Electrical activity of the human cardiomyocytes plated on patterned MEAs was recorded following 4–10 days in vitro (DIV) using a Multichannel Systems 60 channel amplifier (MEA 1040, Multichanel Systems). Prior to recording, the cells were allowed to equilibrate for 15 min in the lab atmosphere at 37°C. Temperature was maintained with a TC02 temperature controller (Multichannel Systems). The cells were stimulated using a STG 1002 stimulator (Multichannel Systems) by applying 1 ms wide bipolar square pulses of 500–700 mV amplitude at increasing frequencies ranging from 0.5 Hz to 2.5 Hz in 0.5 Hz increments. The recording medium was the same as the plating medium. For drug experiments, sotalol, (Sigma, cat#S0278), norepinephrine (Sigma, cat#A7257) or verapamil (Sigma, cat#381195) were added to the bathing medium in increasing concentrations of 10, 30, 100 and 300 μM – sotalol, 0.1, 0.3, 1 and 3 μM – norepinephrine, and 0.3, 1, 3 and 10 μM – verapamil. In all drug experiments, for control and each new drug concentration a total of 5 minutes of recordings were performed, the first 150 seconds with no stimulation, followed by 30 seconds of stimulation, and ended with no stimulation. The data was converted to pClamp (Axon Instruments) format using MC-Data Tool (Multichannel Systems) and analyzed with Clampfit (Axon Instruments) software and software written in Matlab.
Sotalol
Manufactured by Merck Group
Sourced in United States
Sotalol is a type of laboratory equipment used for analytical and research purposes. It is a specialized device designed to measure and analyze various chemical and biological samples. The core function of Sotalol is to provide accurate and reliable data for scientific investigations and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Propranolol, by Merck Group (2 mentions)
Timolol, by Merck Group (2 mentions)
Isoproterenol, by Merck Group (2 mentions)
Verapamil, by Merck Group (2 mentions)
Elastase from porcine pancreas, by Merck Group (1 mentions)
Theophylline, by Merck Group (1 mentions)
Gi254023x, by Merck Group (1 mentions)
α2 3 6 8 9 neuraminidase a, by New England Biolabs (1 mentions)
Isoproterenol iso, by Merck Group (1 mentions)
Forskolin, by Merck Group (1 mentions)
Gm6001, by Merck Group (1 mentions)
Mmp 2 mmp 3 inhibitor 1, by Santa Cruz Biotechnology (1 mentions)
Tapi 2, by Santa Cruz Biotechnology (1 mentions)
Mouse monoclonal anti flag m2 antibody, by Merck Group (1 mentions)
Irdye 680rd goat anti mouse igg, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions)
Gw280264x, by Aobious (1 mentions)
Diclofenac, by Cayman Chemical (1 mentions)
Gabapentin, by Merck Group (1 mentions)
Monuron, by Merck Group (1 mentions)
12 protocols using sotalol
1
Electrophysiological Assessment of Cardiomyocytes on MEAs
2
Antibodies and Reagents for β1-Adrenergic Receptor Analysis
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Antibodies were from the following sources: rabbit polyclonal anti-β1AR (Cat# ab3442, raised against residues 394-408 in human β1-ARs) was from Abcam. Mouse monoclonal anti-FLAG M2 antibody (Cat# F1804) was from Sigma-Aldrich. The Abcam rabbit polyclonal anti-β1AR was validated in our previous publication.3 IRDye 800CW goat anti-rabbit IgG (Cat# 925-32211) and IRDye 680RD goat anti-mouse IgG (Cat# 925-68070) secondary antibodies were from LI-COR Biosciences. Elastase from porcine pancreas (Cat# E0258), GM6001 (Cat# 364206), GI254023X (ADAM-10 inhibitor, Cat# SML0789), matrix metalloproteinase (MMP)-2/MMP-9 inhibitor I (Cat# 444241), MMP-9/MMP-13 inhibitor I (Cat# 444252), forskolin (Cat# F3917), isoproterenol (Iso) (Cat# 420355), propranolol (Cat# P0884), sotalol (Cat# S0278), and theophylline (Cat# T1633) were from Sigma-Aldrich. MMP-2/MMP-3 inhibitor I (Cat# sc-295483) and TAPI-2 (Cat# sc-205851) were from Santa Cruz Biotechnology. GW280264X (ADAM-10/ADAM-17 inhibitor, Cat# 7030) was from AOBIOUS Inc. α2-3,6,8,9 neuraminidase A (Cat #: P0722) was from New England BioLabs Inc. All other chemicals were reagent grade.
3
Analytical Standards for Environmental Analysis
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LC-MS grade methanol and water were obtained from VWR, Darmstadt, Germany. Quercetin dehydrate (≥95%, high performance liquid chromatography (HPLC)) was purchased from Enzo Life Sciences GmbH, Lörrach, Germany. Diclofenac (>99%) was obtained from Cayman Chemical Company, Ann Arbor, Michigan, MI, USA. Glyphosate (100 quality level, HPLC), gabapentin (200 quality level, HPLC), monuron (100 quality level, HPLC), chloridazon (100 quality level, HPLC), carbetamide (100 quality level, HPLC), metobromuron (100 quality level, HPLC), sotalol (≥98%), quinoxyfen (100 quality level, HPLC), metconazol (100 quality level, HPLC) and fenofibrate (≥99%) were obtained from Sigma, Darmstadt, Germany. Metformin (300 quality level, HPLC) was obtained from Fluka, Buchs, Switzerland. Furthermore, chlorbromuron (99.24%) and diazinon (99.53%) were obtained from Dr. Ehrenstorfer, Augsburg, Germany. Carbamazepine, 2,3-dihydro-2,3-dihydroxycarbamazepine, 10,11-dihydro-10,11-dihydroxy-carbamazepine, 10,11-dihydro-10-hydroxy-carbamazepine, 9-acridine carboxaldehyde and carbamazepine-10,11-epoxide were kindly provided by the German Research Center for Environmental Health, Comparative Microbiome Analysis (COMI), Helmholtz Centrum of Munich, Munich, Germany.
4
BPC 157 Dosage and Administration
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Stable gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, molecular weight 1419; Diagen, Slovenia), a partial sequence of the human gastric juice protein BPC, which is freely soluble in water at pH 7.0 and in saline, was prepared as a peptide with 99% high-performance liquid chromatography (HPLC) purity, with 1-des-Gly peptide being the main impurity. The BPC 157 dose and application regimens (10 µg or 10 ng/kg given as an intragastric administration, were as described previously (i.e., without the use of a carrier or peptidase inhibitor) (for review see, i.e., [11 (link),12 (link),13 (link),14 (link),15 (link),16 (link),17 (link),18 (link),19 (link),20 (link),21 (link),22 (link)]). Sotalol was commercially purchased (Sigma, Aldrich, St. Louis, MO, USA).
5
Receptor Signaling Pathway Assay
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Isoproterenol, propranolol, pindolol, carvedilol, sotalol, timolol, anti-FLAG monoclonal antibody (M1 clone) and Hoechst 33,258 were from Sigma–Aldrich (St. Louis, MO, USA) while anti-β2 -AR antibody from Abcam (Cambridge, UK). Cell culture media, fetal bovine serum, G418, and Lipofectamine were from Invitrogen (Milan, Italy).
6
Heterogeneous Cardiac Electrophysiology Mapping
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The heart was paced by 2 ms pulses at 2 Hz, with constant current amplitudes 2× the diastolic threshold, on either the left (LV) or right (RV) ventricular epicardial surface, mimicking ectopic activity, and during RA pacing representing normal sinus rhythm. Regional activation and repolarization heterogeneities were introduced through cooling then heating of the LAD perfusate to various temperature (min 21°C; max 40°C), as well as through local perfusion of Sotalol (10 mg/mL) from Sigma-Aldrich (Zwijndrecht, Netherlands). Recordings were taken in these different states during sinus rhythm and ventricular pacing. In total 55 different sequences were obtained across the three hearts.
Electrical and optical signals were measured simultaneously for each sequence. Tank and sock unipolar electrograms were recorded at 2 kHz (BioSemi, Netherlands) and referenced to a Wilson’s central terminal defined using tank electrodes. Optical mapping signals were acquired simultaneously at a frame rate of 1 kHz
Electrical and optical signals were measured simultaneously for each sequence. Tank and sock unipolar electrograms were recorded at 2 kHz (BioSemi, Netherlands) and referenced to a Wilson’s central terminal defined using tank electrodes. Optical mapping signals were acquired simultaneously at a frame rate of 1 kHz
7
Autonomic Control of Anuran Heart Rate
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To study the autonomic control of heart rate (fH), atropine (cholinergic muscarinic antagonist; 3.0 mg kg−1) and sotalol (β-adrenergic antagonist; 3.0 mg kg−1) were purchased from Sigma-Aldrich (St Louis, MO, United States) and dissolved in amphibian Ringer solution (composition in mmol l–1: 46.9 NaCl; 21.0 KCl; 2.40 CaCl; 1.29 MgCl; 3.14 NaHCO3; according to Zena et al., 2016 (link); Longhini et al., 2017 (link)). Drugs and doses were chosen based on previous studies performed on both tadpole and adult anuran amphibians (Zena et al., 2016 (link); Longhini et al., 2017 (link)).
8
Examining Ion Channel Modulators
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E4031 (500 nM), ATX-II (30 nM; both from Alomone labs), ouabain (0.5–1 μM), sotalol (20 μM), isoproterenol (1 μM), and erythromycin (30 μM) were dissolved in H2O, while chromanol 293B (30 μM), cisapride (100 nM; all from Sigma), and nilotinib (1 μM; Adooq Bioscience) were dissolved in DMSO. Identical DMSO amounts (0.1%) were used as vehicle controls.
9
Pharmaceutical Standards Characterization Protocol
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Atenolol, sotalol, timolol, bezafibrate, pravastatin, tamoxifen, ifosfamide, and etoposide were obtained from Sigma (Switzerland), and metoprolol, propranolol, clofibric acid, fenofibrate, gemfibrozil, carbamazepine, diazepam, fluoxetine, lorazepam, and cyclophosphamide standards were obtained from Fluka (Switzerland). The physicochemical properties of the investigated pharmaceuticals are presented in the supplementary material (Table S1 ). HPLC-grade methanol, hydrochloric acid (37%), formic acid (98%), and ethylenediaminetetraacetic acid disodium salt solution (Na2EDTA) were obtained from Merck (Darmstadt, Germany). While a glass fiber filter with a 1.2 μm pore diameter was acquired from Whatman (USA), a 0.45-μm nylon membrane filter was acquired from Sartorius (Göttingen, Germany). The Oasis HLB cartridge (60 mg, 3 mL) used for solid phase extraction (SPE) was obtained from Waters Corporation. Deionized water was supplied from a Millipore brand ultrapure water device. High-purity nitrogen gas was provided by a nitrogen generator (Peak Scientific).
10
Electrophysiological Measurements of Compound Effects
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For all dosing experiments, the LEAP induction was started 20 minutes after administering the compounds to the plate, such that the LEAP measurements were acquired 30 minutes post-dose. When FP measurements were also acquired post-dose, the FP data was acquired from 15–20 minutes post-dose. In the majority of dosing experiments, the LEAP measurements were only made at 30 minutes post-dose, and the dosing conditions were compared to the vehicle control wells (see Figs 5 and 6 ). In Fig. 8 , however, LEAP induction was performed on a subset of electrodes in each well in baseline and then in a separate subset of electrodes in each well after dosing. The LEAP signals from baseline and dosed conditions were overlaid in Fig. 8 .
The compounds used in this study were Nifedipine (Sigma, cat. no. N7634), E-4031 (Cayman Chemical, cat. no. 15203), Verapamil (Sigma, cat. no. V4629), Astemizole (Sigma, cat. no. 1044301), Terodiline (Sigma, cat. no. T4577), and Sotalol (Sigma, cat. no. S0278).
The compounds used in this study were Nifedipine (Sigma, cat. no. N7634), E-4031 (Cayman Chemical, cat. no. 15203), Verapamil (Sigma, cat. no. V4629), Astemizole (Sigma, cat. no. 1044301), Terodiline (Sigma, cat. no. T4577), and Sotalol (Sigma, cat. no. S0278).
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