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For transfection, siRNAs were delivered to a final concentration of 10 nM using Lipofectamine RNAiMAX transfection reagent (ThermoFisher Scientific) as detailed by the manufacturer

Box 1 Clinical Practice Guidelines for Vitamin D 3 Immunomodulatory Applications Therapeutic Targets: Optimal Range: Maintain serum 25(OH)D levels between 40-60 ng/mL for maximal immunomodulatory benefits (guideline) ( Minimum Threshold: Ensure levels exceed 30 ng/mL for basic immune function support (guideline) ( Safety Ceiling: Avoid exceeding 100 ng/mL to prevent hypercalcemia and adverse effects (guideline) ( Dosing Guidelines: General Population: 15002000 IU Vitamin D 3 daily for adults with deficiency or insufficiency (guideline and consensus statement) (66)

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While there is evidence of disrupted serotonergic neurotransmission in AD (267), the role of neurosteroids, such as DHEAS in the progression of this age-related neurodegenerative disease has not been fully elucidated