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6 protocols using dl norepinephrine hydrochloride

1

Modulation of Neuronal Activity

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All pharmacological agents used were diluted in ACSF for bath application: serotonin hydrochloride (20 µM, Sigma-Aldrich), DL-Norepinephrine hydrochloride (20 µM, Sigma-Aldrich), acetylcholine chloride (100 µM, Sigma-Aldrich), mecamylamine hydrochloride (20 µM, Sigma-Aldrich), atropine (10 µM, Sigma-Aldrich), (-)-Nicotine, Ditartrate (100 µM, Merck KGaA, Darmstadt, Germany), (+)-Muscarine chloride (1 µM, Sigma-Aldrich), 1(S),9(R)-(-)-bicuculline methyl bromide (50 µM, Sigma-Aldrich).
The doses used were chosen based on minimal doses reported to elicit in vitro effects8 (link),26 (link),47 (link),72 (link)–76 (link).
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2

Microbial Atenolol Interaction Assay

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All the chemicals used in this study were of pharmaceutical grade. The microbiological media were ordered from Oxoid (Hampshire, UK). Atenolol (CAS number: 29122-68-7), N-hexanoyl-DL-homoserine lactone (AHL) (CAS Number: 106983-28-2), thiamine (CAS number: 67-03-8), DL-norepinephrine hydrochloride (CAS Number: 55-27-6), and Dulbecco’s Modified Eagle’s Medium (DMEM) medium were obtained from Sigma-Aldrich (St. Louis, MO, USA). It is worthy to mention that used Atenolol was in form of hydrochloride salt and it was dissolved in water and kept in stocks prior to use in all the next experiments.
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3

Investigating Bacterial Response to Signaling Molecules

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All the used chemicals were of pharmaceutical grade. The microbiological media were purchased from Oxoid (Hampshire, UK). N-hexanoyl-DL-homoserine lactone (AHL) (CAS Number: 106983-28-2), prazosin (CAS number: 19237-84-4), DL-norepinephrine hydrochloride (CAS Number: 55-27-6), thiamine (CAS number: 67-03-8) and Dulbecco’s Modified Eagle’s Medium (DMEM) medium were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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4

Antimicrobial Activity Evaluation

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All the chemicals used were of pharmaceutical grade. Mueller Hinton (MH) agar and broth, Luria–Bertani (LB) agar and broth, Tryptic Soy Agar (TSA), and Tryptone soy broth (TSB) were purchased from Oxoid (Hampshire, UK). Dulbecco’s Modified Eagle’s Medium (DMEM) medium, DL-norepinephrine hydrochloride (CAS Number: 55-27-6), and N-hexanoyl-DL-homoserine lactone AHL (CAS Number: 106983-28-2) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Salmonella enterica serovar Typhimurium (NCTC 12023) and Chromobacterium violaceum CV026 (ATCC 31532) were used in this work.
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5

Multiplexed Neurochemical Quantification

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Fluorescein, fluorinert FC-40, dopamine hydrochloride,
acetylcholine chloride, dl-norepinephrine hydrochloride,
serotonin hydrochloride, adenosine (Ado), and γ-aminobutyric
acid (GABA) were purchased from Sigma-Aldrich (St. Louis, MO). Solutions
were prepared with HPLC-grade water. For the multiplexing detection
experiment, a master solution of neurochemical standards was prepared
by dissolving six neurochemicals into deionized (DI) water, including
25 μM Ado, 50 μM acetylcholine (ACh), 50 μM dopamine
(DA), 125 μM norepinephrine (NE), and 125 μM serotonin
(5-HT). Then, this mixture was diluted into a series of concentrations
while GABA was kept at 25 μM in all mixtures.
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6

Neurotransmitter Ratio Preparation

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Dopamine hydrochloride and DL-norepinephrine hydrochloride were purchased from Sigma Aldrich Co. Ltd. Serotonin hydrochloride was purchased from Tokyo Chemical Industry Co., Ltd. The sample molecules were used without further purification. The neurotransmitter samples were dissolved in MilliQ water. The concentration of the solutions was 1 μM. In the mixed sample solutions, the neurotransmitters were present at molecular ratios of DA:NE:5-HT of 1:2:4 (D1N2S4), 2:4:1 (D2N4S1), and 4:1:2 (D4N1S2), and the minimum neurotransmitter concentration was 1 μM.
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