According to a strict protocol, all RTR were asked to collect a 24-hours urine sample during the day before to their visit to the outpatient clinic and on that day fasting blood samples were taken. Serum creatinine was determined using the Jaffé reaction (MEGA AU510, Merck Diagnostica, Germany); plasma glucose by the glucose oxidase method (YSI 2300 Stat Plus, Yellow Springs Instruments, Yellow Springs, OH, USA). uEGF concentration was measured by ELISA (R&D Systems, Minneapolis, MN, USA); the test has a range of detection of 3.9–250 pg/mL and the intra- and inter-plate coefficients of variation were less than 10% and 15%, respectively [15 (link)]. Urinary creatinine concentration was measured by colorimetric detection kit (Enzo, New York, NY, USA). Finally, the concentration of uEGF was normalized by the concentration of urinary creatinine, and a ratio was created and used for all analyses (uEGF/Cr).
Body surface area was calculated according to the Du Bois formula [19 (link)], estimated glomerular filtration rate (eGFR) by the serum creatinine based Chronic Kidney Disease EPIdemiology collaboration equation (CKD-EPI) [20 (link)] and the cumulative dose of prednisolone as the sum of the maintenance dose of prednisolone from transplantation until enrollment.
Body surface area was calculated according to the Du Bois formula [19 (link)], estimated glomerular filtration rate (eGFR) by the serum creatinine based Chronic Kidney Disease EPIdemiology collaboration equation (CKD-EPI) [20 (link)] and the cumulative dose of prednisolone as the sum of the maintenance dose of prednisolone from transplantation until enrollment.