the university does,” Bish said. “The goal with every university technology disclosed to TAO is to get industry to commercialize it. This is what I call the ‘Three-way win.’”
The company wins because they produce a commercial product. The license helps money go to the university, and the university shares some of what they received with the inventors.
Now, Shift is at the commercializing stage.
“Everything before 2016 was to find the lead compound,” Lorson said. “Everything after 2016 is to prove that it’s safe and efficacious.”
Shift still needs to further develop the technology prior to submitting it to the FDA and starting clinical trials.
Luckily, it’s not expected for a company to fully develop a product to the market. Big pharmaceutical companies often partner, acquire a company or license the technology after a start-up like Shift Pharmaceuticals does the legwork to show progress and potential.
Bish attributes the success to the amount of money, or non-diluting capital, Shift has raised and how Lorson created the company.
“Dr. Lorson was very smart in the way he went about this,” Bish said. “He realized he needed partners to get this company on its feet. Some people will say, ‘Well that shows weakness. You didn’t know how to do it on your own.’ To me, that shows wisdom and strength because nobody knows his science like he knows it, but there are other people that know business and what it takes to get successfully launch and develop a biotech company that he may not have.”
Even though Shift has far to go to make the new technology available to those with SMA, Lorson recognizes the work that has gotten him and his team to this point.
“Having been in the field for 20 years, it’s exciting to see it developed,” Lorson said. “Having known many families with SMA and getting to see them every year for 20 years, it’s exciting to see progress being made in addition to all the other stuff that’s being done by many other labs and companies. I think the interactions between SMA researchers and SMA families is one of the most exciting parts of my job. It permeates what I do every day, and I try to instill that into the lab every chance I get.”
This research is an example of how the NextGen Precision Health Institute at Mizzou is expanding collaboration in personalized health care and the translation of interdisciplinary research for the benefit of society. Partnering together with government and industry leaders, the NextGen Institute empowers bold cross-disciplinary innovation and life-changing precision health advancement targeting individual genetic, environmental and lifestyle factors. The work of the institute is a cornerstone of the University of Missouri System’s statewide NextGen Precision Health Initiative.
Medical bacteriologist, George Stewart has had a few stops along the way before he got to Bond LSC in 2004. Having done schooling and research at universities from the midwest to the east coast, it has been a long journey filled with many ups and downs, but a rewarding one at that.
Stewart hadn’t always wanted to be a medical bacteriologist though, “When I was going to college, I actually wanted to be a marine biologist,” he says, “And then my sophomore year in college, I took a microbiology course. And that changed that.”
This microbiology course at the University of North Texas was the first course where he was able to do hands on work in a laboratory and he found that, “culturing organisms and identifying organisms was very appealing.”
“The technological advancements are so fast in this discipline that you just have to change and there are easier ways to do things and better ways to do things now,” he states.
But these changes help Stewart delve deeper in understanding his passion for an organism best known for the white powder derived from its spores and sometimes used in terror attacks. Technically called Bacillus anthracis, the organism causes the disease anthrax. Stewart focuses on the outer spore of the bacteria which first comes in contact with the infected host and is responsible for protein production. He looks at how the spore contributes to anthrax’s ability to persist in soil and how it contributes to resistance properties.
Unfortunately for Stewart, funding for anthrax research has declined over the years, “It’s not considered a high priority organism,” he explains, however, “it still is an organism of concern for bio-threats.” To work around the lack of funding, Stewart has turned in other directions.
“Part of that has actually been the biggest success stories of recent times in my lab,” he said.
The direction? Using Bacillus thuringiensis, a bacterium similar to anthrax, to do research on. One problem Stewart approached using this bacterium was cleaning up Atrazine, a herbicide used by farmers to kill weeds. This herbicide, while helpful to farmers, can contaminate groundwater, potentially causing diseases in humans.
To find a solution, Stewart and Brian Thompson, a postdoc student in his lab, collaborated with Chung-ho Lin, a research associate professor in the MU Center for Agroforestry, “looking at could we possibly have come up with approaches using this basic technology to improve agriculture from a number of different ways to make plants grow faster, better, that sort of thing.”
They found that they could display proteins on Bacillus thuringiensis spore that would break down Atrazine. Farmers would only have to spray their fields with this organism and the soil would be clean of Atrazine.
Taking this a step further, Thompson created the company, Elemental Enzymes, to sell products like this to farmers in order to clean the environment. Located in St. Louis, Elemental Enzymes currently has 11 products in use today on more than 11% of corn acres in the United States in 2019, and that number seems to be increasing.
“It’s kind of interesting in the sense that we started off trying to answer a very basic biological question related to spore biology and involving a significant high consequence pathogen, and we ended up discovering something which has huge benefits for agriculture. So you never know what direction it’s going to take you in these things,” says Stewart.
Stewart hasn’t stopped there. In fact, he was one of the driving forces behind starting a Bachelor of Science degree program in microbiology here at MU. After a microbiology degree program here was discontinued in the early 2000s, Stewart has since found evidence that the degree would be successful here at the university and it was approved by the board of curators and the Missouri Department of Higher Education in 2018. The first students in this program began this past semester and Stewart says, “we’re getting underway.”
Despite these accomplishments, Stewart is getting ready to retire this year, explaining, “part of the reason that I stayed on this year was to make sure this program gets off the ground and help out with that.” While his career is winding down here at MU, he is still keeping busy, “I’m actually teaching two undergraduate courses in microbiology, once called fundamentals of microbiology, it’s an introductory microbiology course. And the other one is public health microbiology, which is more of a junior level type course.”
He also continues to do some work in his lab as he has two graduate students finishing up their degrees this year and recently stepped down as the Department of Veterinary Pathobiology chair in January.
What does Stewart have planned for when he retires? He is not slowing down, that is for sure. Stewart and his wife already have a vacation planned for New Zealand within the next few months, and he plans on spending quality time with his children and grandchildren.
“I won’t have accomplished all the things I wanted to,” he says, “but I think we’ll have gotten the major things done. So that’s satisfying.”
With his wide range of teas laid out along the windowsill, and his small posters still in stacks on the floor, Mel Oliver is still setting up his new office at Bond LSC after arriving in December.
It takes a while to settle into a new space like the office on the south side of Bond LSC’s third floor, especially after spending 30 years in a job like the one he held with the US Department of Agriculture until he retired late last year. On most days, he types away at his computer writing his papers and sits back to think whilst playing with his wedding ring, but even with 40 years of discovery under his belt, Oliver isn’t quite done with his search for answers.
Oliver grew up just north of London — which you can hear in his lingering accent — and received his undergraduate degree from the University of East London. After he earned his master’s and Ph.D. in Calgary Alberta, Canada, he came to St. Louis to work with a top plant molecular biologist. He met his wife of 34 years in St. Louis and after living in New Mexico and Texas, he returned to Missouri.
Oliver was a graduate student in Calgary when he found his calling in plant sciences.
“I was interested in all sorts of different things in plants, and the Ph.D. supervisor I started working for put a piece of [dried] moss in my hand and squirted water on it, and it [came back to life].” Oliver said. “I was hooked.”
Oliver gained an
interest in desiccation tolerance — the
ability of certain plants to dry completely and then fully recover once water was
introduced again.
At a meeting in California, Oliver’s Ph.D. supervisor was talking and kept mentioning Oliver’s work in desiccation tolerance. At that same meeting, there were United States Department of Agriculture recruiters looking for a molecular scientist who understood plant stress.
At 39 years old, Oliver was recruited by USDA and has worked for them for the past 30 years.
“It was good to work for them, but it’s a different thing
than university in the sense that we have a more practical leaning than does a
university researcher,” Oliver said. “So, you’re trying to work towards a more
agricultural goal to improve U.S. farming, whereas here you’re interested in
that, but you’re also more focused on scientific questions and that’s the
difference.”
Even though Oliver retired from the USDA on Nov. 30, he still has two grants that he’s working on — one with a group of MU researchers led by Robert Sharp and is collaborating with Bond LSC researcher Ruthie Angelovici — so moving into Bond LSC seemed like a logical next step.
“I
didn’t want to stay where I was in Curtis [Hall] because then I’m so close to
where I’ve been for the last 15 years,” Oliver said. “I didn’t feel like
there’s been any change.”
One of his grants focuses on plant metabolomics and the effect of drought on nodal root growth in maize, where he collaborates with Robert Sharp, director of MU’s Interdisciplinary Plant Group. The other grant, Oliver’s self-proclaimed “claim to fame,” is on desiccation tolerance and is part of a large multidisciplinary effort. He has been also working with Angelovici on aspects of desiccation tolerance in grasses.
For the past 40 years, Oliver has been trying to figure out how these “resurrection plants” can be completely drained of water and then come back to life. This pursuit of knowledge has all been in an effort to figure out how people can make plants more tolerant of drought.
“We
now understand a lot more about how resurrection plants work. So, we have a
much better understanding of how these plants deal with desiccation on a
metabolic level,” Oliver said. “I’ve been involved in sequencing many genomes
of resurrection plants as well as the mosses and that’s given us insights that
we’ve never had before.”
In his research, Oliver studies these plants through their genomes and gene expression.
“I
think the thing we’ve confirmed more than anything — the ability to survive
drying is a very ancient trait,” Oliver said. “There’s no real evidence for it,
but genomes are indicating that it’s probably the origin of land plants
depended on desiccation tolerance.”
As
part of the plant sciences division, Walter Gassmann — interim director for
Bond LSC — worked with Oliver in coordinating the 2015 IPG symposium at Bond
LSC.
“I
think his work is really fascinating,” said Gassmann. “He doesn’t limit himself
to the easy things. Some of these plants are not standard experimental systems,
so you have to build techniques and learn how to work with them, so he’s been
very pioneering in his work.”
After these two
grants are finished, Oliver will officially retire and put up his lab coat.
“I
don’t think any scientist is ever ready to leave it…” Oliver said. “You have
to hope what you’ve written and what you’ve said in meetings has inspired
someone younger to go that route. And there are! Our field, in the last 10
years, has boomed in the number of people getting into looking at resurrection
plants because of the impact of global warming. Everybody’s interested in how
plants that survive in very extreme environments can survive and how do they do
that and what can we learn from that? That’s what drove me and that’s what’s
driving young people now.”
