Corrosion-driven droplet wetting on iron nanolayers.

The classical Evans' drop describes a drop of aqueous salt solution, placed on a bulk metal surface where it displays a corrosion pit that grows over time producing further oxide deposits from the metal dissolution. We focus here on the corrosion-induced droplet spreading using iron nanolayers whose semi-transparency allowed us to monitor both iron corrosion propagation and electrolyte droplet behavior by simple optical means. We thus observed that pits grow under the droplet and merge into a corrosion front. This front reached the triple contact line and drove a non radial spreading, until it propagated outside the immobile droplet. Such chemically-active wetting is only observed in the presence of a conductive substrate that provides strong adhesion of the iron nanofilm to the substrate. By revisiting the classic Evan's drop experiment on thick iron film, a weaker corrosion-driven droplet spreading is also identified. These results require further investigations, but they clearly open up new perspectives on substrate wetting by corrosion-like electrochemical reactions at the nanometer scale.

DNA-protamine condensates under low salt conditions: molecular dynamics simulation with a simple coarse-grained model focusing on electrostatic interactions.

Protamine, a small, strongly positively-charged protein, plays a key role in achieving chromatin condensation inside sperm cells and is also involved in the formulation of nanoparticles for gene therapy and packaging of mRNA-based vaccines against viral infection and cancer. The detailed mechanisms of such condensations are still poorly understood especially under low salt conditions where electrostatic interaction predominates. Our previous study, with a refined coarse-grained model in full consideration of the long-range electrostatic interactions, has demonstrated the crucial role of electrostatic interaction in protamine-controlled reversible DNA condensation. Therefore, we herein pay our attention only to the electrostatic interaction and devise a coarser-grained bead-spring model representing the right linear charge density on protamine and DNA chains but treating other short-range interactions as simply as possible, which would be suitable for real-scale simulations. Effective pair potential calculations and large-scale molecular dynamics simulations using this extremely simple model reproduce the phase behaviour of DNA in a wide range of protamine concentrations under low salt conditions, again revealing the importance of the electrostatic interaction in this process and providing a detailed nanoscale picture of bundle formation mediated by a charge disproportionation mechanism. Our simulations also show that protamine length alters DNA overcharging and in turn redissolution thresholds of DNA condensates, revealing the important role played by entropies and correlated fluctuations of condensing agents and thus offering an additional opportunity to design tailored nanoparticles for gene therapy. The control mechanism of DNA-protamine condensates will also provide a better microscopic picture of biomolecular condensates, i.e., membraneless organelles arising from liquid-liquid phase separation, that are emerging as key principles of intracellular organization. Such condensates controlled by post-translational modification of protamine, in particular phosphorylation, or by variations in protamine length from species to species may also be responsible for the chromatin-nucleoplasm patterning observed during spermatogenesis in several vertebrate and invertebrate species.

Biocorrosion on Nanofilms Induces Rapid Bacterial Motions via Iron Dissolution.

Stability and reactivity of solid metal or mineral surfaces in contact with bacteria are critical properties for development of biocorrosion protection and for understanding bacteria-solid environmental interactions. Here, we opted to work with nanosheets of iron nanolayers offering arbitrarily large and stable areas of contact that can be simply monitored by optical means. We focused our study on the sediments' bacteria, the strain Shewanella oneidensis WT MR-1, that served as models for previous research on electroactivity and iron-reduction effects. Data show that a sudden uniform corrosion appeared after an early electroactive period without specific affinities and that iron dissolution induced rapid bacterial motions. By extending the approach to mutant strains and three bacterial species, we established a correlation between corrosion onset and oxygen-depletion combined with iron reduction and demonstrated bacteria's extraordinary ability to transform their solid environments.

Recent advances in studying single bacteria and biofilm mechanics.

Bacterial biofilms correspond to surface-associated bacterial communities embedded in hydrogel-like matrix, in which high cell density, reduced diffusion and physico-chemical heterogeneity play a protective role and induce novel behaviors. In this review, we present recent advances on the understanding of how bacterial mechanical properties, from single cell to high-cell density community, determine biofilm tri-dimensional growth and eventual dispersion and we attempt to draw a parallel between these properties and the mechanical properties of other well-studied hydrogels and living systems.

Mechanical Behavior of a Bacillus subtilis Pellicle.

Bacterial biofilms consist of a complex network of biopolymers embedded with microorganisms, and together these components form a physically robust structure that enables bacteria to grow in a protected environment. This structure can help unwanted biofilms persist in situations ranging from chronic infection to the biofouling of industrial equipment, but under certain circumstances it can allow the biofilm to disperse and colonize new niches. Mechanical properties are therefore a key aspect of biofilm life. In light of the recently discovered growth-induced compressive stress present within a biofilm, we studied the mechanical behavior of Bacillus subtilis pellicles, or biofilms at the air-liquid interface, and tracked simultaneously the force response and macroscopic structural changes during elongational deformations. We observed that pellicles behaved viscoelastically in response to small deformations, such that the growth-induced compressive stress was still present, and viscoplastically at large deformations, when the pellicles were under tension. In addition, by using particle imaging velocimetry we found that the pellicle deformations were nonaffine, indicating heterogeneous mechanical properties with the pellicle being more pliable near attachment surfaces. Overall, our results indicate that we must consider not only the viscoelastic but also the viscoplastic and mechanically heterogeneous nature of these structures to understand biofilm dispersal and removal.

A route to self-assemble suspended DNA nano-complexes.

Highly charged polyelectrolytes can self-assemble in presence of condensing agents such as multivalent cations, amphiphilic molecules or proteins of opposite charge. Aside precipitation, the formation of soluble micro- and nano-particles has been reported in multiple systems. However a precise control of experimental conditions needed to achieve the desired structures has been so far hampered by the extreme sensitivity of the samples to formulation pathways. Herein we combine experiments and molecular modelling to investigate the detailed microscopic dynamics and the structure of self-assembled hexagonal bundles made of short dsDNA fragments complexed with small basic proteins. We suggest that inhomogeneous mixing conditions are required to form and stabilize charged self-assembled nano-aggregates in large excess of DNA. Our results should help re-interpreting puzzling behaviors reported for a large class of strongly charged polyelectrolyte systems.

Bacillus subtilis Bacteria Generate an Internal Mechanical Force within a Biofilm.

A key issue in understanding why biofilms are the most prevalent mode of bacterial life is the origin of the degree of resistance and protection that bacteria gain from self-organizing into biofilm communities. Our experiments suggest that their mechanical properties are a key factor. Experiments on pellicles, or floating biofilms, of Bacillus subtilis showed that while they are multiplying and secreting extracellular substances, bacteria create an internal force (associated with a -80±25 Pa stress) within the biofilms, similar to the forces that self-equilibrate and strengthen plants, organs, and some engineered buildings. Here, we found that this force, or stress, is associated with growth-induced pressure. Our observations indicate that due to such forces, biofilms spread after any cut or ablation by up to 15-20% of their initial size. The force relaxes over very short timescales (tens of milliseconds). We conclude that this force helps bacteria to shape the biofilm, improve its mechanical resistance, and facilitate its invasion and self-repair.

Elasticity and wrinkled morphology of Bacillus subtilis pellicles.

Wrinkled morphology is a distinctive phenotype observed in mature biofilms produced by a great number of bacteria. Here we study the formation of macroscopic structures (wrinkles and folds) observed during the maturation of Bacillus subtilis pellicles in relation to their mechanical response. We show how the mechanical buckling instability can explain their formation. By performing simple tests, we highlight the role of confining geometry and growth in determining the symmetry of wrinkles. We also experimentally demonstrate that the pellicles are soft elastic materials for small deformations induced by a tensile device. The wrinkled structures are then described by using the equations of elastic plates, which include the growth process as a simple parameter representing biomass production. This growth controls buckling instability, which triggers the formation of wrinkles. We also describe how the structure of ripples is modified when capillary effects are dominant. Finally, the experiments performed on a mutant strain indicate that the presence of an extracellular matrix is required to maintain a connective and elastic pellicle.

DNA condensed by protamine: a "short" or "long" polycation behavior.

Using centrifugation assay and light scattering measurements, we study the condensation of DNA by the salmon protamine, a highly basic protein carrying 21 positive charges out of 30 amino acids, in the presence of a high amount of monovalent salt. The DNA condensation is followed by a macroscopic phase separation. It occurs while a large amount of polycations remains freely diffusing in the bulk. A similar behavior was described before for small multivalent ions in diluted DNA solution in a lower salt range. Sensitivity to the salt is however amplified when increasing the charge of polycations. Indeed, a high power-law dependence is observed here with an exponent 11. This variation agrees with the power-law dependence that characterizes the binding of small polycations to DNA. In other words, we show that protamines behave like small polycations in the diluted DNA-high salt regime, while they behave like other large polycations in the diluted DNA-low salt regime as shown in a previous study. In addition, instead of the classical view where binding of polycations to DNA is supposed to trigger DNA condensation in low and moderate salt conditions, we propose that, under high salt conditions, the potential presence of a DNA dense phase triggers the binding of protamines to DNA.

Confinement of DNA in water-in-oil microemulsions.

The study of systems that allow DNA condensation in confined environments is an important task in producing cell-mimicking microreactors capable of biochemical activities. The water droplets formed in water-in-oil emulsions are potentially good candidates for such microcompartments. The anionic surfactant AOT was used here to stabilize the droplets. We have studied in detail the DNA distribution and the structural modifications of these microemulsion drops by varying the concentration and molecular weight of DNA and using various techniques such as light, X-ray, and neutron scattering, electrical conductivity, and surface tension. DNA induces the formation of large drops into which it is internalized. The size of these drops depends on the amount of DNA dissolved in water as well as on its molecular weight. The local DNA concentration is very high (>100 mg/mL). The large drops coexist with small empty drops (not containing DNA), similar to those found in the DNA-free microemulsion.

Pressure built by DNA packing inside virions: enough to drive DNA ejection in vitro, largely insufficient for delivery into the bacterial cytoplasm.

Tailed bacteriophage particles carry DNA highly pressurized inside the capsid. Challenge with their receptor promotes release of viral DNA. We show that addition of the osmolyte polyethylene glycol (PEG) has two distinct effects in bacteriophage SPP1 DNA ejection. One effect is to inhibit the trigger for DNA ejection. The other effect is to exert an osmotic pressure that controls the extent of DNA released in phages that initiate ejection. We carried out independent measurements of each effect, which is an essential requirement for their quantitative study. The fraction of phages that do not eject increased linearly with the external osmotic pressure. In the remaining phage particles ejection stopped after a defined amount of DNA was reached inside the capsid. Direct measurement of the size of non-ejected DNA by gel electrophoresis at different PEG concentrations in the latter sub-population allowed determination of the external osmotic pressure that balances the force powering DNA exit (47 atm for SPP1 wild-type). DNA exit stops when the ejection force mainly due to repulsion between DNA strands inside the SPP1 capsid equalizes the force resisting DNA insertion into the PEG solution. Considering the turgor pressure in the Bacillus subtilis cytoplasm the energy stored in the tight phage DNA packing is only sufficient to power entry of the first 17% of the SPP1 chromosome into the cell, the remaining 83% requiring application of additional force for internalization.

A kinetic analysis of DNA ejection from tailed phages revealing the prerequisite activation energy.

All tailed bacteriophages follow the same general scheme of infection: they bind to their specific host receptor and then transfer their genome into the bacterium. DNA translocation is thought to be initiated by the strong pressure due to DNA packing inside the capsid. However, the exact mechanism by which each phage controls its DNA ejection remains unknown. Using light scattering, we analyzed the kinetics of in vitro DNA release from phages SPP1 and lambda (Siphoviridae family) and found a simple exponential decay. The ejection characteristic time was studied as a function of the temperature and found to follow an Arrhenius law, allowing us to determine the activation energy that governs DNA ejection. A value of 25-30 kcal/mol is obtained for SPP1 and lambda, comparable to the one measured in vitro for T5 (Siphoviridae) and in vivo for T7 (Podoviridae). This suggests similar mechanisms of DNA ejection control. In all tailed phages, the opening of the connector-tail channel is needed for DNA release and could constitute the limiting step. The common value of the activation energy likely reflects the existence for all phages of an optimum value, ensuring a compromise between efficient DNA delivery and high stability of the virus.

Aggregate formation in aqueous solutions of carboxymethylcellulose and cationic surfactants.

The addition of cationic surfactants to an aqueous solution of an anionic polymer, carboxymethylcellulose (carboxyMC), causes the spontaneous formation of aggregates in a certain range of concentrations. Here we studied two surfactants, dodecyl and hexadecyl trimethylammonium bromide (DTAB and CTAB, respectively). Using different techniques (light scattering, potentiometry, viscosimetry, and zetametry), we found that a simple lengthening of the surfactant tail length by four CH2 groups drastically changes the aggregate morphology, size, and charge. We explored in detail how the surfactant and polymer concentrations act on these systems.

Solubility and charge inversion of complexes of DNA and basic proteins.

The basic proteins, protamines and histones H1, are known to condense DNA in vivo. We examine here their ability to condense and solubilize in vitro linear DNA [and a synthetic polyanion, Poly(Styrene-Sulfonate) or PSS] at low ionic concentrations by varying the charge concentration ratio. Phase separation is observed in a very narrow range of ratios for short DNA and PSS; on both sides of this range, polydisperse and charged complexes are formed. A charge inversion is detected. For long DNA chains however, a different behavior is observed: the complexes are not soluble in excess of proteins.

Are liquid crystalline properties of nucleosomes involved in chromosome structure and dynamics?

Nucleosome core particles correspond to the structural units of eukaryotic chromatin. They are charged colloids, 101 Angstrom in diameter and 55 Angstrom in length, formed by the coiling of a 146/147 bp DNA fragment (50 nm) around the histone protein octamer. Solutions of these particles can be concentrated, under osmotic pressure, up to the concentrations found in the nuclei of living cells. In the presence of monovalent cations (Na(+)), nucleosomes self-assemble into crystalline or liquid crystalline phases. A lamello-columnar phase is observed at 'low salt' concentrations, while a two-dimensional hexagonal phase and a three-dimensional quasi-hexagonal phase form at 'high salt' concentrations. We followed the formation of these phases from the dilute isotropic solutions to the ordered phases by combining cryoelectron microscopy and X-ray diffraction analyses. The phase diagram is presented as a function of the monovalent salt concentration and applied osmotic pressure. An alternative method to condense nucleosomes is to induce their aggregation upon addition of divalent or multivalent cations (Mg(2+), spermidine(3+) and spermine(4+)). Ordered phases are also found in the aggregates. We also discuss whether these condensed phases of nucleosomes may be relevant from a biological point of view.

DNA ejection from bacteriophage T5: analysis of the kinetics and energetics.

DNA ejection from bacteriophage T5 can be passively driven in vitro by the interaction with its specific host receptor. Light scattering was used to determine the physical parameters associated with this process. By studying the ejection kinetics at different temperatures, we demonstrate that an activation energy of the order of 70 k(B)T must be overcome to allow the complete DNA ejection. A complex shape of the kinetics was found whatever the temperature. This shape may be actually understood using a phenomenological model based on a multistep process. Passing from one stage to another requires the mentioned thermal activation of pressurized DNA inside the capsids. Both effects contribute to shorten or to lengthen the pause time between the different stages explaining why the T5 DNA ejection is so slow compared to other types of phage.

Osmotic pressure inhibition of DNA ejection from phage.

Bacterial viral capsids in aqueous solution can be opened in vitro by addition of their specific receptor proteins, with consequent full ejection of their genomes. We demonstrate that it is possible to control the extent of this ejection by varying the external osmotic pressure. In the particular case of bacteriophage lambda, the ejection is 50% inhibited by osmotic pressures (of polyethylene glycol) comparable to those operative in the cytoplasm of host bacteria; it is completely suppressed by a pressure of 20 atmospheres. Furthermore, our experiments monitor directly a dramatic decrease of the stress inside the unopened phage capsid upon addition of polyvalent cations to the host solution, in agreement with many recent theories of DNA interactions.

An alternative method to the osmotic stressing polymers: the osmomanometer.

Although stressing polymers have been widely and successfully used to determine the osmotic properties of solutes in aqueous media, the osmotic stress method presents some limitations. To overcome these drawbacks, an alternative and more direct method, which has been named the osmomanometer, is described in this letter. The osmotic pressure accessible by this method ranges typically from 1 to 30 kPa using a simple hydrostatic effect and can be extended to higher pressures by using pressurized gas. This method needs neither a pressure sensor nor calibration.