Detection of hydroxyl free radicals in the reperfused primate heart.

Early reperfusion of an ischemic region can result in significant salvage of the area at risk. We show the presence of hydroxyl free radicals at the time of post ischemia reperfusion using electron paramagnetic resonance (EPR) spectroscopy in a macaque model. These free radicals may be formed as a result of reperfusion or may be an un-involved bystander. It is possible that they may be involved in reperfusion injury.

Failure of deferoxamine to reduce myocardial infarct size in a primate model of ischemia-reperfusion injury.

Baboons were subjected to treatment with deferoxamine (DF), a strong iron-chelating agent, to inhibit the iron-dependent production of hydroxyl radicals. Studies were then done to determine if this would result in a reduction in the size of myocardial infarct. Baboons underwent occlusion of the left anterior descending coronary artery for 2 hr followed by reperfusion for the next 22 hr. A treated group (n = 4) received a 2-hr preischemic intravenous infusion of DF (10 mg/kg/hr). This infusion continued throughout the ischemic phase and 2 hr into the reperfusion phase. A control group (n = 8) underwent the identical protocol minus the DF infusion. At the end of the reperfusion period, the hearts were sectioned and stained for histological examination. The treated animals had a 22% larger volume of infarct compared with those of the controls (P = 0.06). There was no statistically significant difference (P > 0.05) in hemodynamic or epicardial ST segment measurements between the two groups. In this primate model, there was no myocardial protection afforded by DF. Baboons are similar to humans in that both have minimal collateral circulation. In the literature, DF has been noted to actually contribute to the production of free radicals in certain circumstances. This experiment appears to indicate that caution should be exercised in the use of DF in the treatment of ischemia-reperfusion injury of the heart.

Antiserum to endotoxin in hemorrhagic shock.

Antiserum to Escherichia coli J5, a mutant endotoxin (LPS) which contains only core determinants, has proven effective in reducing mortality from endotoxic shock due to a wide variety of gram-negative bacteria. Twenty New Zealand white rabbits with coliforms in the gut were subjected to hemorrhagic shock of 36 mm Hg for 3 hr. Treated rabbits were resuscitated with 15 cc of rabbit J5 antiserum (hemagglutinating antibody titer against J5 lipopolysaccharide of 1:1024), remaining shed blood, and lactated Ringer's to achieve a mean arterial blood pressure (MABP) within 20% of baseline. The control group was similarly resuscitated but received 15 cc normal rabbit serum (titer 1:2). Catheters were removed and rabbits were returned to their cages until death or 5 days of survival. Hemodynamic parameters (heart rate, MABP, cardiac output, and total peripheral resistance) did not differ significantly between groups. However, six treated rabbits survived 5 days (60%) and no control rabbit lived past the third postexperimental day (P less than 0.019). Our data suggest that systemic endotoxemia may contribute to morbidity and mortality in severe hemorrhagic shock.

Regional myocardial blood flow in experimental myocardial infarction after pretreatment with aspirin.

The effects of aspirin on myocardial blood flow in an area of ischemia were studied in 12 baboons. In each, a diagonal branch of the left anterior descending coronary artery was ligated. Six of the baboons received aspirin (2 X 600 mg orally, 12 hours and 1 hour before ligation); the other six did not receive aspirin and served as a control group. The extent of myocardial ischemia was delineated with an electrode wire grid on the surface of the anterior left ventricular wall. The maximal area circumscribed by electrodes with 2 mV or more ST segment elevation was compared with the area of reduced myocardial blood flow. Myocardial blood flow was measured with the radioactive microspheres method using strontium-85-labeled carbonized spheres. Two areas of reduced myocardial blood flow were noted, one with severely reduced flow in the center of the myocardial infarct (0 to 49% of noninfarcted myocardium) and another with mild to moderately reduced myocardial blood flow at the border of the myocardial infarct (50 to 90% of noninfarcted myocardium). Myocardial blood flow in the border area (margins of ST elevation area) for the total wall was 85 +/- 8% of normal in the aspirin-treated animals and 40 +/- 4% in the control group (p less than 0.01); for the epicardium it was 67 +/- 10% of normal in noninfarcted myocardium after aspirin and 37 +/- 5% for the control group (p less than 0.05); and for the endocardium it was 78 +/- 8% of normal in noninfarcted myocardium after aspirin and 39 +/- 6% in the control group (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of thoracic aortic cross-clamping and declamping on visceral organ blood flow.

Blood flow was measured using radioactive microspheres in 11 macaque monkeys 1) before hemorrhage shock, 2) after onset of shock, 3) after aortic cross-clamping and resuscitation, and 4) after release of the cross-clamp and stabilization. Hemodynamic parameters (cardiac output, arterial, right atrial and left atrial pressure) and blood gases were also monitored. Total abdominal organ flow fell with hemorrhage and fell further with aortic clamping. Reinfusion of shed volume did not restore abdominal organ flow (4.7% baselines) but increased LAP and cardiac output to the upper body. Release of the cross-clamp produced profound acidosis that was treated effectively with NcHCO3. After stabilization of blood, flow to kidney remained low (49% baseline) although intestinal flow was increased threefold (320% of baseline). It is clear that thoracic aortic cross-clamping in shock further compromises already reduced visceral blood flow and may contribute to the problem of ischemic multiple organ failure after resuscitation from hemorrhagic shock.

Quantitative assessment in infarct size reduction by coronary venous retroperfusion in baboons.

Initial favorable reports in which coronary venous retroperfusion was begun after acute coronary artery occlusion have demonstrated a reversal of ischemic injury and improved left ventricular function. However, little information has been generated to document the extent to which retroperfusion may decrease ultimate histologically determined infarct size. The objective of the present study was to evaluate the effectiveness of retroperfusion in reducing infarct size by using an accurate quantitative method in which infarct size was related to the size of the anatomic perfusion bed of the occluded artery (region at risk for infarction). In an experimental group of 5 baboons, the left anterior descending coronary artery was occluded and coronary venous retroperfusion started 1 hour after occlusion. After a 4-hour period of occlusion, retroperfusion was discontinued and anterograde perfusion was simultaneously restored. A control group of 5 baboons underwent an identical procedure without retroperfusion. Twenty-four hours after occlusion, hearts were excised and the previously occluded left anterior descending coronary artery as well as the adjacent arteries were infected with microvascular dye to delineate the perfusion bed of the occluded artery. Planimetry of serial corss-sections of the left ventricle enabled the size of the perfusion bed of the occluded artery and size of the infarct to be determined. The mean percentage of the perfusion bed infarcted in the control group was 94.1 +/- 0.9 (mean +/- standard error) and in the retroperfused group was 57.4 +/- 3.5 (p less than 0.001). Hence, the results demonstrated that when retroperfusion was initiated after 1 hour of coronary occlusion, the mean percentage of the perfusion bed salvaged was increased by 36.7%.

Failure of nifedipine therapy to reduce myocardial infarct size in the baboon.

The value of nifedipine in reducing the ultimate size of an infarct associated with a period of coronary occlusion followed by reperfusion was assessed. Eight baboons were administered a bolus dose of nifedipine, 5 micrograms/kg intravenously, and then a maintenance dose of 30 micrograms/kg per hour was begun 1 hour before occlusion. This regimen resulted in an 8.5 +/- 1.2 percent (mean +/- standard error) decrease in mean arterial pressure. The left anterior descending coronary artery was occluded for 2 hours and then perfusion restored. At 2 hours after reperfusion the nifedipine infusion was discontinued. Eight control baboons underwent an identical protocol without nifedipine therapy. At 24 hours after occlusion, microvascular dyes were injected into the left anterior descending coronary artery and adjacent arteries to delineate the perfusion bed of the previously occluded artery. The volume of infarction was determined with planimetry and compared with the volume of the perfusion bed of the occluded artery. The area of infarction was always contained within the perfusion bed of the occluded artery. The mean percent of the perfusion bed with infarction was 50.1 +/- 5.8 in the control group and 41.7 +/- 9.5 in the treated group (difference not significant; p greater than 0.05). In both control and treated groups of baboons hemorrhage occurred only within the region of infarction. In both groups electron microscopy revealed large electron-dense granules within the mitochondria. In conclusion nifedipine therapy during a 2 hour period of coronary occlusion followed by reperfusion did not result in any significant reduction in ultimate infarct size in the baboon.

Infarct distribution in subhuman primates after acute coronary occlusion.

The extent of epicardial, endocardial and septal infarct was determined in 24 monkeys and five baboons 1 week after acute left anterior descending or diagonal branch coronary artery ligation. All 24 Macaca cynmologous monkeys had non-dominant left anterior descending coronary arteries. A snare ligature was placed just distal to the first diagonal branch for varying time periods (1, 2, 4, and 6 h followed by reperfusion or left in place permanently). In five baboons a chronic ligature was placed around a diagonal branch. All animals lived and were killed a week later. Histological mapping and planimetry of serial cross-sections were employed to quantify the extent and distribution of the infarct. All infarcts were transmural. The extent of epicardial infarct was significantly greater than the extent of endocardial infarct for 2 h ligations (3.5 +/- 0.87% of the left ventricle versus 2.4 +/- 0.58% of the left ventricle, P less than 0.05), the chronic left anterior descending coronary artery ligations (5.4 +/- 1.06% of the left ventricle versus 4.5 +/- 0.92% of the left ventricle, P less than 0.05 and for the chronic diagonal branch ligations (4.06 +/- 0.66% of the right ventricle + left ventricle versus 2.64 +/- 0.51+ of the right ventricle + left ventricle, P less than 0.02). It is evident, however, that the magnitude of this epicardial preponderance was not great and that, in general, the infarcts were transmural and rectangular in configuration.

Small intestinal blood flow after 48 hours ileus, prostigmin and manual decompression.

In eight piglets small intestinal blood flow (IBF) and motility has been measured after 48 hrs mechanical obstruction, prostigmin application and manual decompression using the microspheres method. Intraenteric basic pressure is in a normal range; the number of tonic contraction waves (type 3 waves) is slightly increased. IBF after 48 hrs ileus is markedly increased compared to normal (218%). Prostigmin does not influence IBF significantly. After manual decompression, IBF decreases to 56% of normal and does not recover within 30 minutes. Whereas IBF in the segment proximal to the obstruction is increased, IBF in the distal part is significantly reduced (48%) after 48 hrs of ileus. It is concluded that 48 hrs of ileus have no harmful effect on intestinal blood flow and function. Prostigmin does not increase IBF, whereas motility is stimulated strongly. Mechanical decompression results in a 50% reduction of IBF, which might be in accordance with clinical observation of retarded recovery and reduced motility after this maneuvre.

Intestinal blood flow at various intraluminal pressures in the piglet with closed abdomen.

The influence of intraluminal pressure on intestinal blood flow was studied in two segments of the small intestine and two of large intestine ligated after insertion of intraluminal catheters in ten piglets. Intestinal segments were inflated in stepwise increments in intraluminal pressures of 15, 30, 45 and 60 mmHg and blood flow was measured with radioactive microspheres using four isotopes (Ce, Cr, Sr, Sc). Other segments were inflated to a pressure of 60 mmHg and then pressure decreased in a stepwise fashion to 30, then 0 mmHg for the last two injections. Small and large intestinal blood flow fell progressively with increasing intraluminal pressure. At 60 mmHg a forward flow of 25% of normal was still present. Furthermore, not only was there an absolute decrease in blood flow with increasing intraluminal pressure but this decrease was disproportionately large in the intestinal mucosa. A hyperemic response lasting approximately 15 minutes was observed after complete decompression. The intestinal blood flow distal to the ligated segments was always moderately increased as compared to intestinal blood flow proximal to the segments. The results reported herein are at some variance from other reported studies performed with the abdomen open and on isolated segment preparations. The reasons for these variations are discussed.

Filtration of debris from banked blood.

Commercially available filters will effectively remove microaggregates from stored blood. The combined results of screen filtration pressure (SFP), debris weight, and particle size analysis offer a reproducible means of evaluating different filters. The effectiveness of three blood filters (Fenwal, Bentley, and Pall) is evaluated using SFP, debris weight measurement, particle size analysis, and determinations of filter capacity. Of the filters studied the Fenwal filter provides the most efficient means of removing debris while maintaining adquate flow rates for relatively large volumes of blood. The filter appears comparable to the Swank filter in overall effectiveness and flow characteristics.

Oxygen consumption changes with stored blood infusions.

Stored blood contains microaggregates, often implicated in the pathogenesis of post-traumatic pulmonary insufficiency. This study was an attempt to further elucidate the effect of autologous stored, filtered and non-filtered blood infusions and homologous stored and fresh blood infusions on pulmonary function and hemodynamics. Inconsistent changes in pulmonary hemodynamics and blood oxygenation were noted. The one significant finding was an increase in oxygen consumption, which occurred with unfiltered autologous or homologous blood but not with fresh or filtered blood. Since an increased oxygen consumption results in an oxygen demand which is difficult to meet in the face of multiple other injuries, it is conceivable that this observation implicates massive stored blood transfusion as a major contributing factor in the development of so-called irreversible shock.

Comparison of filtering efficiency of four new in-line blood transfusion filters.

Efficient removal of debris from stored human blood prior to transfusion has become increasingly important. The debris, consisting largely of microaggregates of platelets and fibrin, is not effectively removed by passage through a standard transfusion filter. This study evaluated the performance of four of the currently available small pore in-line blood transfusion filters. Filters tested included the Bentley PF-127, the Pall Ultipor SQ-40, the Swank In-Line IL-200 and the Fenwal Microaggregate Blood Filter. A standard blood administration filter was also tested, the McGraw V-2950. The rate of blood flow through the filters was recorded using single and multiple units of blood. The screen filtration pressure and debris weight of the filtered blood were studied to compare effectiveness of filtration. The Swank filter was effective in debris removal and maintained good flow rates. The Bentley and Fenwall filters removed debris nearly as well, but had reduction of flow rates after smaller infusions. The Pall filter maintained high flow rates but did not remove debris as effectively, particularly with pressure infusion. The standard 170 mu pore blood transfusion filter does not remove microaggregates.

Maintenance of fiable segments of human artery in vitro.

A system for maintaining viable segments of atherosclerotic human artery in vitro for periods of up to 32 days is described. Viability of the segment has been confirmed from study of lactate metabolism and vessel wall histology. Preliminary lipid studies using TLC on plasma perfusate indicate lipid may be removed from the vessel wall during perfusion.