Dad1p, third component of the Duo1p/Dam1p complex involved in kinetochore function and mitotic spindle integrity.

We showed recently that a complex between Duo1p and Dam1p is required for both spindle integrity and kinetochore function in the budding yeast Saccharomyces cerevisiae. To extend our understanding of the functions and interactions of the Duo1p/Dam1p complex, we analyzed the novel gene product Dad1p (for Duo1 and Dam1 interacting). Dad1p physically associates with Duo1p by two-hybrid analysis, coimmunoprecipitates with Duo1p and Dam1p out of yeast protein extracts, and shows interdependent localization with Duo1p and Dam1p to the mitotic spindle. These results indicate that Dad1p functions as a component of the Duo1p/Dam1p complex. Like Duo1p and Dam1p, Dad1p also localizes to kinetochore regions in chromosomes spreads. Here, we also demonstrate by chromatin immunoprecipitation that Duo1p, Dam1p, and Dad1p associate specifically with centromeric DNA in a manner that is dependent upon Ndc10 and partially dependent upon the presence of microtubules. To explore the functions of Dad1p in vivo, we generated a temperature-sensitive allele, dad1-1. This allele shows spindle defects and a mitotic arrest phenotype that is dependent upon the spindle assembly checkpoint. In addition, dad1-1 mutants undergo chromosome mis-segregation at the restrictive temperature, resulting in a dramatic decrease in viability.

The peripheral vascular response to exercise is impaired in patients with risk factors for coronary artery disease.

BACKGROUND: Abnormal coronary and brachial artery responses have been described in individuals with risk factors for coronary artery disease (CAD). Peripheral arterial tonometry (PAT), a newly developed digital plethysmographic technique was used to assess peripheral vascular response to exercise in healthy controls and individuals with risk factors. METHODS AND RESULTS: Continuous finger PAT during Bruce protocol exercise test was performed in 30 subjects with risk factors for CAD and 30 healthy individuals. Compared with baseline, the PAT wave amplitude at peak exercise decreased in the subjects but increased in the controls: 83 +/- 28% vs. 114 +/- 40% respectively, p < 0.01. CONCLUSIONS: A different pattern of systemic vascular response to exercise was found in individuals with risk factors for atherosclerosis. Since the vascular behavior in these patients is probably related to endothelial dysfunction, it may be that peripheral arterial tonometry can be used as a simple, readily available technique to assess endothelial function.

Irradiation of fresh whole blood for prevention of transfusion-associated graft-versus-host disease does not impair platelet function and clinical hemostasis after open heart surgery.

Since our previous studies suggested that the transfusion of 1 unit fresh whole blood (FWB) after cardiopulmonary bypass (CPB) using a bubble oxygenator may provide hemostatic benefit equivalent to 8-10 units of platelet concentrates, we have routinely used FWB at the termination of CPB. Two patients who received FWB and developed transfusion-associated graft-versus-host disease (TA-GVHD) prompted us to investigate the effect of irradiation of FWB on platelet and clinical hemostasis. Twenty-four patients were randomized to receive either 1 unit FWB (12 patients), or 1 unit irradiated FWB (IrFWB, 1,500 cGy,12 patients) after CPB. Platelet aggregation on extracellular matrix, studied by a scanning electron microscope and graded from 1 to 4 (from poor to excellent aggregation), was similar in both groups preoperatively [3.3 +/- 0.9 (FWB) and 3.5 +/- 0.5 (Ir FWB)], and at the end of CPB [1.8 +/- 1.2 (FWB) and 1.9 +/- 0.9 (IrFWB)]. Platelet aggregation was similar after transfusion of FWB (3.0 +/- 1.0) and after IrFWB (3.2 +/- 0.8), as was the increase in platelet count. Twenty-four hours total postoperative bleeding was similar (560 +/- 420 and 523 +/- 236 ml for FWB and IrFWB, respectively). We conclude that irradiation of FWB for prevention of TA-GVHD does not impair platelet aggregating capacity, and can be used when blood is donated by the patient's next of kin.

"King sized" hearts in infants: comparison of cardiac to body mass in ventricular septal defect in various age-groups.

Heart and body weight were reviewed in reports of 223 human cadavers. Of these, 146 were of control (normal) hearts and 77 of hearts with ventricular septal defects. In each case the heart weight as a percentage of body weight was calculated (defined as pathological heart-body ratio). It was observed that under the age of 4 years, monstrous cardiomegalies may reach 3.2% of the total body weight. These "king sized" hearts were not found above the age of 4.

Platelet protection by low-dose aprotinin in cardiopulmonary bypass: electron microscopic study.

To evaluate the effect of low-dose aprotinin during cardiopulmonary bypass on platelet function and clinical hemostasis, 30 patients undergoing various cardiopulmonary bypass procedures employing bubble oxygenators were randomized to receive either low-dose aprotinin (2 x 10(6) KIU in the cardiopulmonary bypass priming solution, 15 patients [group A]) or placebo (15 patients [group B]). Blood samples were collected before and after cardiopulmonary bypass to assess platelet count and aggregation on extracellular matrix, which was studied by a scanning electron microscope. On a scale of 1 to 4 preoperative mean platelet aggregation grades were similar in both groups (3.8 +/- 0.5 and 3.5 +/- 0.5 for groups A and B, respectively). Postoperatively, platelet aggregation on extracellular matrix decreased slightly in group A (2.8 +/- 1.3; p < 0.01) and significantly in group B (1.3 +/- 0.5; p < 0.001). Eleven of the 15 patients in group A remained in aggregation grade 3 or 4 compared with none of the group B patients. Platelet count was similar in both groups preoperatively and postoperatively. Total 24-hour postoperative bleeding and blood requirement were lower in the aprotinin group (487 +/- 121 mL and 2.3 +/- 1.0 units) than in the placebo group (752 +/- 404 mL and 6.8 +/- 5.1 units; p < 0.01). These results show that the use of low-dose aprotinin during cardiopulmonary bypass provides improved postoperative hemostasis, which might be related to the protection of the platelet aggregating capacity.

Aprotinin prevents cardiopulmonary bypass-induced platelet dysfunction. A scanning electron microscope study.

BACKGROUND: Administration of aprotinin during extracorporeal circulation reduces blood loss and improves platelet function. METHODS AND RESULTS: To evaluate the protective effect of aprotinin on platelets, 50 patients undergoing cardiopulmonary bypass were randomized before surgery to one of three groups. Seventeen patients (group A) received continuous high-dose aprotinin (7 x 10(6) KIU) during cardiopulmonary bypass, 17 (group B) received a single bolus of aprotinin in the pump prime (2 x 10(6) KIU), and 16 (group C) received placebo. Scanning electron microscopy was used to evaluate platelet aggregation on extracellular matrix. The platelet function was graded from 1 to 4, with grade 4 being normal aggregation. Immediately after cardiopulmonary bypass, 16 patients in group A (94%) reached preoperative aggregation grade (mean grade, 3.4 +/- 0.7) compared with nine of 17 in group B (52%) (mean grade, 2.9 +/- 1.2), and none in group C (0%) (mean grade, 1.4 +/- 0.5; p < 0.001). Postoperative platelet count did not differ significantly among the three groups. After surgery, group A bled less than groups B and C (395 +/- 120 versus 488 +/- 135 and 780 +/- 408 ml, respectively; p < 0.01). Patients in the aprotinin groups received fewer red blood cell units (0.9 +/- 1.2 and 1.9 +/- 1.2 versus 3.4 +/- 1.9, respectively; p < 0.01) and were exposed to less homologous blood products (1.3 +/- 1.7 and 2.1 +/- 1.1 versus 6.1 +/- 5, respectively; p < 0.001). CONCLUSIONS: By preserving platelet function, aprotinin improves postoperative hemostasis in all patients who receive high dose and in most who receive low dose.

Synergism between infarct-borne left ventricular dysfunction and cardiomegaly in increasing the risk of coronary bypass surgery.

The effect of cardiomegaly on operative and late mortality in patients with left ventricular dysfunction undergoing coronary bypass operation was investigated. The study group consisted of 178 patients whose left ventricular ejection fraction was below 45% and who were operated on from 1978 through 1985. Forty-five patients (group A) had severe left ventricular dysfunction (ejection fraction < 30%) and 133 (group B) had moderate dysfunction (30% > ejection fraction > 45%). Twenty-four of group A (53%) and 54 of group B (41%) patients had cardiomegaly (cardiothoracic ratio on chest x-ray films > 0.5). There were 10 (6%) hospital deaths, four in group A (9%) and six in group B (4.5%). All four deaths in group A and the six deaths in group B were patients who had cardiomegaly. Regardless of the severity of the left ventricular dysfunction, there was no operative death among patients with normal heart size (p < 0.001). Age over 65, bypass time longer than 2 hours, and incomplete revascularization emerged as risk factors. Follow-up ranged from 5 to 13 years (mean 7.8 years). Overall 5-year actuarial survival, including hospital mortality, was 80% +/- 3%. Reduced 5-year survival was observed in patients with cardiomegaly (67% +/- 5% versus 91% +/- 3%, p < 0.05). Five- and 10-year survival of patients from group A with cardiomegaly was 53% +/- 7% and 18% +/- 13%, respectively.

Clamping of the atherosclerotic ascending aorta during coronary artery bypass operations. Its cost in strokes.

In 632 consecutive primary operations for coronary bypass grafting, the effect of instrumentation of the ascending aorta on the prevalence of stroke was evaluated. There were five surgical pathology groups: group A: soft aorta with no palpable disease, 463 patients; group B: distinctly palpable aorta with focal atheromas necessitating minor surgical modifications, 132 patients; group C, unclampable aorta (no plane for crossclamping the aorta without compression of atheromas was present), 16 patients; group D, untouchable aorta, in which the entire ascending aortic wall was involved by atheromatosis (these aortas were not touched), 14 patients; group E, aneurysmal aorta with soft walls, 7 patients. There were four strokes (0.63%), all related to instrumentations of the aorta. There were no cerebrovascular accidents in patients in whom precautions were taken. Of all risk factors studied, age was the only predictive one for major atheromatosis of the aorta (chi 2 test, p less than 0.001). Of the 30 patients in groups C and D, only one was younger than 60 years.

Fresh blood units contain large potent platelets that improve hemostasis after open heart operations.

Twenty units of fresh whole blood were separated into fresh packed red blood cells (PC) and platelet-rich plasma (PRP) and were transfused to 40 patients immediately after coronary bypass grafting. Patients were preoperatively randomized to receive either PRP (group A, 20 patients) or PC (group B, 20 patients). Platelet number in the PRP group was greater, but not significantly greater, than in the PC group (7.5 +/- 3 versus 5.9 +/- 2.2 x 10(10); p = not significant). However, mean platelet volume in the PC group was significantly greater (8.75 +/- 1.1 versus 6 +/- 0.7 fL). Postoperatively, group A patients bled more than group B (566 +/- 164 versus 327 +/- 41 mL; p less than 0.01) and received more red blood cell units (2.7 +/- 1.2 versus 1.6 +/- 0.7 U; p less than 0.05) and a larger number of blood products (5.9 +/- 3.7 versus 2.6 +/- 1.2 U; p less than 0.05). Transfusion of PRP to group A increased platelet count from 128 +/- 20 to 148 +/- 110 x 10(9)/L; however, platelet functions did not improve. Administration of PC to group B increased platelet count from 139 +/- 22 to 156 +/- 23 x 10(9)/L, improved platelet aggregation (with collagen from 33% +/- 20% to 53% +/- 23%, with epinephrine from 36% +/- 24% to 51% +/- 20%; p less than 0.05), and corrected the prolonged bleeding time. The results suggest that the improved hemostasis observed after fresh whole blood administration is related to the large, potent platelets that remained in the PC and were not separated to the PRP during standard platelet concentrate preparation.

Surgical closed pulmonary valvotomy for critical pulmonary stenosis: implications for the balloon valvuloplasty era.

BACKGROUND: Closed pulmonary valvotomy for critical pulmonary stenosis has no apparent advantage over the percutaneous balloon technique, though it is used when balloon valvuloplasty fails. Experience of this technique at the Heart Institute, Tel Hashomer, since it was first used in 1973 has been reviewed. METHODS: Thirty eight infants up to 1 year old (25 of them neonates--that is, nil to 1 month old) with critical pulmonary stenosis were operated on from 1973 to 1989. All had a transventricular valvotomy, by a modification of the Brock method, and all underwent cardiac catheterisation before surgery. RESULTS: Five of the 25 neonates (20%) died, but none of the other infants, so that the total mortality (five out of 38) was 13%. Three of the 38 required an aortopulmonary shunt. All 38 survivors were followed up--from one month to 14 years (mean 7.5 years). All were symptom free at the last check up. Fifteen of the survivors had required further surgery; this was successful in all cases. CONCLUSIONS: For the balloon valvuloplasty era surgical pulmonary valvotomy provides a good back up for failed attempts at percutaneous valvuloplasty. Review of outcome provides data for comparison with balloon valvuloplasty in the future.

Platelet protection by aprotinin in cardiopulmonary bypass: electron microscopic study.

To evaluate the functional integrity of platelets in patients administered the proteinase inhibitor aprotinin during cardiopulmonary bypass, 20 patients undergoing a complicated and prolonged open heart operation were studied. They were randomized to receive either a high dose of aprotinin (total dose, 6 to 7 x 10(6) KIU) before and during cardiopulmonary bypass (10 patients) or a placebo (10 patients). Blood samples were collected preoperatively, at the termination of bypass, and 90 minutes thereafter to assess platelet count and aggregation on extracellular matrix, which was studied by scanning electron microscopy. On a scale of 1 to 4, mean preoperative platelet aggregation grades were similar in both groups (3.5 +/- 0.5). Postoperatively, at the termination of cardiopulmonary bypass and 90 minutes thereafter, all 10 patients treated with aprotinin revealed normal, unchanged platelet aggregation (grade, 3.5 +/- 0.5), whereas all placebo-treated patients showed severely disturbed aggregation (grade, 1.4 +/- 0.5) (p less than 0.001). The platelet count was similar in both groups before and after operation (preoperatively, 182 +/- 75 x 10(9)/L and 146 +/- 30 x 10(9)/L, and postoperatively, 87 +/- 13 x 10(9)/L and 80 +/- 27 x 10(9)/L for the aprotinin and placebo groups, respectively). Total 24-hour postoperative bleeding and blood requirement were significantly lower in the aprotinin group (371 +/- 84 mL and 2 +/- 0.7 units, respectively) compared with the placebo group (608 +/- 28 mL and 3.4 +/- 1.3 units, respectively) (p less than 0.01). These results demonstrate that improved postoperative hemostasis is directly related to the complete preservation of platelet function achieved by the protective properties of aprotinin.

Correlation between myocardial ischemia and changes in arterial resistance during coronary artery bypass surgery.

The arterial resistometer provides continuous on-line monitoring of changes in arterial resistance. Resistance index (Ri), which bears a direct relationship to systemic vascular resistance (SVR), is defined by the equation Ri = P'/(dP'/dt), where dP'/dt is the peak dP/dt of the arterial waveform, and P' is the pressure at dP'/dt. In 42 patients with unstable angina, changes in Ri were studied at six periods during aortocoronary bypass surgery before tracheal intubation, during tracheal intubation, leg elevation, presternotomy, sternotomy, and dissection of the internal mammary artery. Thirty-four episodes of ischemia (0.1 mV ST segment changes) were observed in 26 patients. All ischemic episodes were associated with increased Ri (mean increase, 102 +/- 52%). Elevation of the pulmonary capillary wedge pressure correlated with ischemia during the preintubation, intubation, and sternotomy periods, but not in the remaining periods. Changes in arterial pressure and heart rate were not good predictors of ischemia. The prevalence of ST segment changes increased markedly during all periods of anesthesia with increase in Ri (P less than 0.05). Ninety-one percent of ST segment changes were associated with a 25% increase from the baseline Ri. Raising the cutoff point to a greater than or equal to 75% increase in Ri improved the specificity of Ri in ischemia detection from 61% to 92%. An increase of greater than or equal to 75% in Ri occurred in only 8% of cases without ST segment changes. It was found that an increase in Ri as depicted by the arterial resistometer was the best hemodynamic correlate of myocardial ischemia.

Prostacyclin and prostaglandin E2 mediate reduction of increased mean arterial pressure during cardiopulmonary bypass by aspiration of shed pulmonary venous blood.

Increased mean arterial pressure during the aortic crossclamp period while on cardiopulmonary bypass was usually treated by us with hypotensive drugs. We noticed, however, that aspirating shed excess pulmonary venous blood from the open pleural cavities causes an immediate reduction in mean arterial pressure, obviating the need for any further pharmaceutical intervention. In this study we investigated the relationship between the reduction in mean arterial pressure and the levels of prostacyclin and prostaglandin E2 in the peripheral and pulmonary venous blood. Ten men undergoing coronary bypass operations had 21 episodes of increased mean arterial pressure (106.9 +/- 11.4 mm Hg) during aortic crossclamping, which was reduced to 67.4 +/- 11.4 mm Hg (p less than 0.001) only by aspirating a mean of 490 ml (range 150 to 1100 ml) of pulmonary venous blood from the pleurae back into the circulation. Mean peripheral prostacyclin level, measured as 6-keto-prostaglandin F1 alpha, and prostaglandin E2 level, both measured by radioimmunoassay technique, were significantly lower at peak mean arterial pressure (419 +/- 180 and 59.5 +/- 21.2 pg/ml) than at lowest mean arterial pressure (632 +/- 271 and 96.7 +/- 52.4 pg/ml for 6-keto-prostaglandin F1 alpha and prostaglandin E2, respectively; p less than 0.001). Prostaglandin F1 alpha and prostaglandin E2 levels in the aspirated pulmonary venous blood were 2309 +/- 3098 pg/ml and 749 +/- 909 pg/ml, respectively. The hypotensive effect of shed pulmonary venous blood that is aspirated back from the pleurae into the circulation seems to be mediated by the high levels of prostacyclin and prostaglandin E2, both powerful vasodilators.

Is transfusion of fresh plasma after cardiac operations indicated?

Patients undergoing cardiac operations constitute the majority of recipients of fresh frozen plasma. In most centers the reason for transfusing fresh frozen plasma is to replace clotting factors. However, the decrease of clotting factors during cardiopulmonary bypass is not sufficient in most patients to cause abnormal bleeding. One of the major causes of nonsurgical bleeding after cardiac operations is acquired platelet dysfunction, which can be corrected by transfusion of 1 unit of fresh whole blood. Because plasmatic factors in fresh whole blood may be responsible for this improvement, a study was designated to evaluate the effect of transfusing fresh plasma on platelet function after cardiac operations. Forty patients undergoing cardiopulmonary bypass were randomized to receive either fresh plasma or the fresh packed cell fraction. Administration of packed cells increased platelet number (118 +/- 8.5 to 154 +/- 7.6 x 10(9)/L, p less than 0.05), shortened bleeding time (7.57 +/- 0.4 to 4.0 +/- 0.3 minutes, p less than 0.05), and improved platelet aggregation in response to collagen and epinephrine (32% +/- 4.7% to 50% +/- 5.6% and 37% +/- 5.8% to 50% +/- 5.8%, respectively, p less than 0.05). Fresh plasma, however, neither increased platelet number nor improved bleeding time or platelet aggregation. Each group later received the remainder of the blood unit, with similar results. The results suggest that improvement of platelet function in patients receiving fresh whole blood after cardiac operations is not related to plasmatic factors. Therefore the massive use of fresh frozen plasma in patients after cardiopulmonary bypass should be reconsidered.

Left ventricular thrombectomy in the early postinfarction period.

Emergency left ventricular thrombectomy was performed on four patients soon after infarction. In three patients surgery was carried out after embolisation had occurred and when a large, residual, protruding, mobile thrombus remained in the left ventricle. Surgery was performed in the fourth patient after a high risk thrombus was detected and initial attempts to lyse it had failed. All four patients had an uneventful recovery and were discharged within two weeks of surgery. These cases indicate that the therapeutic option of left ventricular thrombectomy is feasible for patients with acute infarcts and problematic left ventricular thrombi.

Epidural anesthesia with the Trendelenburg position for cesarean section with or without a cardiac surgical procedure in patients with severe mitral stenosis: a hemodynamic study.

The hemodynamic effects of epidural anesthesia (EA) with the Trendelenburg position were studied in seven patients with severe mitral stenosis undergoing emergency cesarean section (CS) because of hemodynamic deterioration. In six patients, the CS was immediately followed by an open mitral commissurotomy under general anesthesia, whereas in one patient, the CS was performed alone. A significant reduction in heart rate (120 +/- 5 to 83 +/- 7 beats/min; P less than 0.001) was observed after induction of EA. Mean arterial pressure (MAP) decreased (78 +/- 9 to 55 +/- 5 mm Hg; P less than 0.01) simultaneously with reduction of the pulmonary capillary wedge pressure (PCWP) (37 +/- 4 to 15 +/- 4 mm Hg, P less than 0.001) and cardiac index (CI) (2.4 +/- 0.3 to 1.8 +/- 0.32 L/min/m2; P less than 0.001). However, PCWP could be adjusted by selecting the appropriate angle of the Trendelenburg position. When the PCWP was approximately 25 mm Hg, MAP and Cl increased to 72 +/- 7 mm Hg and 3.1 +/- 0.4 L/min/m2, respectively, and a satisfactory hemodynamic state was achieved. Systemic vascular resistance decreased after induction of EA (2,250 +/- 250 to 1,750 +/- 450 dyne.s.cm-5; P less than 0.001), and remained unchanged during the perioperative period. It is concluded that the combination of epidural anesthesia with tilting of the table is a safe method for urgent CS in pregnant women with critical mitral stenosis in whom termination of pregnancy is indicated because of hemodynamic deterioration.

Transfusion of fresh whole blood stored (4 degrees C) for short period fails to improve platelet aggregation on extracellular matrix and clinical hemostasis after cardiopulmonary bypass.

It has recently been shown that the hemostatic effect of 1 unit of fresh whole blood is equivalent to the effect of 8 to 10 platelet units. This study was designed to evaluate the effect of short periods of cold (4 degrees C) storage on the hemostatic effect of fresh whole blood transfusion in 36 patients immediately after cardiopulmonary bypass. Twelve patients (group A) received unrefrigerated fresh whole blood, 12 (group B) received fresh whole blood after 5 hours' storage at 4 degrees C, and 12 (group C) after 24 hours' storage at 4 degrees C. For evaluation of platelet function, a method with an extracellular matrix and an electron microscope was used. The platelet function was graded from 1 to 4, with grade 4 being normal aggregation. Postoperatively, group A patients bled less than groups B and C (267 +/- 42 versus 397 +/- 72 and 601 +/- 172 ml/24 hr, respectively, p less than 0.001) and therefore received fewer blood units (1.4 +/- 0.5 versus 2 +/- 0.9 and 3 +/- 1.4, respectively, p less than 0.01). Five patients of group A (42%) reached grade A aggregation after transfusion of unstored fresh whole blood, compared with two (17%) of group B and none (0%) of group C (p less than 0.01). Posttransfusion platelet count and mean platelet volume were not significantly different in the three groups. We conclude that storage at 4 degrees C, even for a short period of 5 hours, diminishes the hemostatic effect of fresh whole blood by decreasing platelet aggregability.