Isolation and characterization of a tomato Acid phosphatase complementary DNA associated with nematode resistance.

The tomato (Lycopersicon esculentum) acid phosphatase-1 (Apase-1(1), EC 3.1.3.2) isozyme variant, genetically linked to the root-knot nematode resistance locus (Mi) on chromosome 6, has been purified by a rapid procedure from tomato cell suspension cultures. Peptide fragments of the purified enzyme were generated from trypsin and Lys-C endoprotease digests and separated by reverse-phase high-performance liquid chromatography. Amino acid sequences derived from the purified peptide fragments represented >50% of the total amino acid content of the protein and enabled the construction of degenerate oligonucleotide probes that were used to screen a tomato cell culture complementary DNA library. Clones corresponding to full-length coding sequences for Apase-1 have been isolated and sequenced. Southern blot analysis of DNA isolated from a number of tomato cultivars shows that the Apase-1(1) gene (aps1) is present at one copy per genome and that genotypes containing the aps1(1) allele have restriction fragment length polymorphisms that distinguish them from cultivars having the aps1(+) allele. Segregation analysis demonstrates that the restriction fragment length polymorphisms are associated with the aps1 locus. Tomato Apase-1(1) is also found to have significant homology at the amino acid sequence level to a class of vegetative storage proteins characterized in soybean.

The heart and conduction system in acute Kawasaki disease: report of fraternal cases--one lethal, one relapsing.

The heart and conduction system were studied in a 4 1/2-year-old boy with Kawasaki disease who died suddenly in the early phase of illness. Severe perivasculitis, neuritis, pancarditis, and involvement of all parts of the specialized conduction system were found. The boy's brother, born a year later, at the age of 12 months had relapsing Kawasaki disease. He received intravenous gamma globulin and recovered.

UDP-Glucose: (1-->3)-beta-Glucan Synthases from Mung Bean and Cotton: Differential Effects of Ca and Mg on Enzyme Properties and on Macromolecular Structure of the Glucan Product.

A re-examination of the kinetic properties of UDP-glucose: (1-->3)-beta-glucan (callose) synthases from mung bean seedlings (Vigna radiata) and cotton fibers (Gossypium hirsutum) shows that these enzymes have a complex interaction with UDP-glucose and various effectors. Stimulation of activity by micromolar concentrations of Ca(2+) and millimolar concentrations of beta-glucosides or other polyols is highest at low (<100 micromolar) UDP-glucose concentrations. These effectors act both by raising the V(max) of the enzyme, and by lowering the apparent K(m) for UDP-glucose from >1 millimolar to 0.2 to 0.3 millimolar. Mg(2+) markedly enhances the affinity of the mung bean enzyme for Ca(2+) but not for beta-glucoside; with saturating Ca(2+), Mg(2+) only slightly stimulates further production of glucan. However, the presence of Mg(2+) during synthesis, or NaBH(4) treatment after synthesis, changes the nature of the product from dispersed, alkali-soluble fibrils to highly aggregated, alkali-insoluble fibrils. Callose synthesized in vitro by the Ca(2+), beta-glucoside-activated cotton fiber enzyme, with or without Mg(2+), is very similar in size to callose isolated from cotton fibers, but is a linear (1-->3)-beta-glucan lacking the small amount of branches at C-0-6 found in vivo. We conclude that the high degree of aggregation of the fibrils synthesized with Mg(2+)in vitro is caused either by an alteration of the glucan at the reducing end or, indirectly, by an effect of Mg(2+) on the conformation of the enzyme. Rate-zonal centrifugation of the solubilized mung bean callose synthase confirms that divalent cations can affect the size or conformation of this enzyme.

Therapy in cytopenia.

Intravenous immunoglobulin (IVIG) may be considered first-line maintenance therapy for idiopathic thrombocytopenic purpura (ITP) because it has been proven to be the least toxic. In a study of 25 children with acute ITP, treatment with IVIG maintained platelet counts above 40,000/mm3 in all of the children. After 1 year, none of these patients required further therapy. In another study group of 25 pediatric patients with chronic ITP, treatment with IVIG circumvented splenectomy in 60% of the cases. The therapeutic regimens for adults and children are described, as is a strategy to overcome IVIG resistance. Experience with IVIG in hemolytic anemia and neutropenia are discussed. The mechanism of action is explored in some detail, specifically as it relates to reticuloendothelial system (RES) Fc receptor blockade and suppression of antiplatelet antibody synthesis.

Effects of intubation on coronary blood flow and myocardial oxygenation.

Effects on haemodynamics and myocardial oxygenation of endotracheal intubation were examined in 17 patients after halothane induction and 12 after 1 mg X kg-1 of IV morphine. Six patients having each anaesthetic were pretreated with IV propranolol (0.1 mg X kg-1) 45 minutes earlier. Arterial and intracardiac pressures, cardiac output and total coronary sinus blood flow (CSBF), both by thermodilution, were determined plus arterial-coronary differences of oxygen, haemoglobin and lactate. Blood pressure (BP), heart rate and CSBF were recorded continuously during intubation. The subjects were candidates for coronary bypass grafts, but had good ventricular function (mean ejection fraction 0.68 +/- 0.13 SD). From their reduced levels after induction, BP, cardiac index and systemic vascular resistance increased to awake levels following intubation. Mean CSBF in nonbetablocked patients increased to awake level along with BP. More myocardial oxygen was extracted and consumed after intubation, but lactate extraction continued: these data are evidence of adequate oxygen supply. Induction with either halothane or morphine effectively prevented the hypertensive response to intubation. Acute beta blockade led to less increase in heart rate from intubation.

Rate-pressure product correlates poorly with myocardial oxygen consumption during anaesthesia in coronary patients.

In 26 patients having coronary grafts, haemodynamics, coronary sinus blood flow and the arterio-coronary sinus difference of oxygen content were determined, awake and at four intervals during morphine-oxygen or halothane--oxygen anaesthesia. Rate-pressure product (RPP), triple product (TP) and myocardial oxygen consumption (MVO2) were calculated. The correlation of the two indirect indices to MVO2 were tested by repeated measures and regression analyses. No significant correlations were seen at four of five study times, when outlying data points were appropriately excluded. A pitfall of using more than one data point from each patient in the linear regression analysis is pointed out. In addition to the lack of correlation of RPP to MVO2, RPP was an imprecise predictor of myocardial lactate production and of postoperative infarction.

Transient hemodynamic dysfunction after myocardial revascularization. Temperature dependence.

We studied hemodynamics and the effects of right atrial pacing (110 beats/min) following complete myocardial revascularization and hypothermic multidose potassium crystalloid cardioplegia in 12 patients with a normal preoperative left ventricular ejection fraction (LVEF). Measurements were made immediately preoperatively, postoperatively at specified temperatures during the rewarming period (90 degrees F, 94 degrees F, and 98 degrees F), and at 24 hours. No patient had a perioperative myocardial infarction. At 90 degrees F, hemodynamics were characterized by significant decreases in cardiac index, stroke volume index, and left ventricular stroke work index (LVSW) and an increase in systemic vascular resistance index (SVRI) compared to preoperative values (p less than 0.05). Right atrial pacing significantly increased cardiac index preoperatively and 24 hours postoperatively, but not during the rewarming period. Over the entire rewarming period (90 degrees F to 98 degrees F), each of the following variables correlated with temperature: cardiac index (r = 0.71 in sinus rhythm and r = 0.66 with right atrial pacing); stroke volume index (r = 0.33 and 0.66); SVRI (r = -0.80 and -0.64); LVSW (r = 0.37 and 0.73); and heart rate in sinus rhythm (r = 0.51). During the rewarming period, there was an inverse relationship between cardiac index and SVRI (r = -0.87). In conclusion, after myocardial revascularization: (1) transient hemodynamic dysfunction occurs during the rewarming period (90 degrees F to 98 degrees F); (2) this dysfunction is temperature-dependent; and (3) right atrial pacing at 110 beats/min does not improve hemodynamic function during the rewarming period. Temperature must be considered in the evaluation of left ventricular and hemodynamic function following myocardial revascularization.

Myocardial and systemic effects of nitroglycerin, given awake and during anaesthesia in coronary patients.

The haemodynamic and myocardial energy-metabolic effects of intravenous nitroglycerin (NTG) were investigated in seven male patients about to undergo coronary artery bypass grafting. Effects of one mg of NTG given at a rate of 100 micrograms . min-1 to sedated, awake hypertensive patients, were compared to effects of the same dose of NTG in the same patients, after sternotomy. Coronary sinus flow was measured by a thermodilution catheter and myocardial oxygen consumption (MVO2) was calculated using the arterio-coronary sinus difference of oxygen content. With the patients awake, cardiac index and stroke volume and work indices fell during NTG administration. There was only a small fall in arterial pressure as heart rate and systemic resistance increased. Pulmonary capillary wedge pressure decreased but not MVO2. With NTG administration during anaesthesia, arterial pressure fell significantly to normal, as systemic resistance decreased, without a change in heart rate or cardiac index. Calculated MVO2 decreased as the arterial-coronary venous difference of oxygen content increased. Both oxygen and lactate balance of the myocardium were maintained. NTG given during anaesthesia was more effective in reducing blood pressure and MVO2 than when given to awake patients.

Nitrous oxide added to halothane reduces coronary flow and myocardial oxygen consumption in patients with coronary disease.

The haemodynamic and myocardial metabolic effects of adding 50 per cent nitrous oxide to 0.5 per cent halothane were studied in 13 patients, before the surgical incision for coronary artery vein grafts. Cardiac output and coronary sinus blood flow were determined by thermodilution, along with haemodynamic measurements. Measurements 15 minutes after addition of nitrous oxide revealed a significant decrease in heart rate, arterial pressure, cardiac index, coronary sinus blood flow and myocardial oxygen consumption. There was a significant increase in coronary sinus lactate content, and a significant decrease, from 27 to 11 per cent, in myocardial lactate extraction. We conclude that these circulatory changes were likely to be due to a depression of ventricular function by the nitrous oxide. The myocardia of these patients with severe coronary disease were becoming globally ischaemic while they were receiving 50 per cent oxygen, in the presence of hypotension. Nitrous oxide should be turned off when hypotension occurs in coronary patients.

Myocardial metabolism and hemodynamic responses to halothane or morphine anesthesia for coronary artery surgery.

Eighteen patients having coronary artery bypass grafts were randomly anesthetized with morphine (1 mg/kg) or halothane and oxygen. Central and peripheral pressures were measured serially, plus cardiac output and total coronary sinus blood flow, both by thermodilution catheters, starting before induction of anesthesia and continuing until completion of sternotomy. No significant differences in hemodynamic responses were seen between the two anesthetic techniques during induction: blood pressure and peripheral vascular resistance decreased significantly, but not cardiac output or coronary flow. Myocardial oxygen consumption decreased significantly with induction as oxygen content of coronary sinus blood increased, indicating preservation of oxygen balance. Heart rate and blood pressure increased after sternotomy in the patients given morphine, with the myocardium producing lactate in two of six patients and with nitroprusside being required in four patients to decrease arterial pressure. Halothane-oxygen anesthesia effectively controlled autonomic responses to sternotomy, although one of 12 patients had myocardial lactate production at that time. Neither rate-pressure product or ST segment changes were useful predictors of the ratio between myocardial oxygen consumption and supply. Myocardial oxygen balance can be maintained in coronary patients before cardiopulmonary bypass if pulse rate and blood pressure are kept at less than awake levels.

Dobutamine and cardiac oxygen balance in patients following myocardial revascularization.

Dobutamine is frequently used in the early postoperative period following myocardial revascularization to improve cardiac output. Seven postoperative adult patients with low output syndrome were studied before and during intravenous dobutamine (mean +/- SD: 5.1 +/- 2.5 micrograms/kg/min) infusion. The metabolic effects were evaluated and related to hemodynamic changes. Cardiac index increased 40% (p less than 0.05) with an increase in heart rate (p less than 0.05) and decreases in systemic vascular resistance and right atrial pressure (p less than 0.05). No significant changes occurred in arterial or pulmonary capillary wedge pressures or in stroke volume index. Dobutamine produced a 29% increased in myocardial oxygen consumption which, in these revascularized patients, was accompanied by a 35% increase in coronary blood flow. No significant alteration was observed in coronary sinus oxygen content or in global myocardial lactate extraction. Thus, despite the increased metabolic cost of dobutamine, global myocardial ischemia was not observed.

Intravenous nitroglycerin and myocardial metabolism during anesthesia in patients undergoing myocardial revascularization.

Although intravenous nitroglycerin has been used to control the hypertensive response during sternotomy in patients undergoing myocardial revascularization, the effects of the drug on myocardial oxygen supply and demand have not been anesthetized for coronary artery bypass, were studied before and after administration of intravenous nitroglycerin (mean dose 12 microgram/kg in 6 minutes). Evaluation of myocardial metabolism showed an increase in coronary sinus oxygen content (p less than 0.05) and a reduction in myocardial oxygen consumption (p less than 0.05). Although mean myocardial lactate extraction and coronary sinus blood flow were not significantly altered in the group as a whole, variations in individual patient responses were observed and are discussed. These direct observations of global myocardial metabolism observed in this study group are similar to the conclusions reached by other investigators using indirect indices of myocardial oxygen supply and demand.

Effects of calcium on the coronary and systemic circulation in patients after coronary surgery.

Nine patients were studied three hours after aorto-coronary bypass. Before anaesthesia a radial arterial cannula was inserted and a thermodilution catheter placed into the pulmonary artery by fluoroscopy. A special thermodilution catheter was manipulated into the coronary sinus. Haemodynamic measurements were made plus cardiac output and coronary sinus blood flow. Content of oxygen and lactate in arterial and coronary sinus blood was determined. Series of measurements were done before and after 1 gm of CaCl2 given intravenously over 15 minutes. Calcium increased cardiac index and arterial pressure but not systemic vascular resistance. Total coronary sinus blood flow did not change, nor did myocardial oxygen consumption or coronary sinus oxygen content. Content of lactate in arterial and coronary sinus blood was unaltered and lactate extraction by the heart continued, in eight of nine patients. The improved haemodynamics were accomplished without inordinate risk to global ventricular energy metabolism.

Effects of digoxin on myocardial oxygen supply and demand in patients following coronary artery bypass surgery.

Although digoxin is used frequently in patients in the prophylaxis of postoperative supraventricular tachyarrhythmias, the effects of the drug on myocardial oxygen supply and demand after coronary bypass have not been described. Seven adult patients with good ventricular function who underwent myocardial revascularization were studied before and three hours after digoxin (0.5 mg, iv). There were no significant changes observed in any measured systemic hemodynamic variable. Evaluation of global myocardial metabolism showed an increase in myocardial oxygen consumption (P less than 0.05) which was compensated satisfactorily, as no significant alteration was noted in the coronary sinus oxygen content, or in the lactate gradient across the myocardium. Since the authors studied the effects of only one dose of digoxin, the effects of full digitalization in these patients remains to be defined.

Effects of protamine sulfate on myocardial oxygen supply and demand in patients following cardiopulmonary bypass.

The effect of protamine sulfate on myocardial oxygen supply and demand was studied under clinical conditions in nine patients following cardiopulmonary bypass. Before surgery, the patients had severe coronary artery disease with good ventricular function. The patients required no vasoactive drugs, but only blood volume adjustments when weaned off bypass, and were hemodynamically stable at the time of study. The protamine dose of 196 mg (2.5 mg/kg) was infused over 4 +/- 1 minutes. Although modest variation in hemodynamic function occurred in individual patients after administration of protamine, there were no significant hemodynamic alterations for the group. No significant alteration in global myocardial metabolism was observed. Protamine caused a small decrease in measured coronary blood flow, resulting in a corresponding reduction in calculated myocardial oxygen consumption as coronary sinus oxygen content remained unaltered. Myocardial lactate extraction showed no significant alteration. It is concluded that protamine sulfate, given at rapid infusion rates in hemodynamically stable patients, is not associated with an adverse alteration in hemodynamics or global myocardial metabolism.

Cardiovascular effects of morphine in patients with coronary arterial disease.

Large doses of morphine sulfate have been reported to cause myocardial lactate production and reduction in coronary blood flow in animals. Similar effects with clinical doses in man would significantly alter the management of cardiac patients. Eleven adult patients with significant coronary arterial disease and normal left ventricular ejection fraction were studied before and 30 minutes after infusion of morphine (0.25 mg/kg IV). Evaluation of myocardial metabolism showed an increase in coronary sinus oxygen content (p less than 0.001) and a reduction in myocardial oxygen consumption. Myocardial lactate extraction was not altered. No change in coronary sinus blood flow was seen. It is concluded that infusion of morphine sulfate, 0.25 mg/kg IV, does not produce global myocardial ischemia in patients with coronary artery disease and normal ventricular function.