Michael P. Feneley

Cardiac homeobox gene NKX2-5 mutations and congenital heart disease: associations with atrial septal defect and hypoplastic left heart syndrome.

OBJECTIVESnWe sought to examine the importance of mutations in the cardiac transcription factor gene NKX2-5 in patients with an atrial septal defect (ASD), patent foramen ovale (PFO), or hypoplastic left heart syndrome (HLHS).nnnBACKGROUNDnMutations in NKX2-5 have been found in families showing secundum ASD and atrioventricular (AV) conduction block and in some individuals with tetralogy of Fallot. The prevalence of NKX2-5 mutations in sporadic cases of ASD/PFO and other forms of congenital heart disease is unknown.nnnMETHODSnA cohort of 146 individuals with secundum ASD, PFO complicated by paradoxical embolism, or HLHS were evaluated. Patients with ASD or PFO were ascertained irrespective of family history or associated cardiac abnormalities. The coding region of the NKX2-5 locus was amplified by polymerase chain reaction and sequenced.nnnRESULTSnAmong 102 ASD and 25 PFO patients screened, 13 patients (10%) had a positive family history and 5 patients (4%) had AV conduction block. We found one previously documented NKX2-5 missense mutation, T178M, in members of a family with ASD without AV conduction block. One NKX2-5 mutation-positive child from this family had HLHS, although no mutations were subsequently found in 18 patients with sporadic or familial HLHS. In a second ASD family without AV conduction block, we found a missense change, E21Q, previously reported as pathogenic. Because this change did not segregate with disease status, we propose that it is a non-disease-causing polymorphism.nnnCONCLUSIONSnOur findings suggest that NKX2-5 mutations are a relatively infrequent cause of sporadic ASD and HLHS. Screening for NKX2-5 mutations may be warranted in individuals with ASD and a positive family history, irrespective of the presence or absence of AV conduction block.

Accuracy of biplane transesophageal echocardiography in detecting left atrial thrombus

Abstract In summary, biplane TEE had a high sensitivity and specificity, but a low positive predictive accuracy, for detecting left atrial thrombus in 60 patients who underwent mitral valve surgery. The high falsepositive rate observed in this study appears to reflect the difficulty in differentiating severe spontaneous echo contrast from thrombus on TEE.

Inhibition of Red Cell Aggregation Prevents Spontaneous Echocardiographic Contrast Formation in Human Blood

BACKGROUNDnSpontaneous echocardiographic contrast (SEC) is a pattern of blood echogenicity that has been attributed to ultrasonic backscatter from blood cell aggregates that form under low shear conditions. Patients with left atrial SEC have an increased thromboembolic risk. This study examined the role of red cell and platelet aggregates in the pathogenesis of SEC in human blood and the effects on SEC of antithrombotic therapy and red cell disaggregatory agents.nnnMETHODS AND RESULTSnBlood echogenicity was examined with the use of quantitative videodensitometry over a controlled range of flow velocities in an in vitro model characterized by nonlaminar flow conditions. One hundred ninety study samples were prepared from single fresh blood donations (40 to 120 mL) from 24 healthy volunteers and 11 patients. Whole blood echogenicity was unaltered by depletion of platelets, stimulation of platelet aggregation with adenosine diphosphate, or inhibition of platelet aggregation with aspirin. Low flow-related echogenicity increased with increasing hematocrit (P<.001) but was abolished when red cells were lysed selectively with saponin (P<.001). In the presence of red cells, low flow-related echogenicity increased with increasing fibrinogen concentration (P<.001) and with plasma paraproteins. Low flow-related echogenicity in whole blood was unaltered by heparin and warfarin but was reduced in a dose-dependent manner by dextran 40 (40 mg/mL, 70% reduction, P<.001) and poloxamer 188 (8 mg/mL, 47% reduction, P<.001), which inhibited red cell aggregation.nnnCONCLUSIONSnThese results support protein-mediated red cell aggregation as the mechanism of SEC in human blood. Inhibition of red cell aggregation, indexed by resolution of SEC, may provide an alternative to anticoagulant and antiplatelet therapy to reduce cardiac thromboembolic risk.

On-line Synthesis of the Human Ascending Aortic Pressure Pulse From the Finger Pulse

Although systolic pressure in the ascending aorta (AA) can be determined accurately from the radial arterial waveform using a single generalized transfer function (TF) of the upper limb, a better on-line methods is needed for accurate noninvasive synthesis of the AA pressure contour to characterize left ventricular contractile function and ventricular-vascular coupling. AA, tonometric carotid (CA), and photoplethysmographic finger (FA) arterial pressure waveforms were recorded in 12 subjects (10 male, aged 59.1+/-10.3 years, mean+/-SD) during cardiac catheterization. The AA-FA TF was estimated using (1) a single generalized TF (GAA), (2) individualized TFs directly determined from CA-FA recordings in each patient (DAA), and (3) individualized TFs computed from CA-FA recordings in each patient with a mathematical model of the human upper limb (MAA). AA pressure waveforms were synthesized from FA recordings in real time using convolution windows derived from these TFs. Under steady state conditions, the root mean square error (RMSE) between measured and synthesized AA was lower by DAA (3.3+/-1.3 mm Hg) and MAA (3.9+/-1.2 mmHg) than by GAA (4.8+/-2.0 mm Hg, P<.05). During dynamic load alteration induced by the Valsalva maneuver, however, the MAA method performed better (5.4+/-2.8 mm Hg) than both the GAA (5.8+/-3.3 mm Hg, P<.05) and DAA (6.5+/-2.7 mm Hg, P<.01) methods. The beat-to-beat AA contour can be accurately and noninvasively synthesized on-line using individualized TFs. During dynamic load alteration, individualized TFs derived with an upper limb arterial model provide greater accuracy.

Single-beat determination of preload recruitable stroke work relationship: derivation and evaluation in conscious dogs.

OBJECTIVESnTo derive and evaluate a method of estimating the slope (Mw) of the preload recruitable stroke work (PRSW) relationship between left ventricular stroke work (SW) and end-diastolic volume (EDV) from a single beat.nnnBACKGROUNDnMw is a load-insensitive index of contractile function, but its clinical application has been limited by the need to record multiple beats over a wide volume range.nnnMETHODSnPressure-volume loops were recorded over a variable preload and afterload range by vena caval and aortic constrictions in 12 conscious dogs instrumented with epicardial dimension transducers and micromanometers. Single-beat Mw (SBMw) was determined as the ratio SW/(EDV-Vw), where the volume-axis intercept of the PRSW relationship (Vw)(EDV at zero SW) was estimated as k x EDVB + (k - 1)LVwall, k is the ratio of the epicardial shell volumes corresponding to Vw and baseline EDV (EDVB) and LVwall is wall volume.nnnRESULTSnIn the first six dogs, k was found to be essentially constant at 0.7, SBMw estimates were insensitive to wide preload variation, and the relationship between SBMw and multibeat Mw determined during caval and aortic constrictions did not differ significantly from the line of identity. When the same constant k value was applied to SBMw estimation in a different group of six dogs, SBMw did not differ significantly from multibeat Mw (83 +/- 12 erg x cm(-3) x 10(3) and 77 +/- 12 erg x cm(-3) x 10(3), respectively), neither changed significantly during aortic constriction and both increased significantly with calcium infusion (107 +/- 18 erg x cm(-3) x 10(3) and 95 +/- 19 erg x cm(-3) x 10(3), respectively, both p < 0.05). Single-beat Mw was less load-dependent, more reproducible and a more sensitive index of inotropic state than two previously described single-beat indexes, single-beat elastance and maximum power divided by EDV2.nnnCONCLUSIONSnMw can be determined accurately from a single, steady-state beat in the normal canine heart and is sensitive to inotropic alterations while being insensitive to wide variations in preload and afterload. Single-beat Mw estimation should facilitate noninvasive, load-independent assessment of contractile function.

Functional evidence of reversible ischemic injury immediately after the sympathetic storm associated with experimental brain death.

BACKGROUNDnAcute brain death from increased intracranial pressure results in a transient increase in myocardial adenosine and lactate, which indicates that oxygen demand exceeds oxygen delivery during the sympathetic storm. The aim of this study was to determine the functional significance of this period of ischemia.nnnMETHODSnBrain death was inflicted on 40 Westran pigs (36.5-68.0 kg) by inflating a 21-ml subdural balloon over 3 minutes. In 38 animals, micromanometry and sonomicrometry were used to obtain left ventricular pressure-volume loops to determine the preload recruitable stroke work (PRSW) relationship. Data files were recorded before and at 15-minute intervals after beginning balloon inflation. Plasma troponin I was measured before and 60 minutes after beginning balloon inflation in the 38 instrumented and 2 non-instrumented animals.nnnRESULTSnAll animals experienced the classical sympathetic storm. The slope of the PRSW relationship decreased, and the volume-axis intercept shifted to the right 15 minutes after beginning balloon inflation (p < 0.0001). Progressive incremental recovery (leftward shift) occurred between subsequent time points (p < or = 0.0018). In the instrumented animals, the mean plasma troponin I level increased from 1.4 +/- 1.6 microg/liter to 2.8 +/- 2.3 microg/liter (p < 0.001). However, troponin I was not detected before or after induction of brain death in the plasma of either non-instrumented animal (p = 0.001).nnnCONCLUSIONSnThe sympathetic storm produced transient contractile dysfunction, consistent with ischemic injury. However, troponin I release reflected surgical instrumentation and not brain death.

Patterns of Doppler-measured blood flow velocity in the normal and fibrillating human left atrial appendage

Doppler measurement of left atrial appendage (LAA) blood velocity during transesophageal echocardiography has been proposed as a method of assessing LAA contractile function and thromboembolic risk. Clinical and echocardiographic determinants of five LAA Doppler blood velocity patterns were examined in 40 patients with a history of atrial fibrillation (AF), in 10 control subjects, and in 5 patients aged </=60 years having sinus rhythm and left ventricular hypertrophy. In sinus rhythm, two blood velocity patterns were differentiated by the extent of passive emptying of the LAA, which was related to age and left ventricular early diastolic filling properties. In AF, three blood velocity patterns were differentiated by the relative preservation of LAA mechanical function during fibrillatory activity. LAA contractile function is an important but not the sole determinant of blood flow in the normal and fibrillating human LAA.

Effect of inhaled nitric oxide on normal human left ventricular function.

OBJECTIVESnThis study determined the effects of inhaled nitric oxide (NO) on load-independent indexes of normal human left ventricular (LV) function.nnnBACKGROUNDnInhaled NO is a potent and selective pulmonary vasodilator. However, when it is used in patients with congestive heart failure, the decrease in pulmonary vascular resistance (PVR) is often associated with an increase in pulmonary capillary wedge pressure. NO has been shown to have a negative inotropic action, but it is not known whether it affects LV chamber function when delivered by inhalation.nnnMETHODSnEleven subjects (51 to 69 years old) with normal LV function (mean ejection fraction 72% [range 60% to 80%]) were studied. Four patients had concomitant coronary artery disease. Pressure-volume loop recordings were used to determine end-systolic and end-diastolic pressure-volume and preload recruitable stroke work relations. NO was delivered at 20 ppm for 10 min. In an additional group of patients with normal LV function, PVR (n = 5) and NO metabolites (n = 9) were measured.nnnRESULTSnThere was no effect of inhaled NO on steady state LV pressures, volumes, contractility, contraction duration, active relaxation (time constant of relaxation, peak negative first derivative of left ventricular pressure), diastolic compliance or PVR. NO metabolites (methemoglobin and nitrate) were present in the LV cavity at the same concentration as right atrial venous blood, suggesting inactivation of free NO before arrival in the LV chamber. This study had a power of 0.995 to detect a 5% change in contractility (slope of preload recruitable stroke work relation) for alpha = 0.05, based on the multiple linear regression model used.nnnCONCLUSIONSnThese results indicate that 20 ppm of inhaled NO does not have significant effects on normal LV function. This lack of effect may be due in part to rapid inactivation of free NO in transit to the heart.

Stratification of thromboembolic risk of atrial fibrillation by transthoracic echocardiography and transesophageal echocardiography: the relative role of left atrial appendage function, mitral valve disease, and spontaneous echocardiographic contrast.

The role of transesophageal echocardiography (TEE) in thromboembolic risk stratification in atrial fibrillation (AF) has not been established. Left atrial appendage contractile dysfunction in patients with AF predisposes to thrombus formation. The extent of blood stasis and propensity for thrombus can be assessed during TEE by measurement of the peak Doppler velocity of blood outflow from the appendage. Spontaneous echocardiographic contrast (SEC) is a swirling pattern of blood echogenicity that may be detected by TEE in the left atrium in patients with AF. The presence of SEC reflects left atrial blood stasis and a prothrombotic state. SEC is associated with an increased risk of systemic thromboembolic events. Parameters derived from TEE may provide additional prognostic data to clinical history and transthoracic echocardiography in thromboembolic risk stratification in AF.

Lentivirus vector-mediated gene transfer to the developing bronchiolar airway epithelium in the fetal lamb.

Development of effective and durable gene therapy for treatment of the respiratory manifestations of cystic fibrosis remains a formidable challenge. Obstacles include difficulty in achieving efficient gene transfer to mature airway epithelium and the need to stably transduce self‐renewing epithelial progenitor cells in order to avoid loss of transgene expression through epithelial turnover. Targeting the developing airway epithelium during fetal life offers the prospect of circumventing these challenges.