Diabetes and hyperglycemia impair activation of mitochondrial K(ATP) channels.

Hyperglycemia is an important predictor of cardiovascular mortality in patients with diabetes. We investigated the hypothesis that diabetes or acute hyperglycemia attenuates the reduction of myocardial infarct size produced by activation of mitochondrial ATP-regulated potassium (K(ATP)) channels. Acutely instrumented barbiturate-anesthetized dogs were subjected to a 60-min period of coronary artery occlusion and 3 h of reperfusion. Myocardial infarct size (triphenyltetrazolium chloride staining) was 25 +/- 1, 28 +/- 3, and 25 +/- 1% of the area at risk (AAR) for infarction in control, diabetic (3 wk after streptozotocin-alloxan), and hyperglycemic (15% intravenous dextrose) dogs, respectively. Diazoxide (2.5 mg/kg iv) significantly decreased infarct size (10 +/- 1% of AAR, P < 0.05) but did not produce protection in the presence of diabetes (28 +/- 5%) or moderate hyperglycemia (blood glucose 310 +/- 10 mg/dl; 23 +/- 2%). The dose of diazoxide and the degree of hyperglycemia were interactive. Profound (blood glucose 574 +/- 23 mg/dl) but not moderate hyperglycemia blocked the effects of high-dose (5.0 mg/kg) diazoxide [26 +/- 3, 15 +/- 3 (P < 0.05), and 11 +/- 2% (P < 0.05), respectively]. There were no differences in systemic hemodynamics, AAR, or coronary collateral blood flow (by radioactive microspheres) between groups. The results indicate that diabetes or hyperglycemia impairs activation of mitochondrial K(ATP) channels.

Levosimendan, a new positive inotropic drug, decreases myocardial infarct size via activation of K(ATP) channels.

UNLABELLED: We tested the hypothesis that levosimendan, a new positive inotropic drug that activates adenosine triphosphate-regulated potassium (K(ATP)) channels in vitro, decreases myocardial infarct size in vivo. Myocardial infarct size was measured after a 60-min left anterior descending coronary artery occlusion and 3 h of reperfusion in dogs receiving either IV vehicle (0.9% saline) or levosimendan (24 microg/kg bolus followed by an infusion of 0.4 microg x kg(-1) x min(-1)) in the presence or absence of glyburide (a K(ATP) channel antagonist) pretreatment (100 microg/kg). Levosimendan increased (P < 0.05) the maximal rate of increase of left ventricular pressure and decreased myocardial infarct size from 24%+/-2% (control experiments) to 11%+/-2% of the left ventricular area at risk for infarction. Glyburide did not alter the hemodynamic effects of levosimendan but blocked levosimendan-induced reductions of infarct size. Subendocardial collateral blood flow was similar among groups. However, levosimendan increased subepicardial and midmyocardial collateral perfusion in the absence, but not in the presence, of glyburide. Levosimendan exerts cardioprotective effects via activation of K(ATP) channels at a dose that simultaneously enhances myocardial contractility. IMPLICATIONS: Levosimendan may be advantageous in patients requiring inotropic support who are also at risk of myocardial ischemia. Activation of adenosine triphosphate-regulated potassium channels during infusion of levosimendan may produce cardioprotective effects while simultaneously enhancing ventricular contractile function.

Isoflurane-enhanced recovery of canine stunned myocardium: role for protein kinase C?

BACKGROUND: Isoflurane enhances the functional recovery of postischemic, reperfused myocardium by activating adenosine A1 receptors and adenosine triphosphate-regulated potassium channels. Whether protein kinase C is involved in this process is unknown. The authors tested the hypothesis that inhibition of protein kinase C, using the selective antagonist bisindolylmaleimide, attenuates isoflurane-enhanced recovery of stunned myocardium in dogs. METHODS: Fifty dogs were randomly assigned to receive intracoronary vehicle or bisindolylmaleimide (2 or 8 microg/min) in the presence or absence of isoflurane (1 minimum alveolar concentration). Five brief (5 min) coronary artery occlusions interspersed with 5-min reperfusion periods followed by 180 min of final reperfusion were used to produce myocardial stunning. Hemodynamics, regional segment shortening, and myocardial blood flow (radioactive microspheres) were measured at selected intervals. RESULTS: There were no differences in baseline hemodynamics, segment shortening, or coronary collateral blood flow between groups. Isoflurane significantly (P<0.05) decreased heart rate, mean arterial pressure, rate pressure product, and the maximum rate of increase of left ventricular pressure (+dP/dt(max)) in the presence or absence of bisindolylmaleimide. Sustained contractile dysfunction was observed in dogs that received vehicle (recovery of segment shortening to 12+/-8% of baseline), in contrast to those that received isoflurane (75+/-7% recovery). Bisindolylmaleimide at a dose of 2 microg/min alone enhanced recovery of segment shortening (50+/-7% of baseline) compared with vehicle-pretreated dogs, and isoflurane in the presence of 2 microg/min bisindolylmaleimide further enhanced recovery of contractile function (79+/-8% of baseline). In contrast, 8 microg/min bisindolylmaleimide alone (32+/-12%) or combined with isoflurane (37+/-17%) did not enhance recovery of segment shortening compared with vehicle-pretreated dogs. CONCLUSIONS: The results indicate that protein kinase C inhibition using low doses of bisindolylmaleimide alone produces cardioprotection, and isoflurane further enhances this protection. In contrast, high doses of bisindolylmaleimide are not cardioprotective in the presence or absence of isoflurane. A role for protein kinase C during isoflurane-induced recovery of the stunned myocardium cannot be excluded.

Levosimendan enhances cardiac performance after cardiopulmonary bypass: a prospective, randomized placebo-controlled trial.

Levosimendan is a new myofilament calcium (Ca2+) sensitizer that increases myocardial contractility by stabilizing the Ca2+-bound conformation of troponin C. We tested the hypothesis that levosimendan enhances cardiac performance after cardiopulmonary bypass (CPB). Anesthesia was induced and maintained with midazolam, sufentanil, and vecuronium in 18 patients randomly assigned to receive levosimendan (18 or 36 microg/kg loading dose and 0.2 or 0.3 microg/kg/min infusion, respectively) or placebo 15 min before and continued for 6 h after CPB. Significant (p < 0.05) increases in heart rate (HR) and decreases in systemic vascular resistance (SVR) occurred 15 min after CPB in patients receiving placebo. Later increases in mean arterial pressure (MAP) and cardiac output (CO) and decreases in stroke volume (SV) and pulmonary vascular resistance also were observed. HR was greater in patients receiving high- but not low-dose levosimendan versus placebo immediately after CPB. MAP also was lower in patients treated with either dose of levosimendan compared with placebo after CPB. Levosimendan increased CO and decreased SVR (4.2 +/- 0.4 to 7.9 +/- 0.4 L/min and 1,150 +/- 99 to 512 +/- 42 dyn/s/cm5, respectively, 15 min after CPB; mean +/- SEM). CO and SV were higher and SVR was lower in patients receiving levosimendan versus placebo after CPB. No differences in arterial oxygenation and perioperative arrhythmias (Holter analysis) were observed between groups. The results indicate that levosimendan enhances cardiac performance after CPB in humans.

Naturalistic action production following right hemisphere stroke.

An unselected group of right hemisphere, semi-acute stroke patients (n = 30) was run on a laboratory test of naturalistic action production and was found to commit errors of action at a higher rate than what was previously reported for recovering head injury patients [Schwartz et al., Naturalistic action impairment in closed head injury. Neuropsychology, 1997, 8, 59-72]. There were strong similarities in how these two patient groups responded to variations in task demands and in the pattern of errors they produced. Hemispatial biases were evident in the errors of right hemisphere patients with neglect but not those without neglect; and neglect patients also many errors that were unrelated to the spatial layout. We argue that a non-specific resource limitation--which might translate as reduced arousal or effort--is central to the breakdown of naturalistic action production after brain damage, and right hemisphere patients are especially vulnerable to this resource limitation and its behavioral consequences.

Awareness of errors in naturalistic action after traumatic brain injury.

A prospective study was performed to develop a method for assessing "on-line" error detection and correction during performance of naturalistic action, to determine whether traumatic brain injury (TBI) affects error detection and correction, and to compare actual task performance with verbal self-ratings of performance. Participants included 18 persons who had sustained severe TBI from 34 to 186 days prior to study and who were comparable to controls in their rate of naturalistic action error, along with 18 control subjects chosen to be demographically comparable to subjects with TBI. Subjects performed two different tests of naturalistic action in which they completed everyday activities (eg, wrapping a gift, making toast) at different levels of complexity, as manipulated by the addition of distractor objects, the number of tasks that had to be completed per trial, and other demands on planning and working memory. Using a specially developed coding system, each error on these tasks was scored as to whether the subject corrected it and whether the subject otherwise demonstrated awareness of the error. Error scores were also compared to subjects' responses to a questionnaire in which they rated their own performance on the most challenging level of the naturalistic action test. In general, subjects with TBI corrected and showed awareness of proportionally fewer of their errors when compared to controls. Qualitative patterns for some error types also differed between groups. Despite making more errors than control subjects on the most challenging task, subjects with TBI did not rate themselves as performing more poorly with respect to its cognitive demands. However, for subjects with TBI, the number of errors was correlated with performance ratings on certain questionnaire items. This study showed that error detection and correction can be reliably measured during naturalistic action and appear to be impaired in severe TBI even when the base rate of error is controlled. TBI may affect error detection and correction by reducing, or impairing the allocation of, attentional resources needed for the simultaneous execution and monitoring of routine action.

RSR13, a synthetic modifier of hemoglobin-oxygen affinity, enhances the recovery of stunned myocardium in anesthetized dogs.

RSR13 (2-[4-[[(3, 5-dimethylanilino)carbonyl]methyl]phenoxyl]-2-methylpropr ionic acid) is a synthetic allosteric modifier of oxygen (O2)-hemoglobin affinity that increases O2 release to tissue by allosterically stabilizing deoxyhemoglobin. We tested the hypothesis that RSR13 enhances the functional recovery of stunned myocardium in barbiturate-anesthetized dogs instrumented for measurement of left ventricular (LV) and aortic blood pressure, LV +dP/dtmax and subendocardial segment shortening (%SS) in ischemic [left anterior descending (LAD) coronary artery] and normal (left circumflex coronary artery) zones. The partial pressure of oxygen and the Hill coefficient at 50% saturation (P50 and n50, respectively) were determined in arterial blood samples by multiple point tonometry and nonlinear regression analysis. Coronary collateral blood flow in the LAD zone was quantified with radioactive microspheres. Dogs received intravenous vehicle (0.45% saline) or one of two doses of RSR13 (100 or 150 mg.kg-1 bolus followed by a 0.50 or 0.75 mg.kg-1.min-1 infusion, respectively) in a random manner. All dogs were subjected to five 5-min periods of LAD occlusion separated by 5-min periods of reperfusion and followed by 180 min of final reperfusion during which hemodynamics, %SS, arterial blood gases, P50 and n50 were determined at selected intervals. RSR13 caused no hemodynamic effects and coronary collateral blood flow was equivalent among groups. RSR13 increased P50 (+40 +/- 4% for the high dose) and decreased n50 (-31 +/- 2% for the high dose). LAD occlusion caused regional dyskinesia during each 5-min occlusion. Enhanced recovery of %SS by 180 min after final reperfusion was observed in dogs treated with high-dose RSR13 (47 +/- 9% of base line) but not low-dose RSR13 (10 +/- 18% of base line) or vehicle alone (2 +/- 16% of base line). The results suggest that high-dose RSR13 improves the recovery of stunned myocardium throughout reperfusion in open-chest dogs. These findings may be related to increases in O2 availability to ischemic myocardium resulting from RSR13-induced stabilization of the deoxy form of hemoglobin.

Naturalistic action impairment in closed head injury.

The authors sought to determine whether errors of action committed by patients with closed head injury (CHI) would conform to predictions derived from frontal lobe theories. In Study 1, 30 CHI patients and 18 normal controls performed routine activities, such as wrapping a present, under conditions of graded complexity. CHI patients committed more errors even on the simplest condition; but, except for a higher proportion of omitted actions, their error profile was very similar to that of controls. Study 2 involved a subset of patients whose performance in Study 1 was within normal limits. When these high functioning patients were asked to perform the routine tasks under still more taxing conditions, they, too, committed errors in excess of the control group. Accounts based on frontal mechanisms have a difficult time explaining the overall pattern of findings. An alternative based on limited-capacity resources is suggested.

Mental rotation may underlie apparent object-based neglect.

Previous investigations have suggested that object-based neglect may reflect an impairment in attentional allocation that occurs relative to the intrinsic left and right of objects. We report a patient with apparent "object-based" neglect of 90 degrees rotated stimuli for whom the pattern of neglect was a function of task strategy. When the patient was instructed to visualize the rotated stimuli as if they were upright, i.e. mentally rotate them, he showed apparent "object-based" neglect to the left of the principal axes of the stimuli. In contrast, when instructed to refrain from mental rotation, neglect was apparent only with respect to his left, but not the left of the stimuli. Thus, the apparent "object-based" neglect of this patient may be attributed to a process of mental rotation of objects to upright, and subsequent neglect in viewer-centered or environment-centered coordinates. These data suggest a mechanism whereby object-based and viewer/environment-centered reference frames may be aligned, thereby causing viewer/environment-centered neglect to appear as if object-based.

Purification of d'Anjou Pear (Pyrus communis L.) Polyphenol Oxidase.

Polyphenol oxidase (PPO) was extensively purified to homogeneity from d'Anjou pear (Pyrus communis L.) by extraction in the presence of the phenolic binder AG 2-X8 andTriton X-100. Chlorophyll pigment was removed by chromatography resulting in a clear, colorless enzyme extract. Purification of pear PPO was achieved after chromatography on Phenyl Sepharose CL-4B, DEAE-cellulose, and hydroxylapatite columns. Only after the columns were run at room temperature rather than at 4 degrees C were sharp peaks and good resolution obtained. Reproducibility of the entire scheme was excellent with chromatography on the hydrophobic resin as a key to successful purification. Three separate fractions of pear PPO were homogeneous when stained for protein with the silver stain after polyacrylamide slab gel electrophoresis and corresponded to relative mobilities of 0.41, 0.43, and 0.73. The effect of dimethylsulfoxide on enzyme activity was investigated and found to increase significantly the activity of purified pear PPO over the control.

Toxic inhibition of some dehydrogenases by methyl sterculate--a natural occurring substance in cotton seeds.

According to the present study, crude preparations of rainbow trout liver and rabbit muscle lactate dehydrogenase (LDH,EC 1.1.1.27) were not inhibited by methyl sterculate and oleate, while trout liver glucose-6-phosphate dehydrogenase (G6PDH,EC 1.1.1.49) was activated by these esters. Methyl sterculate inhibited purified preparations of trout liver, rabbit muscle, and bovine heart LDH in contrast to methyl oleate which inhibited only a portion of the activity of purified rabbit muscle LDH and had no appreciable effect on the activities of the other purified LDH preparations. Trout liver LDH preparations were not inhibited by p-Chloromercuribenzoate (pCMB), while rabbit muscle and bovine heart LDH were sensitive to the presence of this inhibitor. Trout liver G6PDH was activated at the lower concentrations of pCMB. These data suggest that the reduction of the activities of liver dehydrogenases in the rainbow trout fed a diet containing methyl sterculate was not due to inhibition of these dehydrogenases by this cyclic fatty acid.

Liver dehydrogenase levels in rainbow trout, Salmo gairdneri, fed cyclopropenoid fatty acids and aflatoxin B1.

Cyclopropenoid fatty acids in the diet of rainbow trout caused significant reductions in liver protein and activity of glucose-6-phosphate dehydrogenase, NADP-linked isocitrate dehydrogenase, lactate dehydrogenase, and malate dehydrogenase. Changes in total activity were usually accompanied by similar changes in specific activity. The activity of glucose-6-phosphate dehydrogenase appeared to be more sensitive to the ingestion of cyclopropenoid fatty acids than the other dehydrogenases studied. Feeding 20 ppb aflatoxin B(1) to rainbow trout did not significantly change the activity of the dehydrogenases except for a small increase in the activity of glucose-6-phosphate dehydrogenase after 21 days of feeding. Relationships of these changes to the cocarcinogenicity of cyclopropenoid fatty acids and the carcinogenicity of aflatoxin are discussed.

Ester production by Pseudomonas fragi. IV. Demonstration of esterase activity.

Assay of the esterase activity of sonically treated cell-free extracts, whole cell suspensions, and supernatant fluid of Pseudomonas fragi cultures with a differential respirometer revealed that the esterases were intracellular. Polyacrylamide-gel electrophoresis demonstrated six bands of esterase activity, which revealed substrate specificity differences. Band 1 exhibited slow mobility, bands 2, 3, and 4 moderate mobility, and bands 5 and 6 rapid mobility. Six bands were active with alpha-naphthyl acetate, four bands with alpha-naphthyl propionate, and 5 bands with alphanaphthyl butyrate. These esterases appeared to be more active with aromatic esters than with aliphatic esters.

Activation of coagulase clotting by trypsin inhibitor.

Egg white trypsin inhibitor activated coagulase clotting when added to a final concentration between 2 and 60 mg/ml. The greatest increase in clotting rate was observed in reaction mixtures containing the lowest concentrations of serum and plasma. Maximal activation was reached with 40 mg of trypsin inhibitor per ml when either serum or plasma was used as the source of coagulase-reacting factor (CRF). The increased rate of clotting is partly due to inhibition of plasmin. Freezing and thawing reduced plasma clotting inhibition. Soybean trypsin inhibitor also activated the coagulase reaction. The increased rate of clotting was observed with a coagulase preparation from organisms which produced plasminogen activators and with the culture supernatant fraction from organisms which did not activate plasminogen to plasmin. The tube test for coagulase could be made more sensitive for some strains of staphylococci by increasing the concentration of CRF (added as plasma or serum) by adding trypsin inhibitor, or both.