Morphotype-specific calcium signaling in human microglia.

BACKGROUND: Key functions of Ca2+ signaling in rodent microglia include monitoring the brain state as well as the surrounding neuronal activity and sensing the danger or damage in their vicinity. Microglial Ca2+ dyshomeostasis is a disease hallmark in many mouse models of neurological disorders but the Ca2+ signal properties of human microglia remain unknown. METHODS: We developed a novel genetically-encoded ratiometric Ca2+ indicator, targeting microglial cells in the freshly resected human tissue, organotypically cultured tissue slices and analyzed in situ ongoing Ca2+ signaling of decades-old microglia dwelling in their native microenvironment. RESULTS: The data revealed marked compartmentalization of Ca2+ signals, with signal properties differing across the compartments and resident morphotypes. The basal Ca2+ levels were low in ramified and high in ameboid microglia. The fraction of cells with ongoing Ca2+ signaling, the fraction and the amplitude of process Ca2+ signals and the duration of somatic Ca2+ signals decreased when moving from ramified via hypertrophic to ameboid microglia. In contrast, the size of active compartments, the fraction and amplitude of somatic Ca2+ signals and the duration of process Ca2+ signals increased along this pathway.

Pulmono-atrial shunt and lung assist to treat right ventricular failure.

In right ventricular failure (RVF) a decrease of right ventricular afterload and improvement of left atrial filling could be achieved by a pulmonary artery-left atrial (PA)-shunt. To avoid cyanosis, artificial oxygenation is necessary. In 11 pigs a PA-shunt was created. An interventional lung assist device (ILA) was installed from the femoral artery to vein in 5 pigs (serial in relation to native lung: Group I) and into the PA-shunt in 6 pigs (parallel: Group II). RVF was induced by pulmonary artery banding. Right ventricular performance was determined by pulse contour analysis, pressure - and flow measurements. In both groups a stable RVF was generated. In Group I cardiac output trended to increase but neither right ventricular filling pressures nor arterial pressure changed significantly. The PaO2 decreased significantly. In Group II cardiac output and arterial pressure increased significantly under a shunt flow of 2.3- 2.6 l/min and the animals recovered from cardiogenic shock. In conclusion a PA-shunt with a parallel lung assist can effectively reverse the deleterious effects of RVF.

Video-assisted pericardioscopic surgery: refinement of a new technique for implanting epimyocardial pacemaker leads.

OBJECTIVE: Current alternative approaches for pacemaker lead implantation imply the breach of the pleural space. Recently, the feasibility of experimental lead implantation by rigid endoscopy has been described. The use of flexible endoscopes and a standardised application has not been realised yet. Our main goal was to compare rigid and flexible endoscopy and to establish a standardised protocol for the implementation of a closed-chest subxiphoid approach for epimyocardial lead implantation. METHODS: Rigid and flexible endoscopes were used for placement of screw-in pacing leads (4-F). A total of 17 adult pigs (80 kg) were anaesthetised and a 10-mm subxiphoid axial incision performed. The pericardium was opened and entered under endoscopic vision. Epimyocardial electrodes were implanted through the endoscope onto all four chambers. Standard haemodynamic measurements and pacing measurements were carried out. RESULTS: Both methods were deployed in the first three individuals. Superior endorsement of rigid endoscopy, due to better orientation and stability, led to its exclusive deployment in the remaining 14 individuals. Access to the implantation sites was quick (<10 min). A plastic cover had to be applied to reduce arrhythmia (VentricularExtraSystoles(bare): 17 ± 2.2 min(-1) vs VentricularExtraSystoles(cover): 5 ± 1.9 min(-1); n = 4). Measured pacing parameters were comparable with classic endocardial-derived thresholds. Post-mortem examination revealed no relevant damage/injury and/or bleeding in the heart and circumjacent tissue. There was no evidence of injury at the implantation sites and the corresponding pericardium. The electrodes showed excellent anchorage inside the myocardial tissue (penetration depths: 3 ± 0.2mm) and resisted high tractive forces. CONCLUSION: Flexible endoscopy is not suitable for exclusive deployment inside the pericardial space, whereas rigid endoscopy presented itself as a safe, fast and simple approach for epimyocardial lead implantation using an insulating trocar. Without cover, malignant arrhythmia constrains the implementation of video-assisted pericardioscopic surgery (VAPS). Subxiphoid VAPS permits optimal lead positioning under direct vision without fluoroscopy, without the breach of the pleural space and with a short procedural duration (<60 min). Our standardised minimal-invasive approach allows visualisation and intervention, potentially of all intrapericardial structures.