Comparative study of heart rate variability between healthy human subjects and healthy dogs, rabbits and calves.

Several studies have been performed to assess heart rate variability (HRV) in several species such as humans, dogs, pigs, calves, rabbits and rats. However, haemodynamic parameters are totally different in each animal, and optimal animal models for studying HRV corresponding to human HRV are still unclear. The purpose of this study was to assess HRV in human subjects and to compare those HRV data with canine, bovine and rabbit HRV data. The heart rate in the human subjects (62.8 +/- 7.4 bpm) was significantly lower than that in dogs (124.2 +/- 18.8 bpm, P < 0.001), calves (73.4 +/- 10.5 bpm, P < 0.05), and rabbits (217.3 +/- 21.5 bpm, P < 0.001). The low-frequency waves (LF) (57.9 +/- 65.8 ms(2)/Hz) and high-frequency waves (HF) (33.8 +/- 49.1 ms(2)/Hz) in rabbits were significantly lower than human LF (1216.3 +/- 1220.7 ms(2)/Hz, P < 0.05) and HF (570.9 +/- 581.3 ms(2)/Hz, P < 0.05). Dogs and calves showed similar LF (991.1 +/- 646.1 ms(2)/Hz and 547.0 +/- 256.9 ms(2)/Hz, respectively), HF (702.1 +/- 394.1 ms(2)/Hz and 601.0 +/- 666.6 ms(2)/Hz, respectively) and LF/HF (2.0 +/- 1.3 and 2.5 +/- 1.9, respectively) when compared with the human data. The present study shows that dogs and calves revealed similar HRV values as those which relate to humans. Large deviation of the HRV values in rabbits compared with humans might be considered when conducting animal studies using those animals to reflect human clinical situations.

Hemolysis evaluation of centrifugal pumps using microcapsule suspension.

OBJECTIVES: Bovine and human blood has been widely used for in vitro hemolysis testing to evaluate centrifugal cardiac assist pumps. However, results from such tests are complicated by variations in the susceptibility of individual red blood cells to shear. The objective of this study was to evaluate the use of microcapsule suspension as an alternative to bovine or human blood for hemolysis testing. METHODS: Microcapsule suspensions of 100 micro m maximal diameter (average 79.1 micro m) with a polyurethane membrane were used. Each microcapsule contained a leuco dye, which was used to measure "hemolysis" in the suspension after exposure to mechanical stress. Six centrifugal pumps were used to measure and compare the hemolysis values of microcapsule suspensions, bovine blood and human blood. RESULTS: Correlations were significant between the hemolysis values measured using microcapsule suspensions and those using bovine blood (R = 0.965, p = 0.002) and human blood (R = 0.940, p = 0.005). CONCLUSIONS: Microcapsule suspension can be successfully used instead of blood to compare the relative hemolytic performance of centrifugal blood pumps.

Effects of diltiazem and noradrenaline on extracellular potassium changes in the globally ischaemic rat heart.

OBJECTIVE: The aim was to investigate the effects of a calcium antagonist (diltiazem) and a catecholamine (noradrenaline) on extracellular potassium accumulation during global ischaemia. METHODS: Extracellular potassium concentration ([K+]e) was measured during 30 min global ischaemia in the isolated rat heart using a valinomycin potassium sensitive electrode. Contracture development during ischaemia was measured throughout with an intraventricular balloon inserted into the left ventricle and myocardial adenine nucleotides were measured in separate series of hearts. RESULTS: In control hearts, [K+]e showed a characteristic triphasic change during 30 min global ischaemia. This consisted of an early rising phase followed by a transient falling phase after the initial peak of [K+]e, and then a late rising phase. Diltiazem suppressed the rate of rise of [K+]e during early ischaemia, but extended the time course of the early [K+]e rise with the higher dose, abolishing the transient falling phase of [K+]e. During late ischaemia, the rise in [K+]e was attenuated by diltiazem. Noradrenaline also suppressed the early extracellular potassium accumulation, but in contrast to diltiazem, hastened the time course of the late [K+]e rise. CONCLUSIONS: Although diltiazem suppresses the early potassium loss during ischaemia as previously described, the drug also decreases the [K+]e fall by some as yet unknown mechanism, so that the [K+]e level becomes higher than control during the falling phase.

Abrogation of protein phosphatase 6 promotes skin carcinogenesis induced by DMBA.

Somatic mutations in the gene encoding the catalytic subunit of protein phosphatase 6 (Ppp6c) have been identified in malignant melanoma and are thought to function as a driver in B-raf- or N-ras-driven tumorigenesis. To assess the role of Ppp6c in carcinogenesis, we generated skin keratinocyte-specific Ppp6c conditional knockout mice and performed two-stage skin carcinogenesis analysis. Ppp6c deficiency induced papilloma formation with 7,12-dimethylbenz (a) anthracene (DMBA) only, and development of those papillomas was significantly accelerated compared with that seen following DMBA/TPA (12-O-tetradecanoylphorbol 13-acetate) treatment of wild-type mice. NF-κB activation either by tumor necrosis factor (TNF)-α or interleukin (IL)-1ß was enhanced in Ppp6c-deficient keratinocytes. Overall, we conclude that Ppp6c deficiency predisposes mice to skin carcinogenesis initiated by DMBA. This is the first report showing that such deficiency promotes tumor formation in mice.

Effects of preserving mitral apparatus on ventricular systolic function in mitral valve operations in dogs.

The mitral apparatus can affect left ventricular function through various mechanisms, such as the direct mechanical coupling between the mitral anulus and papillary muscle and the mitral annular contraction. To evaluate the relative contribution of these mechanisms, we investigated in five groups of 35 dogs that underwent open chest operations how preservation of the mitral apparatus affects left ventricular systolic function. We performed atriotomy in the first group. We sutured a prosthetic rigid ring around the mitral anulus in the second group. We replaced the mitral valve and preserved the anterior chordae in the third group, the posterior chordae in the fourth group, and no chordae in the fifth group. The postoperative percentage of recovery of ventricular function (as assessed by the slope of the end-systolic pressure-volume relation) from preoperative control values were 92.2% +/- 4.8%, 89.5% +/- 12.8%, 85.7% +/- 9.5%, 75.1% +/- 12.9%, and 61.3% +/- 8.0%, respectively. Preservation of the mitral apparatus significantly improved left ventricular function compared with that of conventional mitral valve replacement. The average relative contribution of the ventricular muscle to left ventricular function, the mitral anulus-papillary muscle continuity, and the mitral annular contraction were 66.5%, 30.6%, and 2.9%, respectively. We conclude that it is more beneficial to preserve the mitral apparatus in mitral valve replacement. The prosthetic ring does not detract from the functional benefit of the preservation of the mitral apparatus.

The Cleveland Clinic-Nimbus total artificial heart. In vivo hemodynamic performance in calves and preclinical studies.

In vitro function of the Cleveland Clinic-Nimbus electrohydraulic total artificial heart met National Heart, Lung, and Blood Institute hemodynamic guidelines for such devices. In a series of in vivo experiments, we implanted the total artificial heart in eight calves (mean weight 87 kg), one for a short-term experiment and seven for long-term experiments. The mean blood flow during support was 7.7 +/- 1.6 L/min with left atrial pressure 13 +/- 6 mm Hg, right atrial pressure 13 +/- 4 mm Hg, and aortic pressure 97 +/- 9 mm hg. Maximum pump flow (9.6 L/min) occurred after 4 days of support as a result of the high resting cardiac output of the animals. A 10% to 15% right pump stroke-volume limit effectively balanced atrial pressures, and afterload insensitivity was confirmed by the in vivo studies. Calves tolerated treadmill exercise studies well, with an average duration of 22 minutes and an average top speed of 2.1 mph. The experiments were terminated after 1 day to 120 days of support (mean 32 days). Most experiments were terminated as a result of correctable mechanical problems. In a separate study of six adult human patients undergoing orthotopic cardiac transplantation, five showed an excellent fit for the Cleveland Clinic-Nimbus total artificial heart. Further studies using chest roentgenograms, chest measurements, and transesophageal echocardiography should help predict fit of the total artificial heart in potential candidates. Initial candidates for a "vented-electric" version of the Cleveland Clinic-Nimbus total artificial heart are patients for whom univentricular (left ventricular assist device) support is not appropriate, but who require mechanical support as a bridge to cardiac transplantation.

Beneficial effect of pericardial meshing on left ventricular pump performance in dogs.

To evaluate the effects of pericardial meshing (multiple incisions on the pericardium) on cardiac function, we examined left ventricular pump performance before and after pericardial meshing in six open chest dogs. We evaluated left ventricular systolic properties with the slope of end-systolic pressure-volume relation and diastolic properties with end-diastolic pressure-volume relation (chamber compliance). Overall left ventricular performance was assessed with end-diastolic pressure versus cardiac output relation. Left ventricular chamber compliance was increased (31.3%) with pericardial meshing compared with direct closure of the pericardium, and cardiac output was increased (26.7%) for any given left ventricular end-diastolic pressure. The slope of the end-systolic pressure-volume relation was not altered in pericardial meshing. These results suggest that pericardial meshing improves left ventricular pump performance as a result of increasing left ventricular chamber compliance. This technique may benefit cardiac pump performance that is depressed by the direct closure of the pericardium after cardiac operations.

Effects of left heart bypass on right ventricular performance. Evaluation of the right ventricular end-systolic and end-diastolic pressure-volume relation in the in situ normal canine heart.

Although left heart bypass has gained popularity as a powerful technique to assist the severely failed left heart, apparent right heart failure has often developed during the bypass procedure. We investigated whether the coexisting right heart failure is attributable to the left heart bypass in 16 open-chest dogs. We evaluated the effects of left heart bypass on the right ventricular systolic properties by the slope of the end-systolic pressure-volume relation and its effects on the diastolic properties by chamber compliance. Overall right ventricular performance was assessed by the end-diastolic pressure versus cardiac output relationship. The left heart bypass decreased the slope slightly when the assisted flow ratio exceeded 75% (-14% +/- 8% at the assisted flow ratio of 100%, p less than 0.02) and thus had a deleterious influence on right ventricular performance. The left heart bypass, on the other hand, had a counteracting beneficial influence on right ventricular performance through the increase in chamber compliance (38% +/- 5%, p less than 0.01) and the decrease in pulmonary arterial input resistance (-15% +/- 12%, p less than 0.01). The net effect of the left heart bypass was the increase in cardiac output (20% +/- 2%, p less than 0.05) for any given right ventricular end-diastolic pressure. We conclude that in normal hearts the left heart bypass augments right ventricular performance. We ascribe these beneficial effects to diastolic ventricular interdependence and afterload unloading.

A new method of retrograde cardioplegic administration. Right ventricular protection by right atrial perfusion cooling.

Retrograde administration of cardioplegic solution via the right atrium with continuous cooling of the right ventricular cavity (right atrial perfusion cooling) was assessed for its protective effect in 12 dogs with occlusion of the right coronary artery subjected to global ischemia for 60 minutes. After an initial administration of 4 degrees C crystalloid cardioplegic solution by antegrade aortic perfusion, myocardial protection was established either by right atrial perfusion cooling (group I; n = 6) or by antegrade aortic perfusion alone (group II; n = 6). The right ventricular temperature was approximately 15 degrees C in group I and 20 degrees C in group II. After ischemia for 60 minutes, the adenosine triphosphate content of the right ventricular free wall was significantly higher in group I than in group II (24.4 +/- 1.45 versus 13.8 +/- 2.34 mumol/gm dry weight, p less than 0.05). The percent recovery of right ventricular contractility, which was evaluated by end-systolic pressure-volume relationships, was significantly better in group I at each reperfusion period (30 minutes: 130.0% +/- 9.6% versus 86.1% +/- 11.8%, p less than 0.05; 60 minutes: 159.6% +/- 12.9% versus 96.5% +/- 20.1%, p less than 0.05). Postischemic right ventricular stiffness (reciprocal value of compliance) increased in group II compared with group I, although the difference was not statistically significant. There were no major differences in percent recovery of the left ventricular end-systolic pressure-volume relationships between the two groups. The evidence suggests that the right atrial perfusion cooling method produces excellent right ventricular protection.

A possible mechanism of TPA-mediated downregulation of neurotrophin-3 gene expression in rat cultured vascular smooth muscle cells.

We have previously reported that in cultured rat vascular smooth muscle cells (VSMCs), neurotrophin-3 (NT-3) gene expression was suppressed by TPA (12-O-tetradecanoyl phorbol-13-acetate), which induces an AP-1 transcription factor. In the present study, to clarify the mechanism for TPA-mediated downregulation of NT-3 gene expression, effects of cycloheximide and dexamethasone (Dex) on the TPA-mediated downregulation were examined in VSMCs. Pretreatment with cycloheximide, an inhibitor of protein synthesis, or simultaneous treatment with Dex, an inhibitor of AP-1, suppressed the TPA-mediated downregulation of NT-3 gene expression. Furthermore, co-transfection of c-fos and c-jun expression vectors into VSMCs resulted in decrease in the NT-3 gene expression. The present findings suggest that TPA-induced AP-1 de novo synthesis causes the downregulation of NT-3 gene expression in VSMCs.

Device-based change in left ventricular shape: a new concept for the treatment of dilated cardiomyopathy.

OBJECTIVE: We tested a unique new device, the Myosplint device (Myocor, Inc, Maple Grove, Minn), which is designed to change left ventricular shape, reduce left ventricular wall stress, and improve left ventricular systolic function. METHODS: Heart failure was induced in 15 dogs over 27 days by rapid pacing (230 beats/min). Seven animals underwent sham surgery, and 8 animals received 3 transventricular Myosplint devices each. Myosplint devices were tightened to create a symmetric bilobular left ventricular shape and were adjusted to produce a calculated 20% reduction in wall stress. Hemodynamic, 2-dimensional, and 3-dimensional echocardiographic studies were recorded at baseline, immediately after Myosplint placement (acute change), and at 1 month after both groups had a reduced rate (190 beats/min) of pacing designed to maintain heart failure. RESULTS: The Myosplint group had significant sustained improvements in left ventricular ejection fraction from baseline, to the acute change, to 1 month (19% +/- 5%; 36% +/- 8%; 39% +/- 13%) and reductions of left ventricular end-systolic volumes (73 +/- 9 mL; 34 +/- 5 mL; 42 +/- 12 mL) and end-systolic wall stress by 39% (341 +/- 68 10(3) dynes x cm(- 2) to 206 +/- 28 10(3) dynes x cm(-2)) acutely and 31% (372 +/- 83 10(3) dynes x cm(-2) to 250 +/- 40 10(3) dynes x cm(-2)) at 1 month. There were no significant changes in mitral regurgitation. CONCLUSION: Application of a Myosplint device to a dilated impaired left ventricle resulted in reduced wall stress and improved left ventricular systolic function that was sustained at 1 month. Device-based shape change is a promising new opportunity to treat patients with dilated cardiomyopathy.

Novel alternative splicing in the 5' exon of the neurotrophin-3 gene.

The neurotrophin-3 (NT-3) gene has previously been reported to consist of three exons including two 5' short untranslated exons and a 3' long exon encoding the entire protein, and to give rise to two classes of transcripts by alternative splicing of the 5' exons to the 3' coding exon. In the present study, we demonstrated the presence of at least four new classes of transcripts of the NT-3 gene, in addition to the two known transcripts. The present finding proposes the further complexity of the regulational mechanism for NT-3 expression.

In vivo hemodynamic performance of the Cleveland Clinic CorAide blood pump in calves.

BACKGROUND: The Cleveland Clinic CorAide left ventricular assist system is based on a small implantable continuous-flow centrifugal blood pump with a completely suspended rotating assembly designed for long-term circulatory support (5 to 10 years). METHODS: Between June 1999 and August 2000, the CorAide blood pump was implanted in 10 calves for 1 month and in 3 calves for 3 months. RESULTS: The mean pump flow and arterial pressure were 6.1 +/- 1.1 L/min and 97 +/- 5 mm Hg, respectively. The mean plasma free-hemoglobin level after postoperative day 3 was 2.0 +/- 1.8 mg/dL. Renal and hepatic function remained normal in all cases. There was no incidence of mechanical failure, hemolysis, bleeding, or systemic organ dysfunction in any of the cases. Significant findings at autopsy were limited to two cases of renal infarction, one of which was associated with an outflow graft infection. CONCLUSIONS: The CorAide blood pump is easily implanted, reliable, nonhemolytic, and nonthrombogenic, positioning it as a leading third-generation, continuous-flow left ventricular assist system with a completely suspended rotor.

Preoperative risk factors for right ventricular failure after implantable left ventricular assist device insertion.

BACKGROUND: Implantable left ventricular assist device (LVAD) insertion complicated by early right ventricular (RV) failure has a poor prognosis and is generally unpredictable. METHODS: To determine preoperative risk factors for perioperative RV failure after LVAD insertion, patient characteristics and preoperative hemodynamics were analyzed in 100 patients with the HeartMate LVAD (Thermo Cardiosystems, Inc, Woburn, MA) at the Cleveland Clinic. RESULTS: RV assist device support was required for 11 patients (RVAD group). RVAD use was significantly higher in younger patients, female patients, smaller patients, and myocarditis patients. There was no significant difference in the cardiac index, RV ejection fraction, or right atrial pressure between the two groups preoperatively. The preoperative mean pulmonary arterial pressure (PAP) and RV stroke work index (RV SWI) were significantly lower in the RVAD group (p = 0.015 and p = 0.011, respectively). Survival to transplant was poor in the RVAD group (27%) and was 83% in the no-RVAD group. CONCLUSIONS: The need for perioperative RVAD support was low, only 11%. Preoperative low PAP and low RV SWI were significant risk factors for RVAD use.

Mutation of low affinity nerve growth factor receptor gene is associated with the hypertensive phenotype in spontaneously hypertensive inbred rat strains.

We previously reported a missense mutation in the low affinity nerve growth factor receptor (LNGFR) gene of spontaneously hypertensive rats (SHR), proposing this gene as a promising candidate in genetic hypertension. In this study we provide further support for implicating this gene in genetic hypertension using two new inbred strains, WKHT and WKHA rats. These strains originated from crossbreeding SHR rats with normotensive Wistar-Kyoto rats (WKY): WKHT rats are hypertensive but not hyperactive, and WKHA rats are hyperactive but not hypertensive. Nucleotide sequence analysis of the LNGFR gene revealed that WKHT has the same mutation as SHR, whereas WKHA has the normal sequence, as seen in WKY. These results support our original hypothesis that the mutated LNGFR gene is linked to hypertension, since the mutation had co-segregated with the hypertensive trait, and not hyperactivity trait of SHR.

Use of peak systolic strain as an index of regional left ventricular function: comparison with tissue Doppler velocity during dobutamine stress and myocardial ischemia.

OBJECTIVES: The goals of this study were to examine peak systolic strain as an index of regional function in an animal model of inotropic stress and ischemia, and to compare these results with peak systolic myocardial tissue Doppler velocity (MDV). BACKGROUND: Myocardial tissue Doppler velocity is an objective measure of regional left ventricular responses to inotropic stimulation and ischemia, but it is affected by tethering from adjacent segments and translational movement. Myocardial Doppler strain (epsilon, relative change in length) is a more local measure of contractility, which can now be derived noninvasively from MDV. METHODS: Eight dogs underwent graded dobutamine infusion followed by coronary occlusion. Epicardial 2-dimensional echocardiography and color MDV of the left ventricle were obtained and digitized from the short-axis view at baseline and with dobutamine doses of 2, 4, and 8 microg/kg per minute. These were repeated 0, 10, 20, 45, and 90 seconds after occlusion of the left anterior descending artery (LAD) (n = 3) or circumflex coronary artery (n = 5). Dobutamine was continued at 8 microg/kg per minute during coronary occlusion. The peak systolic radial MDV (cm/s) and systolic strain (epsilon(s), percent thickening) in the anterior and posterior walls were measured off-line at each stage. RESULTS: Dobutamine caused an increase in MDV (P =.0001) and epsilon(s) (P =.09) above baseline values. Coronary occlusion caused a reduction in wall motion; after 45 seconds, all nonperfused segments were hypokinetic. There was a corresponding decrease in MDV and epsilon(s), but this occurred earlier for epsilon(s), and the difference between ischemic and nonischemic segments was greater for epsilon(s) than for MDV (P <. 03). Nonischemic regions trended to an increase in epsilon(s) (compensatory hyperkinesis), whereas MDV trended downward, probably reflecting the global decrease in left ventricular function. CONCLUSION: Both MDV and epsilon(s) increase with dobutamine and decrease during ischemia. epsilon(s) appears to respond to local ischemia earlier than MDV, perhaps because it is a more local measure. Thus epsilon(s) may prove to be an accurate parameter for the clinical recognition of regional ischemia.

Use of three-dimensional sonomicrometry to study the motion of the mitral valve.

To develop an anatomically correct mitral valve prosthesis, one needs to study the dynamics of the mitral valve apparatus in vivo. The authors used three-dimensional (3D) sonomicrometry and custom visualization software to develop a system to study the mitral valve. Sixteen ultrasonic transducers each were implanted into the hearts of pigs under cardiopulmonary bypass. Four of these crystals were affixed to the base and apex of both papillary muscles, four were attached to the free edge of the anterior and posterior leaflets where the main chordae attach, six were placed around the mitral annulus, and two were affixed to the epicardial wall. The digital sonomicrometer system sequentially fired each transducer and listened for an ultrasound signal at the other 15. This process was repeated so that all 16 transducers were sequentially fired, and each cycle of 16 was repeated 200 times/sec. The matrix of distances obtained between all the combinations of pairs of the 16 transducers was converted to x, y, z coordinates, the shape of the mitral valve apparatus was reconstructed in 3D on a graphics computer, and the valve's motion was analyzed over several cardiac cycles. The authors conclude that mapping of the mitral valve apparatus in pigs by 3D sonomicrometry provides quantitative measurements that can aid in the design of a mitral valve prosthesis that closely replicates the anatomy and function of the natural valve.

Effect of respiration on the arterial pressure wave in calves with nonpulsatile biventricular bypass.

The possibility of idioperipheral pulsation in calves with chronic nonpulsatile biventricular bypass has been previously reported. To test the hypothesis that both spontaneous respiration and mechanical ventilation are the cause of pulsation in the arterial pressure recording in calves with nonpulsatile biventricular bypass, two protocols were used to generate data. Both right and left ventricles were bypassed using two centrifugal pumps followed by electrical fibrillation in seven calves. In protocol 1, the spontaneous respiratory rate was increased in awake calves by carbon dioxide gas inhalation. In protocol 2, the animals were anesthetized and intubated to control ventilation, and data were collected as the ventilation rate or tidal volume was changed in a stepwise fashion, as well as when the ventilation was stopped for several seconds. The results of protocol 1 showed good correlation between the arterial pulse rate and spontaneous respiratory rate (Y = 1.03 X, r = 0.822, p < 0.001). The results of protocol 2 showed that the relationship between the ventilation rate and the arterial pulse cycle was identical (Y = X, r = 1.000, p < 0.001), and no pulsation was seen when the ventilator was stopped. The authors conclude that the arterial pulsation observed in calves with nonpulsatile biventricular bypass is caused by respiration.

Heat from an implanted power source is mainly dissipated by blood perfusion.

Heat dissipation and its effects on tissue and blood interfaces are common problems associated with the development and increased use of artificial hearts, because all of the implantable actuators for artificial hearts generate waste heat due to inefficiencies of energy conversion. To determine the mechanisms of heat dissipation from artificial hearts, heated disks producing constant heat fluxes of 0.08 watts/cm2 were implanted adjacent to the left lung and the latissimus dorsi muscle in calves for 2 weeks, 4 weeks, and 7 weeks. At the end of each experiment, a series of acute studies was performed in which blood perfusion to the heated tissue was decreased or stopped to observe the contribution of blood perfusion to heat dissipation. The cooling effect of ventilation was also examined to determine its relative contribution to heat dissipation in lung tissue by decreasing the minute ventilation volume. The importance of blood perfusion for heat dissipation was demonstrated by the temperature rise after cessation of blood perfusion to the heated tissue. The contribution of ventilation to heat dissipation in the heated lung tissue was minimal. Contribution of total blood perfusion to heat dissipation was increased with time in the muscle tissue, which has relatively low resting blood perfusion, but not in the lung tissue, which has relatively high blood perfusion. In the heated muscle tissue, the in vivo adaptive response to chronic heat was functionally shown by the increased perfusion. In conclusion, blood perfusion was the main mechanism of heat dissipation from tissues that were adjacent to an implanted power source.

A comprehensive hematologic study in calves with total artificial hearts.

Thromboembolism and infection remain potential threats for long-term circulatory assist and replacement devices. The alteration of the hemostatic system and of blood cell functions caused by device implantation may predispose the recipient to these complications. Many sensitive coagulation assays and the technology of flow cytometry would be powerful tools for this investigation. The availability of such immunologic technologies for animal species other than humans has yet to be established. In a series of in vitro tests we found that the following assays, among others, are usable in calves: TAT, TxB2, platelet surface glycoprotein IIbIIIa, and membrane aminophospholipid. F1.2, D-dimer, beta TG, PF-4, and platelet surface expression of GMP-140 and receptors for fibronectin, thrombospondin, and vWF were not measurable. A sustained mild decrease in hematocrit levels in six calves with the Cleveland Clinic-Nimbus total artificial heart for 11-120 days was attributed to an increase in circulating blood volume, but not to red blood cell damage. Whole blood platelet aggregation was suppressed only for the first 3 post operative days, with decreased GPIIbIIIa expression. Polymorphonuclear phagocytosis, chemotaxis, and superoxide anion production were not altered. Device infection and thromboembolism occurred in one of 13 cases overall.