Adult male Sprague-Dawley rats (Harlan, Indianapolis, IN) weighing between 295 and 355 g were used. Ketamine (65 mg/kg i.p.) and dexdormitor (0.25 mg/kg i.p.) were used for operative anesthesia. For dual probe experiments, concentric microdialysis probes (250 µm diameter) were implanted unilaterally in both the VTA (1 mm long probe) and NAc (1.5 mm long probe) according to following coordinates from bregma and dura: anterior-posterior (AP) −5.3, medial-lateral (ML) ± 0.5, dorsal-ventral (DV) − 8.0 mm and AP + 1.2, ML ± 1.4 and DV − 7.8 mm, respectively. For single probe experiments, probes (3 mm long) were implanted into the mPFC from bregma and dura: AP +3.0, ML ± 0.5, DV − 4.0 (Paxinos and Watson 2007). Probes were secured to the skull by acrylic dental cement and metallic screws. Following surgery, rats were allowed to recover and experiments were performed later. Animals were awake and freely moving with access to food and water throughout the experiment. Microdialysis probes were flushed at 1.5 µL/min with aCSF for 3 h using a Chemyx (Stafford, TX) Fusion 400 syringe pump. Perfusion flow rate was reduced to 0.6 µL/min and samples were collected every 20 min for VTA-NAc experiments. For mPFC experiments, perfusion flow rates were reduced to 1 µL/min to generate 1 µL samples. 1 µL fractions were diluted with 4 µL aCSF, then treated same way as 5 µL sample described below. Following collection of basal fractions, VTA lines were switched to aCSF containing 50 µM bicuculline for the duration (2 h) of experiments. Though 12 µL of dialysate were collected per fraction, only 5 µL of total volume was used for analysis. For mPFC, following 20 min of basal fraction collections, 1 µM neostigmine was perfused through the probe for 5 min.
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Neostigmine
Neostigmine
Neostigmine is a cholinesterase inhibitor used to treat myasthenia gravis, intestinal and urinary tract atony, and to reverse the effects of neurlomuscular blocking agents.
It works by inhibiting the enzyme acetylcholinesterase, which breaks down the neurotransmitter acetylcholine, thereby increasing the concentraton of acetylcholine at the neuromuscular junction and enhancing neurlomascular transmission.
Neostigmine is also employed as a research tool to study cholinergic mechanisms.
Optimal use of Neostigmine in research can be facilitated by PubCompare.ai, an AI-driven solution that helps locate the best protocols and products from literature, preprints, and patents to enhance reproducibility and accuracy, taking the guesswork out of Neostigmine research.
It works by inhibiting the enzyme acetylcholinesterase, which breaks down the neurotransmitter acetylcholine, thereby increasing the concentraton of acetylcholine at the neuromuscular junction and enhancing neurlomascular transmission.
Neostigmine is also employed as a research tool to study cholinergic mechanisms.
Optimal use of Neostigmine in research can be facilitated by PubCompare.ai, an AI-driven solution that helps locate the best protocols and products from literature, preprints, and patents to enhance reproducibility and accuracy, taking the guesswork out of Neostigmine research.
Most cited protocols related to «Neostigmine»
Adult
Animals
Bicuculline
Cranium
Dental Anesthesia
Dental Cements
Dialysis Solutions
Dura Mater
Food
Ketamine
Males
Metals
Microdialysis
Neostigmine
Operative Surgical Procedures
Perfusion
Rats, Sprague-Dawley
Rattus
Syringes
Acetylcholine
Brain
Buffers
Cannula
Cerebrospinal Fluid
Dry Ice
Freezing
Glucose
Immobilization
Isoflurane
isopentane
Microdialysis
Microtomy
Neostigmine
paraform
Phosphates
Saline Solution
Seahorses
Sodium Chloride
Sucrose
Anesthetics
Animals
Atropine
BLOOD
Cerebrovascular Accident
Cornea
Desiccation
Eyelids
Females
Femoral Artery
fMRI
Head
Hydrostatic Pressure
Institutional Animal Care and Use Committees
Intubation
Isoflurane
Ketamine
Mechanical Ventilator
Neostigmine
Ointments
Oral Cavity
Oximetry
Papio
Papio hamadryas
Photic Stimulation
Rate, Heart
Rectum
Respiratory Rate
Saphenous Vein
Vecuronium
Veins
Ellman’s colorimetric method was used [77 (link)] with modifications described previously [78 (link)]. Tested sample (10 μL) at concentration 5 mg/mL was mixed with 20 μL of AChE (or BChE) solution (0.28 U/mL) and completed after 5 min with 35 μL of ATChI (or BTCh) (1.5 mmol/L), 175 μL of 0.3 mmol/L DTNB (containing 10 mmol/L NaCl and 2 mmol/L MgCl2) and 110 μL with Tris-HCl buffer (50 mmol/L, pH 8.0). Samples containing 10 μL of Tris-HCl buffer instead of the studied sample were run in the same way (“blank” samples). The increase in the absorbance due to the spontaneous hydrolysis of the substrate was monitored using “blank” samples containing ATCh (or BTCh) and DTNB completed to 350 μL with Tris-HCl buffer. All samples were incubated at 22 °C (30 min; incubation time was determined after optimization experiments, details not shown), and the absorbance was measured (405 nm, 96-well microplate reader, Tecan Sunrise, Grödig, Austria). The “false-positive” effect of studied compounds was measured according to Rhee et al. [79 (link)] with minor modifications, as described previously [78 (link)]: after mixing of the substrate with the enzyme and buffer, the “false-positive” sample was left for incubation. Then, a studied sample and DTNB were added, followed by an immediate measurement of the absorbance.
Reference cholinesterase inhibitors were used for the calculations of results (eserine, neostigmine, magniflorine, rivastigmine and donepezil). For this purpose, for each compound, 16 dilutions in pure DMSO were prepared (2.57–41.14 μg/mL). These solutions (10 μL) were tested as described above and calibration curves were produced.
Each sample was analyzed in at least eight repeats, and all solutions used in a set of analyses were prepared in the same buffer. For calculations, the background of the sample (10 μL mixed with 340 μL of Tris buffer) was measured at 405 nm and subtracted during calculations. Then, the absorbance of the test sample was subtracted from the absorbance of the “blank” sample.
Reference cholinesterase inhibitors were used for the calculations of results (eserine, neostigmine, magniflorine, rivastigmine and donepezil). For this purpose, for each compound, 16 dilutions in pure DMSO were prepared (2.57–41.14 μg/mL). These solutions (10 μL) were tested as described above and calibration curves were produced.
Each sample was analyzed in at least eight repeats, and all solutions used in a set of analyses were prepared in the same buffer. For calculations, the background of the sample (10 μL mixed with 340 μL of Tris buffer) was measured at 405 nm and subtracted during calculations. Then, the absorbance of the test sample was subtracted from the absorbance of the “blank” sample.
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Buffers
Cholinesterase Inhibitors
Colorimetry
Dithionitrobenzoic Acid
Donepezil
Enzymes
Eserine
Hydrolysis
Magnesium Chloride
Neostigmine
Pain
Rivastigmine
Sodium Chloride
Sulfoxide, Dimethyl
Technique, Dilution
Tromethamine
This is a prospective, single-blinded observational study done with the clearance of Ethical Committee. Informed written consent of 260 patients of either sex, aged 18–65 years, American Society of Anesthesiologist Status I and II undergoing elective surgical procedures under general anesthesia were enrolled for the study. Uncooperative and unwilling patients, history of burns involving head and neck, trauma and airway surgeries, tumor or mass in the neck or airway, patients with restricted mobility at neck and mandible, patients with inability to sit, edentulous or need awake intubation, pregnant females, and patients with body mass index (BMI) ≥35 were excluded from the study. All patients were examined preoperatively to assess airway parameters, the day before surgery by the same anesthesiologist to avoid interobserver variability.
Height and weight were recorded and BMI calculated. Height was measured in centimeters from vertex to heel with the patient standing.
The oropharyngeal view was assessed using:
Table 1 .
Patients were kept nil orally for 8–10 h preoperatively. In operation theater, intravenous (IV) line was secured with 18-gauge IV cannula and Ringer's lactate infusion was started. Electrocardiogram, noninvasive blood pressure, and peripheral oxygen saturation monitor were connected to the patient, and basal heart rate, blood pressure, and oxygen saturation were recorded. Patient was premedicated with injection glycopyrrolate 0.01 mg/kg, injection midazolam 0.05 mg/kg, injection fentanyl 2 μcg/kg intravenously, and preoxygenated with 100% oxygen. Induction of anesthesia was done with injection propofol 2 mg/kg body weight intravenously and injection vecuronium 0.1 mg/kg IV was administered once mask ventilation confirmed. Laryngoscopy was done using Macintosh blade Size 3 or 4 by an experienced anesthesiologist who was blinded to preoperative airway assessment details, and the view was classified as per Cormack-Lehane's Scale:[20 ] Grade 1 - vocal cords visible, Grade 2 - only posterior commissure or arytenoids visible, Grade 3 - only epiglottis visible, and Grade 4 - none of the above visible without any external laryngeal manipulation.
Cormack and Lehane Grade 1 and 2 was considered as easy visualization whereas Grade 3 and 4 was considered as difficult visualization. A maximum of three attempts were allowed with conventional laryngoscope. In case of failure of first two attempts, third attempt was by another senior experienced anesthesiologist. If there was failure to intubate at third attempt, alternate measures such as use of supraglottic device, bougie was done as per the discretion of attending anesthesiologist. External laryngeal manipulation was used to improve visualization after first attempt. Use of additional gadgets/maneuvers during intubation was noted. Oxygenation was ensured in between attempts at intubation. Intubation was done with appropriate sized endotracheal tubes. Confirmation of intubation was done by bilateral auscultation of lung fields and capnography. Number of attempts at intubation was noted. Maintenance of anesthesia was done with oxygen, nitrous oxide, and isoflurane. At the end of surgery, isoflurane disconnected, and after adequate respiratory efforts, injection neostigmine and injection glycopyrrolate were administered to reverse neuromuscular blockade. Patient was extubated after adequate recovery and shifted to the postanesthesia care unit.
Height and weight were recorded and BMI calculated. Height was measured in centimeters from vertex to heel with the patient standing.
The oropharyngeal view was assessed using:
MMT:[17 (link)] Sampson and Young's modification of Mallampati test recorded oropharyngeal structures visible upon maximal mouth opening. Each patient when seated was asked to open mouth maximally and to protrude the tongue without phonation. The view was classified as Grade 0 - epiglottis visualized, Grade 1 - good visualization of palate, fauces, uvula, and tonsillar pillars, Grade 2 - pillars obscured by the base of the tongue but the soft palate, fauces, and uvula visible, Grade 3 - soft palate and base of the uvula visible, and Grade 4 - soft palate not visible
RHTMD:[18 (link)] TMD was measured from the bony point of the mentum to thyroid notch while head was fully extended and mouth closed. RHTMD was calculated as RHTMD = height (in cm)/TMD (in cm) and graded as Grade 1 <23.5 and Grade 2 ≥23.5
Upper lip bite test:[19 (link)] ULBT was done to assess the range of freedom of the mandibular movement and the architecture of the teeth concurrently. It was done by assessing the ability of the patient to touch the vermilion line of upper lip with lower incisors. This test was graded as Class 1 - If the lower incisors could bite the upper lip above the vermilion line, Class 2 - If the lower incisors could bite the upper lip below the vermilion line, and Class 3 - If the lower incisors could not bite the upper lip
IIG:[19 (link)] It was assessed by asking each patient to open the mouth to maximum extent. The distance between upper and lower incisor at the midline is measured, which is usually >3.5 cm
TMD:[15 (link)] TMD was measured from the bony point of the mentum whereas the head is fully extended and mouth closed using a rigid ruler. The distance was rounded to nearest 0.5 cm and graded as Class 1: >6.5 cm, Class 2: 6–6.5 cm, and Class 3: <6 cm
SMD:[15 (link)] SMD was measured from sternal notch to the mentum in centimeter with head fully extended on the neck with the mouth closed which is normally >12.5 cm
Horizontal length of the mandible:[15 (link)] It was measured from angle of the mandible to the mentum. A length of ≥9 cm was considered normal
Maximum range of HNM:[11 (link)] was noted as Grade 1 ≤80° or Grade 2 ≥80°. The patient was first asked to extend the head and neck fully, where a pencil was placed vertically on the forehead and then while the pencil was held firmly in position, the head and neck were flexed.
Patients were kept nil orally for 8–10 h preoperatively. In operation theater, intravenous (IV) line was secured with 18-gauge IV cannula and Ringer's lactate infusion was started. Electrocardiogram, noninvasive blood pressure, and peripheral oxygen saturation monitor were connected to the patient, and basal heart rate, blood pressure, and oxygen saturation were recorded. Patient was premedicated with injection glycopyrrolate 0.01 mg/kg, injection midazolam 0.05 mg/kg, injection fentanyl 2 μcg/kg intravenously, and preoxygenated with 100% oxygen. Induction of anesthesia was done with injection propofol 2 mg/kg body weight intravenously and injection vecuronium 0.1 mg/kg IV was administered once mask ventilation confirmed. Laryngoscopy was done using Macintosh blade Size 3 or 4 by an experienced anesthesiologist who was blinded to preoperative airway assessment details, and the view was classified as per Cormack-Lehane's Scale:[20 ] Grade 1 - vocal cords visible, Grade 2 - only posterior commissure or arytenoids visible, Grade 3 - only epiglottis visible, and Grade 4 - none of the above visible without any external laryngeal manipulation.
Cormack and Lehane Grade 1 and 2 was considered as easy visualization whereas Grade 3 and 4 was considered as difficult visualization. A maximum of three attempts were allowed with conventional laryngoscope. In case of failure of first two attempts, third attempt was by another senior experienced anesthesiologist. If there was failure to intubate at third attempt, alternate measures such as use of supraglottic device, bougie was done as per the discretion of attending anesthesiologist. External laryngeal manipulation was used to improve visualization after first attempt. Use of additional gadgets/maneuvers during intubation was noted. Oxygenation was ensured in between attempts at intubation. Intubation was done with appropriate sized endotracheal tubes. Confirmation of intubation was done by bilateral auscultation of lung fields and capnography. Number of attempts at intubation was noted. Maintenance of anesthesia was done with oxygen, nitrous oxide, and isoflurane. At the end of surgery, isoflurane disconnected, and after adequate respiratory efforts, injection neostigmine and injection glycopyrrolate were administered to reverse neuromuscular blockade. Patient was extubated after adequate recovery and shifted to the postanesthesia care unit.
Anesthesia
Anesthesiologist
ARID1A protein, human
Arytenoid Cartilage
Auscultation
Blood Pressure
Body Weight
Bones
Burns
Cannula
Capnography
Cell Respiration
Chin
Craniocerebral Trauma
Dental Occlusion
Elective Surgical Procedures
Electrocardiography
Epiglottis
Fentanyl
Forehead
General Anesthesia
Glycopyrrolate
Head
Heel
Incisor
Index, Body Mass
Intubation
Isoflurane
Lactated Ringer's Solution
Lamina 3
Laryngoscopes
Laryngoscopy
Larynx
Lung
Mandible
Medical Devices
Midazolam
Muscle Rigidity
Neck
Neck Injuries
Neoplasms
Neostigmine
Neuromuscular Block
Operative Surgical Procedures
Oral Cavity
Oropharynxs
Oxide, Nitrous
Oxygen
Oxygen Saturation
Palate
Palate, Soft
Palatine Tonsil
Patients
Phonation
Pregnant Women
Propofol
Range of Motion, Articular
Rate, Heart
Respiratory Rate
Saturation of Peripheral Oxygen
Sternum
Surgery, Day
Thyroid Gland
Tongue
Tooth
Touch
Uvula
Vecuronium
Vocal Cords
Most recents protocols related to «Neostigmine»
All patients received general anesthesia, either alone or in combined with regional nerve block (including paravertebral nerve block, epidural anesthesia, and intercostal nerve block.) according to the type of surgery. Patients underwent lobectomy or sublobectomy according to surgeon’s comprehensive evaluation based on patient’s condition.
Anesthesia induction used propofol and/or etomidate, sufentanil, and rocuronium or cisatracurium. Anesthesia maintenance used sevoflurane or propofol combined with remifentanil or sufentanil. Rocuronium or cisatracurium was used to maintain muscle relaxation. Supplemental drugs such as flurbiprofen axetil were administered when necessary. The aim was to maintain BIS 40-60, blood pressure within 20% of baseline, and temperature 36-37°C.
Double-lumen endotracheal tube of sizes Ch33-39 was used for lung isolation according to patient height. The ventilation mode was volume control mode with 6-8 ml/kg of tidal volume (TV) during two-lung ventilation and 5-6 ml/kg during one-lung ventilation (OLA), and 0-5 cmH2O of positive end-expiratory pressure (PEEP), and 12-20 breaths/min of respiratory rates. The aim was to maintain PETCO2 35-45 mmHg and SpO2 ≥92%. At the end of anesthesia, neostigmine was used to antagonize muscular relaxant before extubation.
Fluid infusion was administrated with crystalloid at a rate of 4–6 mL/kg-1h-1. Colloids or blood product was used according to anesthesiologist’s comprehensive evaluation based on patient’s condition. Patient-controlled intravenous analgesia was used after surgery for postoperative analgesia to maintain numeric rating scales (NRS) ≤ 3 scores.
Anesthesia induction used propofol and/or etomidate, sufentanil, and rocuronium or cisatracurium. Anesthesia maintenance used sevoflurane or propofol combined with remifentanil or sufentanil. Rocuronium or cisatracurium was used to maintain muscle relaxation. Supplemental drugs such as flurbiprofen axetil were administered when necessary. The aim was to maintain BIS 40-60, blood pressure within 20% of baseline, and temperature 36-37°C.
Double-lumen endotracheal tube of sizes Ch33-39 was used for lung isolation according to patient height. The ventilation mode was volume control mode with 6-8 ml/kg of tidal volume (TV) during two-lung ventilation and 5-6 ml/kg during one-lung ventilation (OLA), and 0-5 cmH2O of positive end-expiratory pressure (PEEP), and 12-20 breaths/min of respiratory rates. The aim was to maintain PETCO2 35-45 mmHg and SpO2 ≥92%. At the end of anesthesia, neostigmine was used to antagonize muscular relaxant before extubation.
Fluid infusion was administrated with crystalloid at a rate of 4–6 mL/kg-1h-1. Colloids or blood product was used according to anesthesiologist’s comprehensive evaluation based on patient’s condition. Patient-controlled intravenous analgesia was used after surgery for postoperative analgesia to maintain numeric rating scales (NRS) ≤ 3 scores.
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Anesthesia
Anesthesiologist
BLOOD
Blood Pressure
cisatracurium
Colloids
Epidural Anesthesia
Etomidate
flurbiprofen axetil
General Anesthesia
isolation
Lung
Management, Pain
Muscle Tissue
Neostigmine
Nerve Block
One-Lung Ventilation
Operative Surgical Procedures
Patient-Controlled Analgesia
Patients
Pharmaceutical Preparations
Positive End-Expiratory Pressure
Propofol
Relaxations, Muscle
Remifentanil
Respiratory Rate
Rocuronium
Saturation of Peripheral Oxygen
Sevoflurane
Solutions, Crystalloid
Sufentanil
Surgeons
Tidal Volume
Tracheal Extubation
In addition to the recordings provided by the hybrid diffuse optical setup, , heart rate (HR) and mean arterial pressure (MAP) were extracted from either an anesthesia monitor (Datex-Ohmeda Aisys™, GE Healthcare, Little Chalfont, United Kingdom) by the open-source VitalSigns capture (VScapture) program60 (link) or from a general monitor (Philips IntelliVue MX800, Koninklijke Philips N.V.). The BIS data were acquired by a BIS sensor (BIS Vista™, Medtronic plc, IRL). These signals were recorded by the same monitor with the same timestamps and were synchronized with the optical signals with a precision of 1 s and according to the beginning of the measurement.
General anesthesia was performed under total intravenous anesthesia with propofol at a concentration of 1% (Propofol Fresenius®, Fresenius Kabi Deutschland GmbH, Bad Homburg, Germany) using the Schneider model of target-controlled infusion anesthesia61 (link) based on age, height, weight, and gender of the patient3 ,62 (link) implemented in a TIVA system (Alaris Asena® PK, Becton, Dickinson and Company, Franklin Lakes, New Jersey). After induction and tracheal intubation, pulmonary volume-controlled ventilation was maintained with a fraction of inspired oxygen ( ) of 0.5, a tidal volume of 6 to , a respiration rate between 12 and 16 breaths per minute, a positive end-expiratory pressure of 4 to 6 mmHg, and a MAP between 60 and 80 mmHg.
It should be noted that the patients received further medications, such as fentanyl for analgesia,63 (link)– 65 (link, link) atropine sulfate, atracurium besylate, or rocuronium bromide to obtain neuromuscular blockade, remifentanil, neostigmine, lidocaine, and midazolam. Some of them are known to influence cerebral hemodynamics (i.e., fentanyl,63 (link)– 65 (link, link) midazolam,66 (link) remifentanil,67 (link) atropine sulfate,68 (link) and lidocaine69 (link)). Our study is not designed to investigate these interactions and it is not trivial to consider these effects since they are not well known. Since the primary goal of the study the direct comparison of the optical and BIS signals, we have decided to remove the duration of their administration from all the signals as described in the following section.
General anesthesia was performed under total intravenous anesthesia with propofol at a concentration of 1% (Propofol Fresenius®, Fresenius Kabi Deutschland GmbH, Bad Homburg, Germany) using the Schneider model of target-controlled infusion anesthesia61 (link) based on age, height, weight, and gender of the patient3 ,62 (link) implemented in a TIVA system (Alaris Asena® PK, Becton, Dickinson and Company, Franklin Lakes, New Jersey). After induction and tracheal intubation, pulmonary volume-controlled ventilation was maintained with a fraction of inspired oxygen ( ) of 0.5, a tidal volume of 6 to , a respiration rate between 12 and 16 breaths per minute, a positive end-expiratory pressure of 4 to 6 mmHg, and a MAP between 60 and 80 mmHg.
It should be noted that the patients received further medications, such as fentanyl for analgesia,63 (link)
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Anesthesia
Anesthesia, Intravenous
Atracurium Besylate
Fentanyl
General Anesthesia
Hemodynamics
Hybrids
Intubation, Intratracheal
Lidocaine
Management, Pain
Midazolam
Neostigmine
Neuromuscular Block
Oxygen
Patients
Pharmaceutical Preparations
Positive End-Expiratory Pressure
Propofol
Propofol Fresenius
Rate, Heart
Remifentanil
Respiratory Rate
Rocuronium Bromide
Saturation of Peripheral Oxygen
Sulfate, Atropine
Tidal Volume
Vision
Both groups were treated with tracheal intubation general anesthesia, propofol 2 mg/kg + cisatracurium 0.15 mg/kg + sufentanil 0.4 μg/kg induction, and all patients received propofol, remifentanil, and sevoflurane as intraoperative maintenance drugs. Propofol was maintained at 2.5–5 mg/kg; remifentanil was maintained at 5–10 μg/kg during the operation, according to the patient’s blood pressure and BIS value (aimed for 40–60); an experimental anesthesiologist controlled the dose and recorded the total dose. Because inhaled anesthetics are a risk factor for PONV, we set the MAC of two groups of sevoflurane to 0.3, and the concentration was less than 1%. If the operation lasted more than one hour, we regularly added atracurium 0.03 mg/kg and sufentanil 5 μg per hour. Neostigmine was administered intravenously to reverse the residual effect of the muscle relaxant when the train-of-four (TOF) ratio was less than 0.9 after the operation at 0.05 mg/kg. Before the end of the surgery, all patients were given flurbiprofen axetil 50 mg intravenously for analgesia.
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A-factor (Streptomyces)
Anesthesiologist
Anesthetics
Atracurium
Blood Pressure
cisatracurium
factor A
flurbiprofen axetil
General Anesthesia
Intubation, Intratracheal
Management, Pain
Muscle Tissue
Neostigmine
Operative Surgical Procedures
Patients
Pharmaceutical Preparations
Postoperative Nausea and Vomiting
Propofol
Remifentanil
Sevoflurane
Sufentanil
Cholinesterase activity was measured by Ellman’s method [47 (link)] adapted for microtiter plates, as previously described by Ristovski et al. [9 (link)]. A complex and its ligands were first screened for the IC50 determination and then the inhibitory constants (Ki) were determined. A stock solution of C1 (1 mg/mL) was prepared in 5% v/v DMSO in deionized water, while a stock solution of chlorido analogue C1′ (1 mg/mL) was prepared in 100% methanol (MeOH). Stock solutions of ligands L1 and pta (1 mg/mL) and the positive control (neostigmine methyl sulphate, 1 mg/mL, Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) were prepared in 100% MeOH. These solutions were added to the wells, and gradually diluted in 100 mM of potassium phosphate buffer (pH 7.4) to the final volume of 50 µL. Acetylthiocholine chloride and 5,5′-dithiobis-2-nitrobenzoic acid were then dissolved in the same buffer to the respective final concentrations of 1 and 0.5 mM and added (100 μL) to the wells of the microtiter plates. C1 was screened against a suite of ChEs of human and animal origin (eeAChE, hrAChE, hsBChE) (all three from Sigma-Aldrich, St. Louis, MO, USA), hrBChE (gift from the research group of Professor Stanislav Gobec, Faculty of Pharmacy, University of Ljubljana), and csBChE. All enzymes were dissolved in the 100 mM potassium phosphate buffer (pH 7.4) to 0.0075 U/mL. Fifty μL of each ChE was added to start the reactions, which were followed spectrophotometrically at 405 nm and 25 °C for 5 min using a kinetic microplate reader (Dynex Technologies Inc., Chantilly, VA, USA). The blank reactions without the inhibitors were run with the appropriate dilutions of the solvents, in which the tested compound and positive control were initially diluted (5% aqueous DMSO or 100% MeOH), and the readings were corrected according to the appropriate blanks. At the end of the experiments, the concentrations of compounds causing 50% inhibition of ChE activity (IC50) were determined. To determine C1 inhibition constants (Ki), the kinetics were monitored using three different final substrate concentrations (0.125, 0.25, 0.5 mM). Each measurement was repeated at least three times. Data were analyzed using OriginPro software (OriginPro 2020, OriginLab Corporation, Northampton, MA, USA).
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2-(N-cyclohexylamino)ethanesulfonic acid
Acetylthiocholine
Animals
Buffers
Chlorides
Cholinesterases
dimethyl sulfate
Enzymes
Faculty, Pharmacy
Homo sapiens
inhibitors
Kinetics
Ligands
Methanol
Neostigmine
Nitrobenzoic Acids
potassium phosphate
Psychological Inhibition
Solvents
Sulfoxide, Dimethyl
Technique, Dilution
The hemidiaphragm with phrenic nerve was pinned on its lateral side into a silicon-coated organ bath containing oxygenated Krebs-Ringer (K-R) solution. Then, the tendon part was linked to the lever of an isometric mechano-electrical transducer (Grass Instruments, West Warwick, RI, USA) via a stainless-steel hook and silk thread. The equal stimulus protocol, equipment, and software were used as previously described by Ristovski et al. [9 (link)]. The concentrations of C1 studied were 30, 60, 90, and 120 μM and their effect on muscle contraction was continuously measured during 60 min after application. The muscle twitch and tetanic contraction blockade produced by C1 was shown as the percentage of the initial maximal response. The well-known reversible AChE inhibitor neostigmine methyl sulphate (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) was used as a positive control, and a corresponding solution of 2.5% DMSO in deionized water as a negative control.
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Bath
dimethyl sulfate
Electricity
Krebs-Ringer solution
Muscle Contraction
Muscular Fasciculation
Neostigmine
Pain
Phrenic Nerve
Poaceae
Silicon
Silk
Stainless Steel
Sulfoxide, Dimethyl
Tendons
Toxoid, Tetanus
Transducers
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Neostigmine is a pharmaceutical compound used as a laboratory reagent. It functions as a cholinesterase inhibitor, which means it helps maintain elevated levels of the neurotransmitter acetylcholine by slowing its breakdown. This property makes Neostigmine a useful tool in various research and experimental applications.
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Neostigmine bromide is a laboratory reagent used as a cholinesterase inhibitor. It functions by reversibly inhibiting the enzyme acetylcholinesterase, which is responsible for the breakdown of the neurotransmitter acetylcholine. This property makes neostigmine bromide a useful tool in various biochemical and pharmacological research applications.
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AF-DX 116 is a muscarinic acetylcholine receptor antagonist. It acts as a selective M2 muscarinic receptor antagonist.
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Indomethacin is a laboratory reagent used in various research applications. It is a non-steroidal anti-inflammatory drug (NSAID) that inhibits the production of prostaglandins, which are involved in inflammation and pain. Indomethacin can be used to study the role of prostaglandins in biological processes.
More about "Neostigmine"
Neostigmine, a cholinesterase inhibitor, is widely used to treat a variety of conditions, including myasthenia gravis, intestinal and urinary tract atony, and to reverse the effects of neuromuscular blocking agents.
This drug works by inhibiting the enzyme acetylcholinesterase, which breaks down the neurotransmitter acetylcholine, thereby increasing its concentration at the neuromuscular junction and enhancing neuromuscular transmission.
Neostigmine is also employed as a valuable research tool to study cholinergic mechanisms.
To optimize the use of Neostigmine in research, the AI-driven solution PubCompare.ai can be leveraged.
This tool helps researchers locate the best protocols and products from literature, preprints, and patents, enhancing reproducibility and accuracy, and taking the guesswork out of Neostigmine research.
In addition to Neostigmine, other related compounds and techniques can be valuable in research, such as Donepezil, a cholinesterase inhibitor used in the treatment of Alzheimer's disease, CMA/11 microdialysis probes for measuring neurotransmitter levels, and Nicotine, a nicotinic acetylcholine receptor agonist.
Furthermore, MRS2500, a P2Y1 receptor antagonist, and statistical software like SPSS 26.0 can be used in conjunction with Neostigmine research.
Researchers can also explore Neostigmine bromide, a salt form of Neostigmine, and AF-DX 116, a selective muscarinic M2 receptor antagonist, to gain a more comprehensive understanding of cholinergic mechanisms.
Additionally, Indomethacin, a non-steroidal anti-inflammatory drug, may be used in combination with Neostigmine to investigate its effects on various physiological processes.
By leveraging the insights gained from the MeSH term description and the Metadescription, researchers can optimize their Neostigmine research and enhance its reproducibility and accuracy.
PubCompare.ai provides an AI-driven solution to streamline this process, making it easier for researchers to navigate the vast amount of available information and choose the best protocols and products for their studies.
This drug works by inhibiting the enzyme acetylcholinesterase, which breaks down the neurotransmitter acetylcholine, thereby increasing its concentration at the neuromuscular junction and enhancing neuromuscular transmission.
Neostigmine is also employed as a valuable research tool to study cholinergic mechanisms.
To optimize the use of Neostigmine in research, the AI-driven solution PubCompare.ai can be leveraged.
This tool helps researchers locate the best protocols and products from literature, preprints, and patents, enhancing reproducibility and accuracy, and taking the guesswork out of Neostigmine research.
In addition to Neostigmine, other related compounds and techniques can be valuable in research, such as Donepezil, a cholinesterase inhibitor used in the treatment of Alzheimer's disease, CMA/11 microdialysis probes for measuring neurotransmitter levels, and Nicotine, a nicotinic acetylcholine receptor agonist.
Furthermore, MRS2500, a P2Y1 receptor antagonist, and statistical software like SPSS 26.0 can be used in conjunction with Neostigmine research.
Researchers can also explore Neostigmine bromide, a salt form of Neostigmine, and AF-DX 116, a selective muscarinic M2 receptor antagonist, to gain a more comprehensive understanding of cholinergic mechanisms.
Additionally, Indomethacin, a non-steroidal anti-inflammatory drug, may be used in combination with Neostigmine to investigate its effects on various physiological processes.
By leveraging the insights gained from the MeSH term description and the Metadescription, researchers can optimize their Neostigmine research and enhance its reproducibility and accuracy.
PubCompare.ai provides an AI-driven solution to streamline this process, making it easier for researchers to navigate the vast amount of available information and choose the best protocols and products for their studies.