Study of adhesion and migration dynamics in ubiquitin E3A ligase (UBE3A)-silenced SYSH5Y neuroblastoma cells by micro-structured surfaces.

During neuronal development, neuronal cells read extracellular stimuli from the micro/nano-environment within which they exist, retrieving essential directionality and wiring information. Here, focal adhesions (FAs-protein clusters anchoring integrins to cytoskeleton) act as sensors, by integrating signals from both the extracellular matrix environment and chemotactic factors, contributing to the final neuronal pathfinding and migration. In the processes that orchestrate neuronal development, the important function of ubiquitin E3A ligase (UBE3A) is emerging. UBE3A has crucial functions in the brain and changes in its expression levels lead to neurodevelopmental disorders: the lack of UBE3A leads to Angelman syndrome (AS, OMIN 105830), while its increase causes autisms (Dup15q-autism). By using nano/micro-structured anisotropic substrates we previously showed that UBE3A-deficient neurons have deficits in contact guidance (Tonazzini et al, Mol Autism 2019). Here, we investigate the adhesion and migration dynamics of UBE3A-silenced SH-SY5Y neuroblastoma cells in vitro by exploiting nano/micro-grooved substrates. We analyze the molecular processes regulating the development of FAs by transfection with EGFP-vector encoding for paxillin, a protein of FA clusters, and by live-cell total-internal-reflection-fluorescence microscopy. We show that UBE3A-silenced SH-SY5Y cells have impaired FA morphological development and pathway activation, which lead to a delayed adhesion and also explain the defective contact guidance in response to directional topographical stimuli. However, UBE3A-silenced SH-SY5Y cells show an overall normal migration behavior, in terms of speed and ability to follow the GRs directional stimulus. Only the collective cell migration upon cell gaps was slightly delayed for UBE3Ash SHs. Overall, the deficits of UBE3Ash SHS-SY5Y cells in FA maturation/sensing and in collective migration may have patho-physiological implications, in AS condition, considering the much more complex stimuli that neurons find in vivo during the neurodevelopment.

Narrowing the Parameter Space of Collapse Models with Ultracold Layered Force Sensors.

Despite the unquestionable empirical success of quantum theory, witnessed by the recent uprising of quantum technologies, the debate on how to reconcile the theory with the macroscopic classical world is still open. Spontaneous collapse models are one of the few testable solutions so far proposed. In particular, the continuous spontaneous localization (CSL) model has become subject of intense experimental research. Experiments looking for the universal force noise predicted by CSL in ultrasensitive mechanical resonators have recently set the strongest unambiguous bounds on CSL. Further improving these experiments by direct reduction of mechanical noise is technically challenging. Here, we implement a recently proposed alternative strategy that aims at enhancing the CSL noise by exploiting a multilayer test mass attached on a high quality factor microcantilever. The test mass is specifically designed to enhance the effect of CSL noise at the characteristic length r_{c}=10^{-7} m. The measurements are in good agreement with pure thermal motion for temperatures down to 100 mK. From the absence of excess noise, we infer a new bound on the collapse rate at the characteristic length r_{c}=10^{-7} m, which improves over previous mechanical experiments by more than 1 order of magnitude. Our results explicitly challenge a well-motivated region of the CSL parameter space proposed by Adler.

Improved Noninterferometric Test of Collapse Models Using Ultracold Cantilevers.

Spontaneous collapse models predict that a weak force noise acts on any mechanical system, as a consequence of the collapse of the wave function. Significant upper limits on the collapse rate have been recently inferred from precision mechanical experiments, such as ultracold cantilevers and the space mission LISA Pathfinder. Here, we report new results from an experiment based on a high-Q cantilever cooled to millikelvin temperatures, which is potentially able to improve the current bounds on the continuous spontaneous localization (CSL) model by 1 order of magnitude. High accuracy measurements of the cantilever thermal fluctuations reveal a nonthermal force noise of unknown origin. This excess noise is compatible with the CSL heating predicted by Adler. Several physical mechanisms able to explain the observed noise have been ruled out.

Superconducting quantum interference device microsusceptometer balanced over a wide bandwidth for nuclear magnetic resonance applications.

Superconducting Quantum Interference Device (SQUID) microsusceptometers have been widely used to study magnetic properties of materials at microscale. As intrinsically balanced devices, they could also be exploited for direct SQUID-detection of nuclear magnetic resonance (NMR) from micron sized samples, or for SQUID readout of mechanically detected NMR from submicron sized samples. Here, we demonstrate a double balancing technique that enables achievement of very low residual imbalance of a SQUID microsusceptometer over a wide bandwidth. In particular, we can generate ac magnetic fields within the SQUID loop as large as 1 mT, for frequencies ranging from dc up to a few MHz. As an application, we demonstrate direct detection of NMR from (1)H spins in a glycerol droplet placed directly on top of the 20 µm SQUID loops.

Nonequilibrium steady-state fluctuations in actively cooled resonators.

We analyze heat and work fluctuations in the gravitational wave detector AURIGA, modeled as a macroscopic electromechanical oscillator in contact with a thermostat and cooled by an active feedback system. The oscillator is driven to a steady state by the feedback cooling, equivalent to a viscous force. The experimentally measured fluctuations are in agreement with our theoretical analysis based on a stochastically driven Langevin system. The asymmetry of the fluctuations of the absorbed heat characterizes the oscillator's nonequilibrium steady state and reveals the extent to which a feedback cooled system departs from equilibrium in a statistical mechanics perspective.

Feedback cooling of the normal modes of a massive electromechanical system to submillikelvin temperature.

We apply a feedback cooling technique to simultaneously cool the three electromechanical normal modes of the ton-scale resonant-bar gravitational wave detector AURIGA. The measuring system is based on a dc superconducting quantum interference device (SQUID) amplifier, and the feedback cooling is applied electronically to the input circuit of the SQUID. Starting from a bath temperature of 4.2 K, we achieve a minimum temperature of 0.17 mK for the coolest normal mode. The same technique, implemented in a dedicated experiment at subkelvin bath temperature and with a quantum limited SQUID, could allow to approach the quantum ground state of a kilogram-scale mechanical resonator.

Upper limits on gravitational-wave emission in association with the 27 Dec 2004 giant flare of SGR1806-20.

At the time when the giant flare of SGR1806-20 occurred, the AURIGA "bar" gravitational-wave (GW) detector was on the air with a noise performance close to stationary Gaussian. This allows us to set relevant upper limits, at a number of frequencies in the vicinities of 900 Hz, on the amplitude of the damped GW wave trains, which, according to current models, could have been emitted, due to the excitation of normal modes of the star associated with the peak in x-ray luminosity.

Caerulein in the treatment of biliary and renal colic.

A randomized controlled study has been carried out in order to check the activity of caerulein in the treatment of biliary and renal colic. In 88% out of 107 patients caerulein, 1 ng/kg IV, relieved biliary colic and had no side effects. To elucidate the mechanism of action of the peptide, 22 cholecystectomized patients, showing a dilation of the common bile duct (CBD), were submitted to caerulein treatment under echo-control. Reduction of CBD caliber was noticed when the dilatation was due to functional obstruction, whereas an organic obstruction of the terminal tract of CBD was found in the non-responding patients. Caerulein appears to be an effective agent in relieving biliary colic through a relaxation of Oddi's sphincter, and may be used in ultrasound differential diagnostics of terminal bile duct obstruction. Caerulein, 75 ng/kg intramuscularly, relieved renal colic in 75% of the examined patients; it is suggested that the effect of caerulein in this syndrome is due to central analgesic action.