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Rallis E, Kintzoglou S, Moussatou V, Riga P

Navarro v

Morning (with breakfast) Vitamin D3 (fat-soluble, needs food) Omega-3 fish oil (fat-soluble, needs food) Vitamin B12 (sublingual, can be taken anytime but morning supports energy) First fiber serving (with 8-12 oz water, at least 30 minutes before or after other supplements) Midday (with lunch or protein shake) Protein powder serving if needed Iron if supplementing (separate from calcium by 2+ hours) Creatine (timing is flexible, consistency matters more) First HMB dose (1 gram) Afternoon Second HMB dose (1 gram) Ginger extract if managing nausea Probiotic (can be taken anytime with food) Evening (with dinner) Magnesium glycinate (supports sleep quality) Calcium (separate from iron and magnesium by 2+ hours is ideal but not critical) Zinc (take with food to prevent stomach upset) Second protein serving if needed Third HMB dose (1 gram) Second fiber serving (with water) Curcumin if supplementing Key timing rules: Iron and calcium should not be taken at the same time

Quantitative real-time PCR (RTqPCR) was performed using SYBR Green PCR master mix (Applied Biosystems, 4309155) with QuantStudio 7 Flex (Applied Biosystems)