Cardiovascular Journal of Africa: Vol 32 No 5 (SEPTEMBER/OCTOBER 2021)

CARDIOVASCULAR JOURNAL OF AFRICA • Volume 32, No 5, September/October 2021 256 AFRICA clamp (Biemer vessel clip, 7-mm jaw length, closing force 30 G, MCI-47-104, Medical Care Instruments, Manchester, UK). The area where the clamp was applied was referred to as the ‘middle’. The location 1 cm proximally was denoted ‘proximal’, the location 1 cm distally was denoted ‘distal’, and both were marked with 4.0 vicryl. At the same level, the femoral vein, femoral nerve and rectus femoris muscle segments were marked in the same fashion. The clamping time was two hours. At the end of two hours, a longitudinal incision was made on the anterolateral side of the leg and a 5-cm segment of tibialis anterior muscle was excised. From the marked lines, the femoral artery, vein, nerve and rectus femoris muscle were excised. The samples were sent to the pathology laboratory for histological analyses. The animals were euthanised after the procedure. In the tourniquet group, a ‘Blue 12 inch for child’ (reference no: 20-54-710, VBM Medizintechnik GmbH, Sulz am Neckar, Germany) tourniquet was used. The standard tourniquet time for all subjects was two hours and the pressure was 200 mmHg. 8 The proximal and distal borders of the cuff were marked with a tissue pen. After two hours, the tibialis anterior muscle sections were excised, as with the clamp group. The tourniquet was released and removed, then the femoral artery, vein, nerve and rectus femoris muscle were excised from the previously marked cuff margins. Fig. 2. (A) Black stars: vacuoles (H&E stain, x 200). (B) Severe injury, black arrow: naked areas (H&E stain, x200). (C) Black star: fibrin plaque (H&E stain, x 200). (D) Black arrow: vacuoles with loss of cytoplasmic structures, blue arrow: cytoplasmic eosinophilia. A C B D