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55 protocols using encore software

1

Body Composition Assessment and Blood Sampling

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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.
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2

Dual-energy X-ray Absorptiometry for Body Composition

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Dual-energy X-ray absorptiometry measurements of participants’ total and segmental fat and lean mass were obtained with a Lunar iDXA with enCORE software version 13.6 (GE Healthcare, Chicago, IL). The accuracy of the densitometer was verified daily by scanning the manufacturer’s hydroxyapatite spine phantom of a known density. When participants did not fit within the scanning area (n=6), the enCORE software estimated missing regions automatically.22 Only the missing regions were estimated; everything inside the scan area was measured directly. For all participants, the left arm was the portion of the body estimated. All DXA scans were completed on the same scanner with the same software by a trained technician.
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3

Whole-body Composition Analysis by DXA

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Whole-body dual energy X-ray absorptiometry (DXA) scans (LUNAR Prodigy; GE Medical systems, Madison, WI, USA) were performed before and after the RES intervention. The rats were anaesthetized in an induction chamber with 3–4% isoflurane (Isoba vet.; Intervet/Shering-Plough, Uxbridge, UK). Anaesthetized rats were placed on the centreline of scanning bed in the prone position. Throughout the measurement, anaesthesia was maintained by gas inhalation through a facemask continuously supplied with 1–2% isoflurane. The facemask was connected to open-circuit gas anaesthesia equipment (Harvard Apparatus with MSS-Vaporizer, Kent, UK). Before the measurements, calibration of DXA scanning equipment was done according to the manufacturer’s guidelines. The small-animal mode of the enCORE software (GE Healthcare, v. 14.10.022) was used to obtain fat and lean mass content in total body. Moreover, lean mass of the right leg was determined by manually adjusting cut positions for the region of interest within the area encompassing the thigh and shank muscles.
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4

Body Composition Assessment by DXA

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BMD, SMI, and body composition were assessed by DXA with EnCore software (Lunar Prodigy, GE Healthcare, UK). Subjects fasted overnight and did not perform any exercise in the morning before measurements. Total body and regional BMD, bone mineral content (BMC), fat mass, and fat-free mass (FFM) were analysed. All scans were performed and analysed by a single trained and licensed technician who was blinded to the group allocation. SMI was calculated as follows: SMIkg/m2=appendicular skeletal muscle massFFMBMC/body height2
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5

Body Composition Assessment Using DXA

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Body weight was checked using a digital scale (Micheletti ®, São Paulo Brazil) with accuracy of 0.01kg. Height was measured using a stadiometer with an accuracy of 0.1cm (Sanny®, São Paulo, Brazil). Afterwards, the participants had their body composition assessed using a DXA (LUNAR®/GE PRODIGY - LNR 41.990, Washington, DC, USA) equipped with enCORE software (GE Healthcare®, version 15.0, Madison, WI, USA). The equipment was properly calibrated before the evaluations and followed the same configuration for all participants (Full Body Evaluation, Voltage (kV): 76.0, Current (mA): 0.150, Radiation dose (µGγ): 0.4 (Very low, no health risk)). Subsequently, the values in Kg of bone, fat, lean and fat-free mass were acquired.
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6

DXA Measurement of Bone Density and Texture

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Measurements of BMD were made by DXA (GE Lunar Prodigy Advance, Lunar Corporation, General Electric, Madison, WI, USA) at three skeletal sites (lumbar spine, femoral neck, and total hip) in all subjects at baseline and follow-up (one year). The same DXA instrument was used for both measurements. Analysis of the DXA date was performed with enCORE Software version 2005 9.30.044 (GE Healthcare, Madison, WI, USA) in accordance with the manufacturer’s recommendations. The L1-4 value was included in the analysis for BMD and TBS. The TBS was calculated using the TBS iNsight Software, version 3.02 (Med-Imaps, Pessac, France) from DXA images of the same vertebrae as the BMD measurements.
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7

Accurate Measurement of Body Composition

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Total body composition was measured using DXA, (Prodigy, General Electric Medical Systems, Madison, WI, USA) and the scans were analyzed using enCore™ software (platform version 13.6, General Electric Medical Systems, Madison, WI, USA). Estimates of abdominal visceral and subcutaneous adipose tissue were obtained using the method described previously for adults (20 (link)). The estimation of VAT using DXA was recently validated in adults (20 (link)) and has been shown to provide a reliable estimate in children (21 (link)). Android fat (abdominal) was measured using a region-of-interest automatically defined with a caudal limit placed at the top of the iliac crest and its height set to twenty percent of the distance from the top of the iliac crest to the base of the skull (22 (link)). The gynoid region (hip/gluteal) is located mid-pelvis to mid-thigh, with the upper limit set below the iliac crest a distance 1.5 times the height of the android region and the lower limit set a distance of 2 times the height of the android region (22 (link)). Subcutaneous fat and visceral fat were estimated within the android region as described previously (20 (link)). All scans were reviewed for accurate placement of the android box by the same technician.
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8

Dual-Energy X-Ray Absorptiometry Body Composition

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Whole and regional body composition were acquired via dual energy x-ray absorptiometry on a Lunar iDXA scanner (GE Healthcare, Chicago, Illlinois). The scanner was phantom calibrated daily prior to data acquisition to ensure instrument reliability. Scans were acquired by one research technician certified in densitometry. Scans were analysed using enCORE software (version 13.6, GE Healthcare) and the android region of interest for quantification of VAT mass was automatically determined by the device software.
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9

Dual-Energy X-Ray Absorptiometry of Hands

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The hand DXR-BMD was calculated using the Sectra DXR software (Sectra, Linköping, Sweden). DXR calculates a mean BMD from metric measures of cortical thickness at the second, third, and fourth metacarpal bones on standardized hand X-rays. The formula includes an estimated porosity aimed to be the fraction of cortical bone not occupied by bone, as described previously [13 (link)]. The hand X-rays for DXR-BMD calculation were acquired with Fuji FCR XG1 (CR; FFD 100 cm; tube voltage 50 kV; exposure dose 5 mA), and the non-dominant hand was used when possible (93/110).
DXA-BMD at the femoral neck, total hip, and spine (L2–4) was assessed using Lunar Prodigy with enCORE software (GE Healthcare, Madison, WI, USA).
Trained nurses performed all the BMD measurements using standardized protocols. The in vivo coefficient of variation (CV%) for DXR at the non-dominant hand was 0.46% and for DXA at the femoral neck 1.68%, total hip 0.88%, and spine (L2–4) 1.26%.
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10

Comprehensive Body Composition Assessment

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Body composition measurements were performed by dual energy X-ray absorptiometry using the iDXA instrument with Encore software (version 13.11.016) (GE Medical Systems, Madison, WI, USA). The whole body scan was utilized to measure total body LM, FM, and BMD. The iDXA has a wider scan field and higher precision that can accommodate individuals up to 181 kg, avoiding the errors typically encountered when measuring overweight/obese individuals, and therefore providing a more accurate assessment of both bone and body composition.[25 (link)] The LM component refers to the total LM (derived from the whole body scan), after excluding bone and FM. BMD of regional skeletal sites, including spine (L2–L4), both left and right femurs (neck and total), and radius (1/3 distal) were measured and analyzed by specialized software for each. The axial skeletal sites, including femoral neck, total femur, and lumbar spine are the sites where fractures usually occur and the ones used to diagnose osteoporosis (based on their T-scores). Therefore, those sites were the ones that we focused our analyses on. The quality analysis for the instrument was conducted on a weekly basis using a standard aluminum spine block (phantom) provided by the manufacturer. Measurements of the phantom were within the manufacturer's precision standard of < 0.05% coefficient of variation.
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