Attenuation of myocardial stunning in isolated rat hearts by a 21-aminosteroid lazaroid (U74389G).

The purpose of this study was to evaluate the effects of reperfusion or in vivo pretreatment with a lipid peroxidation inhibitor, lazaroid (U74389G), on attenuating systolic and diastolic alterations occurring during myocardial stunning in isolated rat hearts. Male Sprague-Dawley rats (350-400 g) were randomized into three groups: control animals (n = 13) received no drugs; hearts from reperfused animals (n = 11) received 5 microM U74389G in the reperfusion solution; pretreated animals (n = 11) received 6 mg/kg U74389G by i.v. infusion 30 min before killing. Isolated, isovolumic rat hearts were subjected to 20 min of ischemia at 37 degrees C and subsequent reperfusion for 30 min. Left ventricular isovolumic developed pressure (LVDP), its first derivative (LVDPdP/dt), end-diastolic pressure (LVEDP), and the time constant of diastolic relaxation (tau) were measured. At baseline, no statistically significant differences were detected in systolic or diastolic function in hearts of rats with or without U74389G treatment. After reperfusion, LVDP stabilized at 87 and 92% in both drug-treated groups compared with 52% in the control group (p < 0.01) and dP/dtmax recovered to 101 and 110% of baseline compared with 58% in the control group (p < 0.01). Diastolic dysfunction showed significant improvement in both U74389G pretreatment groups. The increases in LVEDP and tau were 2.0- and 1.2-fold in pretreated hearts and 2,8-fold and 1.5-fold in drug-reperfused hearts, respectively (compared with 6-fold increases in LVEDP and a 2.5-fold increase in tau in controls; p < 0.05). In conclusion, whether administered before ischemia or during reperfusion, U74389G effectively attenuated the systolic and diastolic dysfunction in this model of myocardial stunning, probably protecting cell membranes from peroxidation by oxygen-derived metabolites.

The role of stressor intensity in influencing the course of heart disease in cardiomyopathic hamsters.

Our earlier work showed that stress had progressively more serious consequences in a hamster model of congestive heart failure as the magnitude of heart failure worsened. Based on that study, we hypothesized that the intensity of the stressor used might play an important part in determining this outcome as well as in influencing coronary reactivity to arginine vasopressin (AVP). Cardiomyopathic (2.5, 6.5, and 10 months) hamsters (CMHs) were stressed with a 2-hr period of supine immobilization for five consecutive days. Stressor intensity was increased by exposing the hamsters to progressively longer periods at 4 degrees C: the low stress group was never put in the cold; the moderate stress group was exposed to cold for 1 hr, and the high stress group for 2 hr. CMHs were anesthetized and sacrificed 5 days after stress, and their hearts were perfused using a modified Langendorff system. Maximum +/- dP/dt, developed pressure, ventricular relaxation time, (T), and coronary vascular resistance (CVR) were recorded, and CVR was also measured following coronary infusion of AVP. Stressor intensity had no effect on cardiac mechanics in 2.5-month CMHs. In 6.5-month CMHs, only the high-intensity stressor impaired ventricular mechanics (decreased maximum +/- dP/dt and developed pressure, increased T; P < 0.05), while low and moderate stress produced no effects. In 10-month CMHs, stress at all intensities exacerbated ventricular dysfunction (decreased maximum +/- dP/dt and developed pressure; P < 0.05). These results support our first hypothesis that stressor intensity interacts multiplicatively with severity of the underlying disease to influence the course of heart failure. However, our second hypothesis was not supported, because stress-regardless of intensity-affected reactivity of the coronary vasculature to AVP only in 2.5-month CMHs. A further test of the relation of stressor intensity and coronary vascular reactivity requires study of additional groups of CMHs during the period of their disease characterized by coronary vasospasm.

Stress triggers different pathophysiological mechanisms in younger and older cardiomyopathic hamsters.

OBJECTIVES: Because cardiomyopathic hamsters (CMHs) in the lesion-forming period of their disease are more susceptible to the lethal effects of stress than older CMHs, we tested the hypothesis that different pathophysiological effects of stress may occur: coronary vasospasm in younger CMHs and congestive heart failure in older ones. METHODS: CMHs aged 2.5 and 6.5 months were stressed with 2 h supine cold immobilization for 5 consecutive days. Three, 5 and 7 days after stress, the hearts were excised and perfused using a modified Langendorff system. Maximum +/- dP/dt, developed pressure, ventricular relaxation time (Tau) and coronary vascular resistance (CVR) were recorded and CVR was also measured following coronary infusion of arginine vasopressin (AVP). RESULTS: Stress produced ventricular dysfunction (decreased maximum +/- dP/dt, developed pressure, and increased Tau) in older CMHs (P < 0.05) but not in younger CMHs. Baseline CVR in younger CMHs was significantly higher than in older CMHs (P < 0.01) and AVP infusion produced a bigger increase in CVR in younger stressed CMHs than in either younger nonstressed or older stressed CMHs (P < 0.05). CONCLUSION: The younger CMH heart exhibits greater resting vascular tone and stress produces coronary vasoconstriction that is consistent with coronary spasm. In contrast, the older CMH experiences a decrease in cardiac function which remains 7 days after stress and indicates an exacerbation of CHF from the mild form existing prior to stress. The lethal effects of stress may occur because of the activation of different pathological processes in younger and older CMHs.

Increased oxygen extraction as adaptation to acute renal ischemia.

Acute renal ischemia is an infrequently encountered clinical entity with occasionally devastating consequences. The renal compensation to acute ischemia is unknown and is the purpose of this report. Eight pigs were anesthetized and ventilated. Left atrial, aortic, CVP, left renal venous, and ureteral catheters were inserted. Renal blood flow (RBF) reduction was accomplished by the graded constriction of the left renal artery using a balloon occluder. Renal oxygen extraction ratio (RER, %), renal oxygen delivery (RO2D, cc/min per 100 gm), renal oxygen consumption (RVO2, cc/min/100 gm), creatinine clearance (CrCl, ml/min), and renal lactate production (delta [L], mg/min per hgm) were measured at baseline and following sequential 90-minute intervals of moderate and then severe left renal flow reduction. Significant increases in renal oxygen extraction were observed when RBF was severely limited (.30 +/- .05 vs .64 +/- .06, p < .01). CrCl decreased precipitously (16.5 +/- 4.6 vs 0.2 +/- 0.07, p < .05). Lactate production by the ischemic organs correlated with blood flow reduction (r = .546, p = .0034). In severe ischemia, healthy kidneys increase oxygen extraction to preserve oxygen consumption.

Enhanced coronary vasoconstriction in the Syrian myopathic hamster supports the microvascular spasm hypothesis.

OBJECTIVE: The heart of the young Syrian cardiomyopathic hamster (CMH) displays discrete patches of active myocytolytic necrosis and it has been hypothesised that these lesions are triggered by arteriolar spasm. Accordingly, we tested the theory that the coronary vasoconstrictor response to arginine vasopressin (AVP) infusion is more vigorous in the actively necrotising young (2-3 month old) CMH than in age matched normal hamsters, 5-6 month old CMHs in the histologically quiescent phase of the disease, or 5-6 month old normal hamsters. METHODS: An isovolumetric isolated heart preparation was used in which the coronary arteries were perfused with Krebs-Henseleit buffer at constant flow by a syringe pump. Coronary vascular resistance was determined by the ratio of measured perfusion pressure to flow rate during extended diastoles. RESULTS: There were no significant differences in baseline coronary vascular resistance among the four groups. The increase in resistance with AVP infusion (0.54 pressor units.min-1) was significantly greater (p < 0.01) in the young CMH [6.66(SEM 4.75) mm Hg.ml-1.min-1] than in the old CMH [1.66(0.78)], the young normal [1.10(1.07)], and the old normal [2.72(1.86)] groups. CONCLUSIONS: There is increased vasoconstrictive responsiveness in the young CMH coronary vasculature. The results suggest a broader coronary abnormality in this myopathic model and are consistent with the microvascular spasm hypothesis of myocytolytic lesion formation.

Protection of the acutely injured heart--deleterious effects of hypothermia.

This experimental study was undertaken to evaluate the hypothesis that cardioplegic arrest at 37 degrees C is superior to 10 degrees C cardioplegic arrest in the myocardial protection of an acutely injured heart. The hypothesis was tested using Sprague-Dawley rat hearts that were excised and retroperfused with Krebs-Henseleit buffer as isolated, isovolumic heart preparations. After 15 min of equilibration and baseline readings, ischemic injury of the myocardium was established by cessation of perfusion for 20 min, followed by 30 min of reperfusion to obtain cardiac measurements and verify uniformity of ventricular dysfunction. All hearts were then arrested for 30 min with continuous cardioplegia (oxygenated crystalloid cardioplegia, 4 ml/min) followed by 30 min of reperfusion. In one group (N = 12), temperature of cardioplegia was 10 degrees C. In the second group (N = 12), cardioplegia temperature was 37 degrees C. The experiments showed that in acutely injured hearts, cardioplegic arrest at 10 degrees C resulted in further deterioration in ventricular dysfunction (P < 0.01). In contrast, similarly injured hearts which underwent cardioplegic arrest at 37 degrees C showed a modest deterioration in ventricular dysfunction which did not reach statistical significance (P > 0.05). The experimental findings suggest that in the presence of acute myocardial injury, cardioplegic arrest at normothermia may provide better myocardial protection.

Coronary alpha-adrenergic stimulation does not alter the heterogeneity of myocardial blood flow and oxygen utilization.

Previous studies have suggested an uneven distribution of alpha-adrenergic receptors within the coronary arterial tree. We hypothesized that stimulation of these receptors would alter the regional and microregional variability of oxygen supply and utilization. Accordingly, we measured the heterogeneity of coronary blood flow to 0.5 g tissue samples and O2 saturation of small cardiac veins during coronary alpha-adrenergic stimulation. In 14 dogs we cannulated and perfused a branch of the left coronary artery. In seven dogs, an intracoronary infusion of the alpha-adrenergic agonist phenylephrine was established after prior beta-adrenergic blockade. Phenylephrine infusion decreased subendocardial venous O2 saturation (41.6 +/- 5.6 vs 45.4 +/- 5.8% saturated, P less than 0.05). It did not change the heterogeneity or distribution of the O2 saturations relative to the normally-perfused region (coefficient of variation = 22.4 +/- 6.7 vs. 19.7 +/- 4.8%, P = 0.20). Regional blood flows were made during control, beta-adrenergic blockade, and phenylephrine infusion conditions in the remaining dogs. The coefficient of variation of blood flow to samples from the perfused region was 27.1 +/- 9.8, 27.8 +/- 10.8, and 24.7 +/- 7.5% (P greater than 0.25), respectively. The results indicate that coronary alpha-adrenergic stimulation does not alter O2 supply/utilization heterogeneity, and suggest a fairly uniform distribution and/or effect of alpha-adrenergic receptors within the coronary resistance vessels.

Ventricular preload alters intravascular and extravascular resistances of coronary collaterals.

Coronary collateral blood flow is determined by both the collateral vessel resistance and a waterfall mechanism. The aim of this study was to determine which of these two mechanisms predominates during alteration of ventricular preload. The left anterior descending coronary artery of 12 anesthetized dogs was cannulated, and the distal vasculature was completely embolized with 25-micron diameter microspheres. Retrograde blood flow (RBF) was collected when the cannula was opened to the atmosphere, and the outflow tubing height was adjusted to provide a variable back pressure. RBF is back pressure-dependent at higher back pressures, and the slope in this region of the constructed pressure-flow relationship determines the collateral conductance. The transition point between the back pressure-dependent and a back pressure-independent region indicates a waterfall pressure impinging on the collateral vessels. At a left ventricular diastolic pressure of 9.3 mmHg, mean RBF, collateral conductance, and the collateral waterfall pressure were 7.3 ml/min, 0.175 ml.min-1.mmHg-1, and 30.1 mmHg, respectively. Corresponding values when preload was reduced to 3.5 mmHg were 9.3 ml/min, 0.186 ml.min-1.mmHg-1, and 23.7 mmHg, all changes being significant. Mean contribution to the overall increase in RBF was 0.5 ml/min for the conductance and 1.2 ml/min for the waterfall mechanism (P less than 0.05), or 29 and 71%, respectively. The results indicate that the extravascular resistance mechanism mediates the collateral flow response to a greater degree than the intravascular resistance during variations in preload. The increase in slope of the conductance portion of the relationship was not accompanied by a concomitant increase in slope of the back pressure-independent region. These data further support a collateral waterfall, and not collateral vessel compliance, as the basis for the back pressure-independent portion of the pressure-flow relationship.

Spatial heterogeneity of local blood flow and metabolite content in dog hearts.

Spatial variation (heterogeneity) of myocardial blood flow was studied under basal conditions in relation to four biochemical markers: creatine kinase (CK), lactate dehydrogenase (LDH), ATP, and glycogen. A total of 508 individual 0.5-g samples from the left ventricular free wall was studied in 12 dogs. Myocardial blood flow was measured by radioactive microspheres (15 micron diam) injected via a pigtail catheter into the left ventricle during light sedation (closed-chest flow measurements); following thoracotomy, a second set of microspheres was injected via a catheter into the left atrium (open-chest flow measurements, n = 5). In 27-54 samples/heart, myocardial blood flow, CK, LDH, protein, ATP, and glycogen were determined, permitting a direct correspondence between local blood flow and metabolic markers in each sample and an assessment of the spatial heterogeneity of flow and metabolite content. Correlations between myocardial blood flow per gram tissue and metabolite concentration per gram tissue in pooled data were weak but significant: partial correlation coefficients were 0.137 for CK (P = 0.024), 0.167 for LDH (P = 0.006), 0.341 for ATP (P less than 0.001), 0.123 for glycogen (P = 0.053), but not significant for protein vs. myocardial blood flow. The coefficient of variation, which defines the extent of spatial heterogeneity, averaged 20% for closed-chest flow measurements, 19% for open-chest flow measurements, 22% for CK, 17% for LDH, 15% for protein, 8% for ATP, and 18% for glycogen; these values are over and above the variability due to the technique error, indicating a definite physiological spatial variability. The correlation between local blood flow and the studied metabolites can only explain a minor portion of the spatial heterogeneity of myocardial blood flow. Although a physiological link between blood flow and metabolite content for small regions of the heart is demonstrated, the true local variability of blood flow may be modulated predominantly by other factors.

Microvascular reactivity of the myopathic Syrian hamster cremaster muscle.

The myopathic Syrian hamster is a genetic model of congestive heart failure that exhibits focal myocytolytic necrosis in both heart and skeletal muscle. Previous investigations of microvascular morphology in heart and skeletal muscle have shown severe arteriolar constrictions without fixed anatomical vessel lesions. This study tested the hypothesis that these constrictions indicate a hyperreactivity of the myopathic microvasculature in vivo and that the reactivity corresponds to the developmental course of myocyte pathology. The microcirculation of the cremaster muscle was studied in eight myopathic and six control hamsters in the active stage of necrosis (39-81 days of age) and five myopathic and six control hamsters in the later stage of muscle healing (150-213 days of age). The internal diameter of second order arterioles was measured during topically applied noradrenaline. The myopathic arterioles of the younger group constricted at significantly lower concentrations of noradrenaline (p less than 0.01) and constricted to 35-50% of their resting internal diameter over a narrower range of noradrenaline (p less than 0.01). This indicated both a reduced threshold to noradrenaline and an enhanced response to the agonist. Active myocytolytic necrosis was found in the contralateral cremaster muscle of each myopathic hamster. The older myopathic and control hamsters showed no significant differences in arteriolar responsiveness to applied noradrenaline and no active necrosis. These results indicate a relation between a hyperreactive microvasculature and active necrosis and a normal reacting microvasculature and diminished necrosis in the two phases of the disease. Thus a general correspondence between vascular responsiveness and myocyte pathology exists in this model of heart failure and muscular dystrophy.

Morphometric study of the total and perfused arteriolar and capillary network of the rabbit left ventricle.

A method has been developed to simultaneously and regionally determine several morphometric indices of both the total and perfused arteriolar and capillary beds of the rabbit heart on a quantitative basis. A high molecular weight fluorescein isothiocyanate (FITC) dextran, was injected into the femoral vein of an anaesthetised open chest rabbit. The hearts were removed and frozen in liquid N2 within 14 s after this injection. Sections, 2 micrometers thick, were obtained from the left ventricular wall and the fluorescence photographed to detect perfused arterioles and capillaries. The sections were then stained to visualise the total arteriolar and capillary network. Standard morphometric techniques were employed to study the total and perfused microvasculature. The method was validated to show that it did not alter the microvasculature of the heart, that both the total and perfused microvasculature could be visualised, and that there was little tissue shrinkage. The study found no significant subepicardial vs subendocardial differences in either the total or perfused capillary of arteriolar networks. The total capillary bed had very much larger morphometric indices than the total arteriolar bed. The percentage of the microvasculature perfused ranged from 55 to 64% of the capillary and 51 to 56% of the arteriolar beds for the various indices. Thus at least a part of the control of capillary perfusions is at the level of the arteriole. There was an almost twofold functional reserve in the microvasculature of the heart which could be mobilised by asphyxia.

Dependence of spatial heterogeneity of myocardial blood flow on mean blood flow rate in the rabbit heart.

The relationship between spatial heterogeneity of myocardial blood flow and the rate of perfusion was examined in anaesthetised-chest rabbits. Blood flow determinations employed the radioactive microsphere technique. Five tissue samples were obtained from each of four regions of the left ventricle: left septum, right septum, subendocardium and subepicardium. Standard deviation (SD) and the coefficient of variation (CV = SD/mean flow x 100), an index of spatial heterogeneity, were computed. A wide range of mean coronary flows was obtained in seven groups by inspiration of room air, 8% O2, 8% O2 and 10% CO2, 100% O2, 100% O2 and hyperventilation, and by administration of adenosine or chromomar HCl. Significant differences in CV were found between treatment groups at the upper and lower ends of the flow range. A significant positive linear correlation between overall SD and mean coronary flow was found (r = 0.760). A significant inverse linear relationship between CV and mean flow was found over the entire range of flows studied (r = -0.482). An improved correlation between CV of calculated coronary vascular resistance and mean vascular resistance (r = 0.742) amongst these treatments suggests that spatial heterogeneity is better described by the variability in calibre of large arterioles. This decreased heterogeneity accompanying high flows or reduced vascular resistance may be beneficial in terms of O2 supply and demand under conditions of myocardial stress.

State of the art in fermentation air filtration.

This article presents several characteristics of a state-of-the-art fermentation air filter. The filter medium is composed solely of PTFE and has an absolute pore size rating of 0.2 mum. Quantitative bacteria and bacteriophage retention is shown based on live organism challenge tests. A nondestructive filter test, correlated to the microorganism challenge tests and called the Forward Flow Integrity Test, is described. This test has a sensitivity of one part in 10(12) and can be performed in situ.

Role of propranolol in improvement of the relationship between O2 supply and consumption in an ischemic region of the dog heart.

Several aspects of the myocardial O(2) supply/consumption relationship were determined after coronary artery occlusion and subsequent beta-adrenergic blockade in 16 anesthetized open-chest dogs. Small artery and vein O(2) saturations, and hence extraction, were obtained microspectrophotometrically and combined with radioactive microsphere blood flow determinations to calculate regional myocardial O(2) consumption. Eight dogs remained untreated after coronary artery ligation while another group was given 2 mg/kg propranolol, 10 min after occlusion. Untreated occlusion resulted in decreased arterial and especially venous O(2) saturations, indicating an increased O(2) extraction. Ischemic O(2) consumption was reduced and the subendocardial/subepicardial consumption ratio was reversed (1.26 vs. 0.37) due to the pattern of occluded area flow. Calculated O(2) supply/consumption also decreased. Propranolol produced no significant changes in volume or distribution of flow within the ischemic region while reducing flow, extraction, and consumption in the unoccluded region. The heterogeneity of arterial and particularly venous O(2) saturations within the ischemic region decreased dramatically. Venous O(2) saturations were elevated relative to the control group resulting in a reduced O(2) extraction. The decrease in heterogeneity of arterial and venous O(2) saturations suggest that propranolol eliminates microregions of relatively high O(2) extraction, consumption, and/or a majority of vessels with extremely low flow. This leads to a significant improvement in the O(2) supply/consumption ratio in the ischemic myocardium of the dog. This may be due to a reduction in the heterogeneity and level of beta(1)-adrenergic receptor activity within the heart.

Effect of papaverine on regional cerebral blood flow and small vessel blood content.

The effect of intravenous infusion of papaverine HCl, (2.0-2.5 mg/kg) on regional cerebral blood flow and small vessel blood content in anesthetized rats was determined. Cerebral blood flow was measured in the cerebral cortex, diencephalon plus midbrain, cerebellum, pons and medulla by means of the uptake of iodo[14C]antipyrine. Small vessel blood content was determined by labelling the plasma siderophilin with 59FeCl3. Cerebral blood flow showed interregional variation under control conditions. Papaverine increased flow in all regions, but the effect was greater in the hind compared to the forebrain. Papaverine did not significantly alter small vessel blood content or calculated open capillary density, indicating it had no functional effect upon cerebral precapillary sphincters, and therefore only affected cerebral arterioles.