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The cells were cultured in a CO2 incubator (5% CO 2 , 37 C) using Dulbeccos modified Eagles medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin

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Common Side Effects When side effects occurred, they were typically mild, transient, and similar in frequency to placebo groups: Reported Adverse Effects Most Common (occurring in small percentage of participants): Mild to moderate headaches (most frequently reported, often resolving with continued use) Injection site reactions (minor redness, swelling, or irritation at injection site) Gastrointestinal effects (particularly at higher oral doses: mild nausea, diarrhea, flatulence) Fatigue (reported occasionally during initial adaptation period) Rare Events: One case of chest tightness was deemed possibly related to treatment in safety trials

This effect demonstrated an L-NAME responsive, L-arginine responsive, opposite effect pattern, indicating bidirectional NO system modulation underlies BPC-157s anti-anhedonic action