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The donor heart was excised and placed in ice-cold cardioplegia solution whilst the recipient rat great abdominal vessels were clamped. End-to-side anastomosis of the donor heart aorta to the recipient infrarenal abdominal aorta, and of the donor heart pulmonary artery to the recipient inferior vena cava was then performed using 10/0 nylon suture (BEAR surgical sutures, Kyowa Precision Instruments Co. Ltd., Chiba, Japan) (Fig. ?(Fig.1b).1b). Clamps and lumbar vessel ties were removed, and the heart reperfused; spontaneous [https://en.wikipedia.org/wiki/Fossariinae Fossariinae] beating was observed in each case. Total cardiac ischemic time was  pulse. Cardiac cine MRI Cardiac cine MRI was performed at 6-24 days postoperatively using a 55?mm [http://www.selleckchem.com/products/Sunitinib-Malate-(Sutent).html Sunitinib nmr] birdcage coil (Rapid Biomedical) in a vertical bore 500 MHz, 11.7 T MR system with a Bruker console running Paravision 2.1.1 (see Online Resource Supplementary Movies). Rats were anaesthetised with 2.0% v:v isoflurane in oxygen and positioned supine in a purpose built cradle. Two ECG signals were obtained, one from the recipient (thoracic) heart and a second, for gating, was obtained from the heterotopic transplanted heart using two subcutaneous needles adjacent to the palpated beat (Fig. ?(Fig.1c).1c). The transplanted heart was located using [http://www.selleckchem.com/screening/pi3k-signaling-inhibitor-library.html selleck] scout images (Fig. ?(Fig.1d).1d). To measure cardiac function, a stack of contiguous 1.5 mm true short axis ECG-gated cine images were acquired to cover the entire left ventricle. The imaging parameters were: field of view 51.2 �� 51.2 mm; matrix size 256 �� 256; TE/TR 1.43/4.6 ms; 17.5�� Gaussian excitation pulse. The epicardial and endocardial borders were outlined in end-diastolic and end-systolic frames using the freehand drawing tool in Image J (17). Cardiac mass and left ventricular volumes were summed over the whole heart. For comparison, scans were also performed of untransplanted (orthotopic) control hearts under identical conditions of anaesthesia and positioning. Statistical analysis Results are means �� SE [range]. Statistical analysis was performed by Student��s t test, and Spearman��s coefficient of rank correlation, as appropriate, using SPSS. Statistical significance was set at P