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Skin steroidogenic potential and production are limited, so skin-derived estrogens play site-specific autocrine and/or paracrine regulatory roles (Taves et al., 2011

Cyanocobalamin has many approved indications (many that lead to a deficiency of cyanocobalamin through an impairment of absorption of cyanocobalamin) including: Cyanocobalamin can be administered via intramuscular (IM) or subcutaneous (SC) routes

Thigh injections are slightly more variable in absorption but provide an alternative when needed

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For the validation cohort (2021), a similar protocol than above was used in 96 well plates with the following modifications: Digestion was made with 0.25 g of trypsin only, acidification was performed with formic acid 1% (FA)