Unraveling lipid/protein interaction in model lipid bilayers by Atomic Force Microscopy

The current view of the biological membrane is that in which lipids and proteins mutually interact to accomplish membrane functions. The lateral heterogeneity of the lipid bilayer can induce partitioning of membrane-associated proteins, favoring protein-protein interaction and influence signaling and trafficking. The Atomic Force Microscope allows to study the localization of membrane-associated proteins with respect to the lipid organization at the single molecule level and without the need for fluorescence staining. These features make AFM a technique of choice to study lipid/protein interactions in model systems or native membranes. Here we will review the technical aspects inherent to and the main results obtained by AFM in the study of protein partitioning in lipid domains concentrating in particular on GPI-anchored proteins, lipidated proteins, and transmembrane proteins. Whenever possible, we will also discuss the functional consequences of what has been imaged by Atomic Force Microscopy

Nanoscale mechanical properties of lipid bilayers and their relevance in biomembrane organization and function

The mechanical properties of biological systems are emerging as fundamental in determining their functional activity. For example, cells continuously probe their environment by applying forces and, at the same time, are exposed to forces produced by the same environment. Also in biological membranes, the activity of membrane related proteins are affected by the overall mechanical properties of the hosting environment. Traditionally, the mesoscopic mechanical properties of lipid bilayers have been studied by micropipette aspiration techniques. In recent years, the possibility of probing mechanical properties of lipid bilayers at the nanoscale has been promoted by the force spectroscopy potentiality of Atomic Force Microscopes (AFM). By acquiring force-curves on supported lipid bilayers (SLBs) it is possible to probe the mechanical properties on a scale relevant to the interaction between membrane proteins and lipid bilayers and to monitor changes of these properties as a result of a changing environment. Here, we review a series of force spectroscopy experiments performed on SLBs with an emphasis on the functional consequences the measured mechanical properties can have on membrane proteins. We also discuss the force spectroscopy experiments on SLBs in the context of theories developed for dynamic force spectroscopy experiments with the aim to extract the kinetic and energetic description of the process of membrane rupture

Observation of the Quantum Zeno Effect on a NISQ Device

We study the Quantum Zeno Effect (QZE) on a single qubit on IBM Quantum Experience devices under the effect of multiple measurements. We consider two possible cases: the Rabi evolution and the free decay. In both cases we observe the occurrence of the QZE as an increasing of the survival probability with the number of measurements

“Kiezdeutsch ist kein gebrochenes Deutsch" L’arricchimento linguistico tra le nuove generazioni

La lingua è un sistema in continua evoluzione ed è costantemente influenzata dalle circostanze sociali che coinvolgono la rispettiva comunità di parlanti. Un esempio lampante è il Kiezdeutsch: si tratta di una varietà del tedesco parlato dai giovani dei quartieri multietnici in Germania, frutto dell'incontro personale e linguistico tra la società ospitante tedesca e le generazioni di migranti e con background migratorio. Questa tesi intende ripercorrere l'evoluzione del Kiezdeutsch dalle sue origini, quindi partendo dalla storia delle migrazioni in Germania nel secondo dopoguerra. Un secondo focus sarà rivolto alle politiche d'integrazione attuate dal governo tedesco a sostegno delle popolazioni migranti, senza le quali forse il Kiezdeutsch non sarebbe fiorito. Infine, il lavoro presenterà le caratteristiche e le novità principali di questo nuovo dialetto, cercando anche di sfatare i falsi miti che si sono diffusi negli anni, a causa di una percezione negativa del Kiezdeutsch

Dynamic Force Spectroscopy on Supported Lipid Bilayers: Effect ofTemperature and Sample Preparation

Biological membranes are constantly exposed to forces. The stress-strain relation in membranes determines thebehavior of many integral membrane proteins or other membrane related-proteins that show a mechanosensitive behavior. Here, we studied by force spectroscopy the behavior of supported lipid bilayers (SLBs) subjected to forces perpendicular to their plane. We measured the lipid bilayer mechanical properties and the force required for the punch-through event characteristic of atomic force spectroscopy on SLBs as a function of the interleaflet coupling. We found that for an uncoupled bilayer, the overall tip penetration occurs sequentially through the two leaflets, giving rise to two penetration events. In the case of a bilayer with coupled leaflets, penetration of the atomic force microscope tip always occurred in a single step. Considering the dependence of the jump-through force value on the tip speed, we also studied the process in the context of dynamic force spectroscopy (DFS). We performed DFS experiments by changing the temperature and cantilever spring constant, and analyzed the resultsin the context of the developed theories for DFS. We found that experiments performed at different temperatures and withdifferent cantilever spring constants enabled a more effective comparison of experimental data with theory in comparisonwith previously published data

Small and large scale segmental motion in polymers: Estimating cooperativity length by ordinary relaxation experiments

We derive a suitable expression for estimating the size of the cooperatively rearranging regions (CRRs) in supercooled polymer melts by fitting data worked out by ordinary relaxation experiments carried out in isothermal conditions. As an example, the average CRR size in poly(n-butyl methacrylate) in proximity to the glass transition temperature is derived from a stress relaxation experiment performed by means of an atomic force microscopy setup. Good agreement is found with results in the literature derived from measurements of temperature fluctuations (the so-called Donth method). The temperature dependence of the CRR size is explored for poly(butadiene); in this case the segmental relaxation function is derived through a novel method for the analysis of the efficiency with which free induction decay echoes are refocused in 1H NMR experiments. It is found that the CRR size increases upon cooling. The results derived from the analysis of the NMR data are found to be in satisfactory agreement with those worked out from broadband dielectric spectroscopy data in the literatur

Method for estimating the cooperativity length in polymers

The problem of estimating the size of the cooperatively rearranging regions (CRRs) in supercooled polymeric melts from an analysis of the α-process in ordinary relaxation experiments is addressed. The mechanism whereby a CRR changes its configuration is viewed as consisting of two distinct steps: a reduced number of monomers reaches initially an activated state, allowing for some local rearrangement; then, the subsequent regression of the energy fluctuation may take place through the configurational degrees of freedom, thus allowing for further rearrangements on larger length scales. The latter are indeed those to which the well-known Donth's scheme refers. Local readjustments are described in the framework of a canonical formalism on a stationary ensemble of small-scale regions, distributed over all possible energy thresholds for rearrangement. Large-scale configurational changes, instead, are described as spontaneous processes. Two main regimes are envisaged, depending on whether the role played by the configurational degrees of freedom in the regression of the energy fluctuation is significant or not. It is argued that the latter case is related to the occurrence of an Arrhenian dependence of the central relaxation rate. Consistency with Donth's scheme is demonstrated, and data from the literature confirm the agreement of the two methods of analysis when configurational degrees of freedom are relevant for the fluctuation regression. Poly(n-butyl methacrylate) is chosen in order to show how CRR size and temperature fluctuations at rearrangement can be estimated from stress relaxation experiments carried out by means of an atomic force microscopy setup. Cases in which the configurational pathway for regression is significantly hindered are considered. Relaxation in poly(dimethyl siloxane) confined in nanopores is taken as an example to suggest how a more complete view of the effects of configurational constraints would be possible if direct measurements of temperature fluctuations were combined with the proposed analysis

γ-Hemolysin oligomeric structure and effect of its formation on supported lipid bilayers: An AFM Investigation.

γ-Hemolysins are bicomponent β-barrel pore forming toxins produced by Staphylococcus aureus as water-soluble monomers, which assemble into oligomeric pores on the surface of lipid bilayers. Here, after investigating the oligomeric structure of γ-hemolysins on supported lipid bilayers (SLBs) by atomic force microscopy (AFM), we studied the effect produced by this toxin on the structure of SLBs. We found that oligomeric structures with different number of monomers can assemble on the lipid bilayer being the octameric form the most stable one. Moreover, in this membrane model we found that γ-hemolysins can form clusters of oligomers inducing a curvature in the lipid bilayer, which could probably enhance the aggressiveness of these toxins at high concentrations

Automated vehicles and the rethinking of mobility and cities

The project CityMobil2 has carried out a forward-looking exercise to investigate a lternative cybermobility scenarios, including both niche and large-market innovations, and their impacts on European cities and their transport systems. The paper describes the current status of and main trends in automated vehicles, a preliminary vision of the future city with mobility supported mainly by automated vehicles, and freight distribution. The expected positive impacts derive from the development of car sharing, the reduction of space required for parking vehicles, the possibilities for older people or those with disabilities to use cars, the enhancement of safety, and the improvement of efficiency of the transport system