ALS-associated FUS mutation reshapes the RNA and protein composition of stress granules.

Stress granules (SG) are part of a cellular protection mechanism where untranslated messenger RNAs and RNA-binding proteins are stored upon conditions of cellular stress. Compositional variations due to qualitative or quantitative protein changes can disrupt their functionality and alter their structure. This is the case of different forms of amyotrophic lateral sclerosis (ALS) where a causative link has been proposed between the cytoplasmic de-localization of mutant proteins, such as FUS (Fused in Sarcoma), and the formation of cytotoxic inclusions.

4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector.

Single-particle tracking techniques enable investigation of the complex functions and interactions of individual particles in biological environments. Many such techniques exist, each demonstrating trade-offs between spatiotemporal resolution, spatial and temporal range, technical complexity, and information content. To mitigate these trade-offs, we enhanced a confocal laser scanning microscope with an asynchronous read-out single-photon avalanche diode array detector.

MA reduction relieves FUS-associated ALS granules.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease due to gradual motoneurons (MN) degeneration. Among the processes associated to ALS pathogenesis, there is the formation of cytoplasmic inclusions produced by aggregation of mutant proteins, among which the RNA binding protein FUS. Here we show that, in neuronal cells and in iPSC-derived MN expressing mutant FUS, such inclusions are significantly reduced in number and dissolve faster when the RNA mA content is diminished.

Single-photon microscopy to study biomolecular condensates.

Biomolecular condensates serve as membrane-less compartments within cells, concentrating proteins and nucleic acids to facilitate precise spatial and temporal orchestration of various biological processes. The diversity of these processes and the substantial variability in condensate characteristics present a formidable challenge for quantifying their molecular dynamics, surpassing the capabilities of conventional microscopy. Here, we show that our single-photon microscope provides a comprehensive live-cell spectroscopy and imaging framework for investigating biomolecular condensation.

Orientational Melting in a Mesoscopic System of Charged Particles.

A mesoscopic system of a few particles can undergo changes of configuration that resemble phase transitions but with a nonuniversal behavior. A notable example is orientational melting, in which localized particles with long-range repulsive interactions forming a two-dimensional crystal become delocalized in common closed trajectories. Here we report the observation of orientational melting occurring in a two-dimensional crystal of up to 15 ions.

Incidence and Long-term Functional Outcome of Neurologic Disorders in Hospitalized Patients With COVID-19 Infected With Pre-Omicron Variants.

Background And Objectives: A variety of neurologic disorders have been reported as presentations or complications of coronavirus disease 2019 (COVID-19) infection. The objective of this study was to determine their incidence dynamics and long-term functional outcome.

Methods: The Neuro-COVID Italy study was a multicenter, observational, cohort study with ambispective recruitment and prospective follow-up.

The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy.

Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access to an entirely new set of single-photon information. Together with the blooming of new SP LSM techniques and the development of tailored SP array detectors, there is a growing need for (i) DAQ systems capable of handling the high-throughput and high-resolution photon information generated by these detectors, and (ii) incorporating these DAQ protocols in existing fluorescence LSMs. We developed an open-source, low-cost, multi-channel time-tagging module (TTM) based on a field-programmable gate array that can tag in parallel multiple single-photon events, with 30 ps precision, and multiple synchronisation events, with 4 ns precision.

Cooled SPAD array detector for low light-dose fluorescence laser scanning microscopy.

The single-photon timing and sensitivity performance and the imaging ability of asynchronous-readout single-photon avalanche diode (SPAD) array detectors have opened up enormous perspectives in fluorescence (lifetime) laser scanning microscopy (FLSM), such as super-resolution image scanning microscopy and high-information content fluorescence fluctuation spectroscopy. However, the strengths of these FLSM techniques depend on the many different characteristics of the detector, such as dark noise, photon-detection efficiency, after-pulsing probability, and optical cross talk, whose overall optimization is typically a trade-off between these characteristics. To mitigate this trade-off, we present, to our knowledge, a novel SPAD array detector with an active cooling system that substantially reduces the dark noise without significantly deteriorating any other detector characteristics.

Design of a Littrow-type diode laser with independent control of cavity length and grating rotation.

We present a novel, to the best of our knowledge, extended-cavity diode laser based on a modified Littrow configuration. The coarse wavelength adjustment via the rotation of a diffraction grating is decoupled from the fine tuning of the external cavity modes by positioning a piezo transducer behind the diode laser, making the laser robust against misalignment and hysteresis even with long external cavities. Two laser prototypes with external cavities of different lengths were tested with a 780 nm laser diode, and locked to an atomic reference.

Exploring early time points of vimentin assembly in flow by fluorescence fluctuation spectroscopy.

Despite the importance for cellular processes, the dynamics of molecular assembly, especially on fast time scales, is not yet fully understood. To this end, we present a multi-layer microfluidic device and combine it with fluorescence fluctuation spectroscopy. We apply this innovative combination of methods to investigate the early steps in assembly of vimentin intermediate filaments (IFs).

A minimalist model to measure interactions between proteins and synaptic vesicles.

Protein dynamics in the synaptic bouton are still not well understood, despite many quantitative studies of synaptic structure and function. The complexity of the synaptic environment makes investigations of presynaptic protein mobility challenging. Here, we present an in vitro approach to create a minimalist model of the synaptic environment by patterning synaptic vesicles (SVs) on glass coverslips.

How and why patients made Long Covid.

Patients collectively made Long Covid - and cognate term 'Long-haul Covid' - in the first months of the pandemic. Patients, many with initially 'mild' illness, used various kinds of evidence and advocacy to demonstrate a longer, more complex course of illness than laid out in initial reports from Wuhan. Long Covid has a strong claim to be the first illness created through patients finding one another on Twitter: it moved from patients, through various media, to formal clinical and policy channels in just a few months.

A comparative analysis of the mobility of 45 proteins in the synaptic bouton.

Many proteins involved in synaptic transmission are well known, and their features, as their abundance or spatial distribution, have been analyzed in systematic studies. This has not been the case, however, for their mobility. To solve this, we analyzed the motion of 45 GFP-tagged synaptic proteins expressed in cultured hippocampal neurons, using fluorescence recovery after photobleaching, particle tracking, and modeling.

Emergency treatment in Lombardy: a new methodology for the pre-Hospital Drugs management on Advanced Rescue Vehicles.

Background And Aim Of The Work: The main objectives of our work were the regional harmonization and standardization of pharmaceutical supplies on MSA in Lombardy.

Methods: The retrospective investigation was articulated in 2 phases: the first was the collection of data in every area of the Region (2012), the second was the analysis and elaboration of the information retrieved.

Results: Beginning with 24 common drugs used by 8 AATs out of 12 an evaluation of the chemical-therapeutic characteristics was performed.

A compact radiofrequency drive based on interdependent resonant circuits for precise control of ion traps.

Paul traps are widely used to confine electrically charged particles like atomic and molecular ions by using an intense radiofrequency (RF) field, typically obtained by a voltage drop on capacitative electrodes placed in vacuum. We present a RF drive realized on a compact printed circuit board and providing a high-voltage RF signal to a quadrupole Paul trap. The circuit is formed by using four interdependent resonant circuits - each of which is connected to an electrode of a Paul trap - fed by low-noise amplifiers, leading to an output voltage of peak-to-peak amplitude up to 200 V at 3.

Human blood platelets contract in perpendicular direction to shear flow.

In their physiological environment, blood platelets are permanently exposed to shear forces caused by blood flow. Within this surrounding, they generate contractile forces that eventually lead to a compaction of the blood clot. Here, we present a microfluidic chamber that combines hydrogel-based traction force microscopy with a controlled shear environment, and investigate the force fields platelets generate when exposed to shear flow in a spatio-temporally resolved manner.

A scalable hardware and software control apparatus for experiments with hybrid quantum systems.

Modern experiments with fundamental quantum systems - like ultracold atoms, trapped ions, and single photons - are managed by a control system formed by a number of input/output electronic channels governed by a computer. In hybrid quantum systems, where two or more quantum systems are combined and made to interact, establishing an efficient control system is particularly challenging due to the higher complexity, especially when each single quantum system is characterized by a different time scale. Here we present a new control apparatus specifically designed to efficiently manage hybrid quantum systems.

Cyclic olefin copolymer as an X-ray compatible material for microfluidic devices.

The combination of microfluidics and X-ray methods attracts a lot of attention from researchers as it brings together the high controllability of microfluidic sample environments and the small length scales probed by X-rays. In particular, the fields of biophysics and biology have benefited enormously from such approaches. We introduce a straightforward fabrication method for X-ray compatible microfluidic devices made solely from cyclic olefin copolymers.

Validation of 1-year predictive score of long-term response to interferon-β in everyday clinical practice multiple sclerosis patients.

Background And Purpose: The Rio score (RS) and the modified Rio score (MRS) are two scoring systems that can identify the early predictive factors of disability progression in relapsing-remitting multiple sclerosis (RRMS) patients treated with interferon-β (IFN-β). The objective of the study was to validate the usefulness of the RS and MRS in a large cohort of multiple sclerosis patients treated with IFN-β in daily clinical practice.

Methods: The analysis included a cohort of RRMS patients treated with different formulations of IFN-β for at least 1 year.

Deep grey matter T2 hypo-intensity in patients with paediatric multiple sclerosis.

Objective: T2 hypo-intensity on magnetic resonance imaging scans is thought to reflect pathological iron deposition in the presence of disease. In this pilot study, we evaluated the utility of the quantification of T2 hypo-intensities in paediatric patients by estimating deep grey matter (DGM) T2 hypo-intensities in paediatric patients with multiple sclerosis (MS) or clinically isolated syndromes (CIS), and their changes over 1 year.

Methods: A dual-echo sequence was obtained from 45 paediatric patients (10 with CIS, 35 with relapsing-remitting MS, 8 with an onset of the disease before the age of 10 and 37 during adolescence) and 14 age-matched healthy controls (HC).

Intercenter differences in diffusion tensor MRI acquisition.

Purpose: To assess the effect on diffusion tensor (DT) magnetic resonance imaging (MRI) of acquiring data with different scanners.

Materials And Methods: Forty-four healthy controls and 36 multiple sclerosis patients with low disability were studied using eight MR scanners with acquisition protocols that were as close to a standard protocol as possible. Between 7 and 13 subjects were studied in each center.

Hepatic resection using a bipolar vessel sealing device: technical and histological analysis.

Introduction: Blood loss and bile leakage are well-known risk factors for morbidity and mortality during liver resection. Bleeding usually occurs during parenchymal transection, and surgical technique should be considered an important factor in preventing intraoperative and postoperative complications.

Objective: Many approaches and devices have been developed to limit bleeding and bile leakage.

Prospective multicenter quality assessment of endotherapy of biliary stones: does center volume matter?

Background And Study Aims: To study the effectiveness of endoscopic treatment for biliary stones in a large case list of patients treated in units with different experience and different workloads in a region of northern Italy.

Patients And Methods: We prospectively studied 700 patients undergoing endoscopic retrograde cholangiopancreatography or sphincterotomy, in 14 units (> or < 200 examinations/year), for their first treatment of biliary stones. The difficulty of the examinations, the results in terms of clearance of the stones, and the late outcomes (24 months) were recorded.