DARTS: an open-source Python pipeline for Ca2+ microdomain analysis in live cell imaging data.

Ca2+ microdomains play a key role in intracellular signaling processes. For instance, they mediate the activation of T cells and, thus, the initial adaptive immune system. They are, however, also of utmost importance for activation of other cells, and a detailed understanding of the dynamics of these spatially localized Ca2+ signals is crucial for a better understanding of the underlying signaling processes. A typical approach to analyze Ca2+ microdomain dynamics is live cell fluorescence microscopy imaging. Experiments usually involve imaging a larger number of cells of different groups (for instance, wild type and knockout cells), followed by a time consuming image and data analysis. With DARTS, we present a modular Python pipeline for efficient Ca2+ microdomain analysis in live cell imaging data. DARTS (Deconvolution, Analysis, Registration, Tracking, and Shape normalization) provides state-of-the-art image postprocessing options like deep learning-based cell detection and tracking, spatio-temporal image deconvolution, and bleaching correction. An integrated automated Ca2+ microdomain detection offers direct access to global statistics like the number of microdomains for cell groups, corresponding signal intensity levels, and the temporal evolution of the measures. With a focus on bead stimulation experiments, DARTS provides a so-called dartboard projection analysis and visualization approach. A dartboard projection covers spatio-temporal normalization of the bead contact areas and cell shape normalization onto a circular template that enables aggregation of the spatiotemporal information of the microdomain detection results for the individual cells of the cell groups of interest. The dartboard visualization allows intuitive interpretation of the spatio-temporal microdomain dynamics at the group level. The application of DARTS is illustrated by three use cases in the context of the formation of initial Ca2+ microdomains after cell stimulation. DARTS is provided as an open-source solution and will be continuously extended upon the feedback of the community. Code available at: 10.5281/zenodo.10459243.

Pulse amplitude adjustment provides immediate pacemaker longevity gain.

OBJECTIVE: Adjusting pacemaker pulse amplitude influences the longevity of the pacemaker. Our aim was to establish the initial longevity gain. METHODS: Forty randomly selected patients with implanted pacemakers were analyzed. Mean age was 65.58+/-13.7 years. All pacemakers were working on factory settings of pulse amplitude 3.5 V and pulse width of 0.4 ms for average of 3 years before the adjustment. Initial mean longevity was projected to 68.61+/-18.86 months, mean battery voltage 2.78 V, and mean battery current 14.21+/-2.61 microA. RESULTS: Pulse amplitude threshold test was performed and average value of 0.632+/-0.22 V was obtained. Pulse amplitude was programmed to 2.5 V and pulse width was left unchanged. New readings of battery data were obtained. Battery voltage did not show immediate changes, and battery current decreased to 11.53+/-1.98 microA. New average longevity was projected to 81.03+/-19.82 months, which presents a 12.42 months of initial longevity gain with statistical significance at 95% confidence interval (p=0.003). Positive correlation was found between the new pulse amplitude and new values of battery current (p<0.01). CONCLUSION: Pulse amplitude decrease of only 1 V provides significant initial longevity gain of more than a year. If found correlations would have any impact on further longevity gains over longer period of time is yet to be established.

Induction of atrioventricular node reentry by simultaneous anterograde conduction over the fast and slow pathways.

Atrio-ventricular node reentry (AVNRT) is typically induced with an anterograde block over the fast pathway (FP) and conduction over the slow pathway (SP), with subsequent retrograde conduction over the FP. Rarely, a premature atrial complex (PAC) conducts simultaneously over the FP and SP to induce AVNRT. Previous publications have reported that conduction over the fast and slow pathway of the atrioventricular node can occur successively one after the other, thus leading to dual ventricular depolarization from what initially was a single atrial impulse. We report a case of an 18-year-old male patient referred for repeated bursts of ectopic activity. Evaluation of the patient's electrocardiographic recordings suggested the presence of dual ventricular activations for each atrial beat. The electrophysiological study revealed that the patient had simultaneous conduction over the fast and slow pathways of the atrioventricular node giving rise to a non-reentrant tachycardia, along with an absence of retrograde (ventriculoatrial) conduction, and a significant atrio-His bundle jump (A-H jump) through the slow pathway from the fast pathway during programmed electrical stimulation from the right atrium. Ablation of the slow pathway at the base of the Koch triangle yielded a cessation of the dual ventricular response, absence of the nonreentrant tachycardia and no A-H jump.