Vitamin D 20

Operations Research Flight Ground Service Education/Outreach Clinical Nutritional Assessment (MedB8.1)

Preflight Inflight Postflight Nutrition SMO Extend MedB 8.1 Inflight Collections CM evaluation 120

100

(nmol/L) 40

25(OH)vitamin D 20

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean 75

60 UPA

15 SV urine Ca, mg/dL

0 0 6 12 18 24 Hour of day Omega-3 Fatty Acids

-5 Whole Body BMD

(% pre-flight) from change -10 0 1 2 3 4 5 Mean Fish Servings (#/wk)

Fish intake may mitigate bone and muscle loss, cardiovascular, and cancer risks. Vitamin K

Vitamin K status does not appear affected by spaceflight (or bed rest). 1750 1500 1250 1000 750 500 250

Bone Breakdown (NTX) 0

0.50 0.55 0.60 0.65 0.70 0.75 Dietary Animal Protein:Potassium Ratio (g/mEq) Preflight Inflight L-180 L-45 FD15 FD30 FD60 FD120 FD180 4-d High 4-d Low 4-d Low 44-d-d Low Low 4-d High 4-d High 4-d Monitored APro:K APro:K APro:K APro:KAPro:K APro:K APro:K R+30 R+180 R+365 Postflight 4-d Monitored 4-d Monitored 4-d Monitored NOTE: the low ratio diet is NOT low protein, and NOT vegetarian Diets are designed to maintain: energy, protein, calcium, sodium Vitamin D 120

100 80 1 Upper quartile of cortisol Lower quartile of cortisol 60

(nmol/L) 2000 IU/d 40 1000 IU/d 25(OH) D vitamin 20 400 IU/d 0 0.01 April/March June/July Sept 120 2000 IU/d

100 10000 IU/wk

Probability of EBV shedding of Probability 0.0001 60 0 20 40 60 80 100 120 (nmol/L) 40 Serum 25-hydroxyvitamin D (nmol/L)

25(OH) D vitamin 20

0 Feb June Sept

800 IU Vitamin D/day maintains vitamin D status during flight. In Antarctic analog: Vitamin D, stress, and viral reactivation are interrelated. Vitamin D 2011 Dietary Reference Intakes

120 120

100 100

80 80

60 60

40 40 Children & Adolescents Young & Middle Aged Adults

Achieved 25OHD (nmol/L)Achieved 20 Older Adults 20 Non-obese Obe se Achieved 25OHD (nmol/L)Achieved 0 0 400 800 1200 1600 2000 2400 2800 0 1000 2000 3000 Total Vitamin D Intake (IU/day) Total Vitamin D intake (IU/d) Vitamin D Dosing Study

10000 IU/wk 50000 IU/wk for 1 mo then monthly 2000 IU/d 15 15 15

10 10 10 mmol/d mmol/d mmol/d 5 5 5

0 0 0 0 7 14 21 28 60 90 0 7 14 21 28 60 90 0 7 14 21 28 60 90 Time (d) Time (d) Time (d)

Increased urinary calcium is more often observed at higher vitamin D doses. Iron Inflammation/Oxidative Damage 20 20

15 15

10 10

5 5 Total iron body (mg/kg) Total iron body (mg/kg) 0 0 0 5 10 15 20 0 500 1000 1500 2000 Oxidative DNA damage (8OHdG, µg/day) SOD (Antioxidant Defense)

15 (pg/mL) α 10

5 Plasma TNF Plasma 0 0 5 10 15 20 Total Body Iron (mg/kg)

Gaps remain in our understanding the interrelationships of iron, oxidative damage, immune function, and radiation. Sodium

Low Na High Na 9000 8000 7000 6000 g/d)

µ 5000 ISS Sodium Intake 4000 3000 10000 (CTX, 2000 Bone Breakdown 1000 8000 0 0 5 10 15 20 25 6000 Study Day mg/day 4000 3500 mg/d = ISS requirement (JSC- 2000 28038); and the “old” RDA) 2300 mg/d = US Dietary Reference Intake 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Tolerable Upper Intake Level (UL)**, and NASA exploration requirement (JSC- Week of Spaceflight 63555)

In 2005-2006, the average US intake of Na was estimated at 3,436 mg Na/d* In 1990-1999, the average US intake of Na was estimated at: 3,377 mg for 31-50 yo M** 3,539 mg for 31-50 yo F * http://www.cdc.gov/media/pressrel/2009/r090326.htm ** IOM, Dietary Reference Intakes, 2004

Biochemistry

Intermediate 3 Intermediate 1 Enzyme B vitamin

Enzyme A vitamin Intermediate 3

X Intermediate 2 C C X One Carbon Transfer

Thymidine synthesis MMA 2MCA 5,10- 5,10-methylenetetrahydrofolate reductase methylenetetrahydrofolate (MTHFR)

cystathionine 5 methyltetrahydrofolate Cystathionine b-synthase (CBS) homocysteine Vitamin B6 tetrahydrofolate Methionine synthase (MS) Vitamin B12

Folic acid methionine S-adenosylhomocysteine (SAH)

Methylated DNA, S-adenosylmethionine proteins, lipids (SAM) DNA, proteins, lipids Enzymes/Polymorphisms

. Proteins • Assembled based on “blueprints” . For many (all?) enzymes, there are small differences in blueprints across the population • These are known as “polymorphisms” –poly = multiple, “morph” = forms • Example: blood types Enzymes/Polymorphisms

• Everybody has 2 copies (one from mom and one from dad), resulting in four possibilities of this MTHFR polymorphism: C/C (~35% of the population) C/T (or T/C)(~50% of the population) C C T C T/T (~15% of the population)

C T T T Thymidine synthesis

5,10- 5,10-methylenetetrahydrofolate reductase MMA 2MCA methylenetetrahydrofolate (MTHFR)

cystathionine 5 methyltetrahydrofolate Cystathionine β-synthase (CBS) homocysteine Vitamin B6 tetrahydrofolate Methionine synthase (MS) Vitamin B12

Folic acid methionine S-adenosylhomocysteine (SAH)

Methylated DNA, S-adenosylmethionine proteins, lipids (SAM) DNA, proteins, lipids 350 300 250 200

MMA 150 (nmol/L) 100 50 0 5,10-methylenetetrahydrofolate reductase MMA R+0 2MCA L-10 FD15 FD30 FD60 R+30 FD120FD180 5,10- Pre Mean (MTHFR) 300 methylenetetrahydrofolate

200

(nmol/L) cystathionine 100

2-methyl citric acid 5 methyltetrahydrofolate 0 Cystathionine β-synthase

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean (CBS) Vitamin B6 700 homocysteine 400 350 tetrahydrofolate Methionine synthase 300 (MS) 250 Vitamin B12 200

(nmol/L) 150

Cystathionine 100 50 0 methionine S-adenosylhomocysteine

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 (SAH) Pre Mean

14 12 Methylated 10 DNA, proteins, 8 S-adenosylmethionine lipids 6 (umol/L) 4 (SAM) Homocysteine DNA, 2 proteins, 0 lipids R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean 16 500 400 15 Homocysteine 300

g/L 14 µ 200 (g/dL) Hemoglobin misincorporation

Ferritin (males) 13 100 0 12 into proteins R+0 L-10 FD15 FD30 FD60 R+30 R+0 FD120FD180 R+30 Pre Mean Pre Mean 350 300 5,10-methylenetetrahydrofolate reductase MMA 2MCA 250 5,10- 200 (MTHFR)

MMA 150

(nmol/L) methylenetetrahydrofolate 100 50 0

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 cystathionine Pre Mean 300 5 methyltetrahydrofolate

200 Cystathionine β-synthase

(nmol/L) (CBS) 100 Vitamin B6 2-methyl citric acid homocysteine 0 tetrahydrofolate R+0 Methionine synthase L-10 FD15 FD30 FD60 R+30 (MS) FD120FD180 Pre Mean Vitamin B12 700 400 350 300 250 200

(nmol/L) 150 methionine Cystathionine 100 S-adenosylhomocysteine 50 0 (SAH)

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean 14 Methylated 12 DNA, 10 S-adenosylmethionine proteins, 8 lipids 6 (umol/L) 4 Homocysteine (SAM) 2 DNA, 0 proteins,

R+0 lipids L-10 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean 16 500

400 15

300

g/L 14 µ 200 (g/dL) Homocysteine Hemoglobin

Ferritin (males) 13 100

0 misincorporation 12

R+0 L-10 FD15 FD30 FD60 R+30 R+0 FD120FD180 R+30 Pre Mean Pre Mean into proteins

350 300 5,10-methylenetetrahydrofolate reductase MMA 2MCA 250 5,10- 200 (MTHFR)

MMA 150

(nmol/L) methylenetetrahydrofolate 100 50 0

R+0 L-10 FD15 FD30 FD60 R+30 FD120FD180 cystathionine Pre Mean 300 5 methyltetrahydrofolate

200 Cystathionine β-synthase

(nmol/L) (CBS) 100 Vitamin B6 2-methyl citric acid tetrahydrofolate homocysteine 0

R+0 Methionine synthase L-10 FD15 FD30 FD60 R+30 (MS) FD120FD180 Pre Mean Vitamin B12 700 400 350 300 250 200

(nmol/L) 150 methionine Cystathionine 100 S-adenosylhomocysteine 50 0 (SAH)

R+0 L-10 FD15 FD30 FD60 R+30 35 FD120FD180 Pre Mean 60 14 30 Methylated

12 mol/d) 25 DNA, 40 µ 10 20 proteins, 8 S-adenosylmethionine (nmol/L) 15 lipids

6 Serum Folate 20

(umol/L) 10 4 Homocysteine (SAM)

Vit B6 (4-PA, B6 Vit 5 2 0 DNA, 0 0 proteins, R+0 FD15 FD30 FD60 Pre R+0 FD120 FD180 R+0 lipids L-10 FD15 FD30 FD60 R+30 FD15 FD30 FD60 R+30 FD120FD180 Pre Mean FD120 FD180 Pre Mean

Polymorphism/Associated Risks

. MTHFR polymorphisms associated with: . Increased risk of ischemic and hemorrhagic stroke . Increased risk of migraine . Increased homocysteine associated with: . Increased risk of vascular events . Increased risk of stroke . Risk factor for retinal venous occlusive disease . Risk factor for narrower retinal vasculature in men . Case study(ies?) exists relating folic acid deficiency and optic neuropathy

Brain Res Bull 71:45-50, 2006; Annals Neurology 59(2): 372-375; Ophthal 109(2):287- 290; Am J Ophthalmol. 2009; 148(6):902-909; J Med Case Reports 2:299, 2008. 1C/VIIP

. We’ve documented 4 intermediates strongly suggesting the existence of polymorphism(s) in the 1-carbon metabolism pathway. . Additional evidence: folate/B6; ferritin, hemoglobin

. Follow-on proposal submitted to HHC/VIIP . Expected results: . Inform risks . Inform therapeutic options . Inform VIIP research Nutrient Requirements Nutrient CountermeasuresPhysiologicalPhysiological Requirements Systems Systems

Vehicle/MissionVehicle/Mission Countermeasures