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Parameterized Framework Inputs: concentration_mcg_per_mL peptide concentration after reconstitution (or convert: mg/mL 1000 = mcg/mL) pump_output_mL_per_actuation delivered volume per spray (from manufacturer specs or direct measurement) Output: amount_mcg_per_actuation = concentration_mcg_per_mL pump_output_mL_per_actuation Optional Volume Mapping (U-100 Convention) If units are used as a volume shorthand (where 100 units = 1.0 mL on a U-100 scale): pump_output_mL_per_actuation = units_per_actuation / 100 amount_mcg_per_actuation = concentration_mcg_per_mL ( units_per_actuation / 100) Nasal Actuation Calculation Table (Placeholders) Worked Examples (Placeholders Only) Example 1 (mL-based pump): If concentration = 3333 mcg/mL (i.e., 10 mg reconstituted in 3.0 mL) and pump output = 0.10 mL/actuation , then amount per actuation = 3333 0.10 = 333.3 mcg/spray

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The same analysis was done in N2a cells co-expressing the phosphatidylserine biosensor mRFP-Lact-C2 25 and the different GFP-fusions, transfected at equal plasmid ratio