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Each sample was then split into two fraction (100 L) and treated as follows: For H 2 S measurement, 100 L of each sample was precipitated with 1 mL of absolute MeOH at 40 C for 2 h, centrifuged (16,000 g at 0 C), and obtained supernatant was evaporated and the residue was resuspended and sonicated in 100 L of 50 mM HEPES buffer (pH 7.4)

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does not resolve with tolerance as most other side effects do Sedation and cognitive slowing: dose-dependent CNS depression that diminishes as tolerance develops over days to weeks of regular use Dizziness and orthostatic hypotension: resulting from opioid-induced vasodilation and blunted baroreceptor reflex responses Pruritus (itching): driven by histamine release from mast cells rather than an IgE-mediated allergic hypersensitivity reaction Miosis (pinpoint pupils): resulting from oculomotor nerve stimulation at the Edinger-Westphal nucleus