Knockout mice are used extensively to explore the phenotypic effects of mammalian gene dysfunction. With the application of RNA-guided Cas9 nuclease technology for the production of knockout mouse lines, the time, as well as the resources needed, to progress from identification of a gene of interest to production of a knockout line is significantly reduced. Here we present our standard methodology to produce knockout mouse lines by the electroporation of Cas9 ribonucleoprotein (RNP) into mouse zygotes. Using this protocol, we have obtained an 80% success rate in the generation of founders for null alleles with a subsequent 93% germline transmission rate. These methods rely on equipment already present in the majority of transgenic facilities and should be straightforward to implement where appropriate embryo handling expertise exists.The synaptogenic hypothesis of major depressive disorder implies that preventing the onset of depressive-like behavior also prevents the loss of hippocampal spine synapses. By applying the psychoactive drugs, diazepam and fluoxetine, we investigated whether blocking the development of helpless behavior by promoting stress resilience in the rat learned helplessness paradigm is associated with a synaptoprotective action in the hippocampus. Adult ovariectomized and intact female Sprague-Dawley rats (n = 297) were treated with either diazepam, fluoxetine, or vehicle, exposed to inescapable footshocks or sham stress, and tested in an active escape task to assess helpless behavior. Escape-evoked corticosterone secretion, as well as remodeling of hippocampal spine synapses at a timepoint representing the onset of escape testing were also analyzed. In ovariectomized females, treatment with diazepam prior to stress exposure prevented helpless behavior, blocked the loss of hippocampal spine synapses, and muted the corticosterone surge evoked by escape testing. Although fluoxetine stimulated escape performance and hippocampal synaptogenesis under non-stressed conditions, almost all responses to fluoxetine were abolished following exposure to inescapable stress. Only a much higher dose of fluoxetine was capable of partly reproducing the strong protective actions of diazepam. Importantly, these protective actions were retained in the presence of ovarian hormones. Our findings indicate that stress resilience is associated with the preservation of spine synapses in the hippocampus, raising the possibility that, besides synaptogenesis, hippocampal synaptoprotection is also implicated in antidepressant therapy. The mechanisms of improvement of left ventricular (LV) function with cardiac resynchronization therapy (CRT) are not yet elucidated. The aim of this study was to characterize CRT responder profiles through clustering analysis, on the basis of clinical and echocardiographic preimplantation data, integrating automatic quantification of longitudinal strain signals. This was a multicenter observational study of 250 patients with chronic heart failure evaluated before CRT device implantation and followed up to 4years. Clinical, electrocardiographic, and echocardiographic data were collected. Regional longitudinal strain signals were also analyzed with custom-made algorithms in addition to existing approaches, including myocardial work indices. Response was defined as a decrease of ≥15% in LV end-systolic volume. Death and hospitalization for heart failure at 4years were considered adverse events. Seventy features were analyzed using a clustering approach (k-means clustering). Five clusters were identified, wT. Automatic quantitative analysis of longitudinal strain curves appears to be a promising tool to improve the understanding of LV mechanics, patient characterization, and selection for CRT. Iatrogenic mitral stenosis is a known limitation of transcatheter edge-to-edge mitral valve repair (TMVr), but determinants of increased postprocedural mean diastolic gradient (MG) are not well defined. The aim of this study was to determine correlates of increased post-TMVr MG or aborted clip implantation due to increased MG. Procedural three-dimensional transesophageal echocardiographic (TEE) data sets of 112 patients who underwent TMVr were retrospectively analyzed. Three-dimensional TEE mitral valve area (MVA) planimetry and mitral annular calcification (MAC) were quantified using multiplanar reconstruction. When MAC extension into the mitral leaflets was present, MAC with leaflet calcification (MAC-LC) length was recorded as the maximum distance from the mitral annulus to the most distal leaflet calcification. Increased MG after TMVr, measured on intraprocedural TEE imaging, was defined as ≥5mm Hg or aborted clip implantation due to increased MG. Baseline MVA was 5.9±1.7cm , baseline MG was 2.1±1.ine MVA (4.0-6.0 cm2) and concurrent MAC-LC length ≥ 6 mm or baseline MG ≥ 4 mm Hg are at moderate risk for increased MG after TMVr. The aim of this study was to present new combination of algorithms for rhythm analysis during cardiopulmonary resuscitation (CPR) in automated external defibrillators (AED), called Analyze Whilst Compressing (AWC), designed for decreasing pre-shock pause and early stopping of chest compressions (CC) for treating refibrillation. Two stages for AED rhythm analysis were presented, namely, "Standard Analysis Stage" (conventional shock-advisory analysis run over 5 s after CC interruption every two minutes) and "AWC Stage" (two-step sequential analysis process during CPR). FK866 AWC steps were run in presence of CC (Step1), and if shockable rhythm was detected then a reconfirmation step was run in absence of CC (Step2, analysis duration 5 s). In total 16,057 ECG strips from 2916 out-of-hospital cardiac arrest (OHCA) patients treated with AEDs (DEFIGARD TOUCH7, Schiller Médical, France) were subjected patient-wise to AWC training (8559 strips, 1604 patients) and validation (7498 strips, 1312 patients). Considering v not interrupting chest compressions for prevalent part of non-shockable rhythms (66-83%); (ii) minimizing pre-shock pause for 92.1% of VF patients. AWC required hands-off reconfirmation in 34.4% of cases. Reconfirmation was also common limitation of other reported algorithms (25.7-100%) although following different protocols for triggering chest compression resumption and shock delivery.