On study days, volunteers reported to our laboratory at ~ 09:00 h fasted (except water) from midnight. Body composition was determined by dual-energy X-ray absorptiometry (DXA; Lunar Prodigy II, GE Medical Systems) with body regions auto-processed (Encore software, GE Healthcare). Fasting blood samples were collected from the antecubital vein into lithium-heparin (for plasma) or fluoride oxalate (for glucose) vacutainers, before centrifugation at 3500 rpm, at 4 °C for 20 minutes, wherein the plasma was then rapidly frozen and stored at − 80 °C.
Lunar prodigy 2
Manufactured by GE Healthcare
The Lunar Prodigy II is a bone densitometry system designed for the assessment of bone mineral density (BMD). It uses dual-energy X-ray absorptiometry (DXA) technology to measure the density of bones, primarily in the spine, hip, and forearm. The device provides precise and reliable measurements to support the diagnosis and monitoring of conditions such as osteoporosis.
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4 protocols using lunar prodigy 2
1
Body Composition Assessment and Blood Sampling
2
Dual-Energy X-Ray Absorptiometry and CPET Assessment
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Testing began with a whole‐body dual‐energy X‐ray absorptiometry scan to assess body composition (Lunar Prodigy II, GE Medical Systems), providing automated data for whole body lean mass, body fat percentage, and lean leg mass.
Blood pressure was then measured in triplicate (mean value used) in the subject’s right arm using oscillometry (Datascope trio patient monitor, Datascope), with the subject in a seated position, after having rested for 5 minutes.18 A ramp incremental CPET was then performed according to the ATS/ACCP guidelines17 using a Lode Corival cycle ergometer (Lode Corival, Lode) and inline gas‐analysis system (ZAN 680, nSpire Health) as previously described16 to achieve a CPET duration of between 8 and 12 minutes.17 Anaerobic threshold (AT) was determined using a combination of the V‐slope and ventilatory‐equivalents methods19 by two blinded, experienced assessors with disagreement resolved by consensus. VO2 peak and exercise tolerance were determined as the oxygen utilization and power output (directly related to test duration) at volitional exhaustion, respectively. All baseline testing procedures were repeated 72 hours after the intervention period.
Blood pressure was then measured in triplicate (mean value used) in the subject’s right arm using oscillometry (Datascope trio patient monitor, Datascope), with the subject in a seated position, after having rested for 5 minutes.
3
Body Composition Measurement by DXA
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Subjects were instructed to refrain from unaccustomed exercise for 48 h and from alcohol and caffeine for 24 h before study days. All subjects fasted from 9 p.m. the night before, with water ad libitum. Studies began at 9 a.m. with measurement of body weight, height, and body composition, measured by DXA (Lunar Prodigy II, GE Medical Systems).34 (link) DXA is well accepted, precise, and reproducible (±5%) in the measurement of whole body and regional (e.g. limb) muscle mass. Although not as precise or sensitive as computed tomography (CT) or magnetic resonance imaging (MRI), DXA is much cheaper and more accessible than CT or MRI and avoids relatively high doses of ionizing radiation in the case of CT.35 (link)–37 (link) DXA also compares well with other recognized methods of body composition measurement such as neutron activation and total body K analysis.28 (link),38 (link)–42 (link)
4
Microvascular Blood Volume Measurement
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CEUS was used to measure microvascular blood volume (MBV) as previously described in detail (21 (link)). Repeated high mechanical-index flash/replenishment cycle recordings were made during intravenous infusion of Sonovue microbubble contrast agent (Bracco) and used to estimate changes in thigh MBV between recording periods indicated in Figure 1 . Leg blood flow (LBF) was measured by using Doppler ultrasound, with a linear 9- to 3-MHz probe positioned over the origin of the left common femoral artery. LBF was estimated as the product of vessel cross-sectional area and mean velocity over 6 cardiac cycles. Mean LBF was calculated at a time corresponding to each CEUS recording episode on the basis of 3 such recordings made on each of 4 occasions distributed across the study day, as indicated in Figure 1 (i.e., flow during 72 cardiac cycles contributed to a mean leg flow measure). Each individual’s LBF was standardized to his own fasting LBF and normalized to leg lean tissue mass as assessed by DXA (Lunar Prodigy II; GE Medical Systems).
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