Twenty-four hours after the final OVA challenge, rats received an intraperitoneal injection of thiopental (20 mg/kg) (Sigma, St. Louis, Missouri, USA) and were euthanized by an injection of potassium chloride (2 mmol/kg) (Sigma, St. Louis, Missouri, USA). After thoracotomy, the lower left lobe of the lungs was removed, washed in PBS and frozen in liquid nitrogen before storage at -80 °C for later determination of eicosanoid concentrations.
Thiopental
Manufactured by Merck Group
Sourced in United States
Thiopental is a barbiturate compound used as a general anesthetic in laboratory settings. It is a white, crystalline powder that is soluble in water and alcohol. Thiopental acts as a central nervous system depressant, inducing sleep and reducing brain activity. Its core function is to provide a controlled and reversible state of unconsciousness for laboratory procedures and experiments.
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Lab products found in correlation
Potassium chloride (kcl), by Merck Group (1 mentions)
Naproxen, by Merck Group (1 mentions)
Heparin, by Merck Group (1 mentions)
Diclofenac, by Merck Group (1 mentions)
Nuclear lysis buffer, by Thermo Fisher Scientific (1 mentions)
Shikonin, by Merck Group (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Formalin buffered saline, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Gentamicin, by Merck Group (1 mentions)
Geraniol, by Merck Group (1 mentions)
Pluronic f 127, by Merck Group (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
Diethyl ether, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Cremophor el, by BASF (1 mentions)
Tween 80, by Merck Group (1 mentions)
9 protocols using thiopental
1
Euthanasia and Lung Tissue Harvesting
2
Pharmacological Evaluation of Compounds
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Naproxen, diclofenac, thiopental and heparin were purchased from Sigma-Aldrich (Prague, Czech Republic). Atorvastatin was purchased from Zentiva Slovakia (Bratislava, Slovakia).
3
Reagent Procurement for Cell Assays
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Shikonin, dimethyl sulfoxide (DMSO), formalin-buffered saline, gentamicin, thiopental, hematoxylin, and eosin were purchased commercially from Sigma, St. Louis, MO, USA. However, phosphate-buffered saline (PBS) was obtained from Gibco, Waltham, MA, USA, yet the lysis buffer and nuclear lysis buffer were obtained from ThermoFisher Scientific, Waltham, MA, USA.
4
Geraniol-based Nanomedicine Formulation
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Geraniol, Pluronic® F127, (Acetonitrile, methanol, and water HPLC grade), thiopental, diethylether and formaldehyde (Sigma-Aldrich, St. Louis, MO, USA); Cremophor® EL (BASF, Ludwigshafen, Germany); Tween 80, hydrochloric acid, potassium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride (Merck-Schuchardt, Hohenbrunn, Germany); synthetic cellulose nitrate membrane 0.22 µm Tuffyrn membrane filter (Sartorius Stedim, Göttingen, Germany).
5
Lung Function Assessment in Small Animals
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Lung function was assessed with a ventilator for small animals (flexiVent™; SCIREQ, Montreal, QC, Canada). After 12 weeks of daily exposure to air or CS, mice were anesthetized with thiopental (250 mg/kg, ip, Sigma-Aldrich, St. Louis, MO, USA) and tracheostomized. Then, a 20G angiocatheter (BD Insyte, Sandy, UT, USA) was inserted into the trachea and a catheter was used to connect the angiocatheter and the ventilator. The tidal volume of the ventilator was set at 10 ml/kg, and respiratory rate was 120 breaths/min with a positive end-expiratory pressure of 3 cm H2O. Lung function was expressed as Newtonian resistance (Rn), respiratory system resistance (Rrs), respiratory system compliance (Crs), and respiratory system elasticity (Ers).
6
Diabetes Induction and Plant Extract Treatment
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The animal experiment was approved by the Ethical Committee of Faculty of Pharmacy, Comenius University in Bratislava (no. 962/10-221). Twenty-four male Wistar rats (DobráVoda, Slovak Republic) weighing 278 ± 5.70 g were used in the experiment. After a brief acclimatization period, rats were randomly divided into 4 groups (6 rats per group): (1) healthy animals—control group; (2) untreated diabetic group (DM was induced by i.p. (intraperitoneal) administration of STZ (Sigma-Aldrich) diluted in citrate buffer (pH 4.5) at a dose of 55 mg/kg); (3) and (4) diabetic animals daily treated for 5 weeks with tested plant extracts. Prior STZ administration blood glucose levels and total body weight of all animals were measured. The blood glucose levels and body weight were subsequently measured 1 and 5 weeks after DM induction.
Following a 5-week period, all rats were killed by anesthetic overdose (thiopental, 100 mg/kg i.p., Sigma-Aldrich). Thoracic aortas and livers were immediately removed, repeatedly rinsed in saline, frozen in liquid nitrogen, and stored at −80 °C until the analysis.
Following a 5-week period, all rats were killed by anesthetic overdose (thiopental, 100 mg/kg i.p., Sigma-Aldrich). Thoracic aortas and livers were immediately removed, repeatedly rinsed in saline, frozen in liquid nitrogen, and stored at −80 °C until the analysis.
7
Electrocardiographic Analysis in Rat 5-FU
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One hour after each 5-FU injection, five to six rats from each group were randomly chosen and anaesthetized with thiopental (50 mg/kg, i.p.; Sigma-Aldrich Co., MO, USA). Rats were kept warm with a heating lamp to avoid the risk of hypothermia. Subcutaneous peripheral limb electrodes were inserted for ECG recording (HPM 7100, Fukuda Denshi, Tokyo, Japan) to determine heart rate (HR), QTc and RR intervals, QRS duration, and to test for ST-segment elevation34 (link).
8
Synthesis and Characterization of Silver Nanoparticles
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In the preparation of the complex, the following reagents were used: βCD hydrate (98%, 1134.98 g/mol), Mel (>95%, 305.2 g/mol), Dimethyl sulfoxide (DMSO)-d6 (99.99% D, 84.17 g/mol), Phosphatidylcholine (99%), Dodecane (99%, 170.33 g/mol), Phosphate buffered saline (PBS) tablets, Thiopental (99%, 242.34 g/mol), and Evans Blue (75%, 960.81) provided by Sigma-Aldrich (Saint Louis, MO, USA). As solvents, ethanol p.a. and water for chromatography LiChrosolv® brand Merck (Darmstadt, Germany) were employed. To obtain the AgNPs via magnetron sputtering, a silver foil of high purity (99.9%) was used as a precursor.
9
Measuring Sciatic Nerve Conduction Velocity in Rats
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After performing the last behavioral assessments (Fig. 1), the rats were anesthetized with thiopental (65 mg/kg, ip; Sigma, St. Louis, MO, USA). Body temperature was monitored and then maintained within normal limits. Furthermore, the motor nerve conduct velocity (MNCV) in the left sciatic nerve was measured using power lab MLT 1030/D device (AD Instruments, Power Lab, Spain) and the same stimulatory and recording pin electrodes (AD Instrument) in terms of the previously described method (Ja'afer et al., 2006) (link). Using a bipolar hook-shaped electrode, in which its cathode was located 2 mm away from the anode, electrical stimulations equal to 10 V were introduced to the proximal section of the sciatic nerve. Compound-muscle action potentials (CMAP) were also recorded by inserting the recording and reference needle electrodes into the gastrocnemius muscle belly and calcaneus tendon, respectively. Finally, the earth electrode was located under the skin. Notably, the velocity was calculated as the ratio between the distances of stimulating and recording pin electrodes (meters) and the corresponding latency difference (seconds).
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