Brett Baker, Desiree Notyce , Julia Gipson, Joan Gomez Community College of Aurora

1. The Effects of Ultraviolet Radiation on Human Skin Normal Flora and Shielding Properties of Minerals Brett Baker, Desiree Notyce, Julia Gipson, Joan Gomez Community College of Aurora Dr. Victor Andersen Professor Tom Dillon

2. Benefits to NASA and Scientific Community • NASA’s Astrobiology Objective 6.2: Adaptation and Evolution of Life Beyond Earth • “Explore the adaptation, survival, and evolution of microbial life beyond Earth”1 • NASA’s The Global Exploration Roadmap: Materials, Structures, Mechanical Systems and Manufacturing (TA12) • “Technology advancements for lightweight structures providing radiation protection, multifunctional structural design and innovative manufacturing.”5 • We designed our experiment to evaluate the effects of a Mars like environment on normal flora and evaluate the protective qualities of various Earth minerals.

3. Mars Analog • Earth is protected from most UV radiation by the O3 layer. • Mars has CO2 as a primary protection from UV radiation. • CO2 provides poor protection from UV radiation. • At 99,000 feet above sea level a satellite would be in the upper portion of the O3 layer. • At this altitude UV protection would be minimal. • Without protection from the O3 layer Earth would receive higher levels of UV radiation. • The amount of UV radiation received is inversely proportional to the square of the distance between the sun and the planet.

4. Mars Atmosphere and Weather • The atmosphere of Mars is quite different from Earths. • The atmosphere of Mars is composed of primarily Carbon dioxide (~95%), a small amount of Nitrogen (~3%), with the reminder being a mixture of other gases. • The conditions on Mars are similar to Earths Stratosphere. • On sol 232 (~ April 1, 2013) the temperature on Mars was -69o C with a pressure of 888 Pa (~0.129 PSI).

5. Mission Overview Background Objective Bacterial Objective: To determine cell viability and detect mutations of human skin normal flora when exposed to a Mars like environment Mineral Objective: To study the protective qualities of the minerals based upon their properties as the pressure, temperature, atmosphere, and ozone change relative to elevation • Alterations in the presence of normal flora may have insidious health risk for long term space travel2,3,4 • Observe the potential protective qualities for naturally occurring minerals against harmful radiation

6. Hypotheses • Staphylococcusspecies exposed to a Mars like environment would survive but acquire nonlethal mutations • There is an expectation that the various minerals will have different shielding abilities. There will also be a distinct difference between the various samples with regards to the shielding. The bacteria samples with the highest mutation determine which shield had the poorest shielding abilities against the UV radiation.

7. Diagram of Box

8. Finished BalloonSat • Test it passed • Whip Test • Drop Test • Stair Test • Conditions Test • Agar Cold Test • Agar Heat Test

9. Methods

10. Mineral Samples Biotite Gypsum Talc Graphite

11. Bacteria Shielding Overlay Plate B Plate A

12. Bacteria Methodology Before Flight • Colonies removed from slant and bacteria stabbed into petri dish agar • 3 to 5 times depending on the Staphylococcus species • Plates incubated for 24 hours • External Control, Internal Control for Flight, and both Experimental Petri Dishes cultured in this matter • External Control were placed on the bench in the lab for parallel conditions Clonal Bacterial Colony Inoculating Needle Inoculating needle 24 hours in incubator Slant

13. Clonal Bacterial Colony Bacteria Methodology After Flight Inoculating Loop • Colonies removed and placed in LB broth for 48 hours • Extracted DNA using QiagenDNeasy Blood and Tissue Kit • DNA will be digested with SmaI or XbaI restriction enzymes • Gel electrophoresis will be performed on digested and undigested DNA New LB broth 48 hours in incubator Sterile inoculating loop to collect bacteria LB Broth LB Broth New agar plate DNA Extraction for Restriction Enzyme Assays

14. Results

15. Temperature Sensors Reading Temperature (Deg C) Time (min)

16. Internal and External Temperature Sensors Benchmark Test Temperature (Deg C) Time (milliseconds)

17. Pressure Sensor Reading Pressure (PSI) Time (min)

18. Accelerometer Sensor Reading Accelerometer (g) Time (min)

19. Speed vs. Time Graph Over the flight the payload experienced a maximum speed of 2.5x10e3 feet per minute. This graph of speed vs. time correlates to the graph from the accelerometer showing a maximum acceleration the decent

20. Atmospheric Density Over Flight The relationship between atmospheric density and altitude is inverse. As the atmospheric density decreased, the altitude of the payload, consequently, increased. The minimum, of the atmospheric density represents the point at which the payload reached a maximum height.

21. Staphylococcus species Survival After Flight S. aureusfrom Plate A S. aureusfrom Plate B

22. Staphylococcus species Survival After Flight S. epidermidis from Plate A S. epidermidis from Plate B

23. Staphylococcus species Survival After Flight S. aureusfrom Internal Flight Control S. epidermidis from Internal Flight Control

24. Preliminary Results DNA Quantification

25. Future Direction • Preliminary DNA extraction may have yielded insufficient DNA for analysis • Currently testing and research methods of increase genomic DNA • Optimizing growth condition for maximum cell density • Start restriction enzyme digestion to detected mutations

26. Predicted Results forGel Electrophoresis • Lane 1 = Ladder • Lane 2 = External Control • Lane 3 = Internal Control (shielded with aluminum foil) • Lane 4 = Shielded with graphite • Lane 5 = Unshielded Bacteria • Lane 6 = Shielded with Talc 1 2 3 4 5 6

27. Possible Directions for Future Studies • Incorporate UVA and UVB sensor during flight • PCR and DNA sequencing to detect mutation within a specific gene • Pulse Field Gel Electrophoresis to improve gel images • Observing Back Mutations

28. Conclusion • The Staphylococcus species survived the Mars like environment • Analysis • Optimizing growth conditions • Start detection of mutations within normal flora • The bacteria may have back mutated

29. Questions?

30. THE END

31. Reference • http://astrobiology.arc.nasa.gov/roadmap/g6.html • GuéguinouN, Huin-Schohn C, Bascove M, BuebJL, Tschirhart E, Legrand-Frossi C, and Frippiat JP. Could spaceflight-associated immune system weakening preclude the expansion of human presence beyond Earth’s orbit? November 2009 Journal of Leukocyte Biology vol. 86 no. 5 1027-1038. • CrucianB, and Sams C. Transcriptional and Proteomic Responses of Pseudomonas aeruginosa PAO1 to Spaceflight Conditions Involve Hfq Regulation and Reveal a Role for Oxygen Appl. Environ. Microbiol. February 15, 2011 77:1221-1230. • Battista N, Meloni MA, Bari M, Mastrangelo N, Galleri G, Rapino C, Dainese E, Agrò AF, Pippia P, Maccarrone M.. 5-Lipoxygenase-dependent apoptosis of human lymphocytes in the International Space Station: data from the ROALD experiment. FASEB J. 2012 May;26(5):1791-8. • http://www.nasa.gov/pdf/591067main_GER_2011_small_single.pdf • http://www.colorado.edu/outreach/BSI/k12activities/interactive/actidhpnf.html • Horneck G, Klaus DM, MancinelliRL. Space microbiology. MicrobiolMolBiol Rev. 2010 Mar;74(1):121-56. • Guéguinou N, Huin-Schohn C, Bascove M, BuebJL, Tschirhart E, Legrand-Frossi C, and Frippiat JP. Could spaceflight-associated immune system weakening preclude the expansion of human presence beyond Earth’s orbit? November 2009 Journal of Leukocyte Biology vol. 86 no. 5 1027-1038. • Crucian B, and Sams C. Transcriptional and Proteomic Responses of Pseudomonas aeruginosa PAO1 to Spaceflight Conditions Involve Hfq Regulation and Reveal a Role for Oxygen Appl. Environ. Microbiol. February 15, 2011 77:1221-1230. • Battista N, Meloni MA, Bari M, Mastrangelo N, Galleri G, Rapino C, Dainese E, Agrò AF, Pippia P, Maccarrone M.. 5-Lipoxygenase-dependent apoptosis of human lymphocytes in the International Space Station: data from the ROALD experiment. FASEB J. 2012 May;26(5):1791-8. • http://spacegrant.colorado.edu/COSGC_Projects/symposium_archive/2010/papers/CUSRS10_01%20Effects%20of%20Near- • http://202.120.143.134/Download/1531f207-c8b9-48d9-8ce9-8eddfd9800e2.pdf • Ryan, Kenneth J and C George Ray ed. Sherris Medical Microbiology: An Introduction to Infectious Disease. 4th Edition. The McGraw-Hill Companies, Inc. 2004. • Huether, Sue E and McCance, Kathryne L. Unerstanding Pathophysiology 5 ed. Elsevier Mosby 2012. • webmineral.com, minerals.net, wikipedia.org, Jim Weedlin (Geology Instructor), refractiveindex.info,

33. Mineral Properties • Luster: the appearance of a metallic shine. • Hardness: How hard the mineral is on average. • Specific gravity: The weight on the Earth. • Cleavage: The ability to reflect light. • Streak: The color of the residue left when a mineral scratches another material. • Refraction: the ability to change the direction of any wave when it goes through changes of density.

34. Talc • Appearance: Pearly to greasy or dull luster. Color pale or shades of gray, hardness 1.0 with perfect basal cleavage. Streak white. Specific gravity 2.58-2.83. Soapy feel, tenacity is sectile. Translucent with biaxial optical properties. Refractive Index 1.538-1.600. • Chemical Composition:Mg3Si4O10(OH)2

35. Gypsum • Vitreous to silky or pearly luster. Colorless to shades of green, gray, or brown. Hardnes2.0. Streak white. Specific gravity:2.67-3.00. Biaxial optical properties with a refractive index listing of 1.519-1.530. • Chemical Composition:CaSO4 * 2(H2O)

36. Functional Block Diagram Temperature Temperature Accelerometer Pressure Camera SD Card Arduino Uno Switch Heater Circuit 9V battery Switch 9V battery 9V battery 9V battery

37. Exterior wall (Foam core) Mineral Mounting Apparatus (Foam core) Mineral Mounting Support (Foam core) Minerals Bacteria and Mineral Mounting Block Diagram Thin Plastic Overlay Insulation 100 mm Petri Dish

38. Sensor Readings From BalloonSat