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Astrocytes are glial cells that exhibit calcium signaling-mediated activity. Here, we present a protocol to monitor and manipulate astrocyte calcium activity from mouse amygdala slices. In the first part of this protocol, we describe the procedure of astrocyte calcium imaging. In the second part, we detail how to disrupt astrocyte calcium activity by patch-clamp-mediated loading of BAPTA. These two approaches are presented separately but they can also be used simultaneously to monitor the effects of disruption on an astrocyte network. For complete details on the use and execution of this protocol, please refer to Wahis et al. (2021).This protocol provides instructions to acquire high-quality cellular contractility data from adult, neonatal, and human induced pluripotent stem cell-derived cardiomyocytes. Contractility parameters are key to unravel mechanisms underlying cardiac pathologies, yet difficulties in acquiring data can compromise measurement accuracy and reproducibility. We provide optimized steps for microscope and camera setup, as well as cellular selection criteria for different cardiomyocyte cell types, aiming to obtain robust and reliable data. Moreover, we use CONTRACTIONWAVE software to analyze and show the optimized results. For complete details on the use and execution of this profile, please refer to Scalzo et al. (2021).We recently demonstrated how lipid droplets can serve as in situ fiducials for correlating cryo-fluorescence microscopy (cryo-FM) and cryo-focused ion beam scanning electron microscopy (cryo-FIB-SEM) datasets of mammalian cells grown on grids. Here we describe a step-by-step protocol for correlative cryo-FM and cryo-FIB-SEM, starting from sample preparation of C2C12 cell line, followed by imaging with cryo-FM and cryo-FIB-SEM. Finally, we detail how to perform the 3D-correlation with sub-micron accuracy. For complete details on the use and execution of this profile, please refer to Scher et al. (2021).We present a unique case of a paravalvular leak through a periannular channel around a bioprosthetic mitral valve. The role of multimodality imaging, in addition to novel technology, helped uncover the complex course of the jet, including its origin and direction, which translated into excellent procedural success. (Level of Difficulty Advanced.).There is very limited experience with simulated virtual implantation of left ventricular assist devices (LVADs) to assess device fitness in pediatric patients. In this clinical vignette, we report the case of a 9-year-old male patient with dilated cardiomyopathy who underwent successful placement of an LVAD after virtual simulated implantation was performed. (Level of Difficulty Advanced.).This case report concerns a 72-year-old-female with severe functional tricuspid stenosis due to phosphoglyceride crystal deposition disease and a history of atrial septum closure and tricuspid valvuloplasty. Phosphoglyceride crystal deposition disease is extremely rare, and percutaneous transcatheter biopsy under intracardiac echocardiographic guidance proved to be useful for its diagnosis. (Level of Difficulty Advanced.).Aortic dissection is very rare in pediatric patients, and associated risk factors include congenital heart disease, previous cardiac surgery, and vasculopathies. Acute postoperative aortic dissection in pediatric patients can be life-threatening. We performed a novel hybrid transcarotid covered stent exclusion of a postoperative ascending aortic dissection in an infant. (Level of Difficulty Advanced.).Transcatheter edge-to-edge valve repair can improve clinical outcomes in otherwise high-risk surgical patients. This is a first-in-human procedure outlining transcatheter edge-to-edge valve repair of a systemic tricuspid valve in an extracardiac Fontan patient born with hypoplastic left heart syndrome with prohibitive surgical risk. (Level of Difficulty Advanced.).A 38-year-old man with a diagnosis of Brugada syndrome had a recurrence of monomorphic ventricular tachycardia 6 years after successful epicardial right ventricular outflow tract ablation by the use of subxiphoid access. Preprocedural and intraprocedural investigations suggested that the pathologic substrate of the right ventricular outflow tract had not been eliminated completely. Therefore, the patient underwent a hybrid electrophysiology-guided video-assisted thoracoscopic ablation. (Level of Difficulty Advanced.).A 59-year-old man presented for implantable cardioverter-defibrillator placement after a wide QRS complex tachycardia cardiac arrest at an outside hospital. In this case report, we discuss the differential diagnosis of this patient's tachyarrhythmia and the electrophysiological studies that established the diagnosis and guided management. (Level of Difficulty Intermediate.).We describe the case of a 14-year-old boy hospitalized for multisystem inflammatory syndrome in children who developed atrial fibrillation during the acute phase and a transient Brugada type 1 pattern in the subacute phase. Eight months later, a provocative test with ajmaline confirmed the suspicion of Brugada syndrome. (Level of Difficulty Intermediate.).Hypertrophic cardiomyopathy (HCM) is a common but an underdiagnosed condition. Fasciculoventricular bypass tract (FVBT) is rare. Galunisertib in vitro Concomitant presence of both conditions is well described in Danon disease. We report a case of familial HCM with FVBT linked to a heterozygous pathogenic variant, c.655G>C (p.Val219Leu), in the cardiac myosin binding protein C3 (MYBPC3) gene. (Level of Difficulty Advanced.).Potential foci for atrial tachycardia have been previously described in various locations including crista terminalis, tricuspid annulus, coronary sinus ostium, pulmonary vein ostia. In this report, we present a case of a focal atrial tachycardia arising from the posterior wall of the left atrium which has not been described before. (Level of Difficulty Advanced.).Secondary prevention of sudden cardiac death in the young patient with arrhythmogenic right ventricular cardiomyopathy and hemodynamically tolerated ventricular tachycardia is still a challenging field. We present a combined approach, including subcutaneous implantable cardioverter-defibrillator (ICD) and catheter ablation, as a promising treatment to prevent both ventricular tachycardia recurrences and ICD shocks. (Level of Difficulty Intermediate.).An 82-year-old woman with nonischemic cardiomyopathy underwent cardiac resynchronization therapy by the use of His bundle pacing. After the procedure, the patient had repolarization abnormality with severely prolonged QTc and anterior inferior T-wave inversions, likely resulting from memory T waves associated with the correction of long-standing left bundle branch block. These changes could be potentially arrhythmogenic. (Level of Difficulty Intermediate.).In this report, we illustrate the different flow-gradient patterns of aortic stenosis associated with discordant grading of stenosis severity at transthoracic echocardiography (TTE). The discordance among TTE parameters (mean gradient and aortic area) can be reconciled and true severity can be confirmed by ruling out potential measurements errors and by using multimodality imaging. (Level of Difficulty Advanced.).A man with recurrent syncope and remote aortic coarctation repair experienced cardiac arrest with exercise stress testing. Critical coronary stenosis was discovered. Further evaluation revealed accessory mitral valve tissue and internal mammary artery occlusion. These rare abnormalities, not previously reported together, presented challenges to treatment. (Level of Difficulty Intermediate.).We present the case of a young woman with systemic sclerosis (SSc) and refractory digital ulceration who was successfully treated with percutaneous revascularization of chronically occluded ulnar and radial arteries. To our knowledge, this is the first detailed report of limb salvage in SSc-induced hand ischemia in which contemporary endovascular techniques were used. (Level of Difficulty Advanced.).Nondominant right coronary artery (NDRCA) occlusion is rare and generally affects a small volume of myocardium. Despite this, NDRCA occlusion can result in dramatic clinical sequelae. These cases demonstrate the characteristic electrocardiographic findings and consequences of NDRCA occlusion, highlighting the importance of recognition of this pathologic condition to institute appropriate management. (Level of Difficulty Intermediate.).A 56-year-old man presented to the emergency department with chest pain. The diagnosis of acute myocardial infarction caused by a left circumflex artery occlusion was made. After conservative treatment, a fistula between the circumflex artery and the left ventricle, and the evolution of the pseudoaneurysm, were noted. (Level of Difficulty Advanced.).Pulmonary arteriovenous malformations (PAVMs) may manifest with bleeding or embolic events necessitating intervention. Transcatheter coil embolization through the pulmonary artery (PA) is an established approach. We present a case of recurrent PAVMs despite numerous PA coil embolizations. PAVM occlusion was achieved through plug placement by a transseptal and pulmonary venous approach. (Level of Difficulty Advanced.).Compression of mediastinal structures by vertebral osteophytes is rare. We report a case of pulmonary vein compression by a vertebral osteophyte that failed stenting. A minimally invasive approach to osteophyte removal with subsequent re-expansion angioplasty yielded an optimal outcome, negating the need for cardiopulmonary bypass, stent removal, and pulmonary venoplasty. (Level of Difficulty Intermediate.).We report the case of a 49-year-old female patient who underwent percutaneous coronary intervention of the right coronary and posterior descending arteries complicated with guidewire-induced coronary artery perforation. We successfully managed and sealed this perforation through the embolization of balloon pieces into the target vessel. (Level of Difficulty Advanced.).Coronary artery perforation is a rare but serious complication during percutaneous coronary intervention. Distal or small vessel perforation is usually treated by coil, fat, or microsphere embolization. We describe 5 cases of distal coronary perforation that were managed successfully by a novel technique that uses absorbable sutures. (Level of Difficulty Advanced.).Muscle bundles in the right atrium are an extremely rare congenital anomaly. We report the case of a patient with 2 atrial septal defects and a large muscle bundle crossing the right atrium. Only 3 comparable cases have previously been published. (Level of Difficulty Intermediate.).Acute cardiorespiratory distress following device closure of ruptured sinus of Valsava (RSOV) aneurysm can have a battery of differentials. We report a case of acute left ventricular dysfunction with aspiration that caused cardiorespiratory distress immediately following RSOV device closure. Supportive medical therapy led to complete recovery. (Level of Difficulty Intermediate.).The 2021 ACC/AHA/SCAI coronary artery disease revascularization guideline highlights the importance of the multidisciplinary heart team in making patient-centered, evidence-based clinical decisions for patients considered for coronary revascularization. We present 2 cases highlighting aspects of heart team decision making for complex patients with coronary artery disease. (Level of Difficulty Intermediate.).
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