The methods utilized to measure EC have previously been described15 (link),19 (link). Briefly, oxygen consumption (ml O2/min) was measured by a ParvoMedics TrueOne 2400 Metabolic Measurement System (Sandy, UT) as participants walked on a treadmill for five minutes at their overground CWS. Data from the 5th minute of walking were subsequently normalized to body weight (kg) and speed (m/min) to yield EC (ml O2/kg/m). For participants that trained at a fast walking speed (see Locomotor Rehabilitation section), EC was also measured at their fast walking speeds.
Trueone 2400 metabolic measurement system
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The TrueOne 2400 Metabolic Measurement System is a lab equipment product designed for measuring metabolic parameters. It provides accurate and reliable data on various metabolic functions.
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21 protocols using trueone 2400 metabolic measurement system
1
Measuring Energy Consumption During Walking
2
Hydrostatic Weighing and V̇O₂peak Assessment
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Body fat was assessed through hydrostatic weighing using an electronic load cell-based system (Exertech, Dresbach, Minn., USA), correcting for estimated residual lung volume. A was conducted on an electronically braked cycle ergometer (Excalibur Sport, Lode, Groningen, Netherlands) using a graded-exercise protocol until volitional fatigue. Subjects began cycling at 95 W and the workload increased by 35 W every 3 min until volitional fatigue. The highest oxygen consumption recorded by a Parvo Medics TrueOne 2400 Metabolic Measurement System (Sandy, Utah, USA) was defined as the . Maximal workload was calculated as the highest completed stage (in watts) + the proportion of time in the last stage multiplied by the 35 W per stage increment. For example, if the subject stopped 30 s into the 235 W stage, the highest completed stage would be 200 W because each stage the workload increases by 35 W. The maximal workload would be 200 W + (35 W × 30 s /180 s) because the subject completed 30 s of the 180-s stage, which would equal 205 W. The workload for the experimental trial was set at 65% of the maximal workload.
3
Measurement of VO2peak via Treadmill
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During the familiarization trial an incremental test to volitional exhaustion was performed on a motorized treadmill (Woodway 4Front™, Waukesha, WI) to measure VO2peak. Open-circuit spirometry (TrueOne 2400® Metabolic Measurement System, Parvo Medics, Inc., Sandy, UT) was calibrated with room air and gases of known concentration, which was used to estimate VO2 (ml∙kg−1∙min−1) by sampling and analyzing breath-by-breath expired gases. VO2peak was determined to be the highest 30-s VO2 value during the test and coincided with at least two of the following three criteria: (a) 90 % of age-predicted maximum heart rate; (b) respiratory exchange ratio > 1.1; and/or (c) a plateau of oxygen uptake (less than 150 mL · min−1 increase in VO2 during the last 60 s of the test). Previous work in our lab has shown the test-retest reliability for VO2peak to be ICC = 0.96 (SEM 1.4 ml.kg.min−1) [23 (link)].
4
Metabolic Measurements Using Indirect Calorimetry
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Metabolism parameters measurements of each participant were completed at the Nutrition and Health Assessment Lab in the Food Science and Nutrition Department of California Polytechnic State University San Luis Obispo. Indirect calorimetry was used to determine each participant’s RMR and RQ. Each morning of testing, the metabolic cart (True One 2400- Metabolic Measurement System, ParvoMedics Inc., Sandy, UT, USA) was calibrated according to manufacture’s specifications. The testing was performed on the same morning as the DXA scans for each participant, after a 10–12 h fast. A canopy system (clear hard plastic breathing hood with drape) was placed over the participant’s head and upper body for measurements. Participants underwent testing on the metabolic cart for 25 min. The mean oxygen uptake and carbon dioxide output for each breath were measured, and the average was taken for every 15 s interval. Data from the last fifteen minutes of each session were used to calculate the RMR and RQ values.
5
Treadmill-Based VO2max Assessment
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Cardiorespiratory fitness was assessed on a treadmill (X-Fit 7 Power
Treadmill), using protocols in accordance with age and progressive
intensities. The ramp protocol was used, which consists of 8 to 12 minutes
of exercise with small and constant increments in speed and gradient based
on the expected maximum oxygen consumption19 . Criteria for the maximum test were: a)
exhaustion or inability to maintain the required speed, b) HR equal to or
above 200 bpm, c) respiratory exchange ratio (RER) equal to or greater than
one. VO2max was determined by the average of the three highest
consecutive values obtained during the maximum test20 .
Oxygen consumption (VO2), volume of expired carbon dioxide
(VCO2), and ventilation (VE) were measured in a gas analyzer
using open-circuit ergospirometry (Parvo Medics TrueOne® 2400
Metabolic Measurement System, Sandy, UT, USA), and the respiratory exchange
ratio (RER) was calculated.
Treadmill), using protocols in accordance with age and progressive
intensities. The ramp protocol was used, which consists of 8 to 12 minutes
of exercise with small and constant increments in speed and gradient based
on the expected maximum oxygen consumption19 . Criteria for the maximum test were: a)
exhaustion or inability to maintain the required speed, b) HR equal to or
above 200 bpm, c) respiratory exchange ratio (RER) equal to or greater than
one. VO2max was determined by the average of the three highest
consecutive values obtained during the maximum test20 .
Oxygen consumption (VO2), volume of expired carbon dioxide
(VCO2), and ventilation (VE) were measured in a gas analyzer
using open-circuit ergospirometry (Parvo Medics TrueOne® 2400
Metabolic Measurement System, Sandy, UT, USA), and the respiratory exchange
ratio (RER) was calculated.
6
Maximal Aerobic Power Protocol
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During visit 3, participants performed a ramp protocol to volitional exhaustion on a cycle ergometer (Lode, Excalibur Sport, Groningen, the Netherlands). Prior to the test, participants completed a warm-up consisting of 5 min of light cycling at an intensity of 50 watts at a self-selected pace, 10 body weight squats, 10 body weight walking lunges, 10 dynamic waking hamstring stretches, and 10 dynamic walking quadricep stretches. The MAP protocol required each participant to maintain a pedaling cadence of 70–80 revolutions per minute (RPM) at an initial workload of 100 watts (W). Seat height was recorded for all participants and standardized for subsequent experimental trials. The workload increased 30 W every two minutes (1 W per 2 s) until participants were unable to maintain a cadence above 70 RPM for ~10 s despite verbal encouragement or volitional fatigue. Expired gasses were analyzed using open-circuit spirometry (True One 2400® Metabolic Measurement System, Parvo-Medics Inc., Sandy, UT, USA). The highest power output achieved was recorded as PPO in watts (W). GET was determined via computerized regression analysis of the slopes of the CO2 uptake (VCO2) vs. the O2 uptake (VO2). Power at the GET was recorded.
7
Assessing Cardiorespiratory Fitness via Treadmill Test
8
Indirect Calorimetry Measurement Approach
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Indirect calorimetry measurements (TrueOne 2400 Metabolic Measurement System, Parvo Medics, UT) were conducted in three physiological states (basal, insulin-stimulated and exercise). Whole-body Fatox and CHOox were calculated using stoichiometric equations, with the assumption that the urinary nitrogen excretion rate was negligible[31 (link)]. The methodological approach adopted has been previously described in detail[5 (link)].
Fatox rates during exercise were estimated from respiratory gazes averaged over the last minute of each exercise stage. Then, the stage at which MFO was achieved was determined, and the corresponding intensity was identified (Fatmax)[32 (link)]. ∆RQ represented the RQ change from basal to hyperinsulineamic state (RQ in the insulin-stimulated condition minus basal RQ).
Testing sessions involving indirect calorimetry measurements were conducted in the morning after a 10-12 h overnight fast and under standardised conditions[5 (link)]. Standardisation of pre-test conditions was in line with previous studies[32 (link)-40 (link)].
Fatox rates during exercise were estimated from respiratory gazes averaged over the last minute of each exercise stage. Then, the stage at which MFO was achieved was determined, and the corresponding intensity was identified (Fatmax)[32 (link)]. ∆RQ represented the RQ change from basal to hyperinsulineamic state (RQ in the insulin-stimulated condition minus basal RQ).
Testing sessions involving indirect calorimetry measurements were conducted in the morning after a 10-12 h overnight fast and under standardised conditions[5 (link)]. Standardisation of pre-test conditions was in line with previous studies[32 (link)-40 (link)].
9
Resting Metabolic Rate Measurement Protocol
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Indirect calorimetry was used to measure resting metabolic rate (RMR) via a ventilated hood and metabolic cart (TrueOne 2400 Metabolic Measurement System, ParvoMedics, Sandy, Utah, USA) at baseline and post-intervention. Participants arrived for a morning visit following an overnight fast (12 h) and having abstained from exercise (including active transport) for at least 14 h [36 (link)]. Participants were asked to lay supine in a comfortable position with their head on a pillow, and to remain motionless and awake. Following a 15 min equilibration period, the participant's head was covered by a transparent ventilated plastic hood, with a drape wrapped around the upper body to avoid leakage of air, for 30 min. During the first 10 min of measurement, the dilution pump flow rate was adjusted (approximately bodyweight (kg) divided by 3) until steady state was reached (≤10% coefficient of variation for VO2 and VCO2). The measurement protocol was developed according to best practice based on a methodological review by Compher et al. [36 (link)]. Resting energy expenditure (kcal/day) is calculated with the system software (TrueOne 32 RMR, version 4.3.4), using the modified Weir equation (5.616 x VO2 + 1.584 x VCO2). The first 10 min of data was discarded, and RMR (kcal/day) was defined as the average of the final 20 min of measurement.
10
Cardiorespiratory Fitness and Cardiometabolic Profile Assessment
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Cardiorespiratory fitness was assessed via indirect calorimetry at baseline, week 12, and 3‐ and 6‐month follow‐up. Participants were required to walk/run on a treadmill, following the Modified Bruce protocol, until volitional fatigue. Peak oxygen uptake (V̇O2peak; TrueOne 2400 Metabolic Measurement System, ParvoMedics, Sandy, Utah) was recorded as the highest mean value attained during two 30‐second periods, before volitional exhaustion [27 (link)]. Participants were asked to avoid exercise and any stimulants (such as caffeine and tobacco) and alcohol in the 24 hours prior to testing and to avoid eating in the 2 hours prior to the test.
Blood pressure, lipid profiles, and blood glucose were sampled at baseline and 12 weeks in a fasted state. Samples of capillary blood were extracted via a contact‐activated lancet and processed using the CardioChek PA (Polymer Technology Services, Inc., Indianapolis, Indiana) and AccuChek Performa (Model NC; Roche, Mannheim, Germany) analyzers, handheld, battery‐operated reflectance spectrophotometers to measure blood lipids (triglycerides, high‐density lipoprotein [HDL], low‐density lipoprotein [LDL], total cholesterol) and blood glucose, respectively.
Blood pressure, lipid profiles, and blood glucose were sampled at baseline and 12 weeks in a fasted state. Samples of capillary blood were extracted via a contact‐activated lancet and processed using the CardioChek PA (Polymer Technology Services, Inc., Indianapolis, Indiana) and AccuChek Performa (Model NC; Roche, Mannheim, Germany) analyzers, handheld, battery‐operated reflectance spectrophotometers to measure blood lipids (triglycerides, high‐density lipoprotein [HDL], low‐density lipoprotein [LDL], total cholesterol) and blood glucose, respectively.
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