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4 protocols using l norepinephrine bitartrate salt monohydrate ne

1

Pharmacological Manipulation of Neurochemicals

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The following drugs were kept at −20°C as stock solutions and dissolved in aCSF to their final concentrations on the day of experiments: L-(−)-norepinephrine (+)-bitartrate salt monohydrate (NE, 10-100 μM, Sigma-Aldrich, shielded from light exposure during preparation, storage and application), picrotoxin (PTX, 50 μM, Tocris), prazosin hydrochloride (10 μM, Sigma-Aldrich), (R)-(−)-phenylephrine hydrochloride (100 μM, Sigma-Aldrich), yohimbine hydrochloride (20 μM, Tocris), tetrodotoxin (TTX, 1 μM, Tocris), TNP-ATP triethylammonium salt (10 μM, Tocris), pyridoxalphosphate-6-azophenyl-2′,4’-disulfonic acid tetrasodium salt (PPADS, 100 μM, Tocris).
Fluorocitric acid (FCA, Sigma-Aldrich) was prepared on the day of experiments with DL-fluorocitric acid barium salt (Paulsen et al., 1987 (link)) dissolved in 1 mL of 0.1 M HCl. Two-to-three drops of 0.1 M Na2SO4 was added to precipitate the Ba2+, after which 2 mL of 0.1 M Na2HPO4 was added and the suspension was centrifuged at 1000 g for 5 min. The supernatant was added to the aCSF to achieve a concentration of 100 μM.
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2

Pharmacological Modulation of Neuronal Activity

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The following drugs were kept at −20°C as stock solutions and dissolved in aCSF to their final concentrations on the day of experiments: L-(−)-norepinephrine (+)-bitartrate salt monohydrate (NE, 1 μM, 100 μM, Sigma-Aldrich), picrotoxin (PTX, 50 μM, Tocris), corticosterone (2 μM, Tocris), 3-hydroxynaphthalene-2-carboxylic acid (3,4-dihydroxybenzylidene) hydrazide (Dynasore, 80 μM, Tocris). 2-Methyl-1,2-di-3-pyridinyl-1-propa-none (metyrapone, Tocris) was dissolved in sterile saline, and injected intraperitoneally at a dose of 100 mg/kg body weight. Clozapine N-oxide (CNO, Sigma-Aldrich) was made fresh daily by dissolving in DMSO (100 mg/50 μL) and then diluting in saline to a working concentration of 5 mg/mL.
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3

Pharmacological Manipulation of Neurochemicals

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The following drugs were kept at −20°C as stock solutions and dissolved in aCSF to their final concentrations on the day of experiments: L-(−)-norepinephrine (+)-bitartrate salt monohydrate (NE, 10-100 μM, Sigma-Aldrich, shielded from light exposure during preparation, storage and application), picrotoxin (PTX, 50 μM, Tocris), prazosin hydrochloride (10 μM, Sigma-Aldrich), (R)-(−)-phenylephrine hydrochloride (100 μM, Sigma-Aldrich), yohimbine hydrochloride (20 μM, Tocris), tetrodotoxin (TTX, 1 μM, Tocris), TNP-ATP triethylammonium salt (10 μM, Tocris), pyridoxalphosphate-6-azophenyl-2′,4’-disulfonic acid tetrasodium salt (PPADS, 100 μM, Tocris).
Fluorocitric acid (FCA, Sigma-Aldrich) was prepared on the day of experiments with DL-fluorocitric acid barium salt (Paulsen et al., 1987 (link)) dissolved in 1 mL of 0.1 M HCl. Two-to-three drops of 0.1 M Na2SO4 was added to precipitate the Ba2+, after which 2 mL of 0.1 M Na2HPO4 was added and the suspension was centrifuged at 1000 g for 5 min. The supernatant was added to the aCSF to achieve a concentration of 100 μM.
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4

Epinephrine and Norepinephrine Impact on Campylobacter Growth

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The effects of Epi and NE on the growth of C. jejuni NCTC 11168 were assessed using standard bacterial growth assay. Briefly, Campylobacter cells were grown in MH broth to log phase and were then pelleted, washed once and resuspended in MEMα medium. The bacterial suspension was inoculated into MEMα medium at a final concentration of 1 × 102 cfu/ml with or without 100 μM of (−)-epinephrine (+)-bitartrate salt (Epi) or L-(−)-norepinephrine (+)-bitartrate salt monohydrate (NE) (Sigma-Aldrich). Cultures were grown in 6-well plates with 5 ml of culture per well for 48 h. The optical density of each well at 600 nm was determined every 12 h of incubation using Eppendorf BioPhotometer. All assays were performed in three independent experiments with triplicate setting for each experiment. Results represent the mean ± the standard deviation (SD) of three independent experiments. The paired Student's t-test was used for statistical analysis at 5% level of significance.
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