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Graphical abstract: Rationale underlying novel drug testing platform development. Pain detected by the central nervous system and following bone fracture cannot be treated or exclusively alleviated using standardised methods. The pain mechanism and specificity/efficacy of pain reduction drugs remain poorly understood. In vivo and ex vivo models are not yet able to recapitulate the various pain events associated with skeletal damage. In vitro models are currently limited by their inability to fully mimic the complex physiological mechanisms at play between nervous and skeletal tissue and any disruption in pathological states. Robust innovative tissue engineering models are needed to better understand pain events and to investigate therapeutic regimes

The genetic elements required to tune gene expression are partitioned in active and repressive nuclear condensates. Chromatin compartments include transcriptional clusters whose dynamic establishment and functioning depend on multivalent interactions occurring among transcription factors, cofactors and basal transcriptional machinery. However, how chromatin players contribute to the assembly of transcriptional condensates is poorly understood. By interrogating the effect of KMT2D(also known as MLL4) haploinsufficiency in Kabuki syndrome, we found that mixed lineage leukemia 4 (MLL4) contributes to the assembly of transcriptional condensates through liquid–liquid phase separation. MLL4 loss of function impaired Polycomb-dependent chromatin compartmentalization, altering the nuclear architecture. By releasing the nuclear mechanical stress through inhibition of the mechanosensor ATR, we re-established the mechanosignaling of mesenchymal stem cells and their commitment towards chondrocytes both in vitro and in vivo. This study supports the notion that, in Kabuki syndrome, the haploinsufficiency of MLL4 causes an altered functional partitioning of chromatin, which determines the architecture and mechanical properties of the nucleus.

It has been recently demonstrated that the exposure of naive neuronal cells to light—at the basis of optogenetic techniques and calcium imaging measurements—may alter neuronal firing. Indeed, understanding the effect of light on nongenetically modified neurons is crucial for a correct interpretation of calcium imaging and optogenetic experiments. Here we investigated the effect of continuous visible LED light exposure (490 nm, $ 0.18 {-} 1.3\;{\rm mW}/{{\rm mm}^2} $0.18−1.3mW/mm^2) on spontaneous activity of primary neuronal networks derived from the early postnatal mouse cortex. We demonstrated, by calcium imaging and patch clamp experiments, that illumination higher than $ 1.0\;{\rm mW}/{{\rm mm}^2} $1.0mW/mm^2 causes an enhancement of network activity in cortical cultures. We investigated the possible origin of the phenomena by blocking the transient receptor potential vanilloid 4 (TRPV4) channel, demonstrating a complex connection between this temperature-dependent channel and the measured effect. The results presented here shed light on an exogenous artifact, potentially present in all calcium imaging experiments, that should be taken into account in the analysis of fluorescence data.

The outcome of an epidemic is closely related to the network of interactions between individuals. Likewise, protein functions depend on the 3D arrangement of their residues and the underlying energetic interaction network. Borrowing ideas from the theoretical framework that has been developed to address the spreading of real diseases, we study for the first time the diffusion of a fictitious epidemic inside the protein nonbonded interaction network, aiming to study network features and properties. Our approach allows us to probe the overall stability and the capability of propagating information in complex 3D structures, proving to be very efficient in addressing different problems, from the assessment of thermal stability to the identification of functional sites.

It is not well understood why severe acute respiratory syndrome (SARS)-CoV-2 spreads much faster than other β-coronaviruses such as SARS-CoV and Middle East respiratory syndrome (MERS)-CoV. In a previous publication, we predicted the binding of the N-terminal domain (NTD) of SARS-CoV-2 spike to sialic acids (SAs). Here, we experimentally validate this interaction and present simulations that reveal a second possible interaction between SAs and the spike protein via a binding site located in the receptor-binding domain (RBD). The predictions from molecular-dynamics simulations and the previously-published 2D-Zernike binding-site recognition approach were validated through flow-induced dispersion analysis (FIDA)─which reveals the capability of the SARS-CoV-2 spike to bind to SA-containing (glyco)lipid vesicles, and flow-cytometry measurements─which show that spike binding is strongly decreased upon inhibition of SA expression on the membranes of angiotensin converting enzyme-2 (ACE2)-expressing HEK cells. Our analyses reveal that the SA binding of the NTD and RBD strongly enhances the infection-inducing ACE2 binding. Altogether, our work provides in silico, in vitro, and cellular evidence that the SARS-CoV-2 virus utilizes a two-receptor (SA and ACE2) strategy. This allows the SARS-CoV-2 spike to use SA moieties on the cell membrane as a binding anchor, which increases the residence time of the virus on the cell surface and aids in the binding of the main receptor, ACE2, via 2D diffusion.

The role and importance of mechanical properties of cells and tissues in cellular function, development and disease has widely been acknowledged, however standard techniques currently used to assess them exhibit intrinsic limitations. Recently, Brillouin microscopy, a type of optical elastography, has emerged as a non-destructive, label- and contact-free method that can probe the viscoelastic properties of biological samples with diffraction-limited resolution in 3D. This led to increased attention amongst the biological and medical research communities, but it also sparked debates about the interpretation and relevance of the measured physical quantities. Here, we review this emerging technology by describing the underlying biophysical principles and discussing the interpretation of Brillouin spectra arising from heterogeneous biological matter. We further elaborate on the technique’s limitations, as well as its potential for gaining insights in biology, in order to guide interested researchers from various fields.

Cells sense and respond to external physical forces and substrate rigidity by regulating their cell shape, internal cytoskeletal tension, and stiffness. Here we show that the combination of micropillar traction force and noncontact Brillouin microscopy provides access to cell-generated forces and intracellular mechanical properties at optical resolution. Actin-rich cytoplasmic domains of 3T3 fibroblasts showed significantly higher Brillouin shifts, indicating a potential increase in stiffness when adhering on fibronectin-coated glass compared to soft PDMS micropillars. Our findings demonstrate the complementarity of micropillar traction force and Brillouin microscopy to better understand the relation between cell force generation and the intracellular mechanical properties.

The experimentally measured vibrational spectrum of glasses strongly deviates from that expected in Debye's elasticity theory: The density of states deviates from Debye's ω2law (“boson peak”), the sound velocity shows a negative dispersion in the boson-peak frequency regime, and there is a strong increase in the sound attenuation near the boson-peak frequency. A generalized elasticity theory is presented, based on the model assumption that the shear modulus of the disordered medium fluctuates randomly in space. The fluctuations are assumed to be uncorrelated and have a certain distribution (Gaussian or otherwise). Using field-theoretical techniques one is able to derive mean-field theories for the vibrational spectrum of a disordered system. The theory based on a Gaussian distribution uses a self-consistent Born approximation (SCBA),while the theory for non-Gaussian distributions is based on a coherent-potential approximation (CPA). Both approximate theories appear to be saddle-point approximations of effective replica field theories. The theory gives a satisfactory explanation of the vibrational anomalies in glasses. Excellent agreement of the SCBA theory with simulation data on a soft-sphere glass is reached. Since the SCBA is based on a Gaussian distribution of local shear moduli, including negative values, this theory describes a shear instability as a function of the variance of shear fluctuations. In the vicinity of this instability, a fractal frequency dependence of the density of states and the sound attenuation ∝ ω1+ais predicted with a≲ 1/2. Such a frequency dependence is indeed observed both in simulations and in experimental data. We argue that the observed frequency dependence stems from marginally stable regions in a glass and discuss these findings in terms of rigidity percolation.

Microglia reactivity entails a large-scale remodeling of cellular geometry, but the behavior of the microtubule cytoskeleton during these changes remains unexplored. Here we show that activated microglia provide an example of microtubule reorganization from a non-centrosomal array of parallel and stable microtubules to a radial array of more dynamic microtubules. While in the homeostatic state microglia nucleate microtubules at Golgi outposts, activating signaling induces recruitment of nucleating material nearby the centrosome, a process inhibited by microtubule stabilization. Our results demonstrate that a hallmark of microglia reactivity is a striking remodeling of the microtubule cytoskeleton and suggest that while pericentrosomal microtubule nucleation may serve as a distinct marker of microglia activation, inhibition of microtubule dynamics may provide a different strategy to reduce microglia reactivity in inflammatory disease.

The experimentally measured vibrational spectrum of glasses strongly deviates from that expected in Debye’s elasticity theory: The density of states deviates from Debye’s ω2law (“boson peak”), the sound velocity shows a negative dispersion in the boson-peak frequency regime, and there is a strong increase in the sound attenuation near the boson-peak frequency. A generalized elasticity theory is presented, based on the model assumption that the shear modulus of the disordered medium fluctuates randomly in space. The fluctuations are assumed to be uncorrelated and have a certain distribution (Gaussian or otherwise). Using field-theoretical techniques one is able to derive mean-field theories for the vibrational spectrum of a disordered system. The theory based on a Gaussian distribution uses a self-consistent Born approximation (SCBA),while the theory for non-Gaussian distributions is based on a coherent-potential approximation (CPA). Both approximate theories appear to be saddle-point approximations of effective replica field theories. The theory gives a satisfactory explanation of the vibrational anomalies in glasses. Excellent agreement of the SCBA theory with simulation data on a soft-sphere glass is reached. Since the SCBA is based on a Gaussian distribution of local shear moduli, including negative values, this theory describes a shear instability as a function of the variance of shear fluctuations. In the vicinity of this instability, a fractal frequency dependence of the density of states and the sound attenuation ∝ ω1+ais predicted with a≲ 1/2. Such a frequency dependence is indeed observed both in simulations and in experimental data. We argue that the observed frequency dependence stems from marginally stable regions in a glass and discuss these findings in terms of rigidity percolation.

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The dynamic structure factors of three glass-formers, o-terphenyl, glycerol and vitreous silica, have been measured at different temperatures Tand exchanged momenta Qusing inelastic x-ray scattering. We find propagating excitations at wavelengths approaching the interparticle separation region. We will show that these excitations are strongly related to the excess of vibrational states in glasses. Moreover, taking as example the measurements performed on o-terphenyl, we demonstrate that the temperature dependence of the dispersion curves allows the determination of the infinite frequency sound velocity C?(T). The derived values of C?(T) in the liquid phase can be used to discriminate among different phenomenological models previously proposed.