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5 protocols using acetylcholine bromide

1

Vasodilator and Vasoconstrictor Evaluation

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The following drugs were used: acetylcholine bromide, noradrenaline bitartrate,
sodium nitroprusside, NG-nitro-L-arginine methyl ester, indomethacin (all
obtained from Sigma Chemical Co., St Louis, Missouri, USA). All drugs were dissolved
in Krebs-Henseleit solution.
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2

Vasopressor Signaling Pathway Modulation

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Drugs and suppliers were as follows: acetylcholine bromide (Sigma, St. Louis, MO, USA); adenosine (Sigma), angiotensin II amide; arginine vasopressin (AusPep, Parkville, Victoria, Australia); enalaprilat (gift from Merck, Rahway, New Jersey, USA); mecamylamine (Sigma); noradrenaline bitartrate (Sigma); and vasopressin V1 receptor antagonist des-Gly-[Phe1,D-Tyr(Et)2,Lys6,Arg8]-vasopressin (Bachem, Bubendorf, Switzerland). All drugs used for in vivo assessment were prepared using sterile 0.9% sodium chloride solution. Angiotensin II was stored as a stock solution at -20 °C until required. All other drugs were made fresh daily.
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3

Synthesis and Characterization of Nicotinic Compounds

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Acetylcholine bromide (CAS No. 66-23-9), (–)-nicotine free base (CAS No. 54-11-5), (–)-cotinine free base (CAS No. 486-56-6), and ( ±)-nornicotine (CAS No. 5746-86-1) were purchased from Sigma-Aldrich (St. Louis, MO, USA). ( +)-Anabasine hydrochloride (CAS No. 53912-89-3), buspirone hydrochloride (CAS No. 33386-08-2), and metanicotine oxalate (CAS No. 220662-95-3) were purchased from Tocris Bioscience (Bio-Techne®, Minneapolis, MN, USA). ( ±)-Anatabine free base, AZD1446, and (S)-N-methylanatabine dihydrochloride were custom synthesized by WuXi AppTec (all: purity ≥ 95%; Shanghai, China).
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4

Cardiovascular Pharmacology Protocols

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The following drugs were used: acetylcholine bromide, DMSA, enalapril maleate,
L-arginine, lead acetate, noradrenaline bitartrate, sodium nitroprusside, and
urethane (all obtained from Sigma Chemical Co., St Louis, Missouri, USA). All drugs
were dissolved in Krebs-Henseleit solution, and the concentrations were expressed in
molarity. Lead and sodium acetate were dissolved in acidified tap water as described
above. L-arginine and enalapril were dissolved in tap water. DMSA solutions were
freshly prepared by dissolving the compound in 5% NaHCO3.
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5

Vasoreactivity Assessment Protocol

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Drugs used were acetylcholine bromide (Sigma, St Louis, MO, USA); ambrisentan (Selleckchem, Houston, TX, USA); bosentan sodium salt (Selleckchem); BQ788 sodium salt (Peptides International, Louisville, KY, USA); endothelin‐1 (Genscript, Piscataway, NJ, USA); macitentan (Selleckchem); Nω‐nitro‐L‐arginine methyl ester hydrochloride (Sigma); (‐)‐noradrenaline bitartrate (Sigma); and sarafotoxin S6c (Auspep, Parkville, Victoria, Australia). All drugs were dissolved in MilliQ water except for endothelin‐1 which was dissolved in 10% dimethylformamide to 10−4 mol L−1, then diluted in MilliQ water, macitentan which was dissolved in DMSO to 10−3 mol L−1, then diluted in MilliQ water, and BQ788 which was dissolved in DMSO to 10−4 mol L−1.
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