Large-Scale Computational Screening Identifies First in Class Multitarget Inhibitor of EGFR Kinase and BRD4.

Inhibition of cancer-promoting kinases is an established therapeutic strategy for the treatment of many cancers, although resistance to kinase inhibitors is common. One way to overcome resistance is to target orthogonal cancer-promoting pathways. Bromo and Extra-Terminal (BET) domain proteins, which belong to the family of epigenetic readers, have recently emerged as promising therapeutic targets in multiple cancers. The development of multitarget drugs that inhibit kinase and BET proteins therefore may be a promising strategy to overcome tumor resistance and prolong therapeutic efficacy in the clinic. We developed a general computational screening approach to identify novel dual kinase/bromodomain inhibitors from millions of commercially available small molecules. Our method integrated machine learning using big datasets of kinase inhibitors and structure-based drug design. Here we describe the computational methodology, including validation and characterization of our models and their application and integration into a scalable virtual screening pipeline. We screened over 6 million commercially available compounds and selected 24 for testing in BRD4 and EGFR biochemical assays. We identified several novel BRD4 inhibitors, among them a first in class dual EGFR-BRD4 inhibitor. Our studies suggest that this computational screening approach may be broadly applicable for identifying dual kinase/BET inhibitors with potential for treating various cancers.

Ultrasonography versus radiography for detection of fluid in the canine tympanic bulla.

The tympanic bullae of 20 previously healthy cadaver animals were randomly filled with sterile saline or air. The heads were positioned using tape ties and a rostrocaudal open mouth radiograph taken. The resulting radiographs were assessed by a blinded ECVDI diplomate, with each tympanic bulla being recorded as being either air or fluid filled. Ultrasound examination of the bullae was performed, by a blinded assessor, using a B-mode 5 mHz sector probe (Sonotron-VingMed Sound A/S, PO Box 141, N3191, Horten, Norway) operating at 3 mHz. Each bulla was recorded as being either air or fluid filled. Results of radiography and ultrasonography were each compared to the actual status of the bulla in a chi square analysis. Sensitivity of radiography was 80%, with specificity of 65%. Ultrasonographic sensitivity was 100% and specificity 100%. Ultrasonography may in future provide a cheap, noninvasive, rapid and widely available method for diagnosing otitis media in clinical cases.

Respiratory variation of mitral and pulmonary venous Doppler flow velocities in constrictive pericarditis before and after pericardiectomy.

The objective of this study was to determine whether Doppler echocardiography is useful in assessing the effects of pericardiectomy in patients with constrictive pericarditis by studying the postoperative change in the respiratory variation of mitral inflow and pulmonary venous Doppler flows. The study population consisted of 35 cases with surgically proven constrictive pericarditis. Thirty-five patients had preoperative Doppler echocardiography, whereas 4 patients died of non-cardiac causes and 1 patient had a heart transplant before follow-up. Postoperative studies were performed at a mean of 1081 +/- 84 days (range 120-2700 days) after pericardiectomy. The mean (+/- SD) respiratory variation changed after surgery from a baseline value of 17% +/- 14% to 8% +/- 8% for peak mitral E velocity (P <.01); from 25% +/- 18% to 7% +/- 13% (P <.001) for pulmonary venous (PV) peak diastolic flow velocity, and from 21% +/- 13% to 11% +/- 13% (P =.009) for PV peak systolic flow velocity. The 23 patients who became asymptomatic after surgery had a significantly lower mean mitral and PV respiratory variation than the 7 patients who were NYHA class II (4% +/- 4% and 6% +/- 4% vs 21% +/- 6% and 19% +/- 10%, respectively, P <.0001 for both). Pulsed Doppler echocardiographic assessment of respiratory variation is useful for evaluating the outcome of pericardiectomy.

Clinical and echocardiographic characteristics of papillary fibroelastomas: a retrospective and prospective study in 162 patients.

BACKGROUND: Cardiac papillary fibroelastoma (CPF) is a primary cardiac neoplasm that is increasingly detected by echocardiography. The clinical manifestations of this entity are not well described. METHODS AND RESULTS: In a 16-year period, we identified patients with CPF from our pathology and echocardiography databases. A total of 162 patients had pathologically confirmed CPF. Echocardiography was performed in 141 patients with 158 CPFs, and 48 patients had CPFs that were not visible by echocardiography (<0.2 cm), leaving an echocardiographic subgroup of 93 patients with 110 CPFs. An additional 45 patients with a presumed diagnosis of CPF were identified. The mean age of the patients was 60+/-16 years of age, and 46.1% were male. Echocardiographically, the mean size of the CPFs was 9+/-4.6 mm; 82.7% occurred on valves (aortic more than mitral), 43.6% were mobile, and 91.4% were single. During a follow-up period of 11+/-22 months, 23 of 26 patients with a prospective diagnosis of CPF that was confirmed by pathological examination had symptoms that could be attributable to embolization. In the group of 45 patients with a presumed diagnosis of CPF, 3 patients had symptoms that were likely due to embolization (incidence, 6.6%) during a follow-up period of 552+/-706 days. CONCLUSIONS: CPFs are generally small and single, occur most often on valvular surfaces, and may be mobile, resulting in embolization. Because of the potential for embolic events, symptomatic patients, patients undergoing cardiac surgery for other lesions, and those with highly mobile and large CPFs should be considered for surgical excision.

Echocardiographic evaluation of valvular stenosis: the gold standard for the next millennium?

Over the past decade, utilization of cardiac catheterization for preoperative hemodynamic assessment of patients with mitral and aortic stenosis has steadily decreased. The reason for this trend is the use of echocardiography, which is emerging as a gold standard for clinical characterization of valvular lesions. Since cardiac catheterization is an invasive procedure that is associated with a significant percentage of complications, echocardiographic evaluation of patients with valvular stenosis is safer and more cost-effective. In the next millennium, echocardiography will probably completely replace the use of catheterization for hemodynamic assessment of the severity of mitral and aortic stenosis.

Comparison of quantitative and semiquantitative methods for assessing mitral regurgitation by transesophageal echocardiography.

Semiquantitative grading of mitral regurgitation (MR) by transesophageal echocardiography (TEE) is widely used for clinical decision making. However, the relation between semiquantitative grading by biplane or multiplane TEE and quantitative measures remains undetermined. Biplane or multiplane TEE was performed in 113 patients in the operating room. MR severity was graded from 1 to 4+ by Doppler color flow mapping. MR was quantified using the thermodilution-Doppler method as mitral regurgitant stroke volume (RSV) derived from the difference between total mitral inflow measured by pulsed Doppler and forward flow measured by thermodilution. Mitral regurgitant orifice area (ROA) was calculated by RSV divided by mitral regurgitant velocity. RSV and ROA were also calculated using the proximal isovelocity surface area method. RSV and ROA significantly correlated with the semiquantitative grading either by TEE or angiogram in a nonlinear fashion, with the best fit being given by an exponential model with correlation coefficients from 0.73 to 0.87 (p <0.001). Substantially increased RSV and ROA were observed in MR grades of > or =3+. In the same grades of 3+ or 4+ MR, the largest RSV was 4 times larger than the smallest (190 to 220 vs 44 to 45 ml), and the largest ROA (1.82 to 2.0 vs 0.26 to 0.27 cm2) was sixfold larger than the smallest. Patients with 2 to 3+ MR had significantly variable RSV and ROA (range 21 to 91 ml and 0.12 to 0.65 cm2, respectively). Color flow mapping by biplane or multiplane TEE or angiography is able to categorize precisely mild (< or =2+) and severe (> or =3+) MR, but cannot accurately determine actual hemodynamic load of MR in more severe degrees of MR.

Qualitative and quantitative effects of harmonic echocardiographic imaging on endocardial edge definition and side-lobe artifacts.

Harmonic imaging is a new ultrasonographic technique that is designed to improve image quality by exploiting the spontaneous generation of higher frequencies as ultrasound propagates through tissue. We studied 51 difficult-to-image patients with blinded side-by-side cineloop evaluation of endocardial border definition by harmonic versus fundamental imaging. In addition, quantitative intensities from cavity versus wall were compared for harmonic versus fundamental imaging. Harmonic imaging improved left ventricular endocardial border delineation over fundamental imaging (superior: harmonic = 71.1%, fundamental = 18.7%; similar: 10.2%; P <.001). Quantitative analysis of 100 wall/cavity combinations demonstrated brighter wall segments and more strikingly darker cavities during harmonic imaging (cavity intensity on a 0 to 255 scale: fundamental = 15.6 +/- 8.6; harmonic = 6.0 +/- 5.3; P <.0001), which led to enhanced contrast between the wall and cavity (1.89 versus 1.19, P <.0001). Harmonic imaging reduces side-lobe artifacts, resulting in a darker cavity and brighter walls, thereby improving image contrast and endocardial delineation.

Reversal of the pattern of respiratory variation of Doppler inflow velocities in constrictive pericarditis during mechanical ventilation.

BACKGROUND: Spontaneous inspiration causes a characteristic decrease of the mitral valve (MV) and pulmonary venous (PV) flow velocities obtained by Doppler echocardiography in patients with constrictive pericarditis (CP). This has been explained by the decrement it causes in the intrathoracic pressure. Positive pressure ventilation (PPV) causes an increment of intrathoracic pressure with mechanical inspiration. Therefore the pattern of respiratory variation produced during PPV may differ from that seen during spontaneous breathing. OBJECTIVE: Our goal was to describe the effect of PPV on the pattern and magnitude of respiratory variation of MV and PV flow velocities in CP. METHODS: We performed intraoperative pulsed Doppler transesophageal echocardiography on 15 patients (13 men, mean age 52+/-15 years) with CP after general anesthesia and before sternotomy and pericardial stripping. The peak velocity and time-velocity integral (TVI) of the mitral inflow E and A waves and the PV systolic and diastolic waves were measured at onset of inspiration and expiration for 3 to 6 respiratory cycles. Respiratory phase was monitored with a heat-sensitive nasal thermistor. The percent change in Doppler flow velocities from mechanical inspiration (INS) to mechanical expiration (EXP) was calculated with the formula %change = INS - EXP / INS x 100. RESULTS: The peak velocity of the mitral inflow E wave was significantly higher during mechanical inspiration than expiration (57 +/-14.5 versus 47+/-13.9 cm/s, P<.001). This represented a percent change of 18%+/-7.9% from expiration to inspiration. The mean TVI of the mitral inflow E was also higher during mechanical inspiration than expiration (P = .02). The peak velocity of the PV D wave was higher during mechanical inspiration than expiration (39+/-17.8 versus 28+/-14.7 cm/s, P<.001). This represented a mean percent change of 28%+/-13.8%. The mean value of the TVI for the PV D wave was also significantly greater during mechanical inspiration than expiration (P <.05). CONCLUSIONS: Positive pressure ventilation reverses the pattern of respiratory variation of the MV and PV flow velocities in CP. The percent change in the peak velocities of the MV and PV flows produced by PPV is the same range reported in CP during spontaneous breathing.

Disk embolization of a Björk-Shiley convexo-concave mitral valve: a cause of sudden cardiovascular collapse and mesenteric ischemia.

Strut fracture and disk embolization of a Björk-Shiley convexo-concave valve is uncommon, but it should always be considered as a cause of sudden cardiovascular collapse in patients with such valves. Recognition of this clinical scenario is essential, given the importance of early diagnosis and the prevalence of these valves worldwide. We present a fatal case of disk embolization of a mitral prosthesis presenting with cardiogenic shock and mesenteric ischemia.

Cosgrove-Edwards Annuloplasty System: midterm results.

BACKGROUND: The Cosgrove-Edwards Annuloplasty System includes a universally flexible band that corrects mitral annular dilatation via measured plication of the posterior annulus. The purpose of this study was to evaluate midterm clinical and functional results in the first 197 patients receiving this flexible annuloplasty band at mitral valve repair. METHODS: From February 1993 to July 1994, 197 consecutive patients with mitral regurgitation had mitral valve repair using this system. Valve disease was degenerative in 73%, rheumatic in 15%, ischemic in 5%, infectious in 2.5%, and other in 4%. RESULTS: Immediately after repair, echocardiographic mitral regurgitation was none or trivial in 92%, 1+ in 5%, and 2+ in 3%. There were no hospital deaths. Late follow-up was available in 195 patients (99%), with 661 patient-years of follow-up available for analysis. Four-year actuarial survival was 93%, freedom from thromboembolism 94%, from endocarditis 98%, and from reoperation 95%. At a mean interval of 18 months, echocardiography in 157 patients demonstrated no or trace mitral regurgitation in 56%, 1+ in 24%, 2+ in 9%, 3+ in 6%, and 4+ in 3%. At a mean of 61 +/- 5 months, reconstruction of the mitral annulus from real-time three-dimensional echocardiographic images in 10 patients confirmed preserved nonplanar shape and sphincter mechanism of the mitral annulus. Annular orifice area decreased 28% +/- 11% during the cardiac cycle from a mean of 10.1 +/- 3.9 cm2 in diastole to 7.2 +/- 2.8 cm2 in systole. CONCLUSIONS: This annuloplasty system is effective for repair of mitral regurgitation secondary to all causes and preserves mitral annular flexibility and function at 5-year follow-up.

Is anterior leaflet repair always necessary in repair of bileaflet mitral valve prolapse?

BACKGROUND: Traditionally, bileaflet prolapse has been treated by posterior leaflet resection combined with one of a number of procedures designed to support the anterior leaflet. However, most patients with bileaflet prolapse do not have important anterior chordal pathology. This study was undertaken to evaluate the effectiveness of a strategy of posterior leaflet resection and annuloplasty alone for patients with bileaflet prolapse and no anterior chordal rupture or severe anterior chordal elongation. METHODS: From 1993 to 1997, 93 patients with transesophageal echocardiography (TEE) demonstrated bileaflet prolapse and without anterior chordal rupture or important anterior chordal elongation had primary isolated mitral valve repair consisting only of posterior leaflet resection (quadrangular in 28 and sliding in 65) and annuloplasty (Cosgrove-Edwards in 83, pericardial in 9, and Carpentier-Edwards in 1). All patients had severe mitral regurgitation documented by intraoperative TEE. Mean age was 55+/-13 years; 60% were men. RESULTS: Postrepair, mitral regurgitation was 0 to trace in 93% and 1+ in 7%. There were no operative deaths. Late follow-up was available in all patients, with 277 patient-years of follow-up available for analysis. Five-year actuarial survival was 95%. At a mean interval of 2.3+/-1.3 (SD) years, echocardiography demonstrated no or trace mitral regurgitation in 65%, 1+ in 28%, and 2+ in 7%. No correlates of late mitral regurgitation were identified by multivariable analysis. No patient has required reoperation. CONCLUSIONS: In the absence of significant anterior chordal pathology, a strategy of posterior leaflet resection and annuloplasty corrects anterior leaflet prolapse and mitral regurgitation, and provides a durable repair without the necessity of additional procedures on the anterior leaflet.

Inferior vena caval masses identified by echocardiography.

The most common cause of an inferior vena caval mass is renal cell carcinoma that extends through the lumen, occurring in 47 of 62 patients (85%). Detection of an inferior vena caval mass affects the surgical approach requiring cardiopulmonary bypass for resection when the mass extends to the heart.

The value of assessing pulmonary venous flow velocity for predicting severity of mitral regurgitation: A quantitative assessment integrating left ventricular function.

Although alteration in pulmonary venous flow has been reported to relate to mitral regurgitant severity, it is also known to vary with left ventricular (LV) systolic and diastolic dysfunction. There are few data relating pulmonary venous flow to quantitative indexes of mitral regurgitation (MR). The object of this study was to assess quantitatively the accuracy of pulmonary venous flow for predicting MR severity by using transesophageal echocardiographic measurement in patients with variable LV dysfunction. This study consisted of 73 patients undergoing heart surgery with mild to severe MR. Regurgitant orifice area (ROA), regurgitant stroke volume (RSV), and regurgitant fraction (RF) were obtained by quantitative transesophageal echocardiography and proximal isovelocity surface area. Both left and right upper pulmonary venous flow velocities were recorded and their patterns classified by the ratio of systolic to diastolic velocity: normal (>/=1), blunted (<1), and systolic reversal (<0). Twenty-three percent of patients had discordant patterns between the left and right veins. When the most abnormal patterns either in the left or right vein were used for analysis, the ratio of peak systolic to diastolic flow velocity was negatively correlated with ROA (r = -0.74, P <.001), RSV (r = -0.70, P <.001), and RF (r = -0.66, P <.001) calculated by the Doppler thermodilution method; values were r = -0.70, r = -0.67, and r = -0.57, respectively (all P <.001), for indexes calculated by the proximal isovelocity surface area method. The sensitivity, specificity, and predictive values of the reversed pulmonary venous flow pattern for detecting a large ROA (>0.3 cm(2)) were 69%, 98%, and 97%, respectively. The sensitivity, specificity, and predictive values of the normal pulmonary venous flow pattern for detecting a small ROA (<0.3 cm(2)) were 60%, 96%, and 94%, respectively. However, the blunted pattern had low sensitivity (22%), specificity (61%), and predictive values (30%) for detecting ROA of greater than 0.3 cm(2) with significant overlap with the reversed and normal patterns. Among patients with the blunted pattern, the correlation between the systolic to diastolic velocity ratio was worse in those with LV dysfunction (ejection fraction <50%, r = 0.23, P >.05) than in those with normal LV function (r = -0.57, P <.05). Stepwise linear regression analysis showed that the peak systolic to diastolic velocity ratio was independently correlated with RF (P <.001) and effective stroke volume (P <.01), with a multiple correlation coefficient of 0.71 (P <.001). In conclusion, reversed pulmonary venous flow in systole is a highly specific and reliable marker of moderately severe or severe MR with an ROA greater than 0.3 cm(2), whereas the normal pattern accurately predicts mild to moderate MR. Blunted pulmonary venous flow can be seen in all grades of MR with low predictive value for severity of MR, especially in the presence of LV dysfunction. The blunted pulmonary venous flow pattern must therefore be interpreted cautiously in clinical practice as a marker for severity of MR.

Intraoperative transesophageal echocardiography in minimally invasive cardiac valve surgery.

The minimally invasive procedure is a new surgical technique that uses a small sternal incision. Because of limited surgical exposure, removal of intracavitary air and visual assessment of cardiac function are not possible. We studied the utility of intraoperative transesophageal echocardiography (IOE) before and after cardiopulmonary bypass in 112 patients (mean age 53.1 +/- 15.2 years, 74 males) who underwent minimally invasive valvular surgery. Surgical procedures included 52 isolated mitral valve procedures (49 repairs, 3 prostheses), 58 isolated aortic valve procedures (16 repairs, 26 prostheses, 16 homografts), and 2 combined aortic and mitral valve repairs. Prepump IOE was useful to confirm valve dysfunction and assist determination of arterial cannulation site. Postpump IOE identified intracardiac air in all patients, which was defined as extensive in 58 (52%) cases. Postoperatively, new left ventricular dysfunction was noted in 22 (20%) patients, more often in the group with extensive air by IOE (17 [30%] of 58 patients) compared with those without extensive air (5 [10%] of 54 patients, P =.01). Second pump runs were required in 7 (6%) of 112 patients: 3 cases of residual aortic regurgitation, 1 case of residual mitral regurgitation, and 3 cases with new ventricular dysfunction. No deaths occurred. We conclude that IOE is essential in minimally invasive valvular surgery because it detects problems that require immediate remedy. IOE allows real-time assessment of ventricular filling, ventricular and valvular function, and intracardiac air.

Safety of coronary arteriography in clinically stable patients following coronary bypass surgery. Post CABG Clinical Trial Investigators.

The frequent use of diagnostic coronary arteriography and its importance in evaluating results of intervention in clinical trials emphasize the necessity of continued assessment of procedural risk. Several studies have described such risks, but they have often included a diverse group of patients with varying levels of clinical stability. Furthermore, this risk has not been well established in a population of patients with saphenous vein bypass grafts. There is need to define the risk of coronary arteriography in a group of patients who are both clinically similar and stable, and to evaluate the influence of improved technology and increased operator experience on the risk of the procedure. The National Heart, Lung, and Blood Institute-funded Post Coronary Artery Bypass Graft Trial offered the opportunity to evaluate the risk of elective diagnostic coronary arteriography in clinically stable patients studied at two points in time: pre-enrollment and 4-5 years after study entry. In this group of 2,635 angiograms from clinically stable patients over 5 years there were no deaths and the risk of myocardial infarction was 0.08%, while 0.7% had clinically important complications. Non-elective, urgent studies (311 angiograms) on unstable patients were more likely to include angioplasty and were associated with a risk of death of 0.6% and myocardial infarction of 1.3%. Complications did not vary with age or gender. Vascular trauma was more likely to occur using the brachial than the femoral artery entry sites. These results indicate that elective angiography on stable patients can be accomplished with a very low risk of mortality (0% in this study) or serious cardiovascular complication. This supports the safety and usefulness of angiography for clinical intervention trials.