Dzmitry Afanasenkau

Accumulation of formamide in hydrothermal pores to form prebiotic nucleobases

Significance The aim of this article is to show that prebiotic nucleobases can be formed in hydrothermal pores, through a significant accumulation of formamide resulting from a combination of thermophoresis and convection. We performed numerical finite element calculations for initial formamide concentrations that correspond to early Earth shallow lake conditions and reveal that formamide accumulates at the bottom of hydrothermal pores in about 45–90 d to high concentrated formamide solutions. The conclusion from these findings is that the combination of thermophoretic mass transport and convection is the missing link, which makes the synthesis of prebiotic nucleobases in porous rocks in contact with shallow lakes under early-earth conditions possible. Formamide is one of the important compounds from which prebiotic molecules can be synthesized, provided that its concentration is sufficiently high. For nucleotides and short DNA strands, it has been shown that a high degree of accumulation in hydrothermal pores occurs, so that temperature gradients might play a role in the origin of life [Baaske P, et al. (2007) Proc Natl Acad Sci USA 104(22):9346−9351]. We show that the same combination of thermophoresis and convection in hydrothermal pores leads to accumulation of formamide up to concentrations where nucleobases are formed. The thermophoretic properties of aqueous formamide solutions are studied by means of Infrared Thermal Diffusion Forced Rayleigh Scattering. These data are used in numerical finite element calculations in hydrothermal pores for various initial concentrations, ambient temperatures, and pore sizes. The high degree of formamide accumulation is due to an unusual temperature and concentration dependence of the thermophoretic behavior of formamide. The accumulation fold in part of the pores increases strongly with increasing aspect ratio of the pores, and saturates to highly concentrated aqueous formamide solutions of ∼85 wt% at large aspect ratios. Time-dependent studies show that these high concentrations are reached after 45–90 d, starting with an initial formamide weight fraction of 10−3 wt % that is typical for concentrations in shallow lakes on early Earth.

Positively charged supported lipid bilayers as a biomimetic platform for neuronal cell culture.

The supported lipid bilayer (SLB) is a well-known system for studying the cell membrane and membrane proteins. It is also promising as a platform for studying cell processes: the cell adhesion, the cell membrane receptors, and the intercellular signaling processes. SLBs made of natural lipids appeared to be protein and cell repellent. Thus, to use the SLB as a substrate for cells, one should functionalize them to provide adhesion. In the present paper, we describe a simple approach to promote adhesion of neuronal cells to the SLB without using proteins or peptides, by introducing positively charged lipids 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) into the SLB made of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). We show that neurons adhere to these bilayers and grow for at least 10 days. The SLBs themselves were found to degrade with time in cell culture conditions, but maintained fluidity (as revealed by fluorescence recovery after photobleaching), demonstrating the possibility of using SLBs for studying neuronal cells in culture.

Reconstitution of Fusion Proteins in Supported Lipid Bilayers for the Study of Cell Surface Receptor–Ligand Interactions in Cell–Cell Contact

Bioactive molecules such as adhesion ligands, growth factors, or enzymes play an important role in modulating cell behavior such as cell adhesion, spreading, and differentiation. Deciphering the mechanism of ligand-mediated cell adhesion and associated signaling is of great interest not only for fundamental biophysical investigations but also for applications in medicine and biotechnology. In the presented work, we developed a new biomimetic platform that enables culturing primary neurons and testing cell surface-receptor ligand interactions in cell-cell contacts as, e.g., in neuronal synapses. This platform consists of a supported lipid bilayer modified with incorporated neuronal adhesion proteins conjugated with the Fc-domain of IgG (ephrin A5 Fc-chimera). We extensively characterized properties of these protein containing bilayers using fluorescence recovery after photobleaching (FRAP), quartz crystal microbalance with dissipation (QCM-D), and immunostaining. We conclude that the Fc-domain is the part responsible for the incorporation of the protein into the bilayer. The biomimetic platform prepared by this new approach was able to promote neuronal cell adhesion and maintain growth as well as facilitate neuronal maturation as shown by electrophysiological measurements. We believe that our approach can be extended to insert other proteins to create a general culture platform for neurons and other cell types.

Influence of temperature and charge effects on thermophoresis of polystyrene beads

Abstract.We study the thermodiffusion behavior of spherical polystyrene beads with a diameter of 25 nm by infrared thermal diffusion Forced Rayleigh Scattering (IR-TDFRS). Similar beads were used to investigate the radial dependence of the Soret coefficient by different authors. While Duhr and Braun (Proc. Natl. Acad. Sci. U.S.A. 104, 9346 (2007)) observed a quadratic radial dependence Braibanti et al. (Phys. Rev. Lett. 100, 108303 (2008)) found a linear radial dependence of the Soret coefficient. We demonstrated that special care needs to be taken to obtain reliable thermophoretic data, because the measurements are very sensitive to surface properties. The colloidal particles were characterized by transmission electron microscopy and dynamic light scattering (DLS) experiments were performed. We carried out systematic thermophoretic measurements as a function of temperature, buffer and surfactant concentration. The temperature dependence was analyzed using an empirical formula. To describe the Debye length dependence we used a theoretical model by Dhont. The resulting surface charge density is in agreement with previous literature results. Finally, we analyze the dependence of the Soret coefficient on the concentration of the anionic surfactant sodium dodecyl sulfate (SDS), applying an empirical thermodynamic approach accounting for chemical contributions.Graphical abstract

Molar mass and temperature dependence of the thermodiffusion of polyethylene oxide in water/ethanol mixtures

In this work, we study the molar mass dependence of the thermodiffusion of polyethylene oxide at different temperatures in ethanol, water/ethanol mixture (c(water) = 0.7), and water in a molar mass range up to M(w) = 180,000 g/mol. Due to the low solubility of polyethylene oxide oligomers in ethanol the measurements are limited up to M(w) = 2200 g/mol. The specific water/ethanol concentration 0.7 has been chosen, because at this weight fraction the thermal diffusion coefficient, D(T), of water/ethanol vanishes so that the system can be treated as a pseudo binary mixture. The addition of ethanol will degrade the solvent quality, so that we expect a change of the interaction energies between polymer and solvent. The analysis of the experimental data within a theoretical model shows the need of a refined model, which takes specific interactions into account.

Chip-Based Heat Stimulation for Modulating Signal Propagation in HL-1 Cell Networks

Signal propagation in cardiac cell networks can be modulated by heat stimulation. Here, the response of a connected HL‐1 cardiomyocyte cell network to the application of confined heat stimuli using Ca2+ imaging is investigated. Localized temperature gradients are generated by resistive heating via microwire arrays on a chip surface, which serves as a substrate for growing a confluent cell network. It is demonstrated that upon heat stimulation, the velocity of the propagating Ca2+ wave in the network is locally increased, leading to a deformation of the wavefront. Furthermore, evidence of a change in the signal propagation direction caused by a relocation of the pacemaker cell is shown. This effect might be used in future applications, where heat is employed as an alternative modality for cell stimulation protocols.