16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro.

Reciprocal copy number variations (CNVs) of 16p11.2 are associated with a wide spectrum of neuropsychiatric and neurodevelopmental disorders. Here, we use human induced pluripotent stem cells (iPSCs)-derived dopaminergic (DA) neurons carrying CNVs of 16p11.2 duplication (16pdup) and 16p11.2 deletion (16pdel), engineered using CRISPR-Cas9. We show that 16pdel iPSC-derived DA neurons have increased soma size and synaptic marker expression compared to isogenic control lines, while 16pdup iPSC-derived DA neurons show deficits in neuronal differentiation and reduced synaptic marker expression. The 16pdel iPSC-derived DA neurons have impaired neurophysiological properties. The 16pdel iPSC-derived DA neuronal networks are hyperactive and have increased bursting in culture compared to controls. We also show that the expression of RHOA is increased in the 16pdel iPSC-derived DA neurons and that treatment with a specific RHOA-inhibitor, Rhosin, rescues the network activity of the 16pdel iPSC-derived DA neurons. Our data suggest that 16p11.2 deletion-associated iPSC-derived DA neuron hyperactivation can be rescued by RHOA inhibition.

Individualized mechanical ventilation in a shared ventilator setting: limits, safety and technical details.

The COVID-19 pandemic has resulted in an increased need for ventilators. The potential to ventilate more than one patient with a single ventilator, a so-called split ventilator setup, provides an emergency solution. Our hypothesis is that ventilation can be individualized by adding a flow restrictor to limit tidal volumes, add PEEP, titrate FiO2 and monitor ventilation. This way we could enhance optimization of patient safety and clinical applicability. We performed bench testing to test our hypothesis and identify limitations. We performed a bench testing in two test lungs: (1) determine lung compliance (2) determine volume, plateau pressure and PEEP, (3) illustrate individualization of airway pressures and tidal volume with a flow restrictor, (4a) illustrate that PEEP can be applied and individualized (4b) create and measure intrinsic PEEP (4c and d) determine PEEP as a function of flow restriction, (5) individualization of FiO2. The lung compliance varied between 13 and 27 mL/cmH2O. Set ventilator settings could be applied and measured. Extrinsic PEEP can be applied except for settings with a large expiratory time. Volume and pressure regulation is possible between 70 and 39% flow restrictor valve closure. Flow restriction in the tested circuit had no effect on the other circuit or on intrinsic PEEP. FiO2 could be modulated individually between 0.21 and 0.8 by gradually adjusting the additional flow, and minimal affecting FiO2 in the other circuit. Tidal volumes, PEEP and FiO2 can be individualized and monitored in a bench testing of a split ventilator. In vivo research is needed to further explore the clinical limitations and outcomes, making implementation possible as a last resort ventilation strategy.

A novel inline PEEP valve design for differential multi-ventilation.

BACKGROUND: Ventilator sharing is one option to emergently increase ventilator capacity during a crisis but has been criticized for its inability to adjust for individual patient needs. Newer ventilator sharing designs use valves and restrictors to control pressures for each patient. A key component of these designs is an inline Positive End Expiratory Pressure (PEEP) Valve but these are not readily available. Creating an inline PEEP valve by converting a standard bag-valve-mask PEEP valve is possible with the addition of a 3D printed collar. METHODS: This was a feasibility study assessing the performance and safety of a method for converting a standard PEEP valve into an inline PEEP valve. A collar was designed and printed that covers the exhaust ports of the valve and returns exhaled gases to the ventilator. RESULTS: The collar piece was simple to print and easily assembled with the standard PEEP valve. In bench testing it successfully created differential pressures in 2 simulated expiratory limbs without leaking to the atmosphere at pressures greater than 60 cm of H2O. CONCLUSION: Our novel inline PEEP valve design shows promise as an option for building a safer ventilator sharing system.