*4.2. Animal Echocardiography*

Transthoracic echocardiography in both groups (control *n* = 8, PE *n* = 8) was performed postpartum in anesthetized animals (1.5% isoflurane via an oxygen mask). Temperature, ECG, and respiration were monitored. By a heated platform, rectal temperature was maintained at 37 ± 2 ◦C. Abdominal hair was removed by depilatory cream. Pre-warmed gel was used as an ultrasound-coupling medium. For imaging, a Vevo3100 high-resolution imaging system (Fujifilm, VisualSonics Inc., Toronto, ON, Canada) mounted with a 21 MHz transducer (MS250) was used. All images were acquired and stored for o ffline analysis. Analysis was done by two blinded observers using VisualSonics VevoStrain software (Version 2.2.0, Toronto, ON, Canada). In parasternal long and short axis view, B-Mode cine loops were used to assess speckle tracking analysis. Images were checked for quality with regard to absence of artifacts and di fferentiation of wall borders. The endocardium of the left ventricle was traced manually in end-diastole. The epicardium was automatically traced by the software, checked and manually adjusted if necessary, for maximum tracking accuracy. Global strain values were obtained from the average of the six segments of the left ventricle. M-mode was obtained to measure cardiac wall and chamber dimensions. Relative wall thickness was calculated by the formula (2\*PWd)/LVEDD. Analysis was performed on three consecutive cardiac cycles; mean values from three measurements were calculated.
