THE EFFECTS OF ULTRAVIOLET RADIATION ON HUMAN SKIN NORMAL

FLORA AND SHIELDING PROPERTIES OF MINERALS

Brett Baker, Desiree Notyce, Julia Gipson, Joan GomezCommunity College of Aurora

Dr. Victor AndersenProfessor Tom Dillon

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.

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.

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).

Mission OverviewBackground

• 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

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

Hypotheses

1. Staphylococcus species exposed to a Mars like environment would survive but acquire nonlethal mutations

2. 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.

Diagram of Box

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

METHODS

Mineral SamplesBiotite Gypsum

TalcGraphite

Bacteria Shielding Overlay

Plate B

Plate A

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

Slant

Inoculating needle

Clonal Bacterial Colony

Inoculating Needle

• 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

24 hours in incubator

Bacteria Methodology After Flight

• Colonies removed and placed in LB broth for 48 hours

• Extracted DNA using Qiagen DNeasy 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

LB Broth

New

agar

plate

48 hours in incubator

LB Broth

Sterile inoculating

loop to collect

bacteria

DNA Extraction for Restriction

Enzyme Assays

Clonal Bacterial Colony

Inoculating Loop

RESULTS

Temperature Sensors Reading

0 20 40 60 80 100 120 140

-20

-10

0

10

20

30

40

50

External Temperature (DEG C) Internal temperature (Deg C)

Tem

pera

ture

(Deg

C)

Time (min)

Internal and External Temperature Sensors Benchmark Test

0 200000 400000 600000 800000 1000000 120000019

20

21

22

23

24

25

26

27

External Temperature (Deg C)Internal Temperature (Deg C)

Tem

pera

ture

(Deg

C)

Time (milliseconds)

Pressure Sensor Reading

0 20 40 60 80 100 120 1400

2

4

6

8

10

12

14

Pressure (PSI)

Pre

ssur

e (P

SI)

Time (min)

0 20 40 60 80 100 120 140

-6

-4

-2

0

2

4

6

Accelx(g) AccelY (g) AccelZ (g)

Acc

eler

omet

er (g

)

Time (min)

Accelerometer Sensor Reading

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

Speed vs. Time Graph

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.

Atmospheric Density Over Flight

Staphylococcus species Survival After Flight

S. aureus from Plate A S. aureus from Plate B

Staphylococcus species Survival After Flight

S. epidermidis from Plate A S. epidermidis from Plate B

Staphylococcus species Survival After Flight

S. aureus from Internal Flight Control

S. epidermidis from Internal Flight Control

Preliminary Results DNA Quantification

Sample Identification Quantity of DNA in ng/mL

S. Epidermidis Internal Control Flight

40

S. Epidermidis Plate A, Unshielded

20

S. Epidermidis Plate A, shielded with Talc

20

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

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

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

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

Questions?

THE END

Reference1. http://astrobiology.arc.nasa.gov/roadmap/g6.html 2. Guéguinou N, Huin-Schohn C, Bascove M, Bueb JL, 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.

3. 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.

4. 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.

5. http://www.nasa.gov/pdf/591067main_GER_2011_small_single.pdf

6. http://www.colorado.edu/outreach/BSI/k12activities/interactive/actidhpnf.html7. Horneck G, Klaus DM, Mancinelli RL. Space microbiology. Microbiol Mol Biol Rev. 2010 Mar;74(1):121-56.8. Guéguinou N, Huin-Schohn C, Bascove M, Bueb JL, 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.

9. 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.

10. 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.

11. http://spacegrant.colorado.edu/COSGC_Projects/symposium_archive/2010/papers/CUSRS10_01%20Effects%20of%20Near-12. http://202.120.143.134/Download/1531f207-c8b9-48d9-8ce9-8eddfd9800e2.pdf13. Ryan, Kenneth J and C George Ray ed. Sherris Medical Microbiology: An Introduction to Infectious Disease. 4 th Edition. The McGraw-

Hill Companies, Inc. 2004.14. Huether, Sue E and McCance, Kathryne L. Unerstanding Pathophysiology 5 ed. Elsevier Mosby 2012.15. webmineral.com, minerals.net, wikipedia.org, Jim Weedlin (Geology Instructor), refractiveindex.info,

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.

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

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)

Functional Block Diagram

HeaterCircuit

Switch

9V battery

9V battery

9V battery

 

 

Camera

ArduinoUno

Temperature

9V battery

Pressure AccelerometerTemperature

Switch

Petri Dish100mm

Petri Dish100mm

Petri Dish

60mm

SD Card

Exterior wall (Foam core) Mineral Mounting

Support (Foam core)Mineral Mounting

Apparatus (Foam core) Minerals

Thin Plastic Overlay Insulation 100 mm Petri Dish

Bacteria and Mineral Mounting Block Diagram

Sensor Readings From BalloonSat