Mylab 25

Muscle thickness of four trunk flexors (rectus abdominis, internal/external oblique, transversus abdominis), one trunk extensor (erector spinae longissimus) and two knee extensors (vastus lateralis and vastus intermedius) were measured unilaterally on the dominant side using B-mode ultrasound (Mylab 25, Esaote, Florence, Italy) equipped with a linear array transducer (frequency band 7.5-12 MHz) (Dupont et al., 2001) . The measurements were conducted according to methodology described elsewhere (Fukunaga, Ichinose, Ito, Kawakami, & Fukashiro, 1997; Ikezoe, Mori, Nakamura, & Ichihashi, 2012) . In short, recordings of trunk flexor thickness were made in the supine position and at the end of a relaxed expiration and participants were in the prone position for trunk extensor measurements. For knee extensors, participants were asked to sit with the hip and knee joints flexed at 90° and to relax the quadriceps muscles. Muscle thickness was analysed with an imageediting program (ImageJ 1.36b, National Institutes of Health, Bethesda, USA) and defined as the largest Euclidean distance between two points that were placed orthogonally to the centreline between the superficial and the deep aponeurosis (Minetto et al., 2016) . A total of three images were acquired for each muscle and the mean peak distance was retained for further analysis.

Echocardiographic data were measured by MyLab 25 instrument (Esaote S.P.A., Genoa, Italy) according to the recommendations of the American Society of Echocardiography by an observer unaware of other study outcomes. LVM was calculated according to the Devereux formula and indexed to height 2•7 (LVMi) [20] in order to minimize any potential distortion attributable to extracellular volume expansion [21] . LVH was defined by a LVMi > 47 g m -2.7 in women or > 50 g m -2.7 in men. RWT was calculated as follows: 2*posterior wall thickness/left ventricular end diastolic diameter, as an index of the left ventricular geometric pattern.
Values indicative of concentric and eccentric left ventricular geometry were established on the basis of age-specific reference standards [22] .

Cross-sectional area (CSA) of the right RF was measured with ultrasonography (LA523, 50 mm array, 10-to 15-MHz transducer, MyLab25, Esaote, Genoa, Italy). Subjects were lying in a supine position and muscle scans were taken at a distance corresponding to 40% of femur length (measured manually as the distance between the lateral femoral condyle and the trochanter major), relative to the lateral femoral condyle. At this location, the width of the RF did not exceed that of the ultrasound transducer.

The vessels of the graft were then disposed on the terminal part of the device by a limited spatulation in their longitudinal axis, as described by the instructions for use provided by the manufacturer. A small incision was subsequently made on the iliac vein to allow the tangential introduction of the anvil. Once the correct positioning of the device was checked, it was armed and the end-to-side anastomosis performed. It was necessary to give a 6/0 Prolene stitch to close the vasotomy. No further stitches were needed to fix the anastomosis (Fig. 4). The same procedure was performed for the renal artery.
Abdominal wall was then closed with interrupted 2/0 Prolene stitches.
The animals were followed up with blood tests and Doppler US scanning to evaluate renal resistive index (RI), shortly after the operation and at 10 days (Table 1). US was performed by the same operator for all the animals. The US scan (MyLab 25, Esaote, Genova, Italy) was performed with a 3.5 MHz probe. In each animal, RI at the interlobular or arcuate artery near the border of the central echo complex was measured three times in the upper, middle, and lower portions of the transplanted kidney. The mean RI value was used for analysis. An RI < 0.6 was considered normal. The statistical analysis was performed with Student's t-test comparing the mean value preoperatively and at 10 days.