Regulation of the intermittent release of giant unilamellar vesicles under osmotic pressure

Osmotic pressure can break the fluid balance between intracellular and extracellular solutions. In hypo-osmotic solution, water molecules, which transfer into the cell and burst, are driven by the concentrations difference of solute across the semi-permeable membrane. The complicated dynamic processes of the intermittent burst have been previously observed. However, the underlying physical mechanism has yet to be thoroughly explored and analyzed. Here, the intermittent release of inclusion in giant unilamellar vesicles was investigated quantitatively, applying the combination of experimental and theoretical methods in the hypo-osmotic medium. Experimentally, we adopted highly sensitive EMCCD to acquire intermittent dynamic images. Notably, the component of the vesicle phospholipids affected the stretch velocity, and the prepared solution of the vesicle adjusted the release time. Theoretically, we chose equations numerical simulations to quantify the dynamic process in phases and explored the influence of physical parameters such as bilayer permeability and solution viscosity on the process. It was concluded that the time taken to achieve the balance of giant unilamellar vesicles was highly dependent on the structure of the lipid molecular. The pore lifetime was strongly related with the internal solution environment of giant unilamellar vesicles. The vesicle prepared in viscous solution accessed visualized long-lived pore. Furthermore, the line tension was measured quantitatively by the release velocity of inclusion, which was in the same order of magnitude as the theoretical simulation. In all, the experimental values well matched the theoretical values. Our investigation clarified the physical regulatory mechanism of intermittent pore formation and inclusion release, which had an important reference for the development of novel technologies such as gene therapy based on transmembrane transport as well as controlled drug delivery based on liposomes.

Nucleation of Surfactant–Alkane Mixed Solid Monolayer and Bilayer Domains at the Air–Water Interface

We investigated the wetting transitions of tetradecane and hexadecane droplets in dodecyltrimethylammonium bromide (C12TAB), tetradecyltrimethylammonium bromide (C14TAB), and hexadecyltrimethylammonium bromide (C16TAB) aqueous solutions. By varying the surfactant concentration, the formation of mixed monolayers of a surfactant and an alkane was observed at the air–water interface. Depending on the combination of surfactant and alkane, these wetting monolayers underwent another thermal phase transition upon cooling either to a frozen mixed monolayer (S1) or a bilayer structure composed of a solid monolayer of a pure alkane rested on a liquid-like mixed monolayer (S2). Based on the phase diagrams determined by phase modulation ellipsometry, the difference in the morphology of the nucleated S1 and S2 phase domains was also investigated using Brewster angle microscopy. Domains of the S1 phase were relatively small and highly branched, whereas those of the S2 phase were large and circular. The difference in domain morphology was explained by the competition of the domain line tension and electrostatic dipole interactions between surfactant molecules in the domains.

Research on Wave Attenuation Performance of Floating Breakwater

In this study, a new type of double-pontoon floating breakwater was designed to improve the wave attenuation performance through the addition of suspended Savonius propeller-blade. Its hydrodynamic characteristics were studied through numerical simulations and performance-testing experiment. The following investigations were performed in this study: Firstly, wave theory and hydrodynamic theory were combined to calculate the wave attenuation performance and motion response of double-pontoon floating breakwater under linear wave conditions. The numerical results showed that the wave attenuation performance was better under a specific wave period and height, the transmission coefficient reached a relatively small value, and the mooring line tension responded periodically and satisfied the condition of maximum breaking force. Secondly, three key geometric parameters of breakwater were researched, including the relative spacing of pontoons, the relative spacing between pontoons and blades, and the height–diameter ratio of Savonius blades. The calculation results showed that the pontoon spacing was closer to the wavelength and the breakwater wave attenuation performance was better. Lastly, experimental tests were also performed on the new double-pontoon floating breakwater and the results showed that the wave attenuation performance and numerical projections were basically the same, which verified the validity and effectiveness of the design method.

Wetting between Cassie–Baxter and Wenzel regimes: a cellular model approach

The cellular model with periodic boundary conditions was proposed for the study of liquid–solid interface properties of solid surfaces decorated by a regular pattern. The solid surface was represented by a mosaic of truncated pyramids of two different slopes of side walls equivalent to a surface covered with triangular grooves of different dihedral angles. On the basis of the computations performed for a single elementary cell, the components of the interfacial energies and the apparent contact angles have been found for different Young contact angles and different tilting angles of the pyramid walls. It was found that at certain sets of angles, the wetting takes place with the partial coverage of the pyramid sidewalls—in between the Cassie–Baxter and Wenzel regimes. The influence of the line tension on the studied surface wettability was also examined. Graphic abstract

Structural Diversity of Photoswitchable Sphingolipids for Optodynamic Control of Lipid Raft Microdomains

Sphingolipids are a structurally diverse class of lipids predominantly found in the plasma membrane of eukaryotic cells. These lipids can laterally segregate with other saturated lipids and cholesterol into lipid rafts; liquid-ordered (Lo) microdomains that act as organizing centers within biomembranes. Owing the vital role of sphingolipids for lipid segregation, controlling their lateral localization is of utmost significance. Hence, we made use of the light-induced trans-cis isomerization of azobenzene-modified acyl chains, to develop a set of photoswitchable sphingolipids, with different headgroups (hydroxyl, galactosyl, phosphocholine) and backbones (sphingosine, phytosphingosine, tetrahydropyran (THP)-blocked sphingosine), able to shuttle between liquid-ordered (Lo) and liquid-disordered (Ld) regions of model membranes upon irradiation with UV-A (λ = 365 nm) and blue (λ = 470 nm) light, respectively. Using combined high-speed atomic force microscopy, fluorescence microscopy, and force spectroscopy, we investigated how these active sphingolipids laterally remodel supported bilayers upon photo-isomerization, notably in terms of domain area changes, height mismatch, line tension, and membrane piercing. Hereby, we show that all sphingosine-(Azo-β-Gal-Cer, Azo-SM, Azo-Cer) and phytosphingosine-based (Azo-α-Gal-PhCer, Azo-PhCer) photolipids behave similarly, promoting a reduction in Lo domain area when in the UV-adapted cis-isoform. In contrast, azo-sphingolipids having THP groups that block H-bonding at the sphingosine backbone (Azo-THP-SM, Azo-THP-Cer) induce an increase in the Lo domain area when in cis, accompanied by a major rise in height mismatch and line tension. These changes were fully reversible upon blue light-triggered isomerization of the various lipids back to trans, pinpointing the role of interfacial interactions for the formation of stable Lo lipid raft domains.

Force Mapping Study of Actinoporin Effect in Membranes Presenting Phase Domains

Equinatoxin II (EqtII) and Fragaceatoxin C (FraC) are pore-forming toxins (PFTs) from the actinoporin family that have enhanced membrane affinity in the presence of sphingomyelin (SM) and phase coexistence in the membrane. However, little is known about the effect of these proteins on the nanoscopic properties of membrane domains. Here, we used combined confocal microscopy and force mapping by atomic force microscopy to study the effect of EqtII and FraC on the organization of phase-separated phosphatidylcholine/SM/cholesterol membranes. To this aim, we developed a fast, high-throughput processing tool to correlate structural and nano-mechanical information from force mapping. We found that both proteins changed the lipid domain shape. Strikingly, they induced a reduction in the domain area and circularity, suggesting a decrease in the line tension due to a lipid phase height mismatch, which correlated with proteins binding to the domain interfaces. Moreover, force mapping suggested that the proteins affected the mechanical properties at the edge, but not in the bulk, of the domains. This effect could not be revealed by ensemble force spectroscopy measurements supporting the suitability of force mapping to study local membrane topographical and mechanical alterations by membranotropic proteins.

Melatonin: Regulation of Biomolecular Condensates in Neurodegenerative Disorders

Biomolecular condensates are membraneless organelles (MLOs) that form dynamic, chemically distinct subcellular compartments organizing macromolecules such as proteins, RNA, and DNA in unicellular prokaryotic bacteria and complex eukaryotic cells. Separated from surrounding environments, MLOs in the nucleoplasm, cytoplasm, and mitochondria assemble by liquid–liquid phase separation (LLPS) into transient, non-static, liquid-like droplets that regulate essential molecular functions. LLPS is primarily controlled by post-translational modifications (PTMs) that fine-tune the balance between attractive and repulsive charge states and/or binding motifs of proteins. Aberrant phase separation due to dysregulated membrane lipid rafts and/or PTMs, as well as the absence of adequate hydrotropic small molecules such as ATP, or the presence of specific RNA proteins can cause pathological protein aggregation in neurodegenerative disorders. Melatonin may exert a dominant influence over phase separation in biomolecular condensates by optimizing membrane and MLO interdependent reactions through stabilizing lipid raft domains, reducing line tension, and maintaining negative membrane curvature and fluidity. As a potent antioxidant, melatonin protects cardiolipin and other membrane lipids from peroxidation cascades, supporting protein trafficking, signaling, ion channel activities, and ATPase functionality during condensate coacervation or dissolution. Melatonin may even control condensate LLPS through PTM and balance mRNA- and RNA-binding protein composition by regulating N6-methyladenosine (m6A) modifications. There is currently a lack of pharmaceuticals targeting neurodegenerative disorders via the regulation of phase separation. The potential of melatonin in the modulation of biomolecular condensate in the attenuation of aberrant condensate aggregation in neurodegenerative disorders is discussed in this review.

The Quasi-Static Response of Moored Floating Structures Based on Minimization of Mechanical Energy

It is essential to design a reasonable mooring line length that ensures quasi-static responses of moored floating structures are within an acceptable level, and that reduces the cost of mooring lines in the overall project. Quasi-static responses include the equilibrium position and the line tension of a moored floating structure (also called the mean value in a dynamic response), etc. The quasi-static responses derived by the classic catenary equation cannot present mooring–seabed interaction and hydrodynamic effects on a mooring line. While a commercial program can predict reasonable quasi-static responses, costly modeling is required. This motivated us to propose a new method for predicting quasi-static responses that minimizes the mechanical energy of the whole system based on basic geometric parameters, and that is easy to implement. In this study, the mechanical energy of moored floating structures is assumed to be the sum of gravitational–buoyancy potential energy, kinetic energy induced by drag forces, and spring potential energy derived by line tension. We introduce fundamental theoretical background for the development of the proposed method. We investigate the effect of quasi-static actions on mooring response, comparing the proposed method’s results with those from the catenary equation and ABAQUS software. The study reveals the shortcomings of the catenary equation in offshore applications. We also compare quasi-static responses derived by the AQWA numerical package with the results calculated from the proposed method for an 8 MW WindFloat 2 type of platform. Good agreement was drawn between the proposed method and AQWA. The proposed method proves more timesaving than AQWA in terms of modeling of mooring lines and floaters, and more accurate than the catenary equation, and can be used effectively in the early design phase of dimension mooring lengths for moored floating structures.