In all participants, a detailed two-dimensional transthoracic echocardiography was performed by a single certified cardiac sonographer using a commercially available system (Vivid T8, GE Health care, Freiburg, GERMANY) equipped with a 3.5 MHz transducer. Left ventricular measurements were obtained, according to the 2015 American Society of Echocardiography and the European Association of Cardiovascular Imaging updated guidelines for cardiac chamber quantification [28]. Measurements of left ventricular diameter (LVED), interventricular septum thickness (IVS), and posterior wall thickness (PWT) were measured at the end of diastole. Simultaneous ECG was performed to correlate the left ventricular measurements with the cardiac cycle. Left ventricular mass (LVM) was calculated according to the American Society of Echocardiography simplified cubed equation linear method, using the following equation: LVM (grams) = 0.8 × 1.04 × [(LVED + IVS + PWT)3 − (LVED)3] + 0.6 g, where LVED is the left ventricular end-diastolic diameter, IVS is the interventricular septal thickness, and PWT is the left ventricular posterior wall thickness. Left ventricular mass was indexed against BSA and height [2, 7]. The relative wall thickness (RWT) was calculated as follows: (2 × PWT) / LVED.

In accordance with international recommendations [29], LV diastolic function was assessed from the apical four-chamber view, which included transmitral, pulsed-wave Doppler and mitral annular velocities with tissue Doppler echocardiography. The transmitral peak early (E) and peak late (A) diastolic velocities were recorded. The mitral annular early diastolic velocity (e’) was measured at the lateral mitral annulus using pulsed-wave tissue Doppler in the apical four-chamber view with gains minimized to allow for a clear tissue signal.

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