Team:Georgia State/Engagement

Team:Georgia State - 2016.igem.org

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Team: Georgia State IGEM

PROJECT

This year the Georgia State iGEM team reached out and attended the 2016 Maker Faire. The team volunteered with Lambert High School iGEM team in promoting and informing people about synthetic biology. Our team and Lambert High School’s team exchanged project ideas and we were very interested in their CRISP-Cas9 system inspired by Georgia Tech. Not only promoting one idea, but our team was able to survey about 40+ people on Cannibidiol oil. The Maker Faire allowed us to answer questions that people in our community had about science, while also educating them on CBD oil and how it's used. We were able to survey people's knowledge of CBD oil and then in turn provide more information on it as the survey takers became curious. Our team took part in several science oriented events that allowed us to share our knowledge with those who might not otherwise be exposed to such information.

Survey Results

Atlanta Science Fair.

We were requested to come back to ASF for 2016 due to how successful our BioBrick activity was last year. This year we educated many students and parents on synthetic biology and how to create a BioBrick construct using legos.

Next Generation Men

Our team connected with our local high schools by bringing them into the lab and to provide them a deeper understanding of what we do as an iGEM team. We partner with Next Generation Men Inc., a non profit that was established by former high school students to provide underrepresented students with professional exposure, leadership development, community service and personal support. This experience allowed the students to get exposure to college and how life is working in a lab. We provided a brief tour for students of the iGem lab spaces by showing them our lab equipment and their basic usage. Students learned about iGem and what synthetic biology is. Students got an opportunity to learn about molecular techniques. We educated students on PCR and gave them an opportunity to run PCR and conduct gel electrophoresis. Many of the students who attended were either interested in non-science majors and or undecided. Their experience in the lab allowed them to explore and ask questions about careers in science.

Synthetic Biology Club Reached Out to Georgia State Students

We also had open lab days in which other college students came into our lab during the school year, and we provided them with information on how to get involved with research as an undergraduate.

STEM Friendly Eyes Conference

Students from our iGem Team participated in a conference that undergraduates, graduate students and faculty attended. Our team members divided into groups and created poster presentations and oral presentations about techniques in synthetic biology. Guests asked questions about our posters and provided feedback on presenting findings and communicating in science.

The Zoo Atlanta

Students from our iGem Team participated in a conference that undergraduates, graduate students and faculty attended. Our team members divided into groups and created poster presentations and oral presentations about techniques in synthetic biology. Guests asked questions about our posters and provided feedback on presenting findings and communicating in science.

Legislative Outreach

Mayor of Clarkston

We have a meeting scheduled with Annette Davis Jackson, State Senate Candidate for District 55 and Tim McCall, State Representative Elberton District and is the Chairperson of the Agriculture and Human Consumption Committee. He also serves as a member of the Natural Resources and Environment Committee.

Questions that we have developed questions for our Legislatives Interviews

How can this update to the bill support the veteran population in regards to PTSD?

What areas in georgia can be associated with growth and cultivation?

How can this update to the bill support the veteran population in regards to PTSD?

If so, will it be seen more as recreational use by the masses if expanded to the general population or do you foresee it always being used for medicinal purposes in Georgia?

Do you foresee any new conditions added to list to increase the number of eligible patients?

Current Treatments

Snake Bites

The CDC has reported that about 8,000 snake bites occur in the U.S. every year. Many of the snakes are known to not be harmless. The snake bites can cause an allergic reaction or an infection. Both are known to cause serious problems for patients. It is imperative for victims to get to the nearest emergency as soon as possible. Unfortunately, many snake bites happen during activites like, camping, hiking or woodsy areas. For many residents who are not inside of the metropolitan area, have to get transported to facilities that are not very close in proximity. Sometimes victims may have to travel 45 minutes to an hour to get to a hospital. This can cause issues for patients receiving prompt and excellent care.

Current priorities for patients are:

Cleaning and immobilizing the site

Cooling the area to minimize swelling

Mainting Heart Rate and Respirations

Recognizing the type of snake if possible.

There isn’t a way to tell if you have been injected with venom but it is important to treat every bite as if you have been injected with venom. Snakes injected venom on about 30%-50% of bites.

@. (n.d.). Grady Hospital | Atlanta, GA | Grady Health. Retrieved from http://gradyhealth.org/

Current Treatment through Emergency Medical Services

Treat for Shock

Keeping area immobilized and possibly using a splint

Ace bandages are good for managing the spread of venom

Limmer, D., O'Keefe, M. F., & Dickinson, E. T. (2009). Emergency care. Upper Saddle River, NJ: Pearson Prentice Hall.

Current Military Treatment

· IV line · Monitor vitals · Clean wound · Place patient in comfortable position · Identify snake · Suction extractor · Splint the area at level of heart

Navy Medicine Manpower, Personnel, Training and Education Command (2009). Hospital Corpsman (NAVEDTRA 14295B). Washington, DC: U.S. Government Printing Office.

Investigating How Snake Bites are managed in India

Management protocol for snake bite poisoning

Local

Specific

Supportive

Check for airway, breathing and circulation of patient

Local:

Tourniquet: should be tied between wound & heart, apply pressure adequate to occlude only lymphatics releasing for few seconds every 10mins

Immobilize: immobilize the effected limb, elastocrepe bandage may be applied (except for viper bites with lots of local reaction)

Clean with normal saline if bite <1hr old: short skin incision is made and suction of discharge at the bite should be done

Immobilize: immobilize the effected limb, elastocrepe bandage may be applied (except for viper bites with lots of local reaction)

If any local reactions present like bullae-they are left intact, if necrosis of tissue or ulcer is present-debridement should be done, compartment syndrome of limbs - fasciotomy is done

Immobilize: immobilize the effected limb, elastocrepe bandage may be applied (except for viper bites with lots of local reaction)

Specific

Anti-snake venom- it is given based on assessment of severity scoring of envenomation and type of poison

Mechanical ventilation

Supportive care:

Antibiotics

Methylprednisolone injection

Fresh frozen plasma , fresh blood transfusion

Prevention and treatment of hypotension

Cancer Patient

We interviewed a cancer patient who just became eligible for CBD oil in georgia, Our interview gave us insight on how difficult it is to obtain the oil and the laws that are in place to regulate CBD oil. This shaped how we proceeded with future directions moving forward with our project. Some studies say that having some THC is helpful for patients, we are seekings to synthesize a 0% THC containing CBD oil.

Following our protein expression of the Mambalgin-1 protein , our next steps will include testing the analgesic properties in an animal model. Mambalgin can provide a non-addictive opioid alternative to morphine.We know that Acid-sensing ion channels (ASICs) are involved with nociception, depression and inflammation. ASICS are activated by lowering of pH levels in the extracellular space. We also know that ASIC1 & ASIC2 are expressed in the central nervous and peripheral nervous system. (Salinas, et. al 2014) These neurons are connected to the dorsal root ganglion, which we know to bring in sensory information. We will be partnering with the Neuroscience Institute at Georgia State using Sprague-dawley rats to test our introduction of mambalgin to the ASICs. We will inject rats with the expressed mambalgin intra planarly and introduce thermal pain. We will be using the paw thermal stimulator to assess nociception. We will assess results by measuring paw withdrawal latency. We will have one experimental group and two control groups. Saline/vehicle will be our negative control, and our positive will be manufactured mambalgin. Our experimental group will be for our expressed mambalgin in Pichia pastoris.

Salinas, M., Besson, T., Delettre, Q., Diochot, S., Boulakirba, S., Douguet, D., & Lingueglia, E. (2014). Binding Site and Inhibitory Mechanism of the Mambalgin-2 Pain-relieving Peptide on Acid-sensing Ion Channel 1a. The Journal of Biological Chemistry, 289(19), 13363–13373. http://doi.org/10.1074/jbc.M114.561076

Experimental Design

We expect mambalgin to provide analgesic properties in the presence of the thermal stimulus. We should see a longer Paw Withdrawal Latency (PWL) for the rats that were administered mambalgin.

Sprague Dawley Rats subjects ( Subjects are acclimated to device twice 30 minutes a day for 3 days.)

Administer expressed Mambalgin (.34 nmoles) every 20 minutes

Negative control rats given saline vehicle Intraplanar injection to hindpaw

Positive control rats given mambalgin protein provided by manufacturer

A paw thermal stimulator will be used to measure paw withdrawal latency 15 minutes after each injection

PWL baseline will be measured after the first 15 minutes

PWL is the time that it takes for the rat to remove its paw from the thermal stimulus

PWLs will be quantified using Graph Pad Software

ANOVA and Fisher Post Hoc test will be used to determine specific group differences. ( Edison and Murphy 2013)

Eidson, L. N., & Murphy, A. Z. (2013). Blockade of Toll-Like Receptor 4 Attenuates Morphine Tolerance and Facilitates the Pain Relieving Properties of Morphine. The Journal of Neuroscience, 33(40), 15952–15963. http://doi.org/10.1523/JNEUROSCI.1609-13.2013