Ca2+ increase and pH decrease induced by hypochlorous acid in single quiescent myocytes isolated from rat ventricles.

Cytosolic free calcium concentration ([Ca2+]i) and intracellular pH (pHi) were simultaneously measured with a fluorescence technique using fura-2 and BCECF. Hypochlorous acid (HOC1) increased [Ca2+]i and decreased pHi of single quiescent myocytes isolated from rat ventricles. The HOC1-induced changes in [Ca2+]i and pHi (delta [Ca2+]i and delta pHi) were 129 +/- 18nM and 0.18 +/- 0.02 (mean +/- SD), respectively, with 200 microM HOC1. A positive linear correlation was obtained between delta [Ca2+]i and delta pHi under various extracellular Ca2+, pH, and Na+ conditions. Chelation of extracellular Ca2+ depressed only one-fourth of delta [Ca2+]i, and Ca2+ antagonists (verapamil and nifedipine) did not reduce delta [Ca2+]i, indicating that delta [Ca2+]i originates mainly from intracellular Ca2+ stores. A disulfide-reducing reagent, 1,4-dithiothreitol (DTT), recovered the increased [Ca2+]i and decreased pHi to the control levels. The simultaneous changes in [Ca2+]i and pHi induced by HOC1 and the simultaneous restoration of the normal [Ca2+]i and pHi in cardiac myocytes.

Sensitivity to oxidants of mitochondrial and sarcoplasmic reticular calcium uptake in saponin-treated cardiac myocytes.

Calcium transport functions of mitochondria and sarcoplasmic reticulum (SR) were studied without prior extraction using isolated rat heart myocytes permeabilized with saponin. Calcium uptake by SR was rapid and its affinity was high in comparison to calcium uptake by mitochondria, which had a higher capacity. The sensitivity of uptake to two oxidants, H2O2 and HOCl (hypochlorous acid), depended on the cytosolic calcium concentration; when this was similar to the concentration in diastole (180 nM), HOCl inhibited calcium uptake by mitochondria and SR, whereas when the calcium concentration was 750 nM, mitochondrial calcium uptake showed relatively high resistance, although SR uptake was still markedly inhibited by HOCl. Calcium uptake of both mitochondria and SR was less sensitive to the action of H2O2 than to HOCl, and the H2O2 effect was less dependent on the cytosolic calcium concentration. Therefore, HOCl, when produced by activated leukocytes and supplied to the heart cells, may seriously impair the excitation-contraction coupling function of SR, whereas H2O2, possibly generated directly by mitochondria or generated from superoxide anions, may be tolerated relatively well by heart SR and mitochondria.

[Extension of measurable range of film dosimetry by a low energy X-ray absorption method].

The availability of an X-ray absorption technique employing a very low power X-ray tube was examined to extend measurable range for photographic film dosimetry. The X-ray tube having titanium target was operated at 8 kV and 0.2 microA to emit Ti KX-rays of moderate intensity. The degree of the Ti KX-ray absorption, defined as similar to photographic density, was measured for the two kinds of badge film, Fuji gamma-ray badge film and Kodak personal monitoring film, type 2 exposed for 60Co or 137Cs gamma-rays and developed by the respective standard procedures. The experimental results show that the dosimetric range of 0.01-100 R for the Fuji film and 0.03-1,000 R for the Kodak film may be easily measured by 1 minute counting with the relative statistical error (sigma) of 10%.

Increase vs. decrease of calcium uptake by isolated heart cells induced by H2O2 vs. HOCl.

Adult rat heart myocytes were labeled rapidly with exogenous [45Ca2+]. Addition of 2.5 mM H2O2 to the heart cell suspension raised the content of rapidly exchangeable intracellular Ca2+ twofold, whereas addition of 1-30 mM HOCl decreased the Ca2+ content. The H2O2-induced increase in Ca2+ content was dependent on the medium Na+, pH, and temperature but was not significantly affected by addition of verapamil, diltiazem, amiloride, or 3-aminobenzamide. The [3H]ouabain binding to myocytes was suppressed by H2O2, whereas the Ca2+ efflux from myocytes was not influenced. An uncoupler, carbonyl cyanide m-chlorophenylhydrazone, reduced Ca2+ content, implying that the H2O2-induced change in Ca2+ content was not directly related to ATP depletion. On the other hand, the H2O2-induced Ca2+ accumulation in myocytes was prevented by deferoxamine or o-phenanthroline. These results suggest that H2O2 inhibited Na+-K+-ATPase, resulting in an increase in intracellular Na+ concentration and stimulation of sarcolemmal Na+-Ca2+ exchange activity, which caused a transient net Ca2+ influx into myocytes. By contrast, HOCl decreased the Ca2+ content of the rapidly exchangeable pool below control levels and this action of HOCl was antagonized by 1,4-dithiothreitol. HOCl accelerated Ca2+ efflux from myocytes. Ca2+ uptake and Ca2+-ATPase of the isolated sarcoplasmic reticular (SR) fraction were highly sensitive to the action of HOCl. Ca2+ uptake by intracellular sites, studied with myocytes permeabilized with digitonin, was inhibited by both H2O2 and HOCl. Thus these results suggest that HOCl inhibits the SR Ca2+ pump, resulting in the observed acceleration of Ca2+ efflux from and decline in Ca2+ content of myocytes.

Effect of diltiazem on coronary blood flow distribution in dog heart under ischemia.

Effect of intracoronary infusion of diltiazem (1 microgram/min) on regional myocardial blood flow (RMBF) was studied using 15-microns radioactive microspheres in 11 excised cross-circulated canine left ventricles. With total coronary blood flow (CBF) and heart rate (HR) held constant, regional ischemia was induced by ligating the left anterior descending coronary artery (LAD). Diltiazem at the dose used had no effects on ventricular Emax before and after LAD ligation. RMBF expressed by the counts divided by the counts averaged in all segments in each layer significantly (p less than 0.05) increased under diltiazem only in the low-flow region that had less than 50% RMBF before diltiazem; from 21% (+/- 12%) to 35% (+/- 18%) in the epicardial, from 22% (+/- 12%) to 32% (+/- 18%) in the midwall, and from 24% (+/- 10%) to 31% (+/- 12%) in the endocardial layers. We conclude that the beneficial effect of diltiazem on the ischemic heart involves a direct action on the coronary vascular system and does not necessarily depend on the concomitant changes in hemodynamics.

Ventricular wall motion and NE release in post-ischemic reperfused myocardium.

To clarify the relationship between post-ischemic myocardial dysfunction and local cardiac sympathetic nerve function, we measured regional myocardial length and norepinephrine (NE) release during sympathetic nerve stimulation in 32 mongrel dogs. Coronary occlusion was produced by balloon occluder for 15 min and reperfused for 60 min. Dogs were divided into 3 groups as follows; Group 1 (n = 14): Sympathetic nerve stimulation, Group 2 (n = 9): Pre-treatment with yohimbine hydrochloride (0.2 mg/kg) and sympathetic nerve stimulation, Group 3 (n = 9): Exogenous NE administration. Sympathetic nerve stimulation or NE infusion were performed before occlusion and after reperfusion. In group 1, the extent of the increase in systolic shortening during sympathetic nerve stimulation (delta - shortening) lowered at 5 min after reperfusion and augmented progressively. But, delta-shortening at an early reperfusion period did not reduce in group 2 and 3. NE release from the ischemic myocardium decreased in group 1 and did not recover for 60 min. When the cardiac sympathetic nerve was denervated with 90% phenol solution, NE release further decreased in group 1. On the other hand, NE release did not decrease in group 2. These results indicate that the response to sympathetic nerve stimulation decreased in post-ischemic reperfused myocardium and this was due to diminished NE release. It was considered that sympathetic nerve conduction was not completely impaired in post-ischemic myocardium and pre-synaptic alpha-2 receptor mediated negative feedback mechanism would play an important role in these diminished NE release.

Hyperkinetic contraction of a nonischemic segment of ischemic left ventricle in anesthetized dogs.

Regional myocardial function during acute coronary artery occlusion was studied with ultrasonic dimension gauges in 20 open-chest anesthetized dogs. Two pairs of ultrasonic crystals were implanted in the left ventricular free wall near the epicardium in an ischemic segment and in a control nonischemic segment, and the segment length (SL) and maximum velocity of systolic shortening (max dL/dt) were measured. In six dogs, the wall thickness (WT) was measured simultaneously in the same regions with sonomicrometry. Left ventricular pressure (LVP), aortic pressure (AoP), and plasma norepinephrine concentration in the coronary sinus (NECS) were also measured. The heart rate was kept constant (180 beats/min) with atrial pacing. The left anterior descending coronary artery was occluded at its distal portion without propranolol in 12 dogs (group 1) and 30 min after propranolol in eight dogs (group 2). In the ischemic region, coronary artery occlusion resulted in an increase in end-diastolic SL (50% at 3 min after occlusion in group 1, P less than 0.005), and a decrease in max dL/dt in systole (36% at 5 min after occlusion in group 1, P less than 0.02). In the nonischemic region, end-diastolic SL did not change significantly, but an increase in max dL/dt (29% at 10 min after occlusion in group 1, P less than 0.005) was observed in systole. Under propranolol (group 2), the results were similar to those of group 1. There were no significant changes in LVP, AoP, AoP, and NECS during occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Treadmill stress test using body surface mapping in coronary artery disease--the clinical significance of ST depression.

In this study we assessed in 27 patients with coronary artery disease whether the size of the ST-depression area, which was measured by body surface maps recorded before and after treadmill exercise, could be a useful indicator for evaluating coronary artery disease (CAD) quantitatively. The patients were divided into 3 groups on the basis of the findings of the left ventriculograms: patients with anterior asynergy (n = 6), those with inferior asynergy (n = 6) and those with no asynergy (n = 15). Coronary arteriograms were evaluated according to Pujadas, and epsilon Grade, as an index of the severity of CAD, was developed by adding the grade numbers of the 4 main coronary stems (right coronary artery, main trunk of the left coronary artery, left anterior descending artery and left circumflex artery). Patients with inferior asynergy and with no asynergy have ST-depression areas in proportion to their epsilon Grade (r = 0.845, p less than 0.001), whereas none of the patients with anterior asynergy showed ST-depression areas regardless of their epsilon Grade. Of 6 patients who had anterior asynergy, 5 (83%) had pathologic Q waves in the left anterior chest leads. These findings emphasize the clinical value of the ST-depression are for the quantitative and non-invasive diagnosis of CAD, especially in patients without pathologic Q waves in the left anterior chest leads.

[Severity and extent of coronary artery disease and their relationship to left ventricular functional reserve in the chronic disease state].

Our previous observations showed that left ventricular wall motion abnormality (asynergy) induced by stress was observed in the phase response of radionuclide technique, and the severity and extent of coronary artery disease (CAD) assessed by the Pujadas score (PS) correlated well with the maximal phase delay response (delta MPD) but not with the ejection fraction response (delta EF) in patients with CAD without old myocardial infarction (MI). This study evaluated the usefulness of EF, MPD and the first-third filling fraction (FF, divided by the volume accrued throughout diastole) at rest and during stress, using ergometer and first-pass radionuclide angiocardiography (RNA), to determine the severity of CAD, and to evaluate any abnormalities in the systolic and diastolic coupling in left ventricular function during stress. Seventy-four patients with significant CAD, including 41 with previous transmural MI (MI group) and 33 without MI (angina group) were the subjects of this study. EF at rest and during stress inversely correlated, and MPD on stress linearly correlated with PS in angina group with PS greater than 10, while no such correlations were found in MI group. A normal EF response (delta EF greater than or equal to 5%) was accompanied by a greater-than-normal response in FF (delta FF) in both groups. A lower EF response was accompanied by a smaller delta FF in angina group, but by a larger delta FF in MI group. The difference was statistically significant (p less than 0.03), without significant differences by age, PS, peak heart rate, systolic blood pressure, and ischemia on ECG during stress. Resting EF and FF by the RNA method correlated with those by left ventriculography (LVG), respectively. It was suggested that RNA is an accurate method for determining EF and FF, while phase analysis may provide some additional information different from that provided by LVG. We concluded that EF and MPD during stress are as useful as delta MPD with the exception of a few cases in predicting the severity of CAD, and that scar tissue within the ventricular wall in MI may play an important role in determining the ventricular diastolic mechanical property during stress.

[Functional reserve of the ischemic left ventricle with ventricular aneurysm to afterload stress: digital subtraction angiographic assessments].

Digital subtraction angiography (DSA) has been confirmed to be an accurate method for determining left ventricular function. It is a relatively non-invasive technique without inducing premature ventricular complexes. The response of left ventricular function to afterload stress was assessed using DSA for eight patients with old anterior myocardial infarction and ventricular aneurysm including that of the anterior wall (averaging 30.3 months after the acute episodes). Their ages ranged from 36 to 65 years and one patient was a woman. Prior to the investigation, we confirmed that a single DSA procedure did not alter left ventricular function in a pilot study of one patient (No. 8). After initial DSA in the basal state, methoxamine was infused intravenously (1 to 2 mg/min). When aortic systolic blood pressure increased by 30 to 50 mmHg, a second DSA was performed for each patient. Left ventricular volumes and ejection fractions were calculated by the area-length method, and regional wall motion was assessed by the visual method according to the AHA classification and the curvature radius of the apical ventricular aneurysm was calculated. Methoxamine induced neither acute heart failure nor angina pectoris in the present series. The heart rates decreased, and there were a significant increase in end-systolic volumes (p less than 0.05), end-systolic radii (p less than 0.05), and a significant decrease in ejection fractions (p less than 0.02) after methoxamine infusion. In 32 of 40 segments, regional wall motion was unchanged by methoxamine as assessed by the visual method; whereas, in the other eight, there was a deterioration.(ABSTRACT TRUNCATED AT 250 WORDS)