AHR to MCh was determined on day 33 by single-chamber body plethysmography as described previously (20 (link)). In brief, each mouse was placed in the Buxco single chamber, acclimatized for 5–10 min and baseline recordings were made for 5 min. The mice were then challenged with PBS or various concentrations of MCh aerosol and signals were recorded with in-built software (Buxco) to determine the Penh values, which are reliable in BALB/c mice (20 (link)). MCh PC200, which is the partial concentration of MCh required to double the baseline Penh, Penh0, was then calculated. To confirm the findings of the non-invasive body plethysmography, respiratory mechanics were determined during mechanical ventilation (FlexiVent System; Scireq, Montréal, Canada) as previously described (20 (link)). In brief, following anesthetisia via intraperitoneal administration of sodium pentobarbital (90 mg/kg; Wuhan Dinghui Chemical Co., Ltd.) mice were intubated after tracheostomy, ventilated with a computer-controlled ventilator, and airway resistance with various concentrations of MCh was estimated using the FlexiVent System (Scireq) that integrates the ventilator with the respiratory mechanics.
Flexivent system
Manufactured by SCIREQ
Sourced in Canada, Macao, United States
The FlexiVent system is a precision lung function measurement device. It is designed to assess the mechanical properties of the respiratory system in small laboratory animals. The FlexiVent system uses the forced oscillation technique to provide detailed measurements of lung function parameters.
Automatically generated - may contain errors
Lab products found in correlation
Methacholine, by Merck Group (27 mentions)
Pentobarbital sodium, by Merck Group (5 mentions)
Pancuronium bromide, by Merck Group (4 mentions)
Hdm extract, by Stallergenes Greer (3 mentions)
Flexiware software, by SCIREQ (3 mentions)
Methacholine, by SCIREQ (3 mentions)
Rompun, by Bayer (3 mentions)
Flexivent, by SCIREQ (3 mentions)
Whole body plethysmography, by EMKA Technologies (3 mentions)
Flexivent software, by SCIREQ (3 mentions)
Acetyl β methylcholine chloride, by Merck Group (3 mentions)
C57bl 6 mice, by Charles River Laboratories (3 mentions)
Flexiware, by SCIREQ (2 mentions)
Ovalbumin (ova), by Merck Group (2 mentions)
Flexivent, by Merck Group (2 mentions)
Methacholine mch, by Merck Group (2 mentions)
Acetyl β methylcholine chloride methacholine, by Merck Group (2 mentions)
Mir 206 agomir, by RiboBio (1 mentions)
Flexivent 7, by SCIREQ (1 mentions)
Pancuronium bromide, by Pfizer (1 mentions)
223 protocols using flexivent system
1
Airway Hyperresponsiveness Measurement
2
HDM-Induced Asthma Model in Mice
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Female C57BL/6 mice were obtained from the Experimental Animal Center of Hubei Province (Wuhan, China). The model was established by HDM sensitization and challenge. Briefly, female C57BL/6 mice received an i.p. injection of 100 μL of a solution of lyophilized HDM extract (0.1 mg/mL; Greer Laboratories) and Al(OH)3 as an adjuvant on days 0, 7, and 14, and received 40 μL of HDM solution (3 mg/mL) or saline intranasally on days 21, 22, and 23. miR-206 agomir (5 nmol in 40 μL saline; RiboBio), control agomir, miR-206 antagomir (20 nmol in 40 μL saline), or control antagomir were administered intranasally on days 20 and 22. Twenty-four hours after the last HDM challenge, respiratory resistance in response to a range of concentrations of i.v. acetylcholine was measured using the forced oscillation technique with the FlexiVent system (SCIREQ) as previously described (73 (link)).
3
Rabbit Pulmonary Function Testing
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On PND1, pressure–volume and forced oscillation maneuvers were performed using the FlexiVent system (SciReq; FlexiVent, Montreal, QC, Canada). After sedation with ketamine (35 mg/kg) and xylazine (6 mg/kg), a tracheostomy was performed, allowing the insertion of an 18-gauge metal cannula into the trachea. Rabbits were ventilated with a tidal volume of 10 mL/kg and positive end-expiratory pressure of 3 cmH2O at a rate of 120 breaths/min. In order to maximally inflate the lungs and standardize lung volume, two deep inflation maneuvers were performed prior to PFT until reaching a pressure of 30 cmH2O. Both pressure–volume (inspiratory capacity, static compliance, and static elastance) and forced oscillation tests (tissue damping, tissue elastance, central airway resistance, respiratory system resistance, dynamic compliance, and dynamic elastance) were performed as previously described [24 (link)]. The mean of 3 measurements for each maneuver, with a coefficient of determination > 95%, was calculated and used as a single data point for analysis.
4
Pulmonary Function Testing in Kittens
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On PND 1, pressure-volume and forced oscillation maneuvers were performed using the FlexiVent system (SciReq; FlexiVent, Montreal, QC, Canada) in the first group of kittens. After sedation with ketamine (35 mg/kg) and xylazine (6 mg/kg), a tracheostomy was performed, enabling the insertion of an 18-gauge metal cannula into the trachea. Rabbits were ventilated with a tidal volume of 10 mL/kg and positive end-expiratory pressure of 3 cmH2O at a rate of 120 breaths/min. To maximally inflate the lungs and standardize lung volume, two deep inflation maneuvers were performed prior to PFT until reaching a pressure of 30 cm H2O. Both pressure-volume (inspiratory capacity, static compliance, and static elastance) and forced oscillation tests (tissue damping, tissue elastance, central airway resistance, respiratory system resistance, dynamic compliance, and dynamic elastance) were performed as previously described [24 (link)]. The mean of three separate measurements for each maneuver, with a coefficient of determination >95%, was calculated and used as a single data point for analysis.
5
Evaluating Lung Function in Irradiated Mice
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Lung function in irradiated mice was evaluated using a flexiVentTM system (SCIREQ, Montreal, QC, Canada), which measures flow-volume relationships in the respiratory system—including forced oscillation—to discriminate between airway and lung tissue variables27 (link). This system was used according to the manufacturer’s instructions.
6
Methacholine-Induced Lung Function Evaluation
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The lung function was determined at 10 and 21 dpi using a FlexiVentTM system (SCIREQ). As a control, naïve mice were instilled intratracheally with 10 µg LPS in 50 µL PBS and subjected to lung function measurements 48 h later. In short, mice were sedated with 150 mg/kg pentobarbital, intubated with a 19 G blunt tip canula and attached to the FlexiVentTM ventilator. Mice were challenged via nebulization with an increasing dose of methacholine (0–50 mg/mL in PBS). Pre-set ventilation protocols of the FlexiVentTM apparatus were applied to characterize resistance, elastance, airway resistance, tissue damping and tissue elastance.
7
Lung Function Measurement via FlexiVent
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Lung function measurements were obtained using FlexiVentTM system (Scireq, Canada) equipped with FlexiVent 7 operating software. Mice were mechanically ventilated at a rate of 150 breaths/min, tidal volume of 10 ml/kg, and a positive end‐expiratory pressure (PEEP) of 3 cmH2O for 3 min. Deep inflation perturbation (slow inflation of the lung to 30 cmH2O) was initiated to measure the total lung capacity. Snapshot perturbation maneuver was imposed to measure the lung compliance, which is obtained by fitting the maneuver signals to a single compartment model of the lung by the operating software. This perturbation was repeated for 3–4 times every 30 s. Then, broadband low‐frequency forced oscillation measurements (1–20.5 Hz) were performed using “Quick Prime‐3 perturbation”. The resulting respiratory input impedance was fit to the constant phase model of the lung by the operating software, and lung tissue resistance was obtained. Similarly to snapshot perturbation, Quick Prime‐3 perturbation was repeated three to four times every 30 s. A coefficient of determination of 0.95 was the lower limit for accepting perturbation outcomes.
8
Comprehensive Lung Function Assessment
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Lung function were measured using FlexiVentTM system (Scireq, Canada). Mice underwent mechanical ventilation in the condition of 150 breaths/min, 10 ml/kg tidal volume, and 3 cm H 2 O positive end-expiratory pressure. Total lung capacity was examined at deep inflation perturbation (30 cm H 2 O). Lung compliance was measured using a snapshot perturbation maneuver and repeated 3-4 times per 30 s. Then lung tissue resistance was detected by broadband low-frequency forced oscillation measurements (1-20.5 Hz) and repeated 3-4 times per 30 s.
9
Methacholine-Induced Airway Responses Post-Infection
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Airway responses to methacholine (Sigma Chemical, St. Louis, MO) were assessed with the Scireq Flexivent system (Scireq, Montreal, QC, Canada) at 7 days post infection (dpi). Animals were anesthetized with ketamine/xylazine (10 mg/kg) and paralysis was induced with 1 mg/kg pancuronium bromide IP (Sigma). The linear single-compartment model was used to assess total respiratory system resistance (Rn). methacholine dose responses were determined as previously described [13 (link), 21 (link)].
10
Airway Responses to Methacholine
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Airway responses to methacholine (Sigma Chemical, St. Louis, MO) were assessed with the Scireq Flexivent system (Scireq, Montreal, QC, Canada) 1-week post RSV challenge and IP injection of control IgG or anti-LIF IgG. Animals were anesthetized with ketamine/xylazine (10 mg/kg) and paralysis was induced with 1 mg/kg pancuronium bromide IP (Sigma). The linear single-compartment model was used to assess total respiratory system resistance (Rn). methacholine dose responses were determined.
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