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111 protocols using labchart pro

1

Fetal Breathing Movements Monitoring

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Continuous recordings of tracheal pressures (as a measure of FBMs) and amniotic pressure began 24 h prior to the first saline/LPS infusion (at 129 days of gestation) and continued until the end of the experiment (at 134 days of gestation) using LabChart pro software (Version 8, ADInstruments, Castle Hill, NSW, Australia). Amniotic and tracheal pressures were measured using pressure transducers (cat# ADInstriuments, cat# MLT0699) connected to a quad bridge amp and powerlab (ADInstriuments, cat# FE224 and PL3508). The tracheal and amniotic pressure signals were collected at 1 kHz using a mains filter and stored offline for analyses using LabChart pro (version 8, ADInstruments).
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2

Arterial Pressure and Heart Rate Monitoring in Rats

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At 90 days of age, male offspring rats (n = 5-6 per group) were anesthetized with ketamine (80 mg kg -1 , i.p.) and xylazine (10 mg kg -1 , i.p.) for the insertion of polyethylene catheters into the femoral artery and vein. The catheters were tunneled through the back of the neck and ketoprofen (5 mg kg -1 ) was injected subcutaneously. Rats underwent a period of surgical recovery for 24 h. Arterial pressure (AP) and heart rate (HR) were recorded in conscious animals connected with the arterial catheter to a pressure transducer (LabChartTM Pro, ADInstruments, Bella Vista, NSW, Australia).
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3

Pulsatile Arterial Pressure Analysis

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The pulsatile arterial pressure (PAP) and HR were recorded for 50-60 min under baseline conditions. Values for systolic arterial pressure (SAP), diastolic arterial pressure (DAP) and mean arterial pressure (MAP) were calculated offline by selection of 10 min intervals for each animal (LabChartTM Pro, ADInstruments, Bella Vista, NSW, Australia).
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4

Isometric Aortic Ring Force Measurement

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Isometric force of aortic rings was recorded in organ bath filled with Krebs-Ringer solution as described29 (link) and the data were acquired using LabChart Pro software (ADInstruments, Colorado Springs, CO). Aortas were progressively stretched to their optimal passive tension as assessed by response to 80 mM KCl. The optimal points on length/tension curve were 1.48±0.2g (wild-type), 1.47±0.2g (wild-type + EPO), 1.43±0.3g (hph1), and 1.47±0.3g (hph1 + EPO; n=7). Concentration-dependent response curves to acetylcholine (10-9 - 10-5 M), and diethylammonium (Z)-1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA-NONOate; 10-10 - 10-5 M) were cumulatively obtained during submaximal contractions to phenylephrine. Concentrations of phenylephrine (1-3×10-7 M) were selected in order to obtain the same submaximal contraction in aortic rings from both wild-type (32±3%) and hph1 mice (28±3%) and those treated with EPO (29±3% and 26±4%, respectively).
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5

Cardiac Pressure-Volume Measurements

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A 2F microtip pressure‐volume catheter (SPR‐869, Millar, Houston, TX, USA) was inserted into the LV from the right carotid artery to obtain the pressure‐volume (P‐V) data. The Lab‐Chart Pro software (AD Instruments, Australia) was used to evaluate the LV end‐systolic pressure (LVESP), LV end‐diastolic pressure (LVEDP), maximal slope of the LV systolic pressure increment (dp/dtmax), diastolic pressure decrement (dp/dtmin), and LV ejection fraction (EF). At the end of the experiment, the volume was calibrated using the relative volume unit (RVU) calibration method with fresh heparinized warm blood as previously reported.61
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6

Measuring Intra-Amniotic Pressure Dynamics

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Intra-amniotic pressure was recorded, digitized and analyzed using PowerLab system and LabChartPro (ADInstruments, Colorado Springs, CO) 10 (link). The integrated area under the curve for intra-amniotic pressure was used as the measure of uterine activity and reported as the mean of the cumulative hourly contraction area (HCA, mmHg.sec/hr). The average peak HCA per day prior to inoculation (with U. parvum or media control) represents baseline contractility and was compared to the peak HCA per day from inoculation until delivery (post-inoculation).
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7

Measuring Intra-Amniotic Pressure Dynamics

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Intra-amniotic pressure was recorded, digitized and analyzed using PowerLab system and LabChartPro (ADInstruments, Colorado Springs, CO) 10 (link). The integrated area under the curve for intra-amniotic pressure was used as the measure of uterine activity and reported as the mean of the cumulative hourly contraction area (HCA, mmHg.sec/hr). The average peak HCA per day prior to inoculation (with U. parvum or media control) represents baseline contractility and was compared to the peak HCA per day from inoculation until delivery (post-inoculation).
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8

Cardiovascular and Cerebrovascular Assessment

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All measurements recorded were converted from analog to digital data at 1 kHz (PowerLab, 16/30; ADInstruments, Dunedin, New Zealand)
and stored for offline analysis (LabChart Pro; ADInstruments).
Respiratory data were extracted on a breath-by-breath basis, while cardiovascular and cerebrovascular data were extracted on a beat-bybeat basis. The last 5 min of each condition was used for analysis and averaged for data representation.
Mean arterial blood pressure (MAP) was calculated as:
Cardiac output (CO) was calculated as SV × HR and total peripheral resistance (TPR) calculated as MAP∕CO. Cerebrovascular conductance index (CVCi) was calculated as MCA V mean /MAP (Flück et al., 2017) .
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9

Femoral Artery MAP Monitoring during Nitroprusside

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The mean arterial pressure (MAP) was measured with a catheter in the femoral artery, and the drug delivery was administrated in the femoral vein. n=6 rats were used for this study, 1 to 3 physiological tests per rat. A 1.0-1.5 cm incision in the medial aspect of the leg was made to expose both vessels, and a cannula (0.6mm outer diameter) previously filled with heparinized saline (20 IU/mL) was inserted and secured to the muscle using 4.0 silk sutures. The arterial cannula was connected to a previously calibrated pressure transducer (AD-Instruments, MLT1199) coupled to a bridge amplifier and power supply modulus (AD-Instruments, FE221 and ML826, respectively) for continuously MAP evaluation. The venous cannula was coupled to an infusion system to administrate the vasoactive drug, nitroprusside (5mg/mL, a bolus of 2.5 µg/g weight; 71778 Sigma Aldrich). Simultaneous nerve activity was recorded with the sutrodes placed on the cVN and SNVP-1 to 4. PowerLab data acquisition system (AD Instruments, Colorado Springs, CO) and LabChart Pro (AD Instruments, Colorado Springs, CO) softwares
were used to process and analyze the mean arterial pressure data.
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10

Proton Activation and Ethanol Modulation

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Data are represented as mean ± s.e.m. Proton activation curves represent nonlinear regression fits to the equation R = Rmax / (1 + 10 ^ {[log(EC50 − P)] · nH}), where R is the peak channel response, Rmax is the maximal response, P is the concentration of protons, EC50 is the concentration producing 50% maximal response, and nH is the Hill coefficient; data for each receptor were normalized to its fitted maximal response. Ethanol modulation was calculated as [(RA0,R1>)/0,R1>] × 100, where RA represents the peak EC10 channel response in the presence of ethanol, and 0,R1> represents the mean of the pre- and post-ethanol EC10 responses. Results were analyzed with one-way analysis of variance and Dunnet's post hoc test, with significant effects set at p < 0.05. Nonlinear regression analysis and all statistical analyses were performed with Prism 5 for Mac (GraphPad Software). For clarity, sample traces were smoothed using the median filter in LabChart Pro (AD Instruments) and exported in the target color using the maximum line width. Protein structures were represented using the University of California, San Francisco Chimera package (Pettersen et al., 2004 (link)).
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