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The dried bacterial pellet was solubilized in 20 ml of B-PER (78243, Thermo Fisher Scientific), supplemented with protease inhibitors (1 µg ml −1 of leupeptin, 1 µg ml −1 of AEBSF HCl and 1 µl of benzonase) and 5 mM imidazole and then transferred to a 50-ml centrifuge tube

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The rationale is that deeper sleep creates a better environment for GH release, and GH secretagogues ensure the pituitary responds maximally during those deep-sleep windows