Rats were anesthetized with sodium pentobarbital (40 mg/kg, i.p.) (n = 8 per group), and hearts were quickly removed and mounted on a Langendorff apparatus via the aorta for retrograde perfusion with Krebs-Henseleit (K-H) buffer at a constant pressure (10 kPa) and constant temperature (37 °C). The K-H buffer (in mmol/l) was composed of the following: NaCl 118, KCl 4.7, MgSO 4 1.2, CaCl 2 2.5, KH 2 PO 4 1.2, NaHCO 3 25, and glucose 11. The buffer was saturated with 95% O 2 /5% CO 2 (pH 7.4). A water-filled latex balloon-tipped catheter was placed into the left ventricle through the left atrium and adjusted to a LVEDP of 3-10 mm Hg during the initial equilibration. The distal end of the catheter was connected to a PowerLab system (AD Instrument, Castle Hill, Australia) via a pressure transducer (model Gould P23Db). After 20 min of stabilization with K-H buffer, hearts were subjected to 30 min of no-flow global ischemia, followed by 60 min of reperfusion. LVDP, LVEDP, and +(LVdP/dt) max , -(LVdP/dt) max were continuously recorded by the PowerLab system. Data were analyzed using Chart software (AD Instrument).
Chart software
Manufactured by ADInstruments
Sourced in Germany, United States, Australia, United Kingdom
The Chart software is a data analysis and visualization tool developed by ADInstruments. It is designed to acquire, analyze, and present data from various scientific and research experiments. The software provides a user-friendly interface for recording, displaying, and processing data in real-time.
Automatically generated - may contain errors
Lab products found in correlation
Powerlab, by ADInstruments (8 mentions)
Powerlab system, by ADInstruments (5 mentions)
Powerlab 8 30, by ADInstruments (4 mentions)
Mlt125 r pulse transducer, by ADInstruments (3 mentions)
Methoxamine, by Merck Group (2 mentions)
Linopirdine, by Merck Group (2 mentions)
Epigallocatechin gallate (egcg), by Merck Group (2 mentions)
Male wistar rats, by Janvier Labs (2 mentions)
Si mb4 tissue bath system, by World Precision Instruments (2 mentions)
Tbm 4 m transbridge, by World Precision Instruments (2 mentions)
Mlt1l, by ADInstruments (2 mentions)
Powerlab 8sp, by ADInstruments (2 mentions)
Powerlab data acquisition system, by ADInstruments (2 mentions)
Inverted microscope, by Zeiss (2 mentions)
Powerlab 4sp, by ADInstruments (1 mentions)
Fleisch pneumotachograph, by Hans Rudolph (1 mentions)
Grass s48 field stimulator, by Natus (1 mentions)
Pcu 2000, by ADInstruments (1 mentions)
Bioamp amplifier, by ADInstruments (1 mentions)
Gould p23db, by ADInstruments (1 mentions)
60 protocols using chart software
1
Isolated Rat Heart Ischemia-Reperfusion
2
Patch Clamp Experiments on Cultured Cells
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Cells were patch clamped after growing them on coated glass coverslips for 2 days. Patch clamp experiments were performed in the fast whole-cell configuration. Patch pipettes had an input resistance of 3–6 MΩ, when filled with a solution containing in (mM) KCl 30, K+-gluconate 95, NaH2PO4 1.2, Na2HPO4 4.8, EGTA 1, Ca2+-gluconate 0.758, MgCl2 1.034, D-glucose 5, ATP 3, pH 7.2 and Ca2+ activity 0.1 μM. The bath was perfused continuously with a standard bicarbonate-free Ringer’s solution, composed of (in mM) NaCl 145, KH2PO4 0.4, K2 HPO4 1.6, MgCl2 1, Ca-gluconate 1.3, glucose 5, pH 7.4) at a rate of 4 mL/min. The access conductance was continuously measured and was 40–100 nS. Currents were recorded with an EPC-7 patch clamp amplifier (List Medical Electronics, Germany), the LIH1600 interface and PULSE software (HEKA, Germany) as well as Chart software (AD-Instruments, Germany). Data were stored continuously on a computer hard disc and analysed using PULSE software. In regular intervals, membrane voltages (Vc) were clamped in steps of 20 mV from − 100 to + 100 mV relative to resting potential.
3
Measuring Respiratory Parameters and EAdi
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Flow was measured with a heated Fleisch pneumotachograph, dead space 51 ml (Hans Rudolph, Kansas City, MO, USA) and airway pressure was measured by a pressure transducer (DP 15–32, Validyne, Northridge, CA, USA) for all modes. Digital EAdi signal was converted into an analog signal (National Instruments, Austin, TX, USA). During all three modes of ventilation, the EAdi waveform was simultaneously recorded with flow and airway pressure from the respective ventilator (see Additional file 1 ). All signals were digitized at a 100-Hz sampling rate (PowerLab/4SP, ADInstruments, Castle Hill, Australia) and recorded on a personal computer for subsequent analysis (Chart software, ADInstruments, Castle Hill, Australia).
4
Gastric Smooth Muscle Contractility Protocol
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Gastric content was flushed out in precooled Kreb’s solution. Smooth muscle strip (2 mm × 10 mm) was prepared and suspended vertically in an eight-channel organ bath filled with Kreb’s solution and oxygenated with 95% O2 and 5% CO2 at 37°C. One pole of the strip was fixed to an organ holder and the other pole was connected to an isometric tonotransducer. Mechanical activity of the smooth muscle strip was recorded with a computer-aided data acquisition system (Power lab biology signal recording system, AD Instruments). The initial tension was set as 1 g. The amplitude and frequency of the spontaneous contraction of the smooth muscle strip were analyzed by chart software (AD Instruments).
5
Langendorff-Perfused Rat Heart Assay
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Rats were anaesthetized with ethyl carbamate (2 g/Kg rat, i.p.) and the hearts rapidly excised and transferred in ice-cold buffered Krebs-Henseleit solution (KHs). The aorta was immediately cannulated with a glass cannula and connected with the Langendorff apparatus to start perfusion at a constant flow-rate of 12 mL/min as previously described [17 (link)]. Hemodynamic parameters were assessed using a PowerLab data acquisition system and analysed using chart software (AD Instruments, Basile, Italy). Exposure to pharmacological agents (sodium nitrite, GSNO, NONOATE, PTIO) was obtained by perfusing the cardiac preparations with KHs containing the specific substance at the desired concentration. After 20 minutes perfusion was stopped and the hearts were rapidly immersed in liquid nitrogen to be stored at −80°C.
6
Mesenteric Artery Tension Measurements
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Male wistar rats, 12 weeks old (Janvier Labs, France) were euthanized by cervical dislocation and used in accordance with Directive 2010/63EU on the protection of animals used for scientific purposes, approved by the national ethics committee, Denmark. Rats were group-housed with regular 12-hour light/dark cycles, in clear plastic containers with ad libitum access to food and water and underwent at least one week of habituation. The intestines were removed, and third-order mesenteric arteries were dissected in ice-cold physiological saline solution containing (in mM): 121 NaCl, 2.8 KCl, 1.6 CaCl2, 25 NaHCO3, 1.2 KH2HPO4, 1.2 MgSO4, 0.03 EDTA, and 5.5 glucose. Segments, 2 mm in length, of mesenteric artery were mounted on 40 μm stainless steel wires in a myograph (Danish Myo Technology, Aarhus, Denmark) for isometric tension recordings. The chambers of the myograph contained PSS maintained at 37°C and aerated with 95% O2/5% CO2. Changes in tension were recorded by PowerLab and Chart software (ADInstruments, Oxford, United Kingdom). The arteries were equilibrated for 30 minutes and normalized to passive force. Artery segments were precontracted with 10 μM methoxamine (Sigma; Copenhagen, Denmark) in the absence or presence of linopirdine (10 μM) (Sigma; Copenhagen, Denmark), before application of ECG, EGCG or EC (Sigma; Copenhagen, Denmark).
7
Mesenteric Artery Tension Measurements
8
Bladder Smooth Muscle Contractility Assay
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Bladders were pinned on a small Sylgard block, and the muscle was dissected free of the mucosal tissue. BSM strips were then cut longitudinally (two mm wide and seven mm long). Muscle strips were mounted in an SI-MB4 tissue bath system (World Precision Instruments, FL, USA). Force sensors were connected to a TBM 4M transbridge (World Precision Instruments), and the signal was amplified by PowerLab (AD Instruments, CO, USA) and monitored through Chart software (AD Instruments). BSM strips were gently prestretched to optimize contraction force and then pre-equilibrated for at least 1 h. All experiments were conducted at 37 °C in physiological saline solution (in mM: 120 NaCl, 5.9 KCl, 1.2 MgCl2, 15.5 NaHCO3, 1.2 NaH2PO4, 11.5 glucose, and 2.5 mM CaCl2) with continuous bubbling of 95% O2/5% CO2. Contraction force was sampled at 2000/s using Chart software. BSM tissue was treated with agonists or antagonists and/or subjected to electrical field stimulation (EFS). BSM strip EFS was carried out using a Grass S48 field stimulator (Grass Technologies, RI, USA) using previously described standard protocols (Rajandram et al., 2016 (link); Chen et al., 2020 (link)).
9
Contractile Properties of Detrusor Muscle
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Following removal of the urothelium, whole detrusor muscle was cut into half and strips were mounted in a 10 ml organ bath (filled with gassed Krebs-bicarbonate solution at 37°C) and connected to a UF1 force transducer. Resting tension was adjusted to 1 g for 30 min and the Krebs solution was changed every 10 min. ATP (10 μm , 100 μm and 1 mm ) and KCl (25–65 mm ) were bath applied with a 20–30 min washout period between concentrations. Bethanechol was bath applied cumulatively (1, 10 and 100 μm ). Tension recordings were obtained using a PowerLab data acquisition system and Chart software (ADInstruments, Colorado Springs, CO, USA). Contraction amplitude was normalised to tissue weight.
10
Measuring Right Ventricular Pressure in Mice
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The mice were anesthetized with an i.p. injection of pentobarbital sodium (50 mg/kg). The mice were orally intubated, and the lungs were ventilated using a mouse ventilator (Shinano Manufacturing Co., Ltd.). The ventilator settings were adjusted (tidal volume, 6 μL/g; frequency, 170–190/min). A 1.4F pressure–volume catheter (catalog SPR-671NR; Millar Instruments) was inserted into the right external jugular vein and advanced into the right ventricle to measure right ventricular pressure (RVP). RVP signals were relayed to pressure amplifiers (PCU-2000; AD Instruments) and then were continuously sampled using a PowerLab system (AD Instruments) and recorded on a computer using Chart software (AD Instruments). Heart rate was typically between 300 and 500 bpm under these conditions. If the heart rate fell below 300 bpm, the measurements were excluded from the analysis.
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