Participants’ body compositions were obtained from dual-energy X-ray absorptiometry (DXA) using the iDXA instrument (GE Medical Systems, Madison, WI, USA) with Encore 2011 software (version 13.6). On the days when data were collected, all participants had fasted overnight for at least 12 h and the T1D group did not administer their morning insulin injections. We took fasting capillary (fingertip) blood samples for analysis of HDL-C, LDL-C, total cholesterol, and triglycerides, and these lipid profiles were analyzed using the Cardiochek PA Blood Analyser (Polymer Technology Systems Inc., Indianapolis, IN, USA). The correlation between the CardioChek PA Blood Analyzer from fingerstick blood samples and venous blood samples was high for HDL-C (r = 0.95), triglycerides (r = 0.95), and total cholesterol (r = 0.88) [26 (link)]. In addition, we used the A1cNow+ (Metrika Inc., Sunnyvale, CA, USA) to analyze HbA1c from a fingerstick blood sample, which has been demonstrated to be highly correlated with the high-performance lipid chromatography test of using a venous blood sample (r = 0.95) [27 (link)]. These devices have been previously validated as well [28 (link),29 (link)].
Encore 2011
Manufactured by GE Healthcare
Sourced in United States
The EnCORE 2011 is a bone densitometry system designed for clinical use. It measures the bone mineral density (BMD) of the patient's body. The EnCORE 2011 uses dual-energy X-ray absorptiometry (DXA) technology to assess bone health.
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12 protocols using encore 2011
1
Body Composition and Lipid Profiles in T1D
2
Comprehensive Anthropometric Evaluation
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A stadiometer with 0.1 cm intervals will be used to measure height, and a digital scale accurate to 0.1 kg, and a maximum capacity of 150 kg, will be used to evaluate weight (Seca, Hamburg, Germany). Waist circumference (WC) will be measured with an inelastic tape placed at the midpoint between the anterior-superior iliac crest and the last rib, as hip circumference (HC) will be measured at the widest portion of the buttocks, with the tape parallel to the floor (Norton, 2018 ). The ratio between WC and HC will be obtained and expressed in cm. Body mass index will be calculated by the ratio between weight and squared height and expressed in kg/m2 (Norton, 2018 ). Lean mass, fat mass, android and gynoid fat depots, and bone mineral density will be assessed by dual-energy X-ray absorptiometry (DEXA; GE® Lunar densitometer, DPX NTVR, with ENCORE 2011 software, version 13.60, GE Healthcare). Participants will be instructed not to perform other procedures with contrast or radiation the day before the measurement. The procedure will be performed after 12 h of fasting, in the supine position, with bare feet, light clothes and no metallic objects.
3
Body Composition Assessment by DXA
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Body composition was measured at baseline and after 5 weeks of intervention by dual-energy X-ray absorptiometry (DXA), using a DPX NT densitometry equipment (General Electric Medical Systems Lunar, Madison, EUA) with Encore 2011 software (version 13.60, GE Healthcare). Participants were instructed not to exercise in the 24 hours before the test, and all tests were conducted at the same time of the day (between 7 a.m. and 12 noon). There was no specific recommendation for hydration, except to maintain the same habits for retesting. During the tests, the participants were barefoot and wearing light clothing. All tests were supervised by professionals with experience in this type of procedure who were blinded to group allocation. The same operator performed all the measures and calibrations. All the necessary adjustment and calibration was taken for measurements by the technician. The coefficient of variation for the DXA tests of lean and fat mass were 0.75% and 1.03%, respectively.
4
Whole-Body DXA Scan Protocol for Body Composition
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Body composition was determined with a whole body DXA scan (Lunar Prodigy Advance; GE-Medical Systems, Madison, WI, USA) and corresponding analysis software (enCore 2011, version 13.60, GE-Healthcare). Scans and analysis were performed according to the manufacturer’s instructions. A urine sample was taken prior to the scan to assess urine specific gravity, and females were subjected to a pregnancy test. Once hydration and pregnancy status were established, participants were instructed to lie in a supine position with their arms resting against the sides of the body approximately 6.5–7 cm away from their torso. A block was placed between their feet and the participants were instructed to maintain a stationary position for the duration of the scan. DXA equipment was calibrated on a daily basis according to the protocol provided by the manufacturer. The operators defined the regions of interest (ROI) for the limb specific measures and custom ROI boxes were draw for the two specific sites. The ROI for the legs followed the inguinal line (cutting through the neck of the femur) to the bottom of the femur and the ROI for the arms followed the head of the radius (at the elbow) and dissected at the radiocarpal joint (of the wrist).
5
Dual-Energy X-Ray Absorptiometry for Body Composition
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A whole-body dual energy X-ray absorptiometry scanner (Lunar Prodigy, GE Medical Systems, Madison, WI, USA) was used to measure body composition. Scans were performed under standardised conditions (early morning, overnight fasted, bladder/bowel voided, and standardised body positioning on the scanning bed). Following the scan, in-built analysis software (version 13.60.033; enCORE 2011, GE Healthcare, Madison, WI, USA) allowed the calculation of total and regional (predefined by the software) lean body mass (excluding bone mineral content), fat-free mass and fat mass. Skeletal muscle mass was calculated using an equation described by Kim and colleagues [20 (link)]. The equation is as follows:
where females are represented with a ‘zero’ and males are represented with a ‘one’ in the equation.
where females are represented with a ‘zero’ and males are represented with a ‘one’ in the equation.
6
Standardized DEXA Body Composition Assessment
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Participants had their body composition assessed by a whole-body DEXA scanner (Lunar Prodigy, GE Medical Systems, Madison, WI, USA). Conditions were standardised to ensure accurate results. This included being in a fasted state (10–12 h prior to scan), minimising fluid intake (no more than 200 mL of water prior to scan), bladder/bowel being voided, jewellery removed, and clothes removed down to the underwear with a hospital gown worn. Prior to every DEXA scan, the machine was calibrated (within 24 h). Body composition results were obtained by in-built analysis software (version 13.60.033; enCORE 2011, GE Healthcare, Madison, WI, USA).
7
Body Composition Assessment Protocol
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Body mass and height were measured with a digital scale (Tanita DC-360 S, Tanita Corp., Tokyo, Japan) and a wall-mounted measuring tape (Seca206; Seca GMBH, Hamburg, Germany), respectively. Whole-body DXA (Lunar Prodigy, enCORE 2011, General Electric, software 14.10.022, Madison, WI) was used to assess the participants' body composition, including fat and lean mass, and android and gynoid fat. The DXA measurements used for subsequent calculations were carried out in one center in the morning, 2 hr after a standardized 350-kcal breakfast, 1 day before the start, and within 7-10 days after the completion of the protocol. This time period was chosen to avoid misinterpretation due to acute changes in tissue glycogen and hydration resulting from the protocol (Bone et al., 2017) (link).
8
Whole-Body DXA Scan for Body Composition
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A whole-body dual energy x-ray absorptiometry scanner, (Lunar Prodigy, GE Medical Systems, Madison, WI) was used to measure body composition under standardised conditions (early morning, overnight fasted, bladder/bowel voided, and standardised body positioning on the scanning bed). Following the scan, in-built analysis software (version 13.60.033; enCORE 2011, GE Healthcare, Madison, WI) allowed the calculation of total, fat and fat-free mass. To eliminate differences in fat-free mass associated with height fat-free mass index (FFM index) was calculated. This was derived as FFM (kg) divided by height (m) squared (kg/m 2 ). 31
9
Bone Density Assessment in Rats
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Bone mineral density was measured under general anesthesia. The anesthetized rats were positioned in ventral recumbency on the scan table. All scans were performed using dual-energy X-ray absorptiometry (DXA) to evaluate the BMD in rats at week 16. All DXA scans were analyzed using the manufacturer’s recommended software (Encore 2011; GE Healthcare). The right femur metaphysis was used for structural analysis of the bone-implant interface (n = 3). Bone tissue was subjected to micro-CT analysis (SkyScan1272; Bruker Micro-CT, Germany). Scanning was set to have the entire sample within the field of view throughout a full 360° rotation. The reconstruction was conducted using CTvox software (Data Viewer Version 1.6.0.0). Tissue volume, bone volume, percent bone volume, tissue surface, bone surface, intersection surface, bone surface/volume ratio, bone surface density, trabecular thickness, trabecular separation, trabecular number, and trabecular pattern factor were determined according to guidelines (CT-Analyzer Version 1.20.8).
10
DEXA-Based Skeletal Muscle Assessment
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Body composition was assessed using a dual-energy x-ray absorptiometry (DEXA) scan (Lunar Prodigy, GE Medical Systems, Madison, WI). All scans were performed under standard conditions ( 18) by a licensed coinvestigator. Subjects were instructed to arrive for the scan after a minimum 12-hour fast from solid food, a minimum 6-hour fast from liquids, and no exercise for a minimum of 24 hours. In-built analysis software (version 13.60.033; enCORE 2011, GE Healthcare) immediately calculated total lean and fat mass and regional subsites to estimate appendicular masses (i.e., upper limb and lower limb). Skeletal muscle mass was calculated using an equation described by Kim et al. ( 28). The equation is as follows: Total Skeletal Muscle Mass = (1.13 × appendicular lean mass) -(0.02 × age) + (0.61 × sex ) + 0.97; where women are represented with a "zero" and men are represented with a "one" in the equation.
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