Conventional measurements and posterior wall thickness and thickening) were obtained from grayscale M-mode tracings. LV end-systolic and end-diastolic volumes and LV ejection fraction were measured by Simpson��s method from two-dimensional parasternal long- and short-axis views. After four weeks of pGz or Control the animals where anesthetized and catheterization was performed with a Millar Catheter SPR-869. The conductance catheter was calibrated by a cuvette calibration method using an actual blood sample in cuvette between 50 and 300 ��l. In vivo the conductance signal was calibrated using hypertonic saline. An intravenous bolus of 50 ��l of 20% saline was used to perform calibration. In order to decrease preload, a small abdominal incision was performed to localize and perform inferior vena cava occlusions. PVL where continuously recorded at baseline, after saline infusion, and during and after IVC occlusions. Recording and calculations were performed using data acquisition software. Validation of coronary occlusion was performed by our laboratory according to the procedure previously described. In a separate cohort of animals infarct size was determined after 24 hr. of coronary occlusion to determine infarct size. At the end of the study and after all hemodynamic measurements, the aorta was clamped and the hearts were perfused with 10mL of saline through a cannula in the ascending aorta to wash out the blood from the myocardium. After saline Temozolomide perfusion, Evans Blue was TH-302 abmole injected into the ascending aorta to separate the non-at-risk area from the risk area. The hearts were cut out and cut in 3, 3mm segments from apex to base parallel to the atrioventricular groove. The segments were incubated for 30 minutes in 2,3,5-triphenyltetrazolium chloride at 37��C in the dark. The segments were fixed between two glass sheets and non-at-risk area, the area-at-risk and the necrotic area were determined by planimetry. The basal side of the segments was measured to better distinguish between myocardium stained by EB and TTC. Segments for comparison were chosen on the basis of reproducibility of area-at-risk to perfused myocardium ratio between animals. Images of the segments were taken with a digital camera set to 60 x magnifications through a dissecting microscope. Viable myocardium and infarcted areas non-at-risk area were measured using a computer program. The percentage share of each the preceding areas was calculated. Area at risk measured by the Evans Blue perfusion-staining and expressed as percent of whole heart. Necrosis was measured by TTC staining and expressed as percent of each myocardial segment. To determine transmurality of the infarct scanned images of the segments were geometrically divided into a 6-sector model using the anterior and inferior insertion of the right ventricle to the left ventricle as markers. Apical, middle and basal necrosis was defined. The sectors were divided into the following groups on the background of the distribution of necrotic myocardium: transmural necrotic, subendocardial necrotic and viable. The combination of the two latter groups is referred to as predominantly viable. Transmural necrotic sectors display thinning of the myocardial circumferential areas of left ventricular wall. Transmurality of the infarct was defined as the sum of the epicardial and endocardial infarct circumference divided by the sum of the total LV epicardial and endocardial circumferences using computer-based planimetry. Hearts were then cut into three transverse segments. Each segment was fixed in 10% para-formaldehyde and embedded in paraffin.