My Goal in Epidemiology

What am I Doing?

        Hello my name is Armando Cancino. I am a senior from Basis Phoenix who wants to share to you what my senior project is all about. In my senior project, I am researching how bio engineered plants, Nicotiana benthamiana, can produce alternate antibodies that can be used to combat the dengue virus. These transgenic plants act like "test reservoirs," where I am able to genetically engineer these plants to produce different variants of antibodies. Normally, I don't know if everyone knows this but plants don't produce antibodies so by changing the test subject's genetic structure I will be able to convince the plants to produce antibodies.

         The reason why I am doing this project is to understand how researchers and epidemiologists  work in their every day career. I want to also receive experience working in a laboratory environment, because then I will be able to determine whether this field is the right one for me. Epidemiology is a very intriguing science, because like biology it is a combination of multiple sciences. There is no way for this science to be enclosed in one area of study, because this science works with other sciences such as biotechnology to fulfill its goals. For instance, in my project I want to understand how antibodies can be expressed from the plants machinery, which the genetic coding for the antibodies come from tobacco-mosaic virus, to prevent the dengue virus from replicating. I understand that one way antibodies prevent a pathogen from reproducing is through the process opsonization, which is the ability for multiple antibodies to bind to the receptors of the pathogen causing it to not be able to bind to other target cells. By this intuition I plan to find a way for these antibodies to be able to attach to dengue virions in mammalian cells, so that hopefully in the future people all over the world won't be harmed from the dengue virus. Hopefully my research will work out. Wish me luck!

This is the dengue virus

Why Use the Nicotiana benthamiana Plants?

Why Are Nicotiana benthamiana Plants Applicable?

After reading the article, The potential of plants as a system for the development and production of human biologics, I have found out that the reason why many researchers use Nicotiana benthamiana plants as a test subject is because of its many advantages. These plants provide low production costs for producing more test subjects and development of antibodies, high scalability in protein expression, low safety precautions, and like any plant there are very few plant pathogens that can be transmitted from the plant to a mammal. What else is more appealing about these plants is that at the Biodesign Institute there is no need for capital prohibitive facilities to store the plants, no need for any bioreactors, fermenters, or sterile delivery methods. Like any other plant these plants require carbon dioxide, water, light, and fertilizers to help them grow and develop. However a key point about these plants are that at the Biodesign Institute these Nicotiana benthamiana plants are considered transgenic, meaning that they are bio-engineered to express the production of antibodies by receiving viral genetic material, that codes for the antibodies structure, from tobacco-mosaic viruses such as Potato virus X. Normally, plants don't have any antibodies to protect their cells from pathogenic infections, as a result the plants that I am working with are synthesized to express the antibodies that can be used to fight off viruses such as dengue. The antibodies produced from the transgenic plants are tested in petri dishes where the researchers testing the antibodies can insert the dengue virus with mammalian cells into the dish in order to examine if the synthesized antibodies are able to disrupt the dengue virus from replicating. In the article, I have found out that the researchers insert mammalian glycosylation genes in order to synthesize antibodies that can coexist in both the plant's and the mammal's immune system. The researchers had to turn to this route, because the initial antibodies produced from the plants were destroyed by the mammalian immune cells, that were found in a separate petri dish. Through the process called N-glycosylation, which is basically attaching an extra sugar molecule called a glycan to a nitrogen atom found in the amino acids of the proteins in the plant cells that produce the antibodies. Through this process the results were successful as the plant-produced antibodies were able to be transported into the mammalian cells to help defend the mammalian cells from the dengue virus.     

Here is a picture of potatoes having Potato Virus X



Meet Potato Virus X

Intro to the Dengue Virus and the Nicotiana benthamiana

What is the Dengue Virus?

The dengue virus is a RNA positive flavivirus that is transmitted to humans through mosquito bites. By the definition of a flavivirus it has a protein envelope and a coccus (circular) shape while also being 40 to 60 nm in diameter. This virus' genome is composed of single-stranded positive RNA, allowing this type of virus to produce its proteins directly from the template RNA strand. The template strand for any organism's genome is the coding sector for the production of proteins in any living or nonliving (viruses) organisms. According to the CDC around 400 million people have been infected by this virus particularly affecting people at the tropical regions. The dengue virus is known to cause a severe fever, headaches, joint pain, low white blood cell counts, and bleeding manifestations around the gums, nose, bruising, and vomiting of blood. The dengue hemorrhagic fever (DHF) is the key symptom that is associated with the dengue virus, where the fever lasts between two to seven days. The fever can cause blood vessels around the infected area(s) to become permeable or "leaky," which causes blood to escape these vessels and leak into the peritoneum, the membrane that surrounds\covers the abdominal cavity and the abdominal organs. Once the blood enters the peritoneum an accumulation of fluid (in this case blood) can envelop in the abdominal cavity, causing swelling of the abdominal region. Over time as the abdominal region becomes swollen up somehow the circulatory system can shut down, causing the infected organism to not be able to transport oxygen around its entire body, which ultimately leads to the organism to its death.
The dengue virus epidemic has been occurring since the 1950's, due to increased mosquito populations at the tropics. I have found that the reason why there are increased mosquito populations around the tropics is because during the 1950's there has been increased rainfall precipitation around the tropics due to rising sea level. I believe that the rising sea level is caused by global warming because throughout the 20th century the Earth's temperature has been increasing, due to the destruction of the ozone layer. The ozone layer is the section of the atmosphere that acts as a sheath, surrounding the Earth and reflects UV radiation from the sun from impacting the Earth. However due to the 20th century technological changes; the ozone layer has been getting destroyed due to the accumulation of green house gases such as methane, DDT, nitrous oxide, carbon dioxide, and etc. Greenhouse gases such as carbon dioxide and methane are accumulating in the world due to increase in industrialization such as the production and operation of cars, factories, generators, and so on. As the ozone layer is depleted, more radiation can enter the Earth causing the Earth to become warmer. Glaciers found in the Artic and at Antarctica are then melted from this increased heat wave, which ultimately causes the sea levels to increase.
In my project, I will be testing Nicotiana benthamiana plants to see if these plants are able to express effective antibodies that can be used to stop the dengue virus. Nicotiana benthamiana plants are common herbs that can be found at Australia and grow from 0.65-5 ft. tall. These plants are very useful in my project, because they are easy to maintain and they have minimal contamination pathways. In my project, I plan to use these plants as a "tool" for me to develop antibodies in my genetically engineered plants. I want to produce antibodies that are suitable to prevent the dengue virus from infecting mammalian cells, so that the dengue virus isn't able to reproduce. If I am able to produce these suitable antibodies, then I can test them on a petri dish filled with grown, incubated mammalian cells where I inject a dengue virus strain into the dish. If the antibodies are working, then what I should observe is a low dengue virus count. Imagine if there are antibodies that can stop every dengue virus strain. Scientists would be able to inject these modified antibodies into mammalian organisms such as humans to fight off the dengue virus. Who knows how much lives can be saved from this discovery?