Interfacial Viscoelasticity and Structure of Self-Assembled Semifluorinated Alkanes on Water

Semifluorinated alkanes self-assemble spontaneously into uniform nanometer-sized domains on water. The order of such “surface micelles” at the air/water interface is achieved by the counter balance of attractive and repulsive interactions. In this thesis, the viscoelastic properties of semifluorinated alkane monolayers are investigated by using interfacial shear and dilational rheology under oscillatory strain. The obtained response function implies a predominantly elastic character of the monolayers, suggesting repulsive interactions between the surface micelles. Both the structure and the form factor of the surface micelles are determined by the quantitative analysis of grazing-incidence small-angle X-ray scattering data. A systematic variation of the length and of the number of fluorocarbon and hydrocarbon chains unravel how a subtle change in the molecular structure modulates the size, shape and correlation of the surface micelles. A promising application of semifluorinated alkanes are contrast agent microbubbles for sonographic imaging. This thesis further shows that perfluorohexane vapor - commonly used to increase the lifetime of microbubbles - reduces the elasticity of the monolayers from semifluorinated alkanes whereas their structure is not influenced. The obtained results contribute to the fundamental understanding of the formation and mechanics of mesoscopic molecular assembly at the interface

Effect of cholesterol on the mechanical stability of gel-phase phospholipid bilayers studied by AFM force spectroscopy

The remarkably low sliding friction of articular cartilage in the major joints such as hips and knees, which is crucial for its homeostasis and joint health, has been attributed to lipid bilayers forming lubricious boundary layers at its surface. The robustness of such layers, and thus their lubrication efficiency at joint pressures, depends on the lipids forming them, including cholesterol which is a ubiquitous component, and which may act to strengthen of weaken the bilayer. In this work, a systematic study using an atomic force microscope (AFM) was carried out to understand the effect of cholesterol on the nanomechanical stability of two saturated phospholipids, DSPC (1,2-distearoyl-sn-glycero-3-phosphatidlycholine) and DPPC (1,2-dipalmitoyl-sn-glycero- phosphatidylcholine), that differ in acyl chain lengths. Measurements were carried out both in water and in phosphate buffer solution (PBS). The nanomechanical stability of the lipid bilayers was quantitatively evaluated by measuring the breakthrough force needed to puncture the bilayer by the AFM tip. The molar fractions of cholesterol incorporated in the bilayers were 10% and 40%. We found that for both DSPC and DPPC, cholesterol significantly decreases the mechanical stability of the bilayers in solid-ordered (SO) phase. In accordance with the literature, the strengthening effect of salt on the lipid bilayers was also observed. For DPPC with 10 mol % cholesterol, the effect of tip properties and the experimental procedure parameters on the breakthrough forces were also studied. Tip radius (2–42 nm), material (Si, Si3N4, Au) and loading rate (40—1000 nm/s) were varied systematically. The values of the breakthrough forces measured were not significantly affected by any of these parameters, showing that the weakening effect of cholesterol does not result from such changes in experimental conditions. As we have previously demonstrated that mechanical robustness improves the tribological performance of lipid layers, this study helps to shed light on the mechanism of physiological lubrication

pH-Dependent Interactions in Dimers Govern the Mechanics and Structure of von Willebrand Factor

Von Willebrand factor (VWF) is a multimeric plasma glycoprotein that is activated for hemostasis by increased hydrodynamic forces at sites of vascular injury. Here, we present data from atomic force microscopy-based single-molecule force measurements, atomic force microscopy imaging, and small-angle x-ray scattering to show that the structure and mechanics of VWF are governed by multiple pH-dependent interactions with opposite trends within dimeric subunits. In particular, the recently discovered strong intermonomer interaction, which induces a firmly closed conformation of dimers and crucially involves the D4 domain, was observed with highest frequency at pH 7.4, but was essentially absent at pH values below 6.8. However, below pH 6.8, the ratio of compact dimers increased with decreasing pH, in line with a previous transmission electron microscopy study. These findings indicated that the compactness of dimers at pH values below 6.8 is promoted by other interactions that possess low mechanical resistance compared with the strong intermonomer interaction. By investigating deletion constructs, we found that compactness under acidic conditions is primarily mediated by the D4 domain, i.e., remarkably by the same domain that also mediates the strong intermonomer interaction. As our data suggest that VWF has the highest mechanical resistance at physiological pH, local deviations from physiological pH (e.g., at sites of vascular injury) may represent a means to enhance VWF’s hemostatic activity where needed

Force sensing by the vascular protein von Willebrand factor is tuned by a strong intermonomer interaction

The large plasma glycoprotein von Willebrand factor (VWF) senses hydrodynamic forces in the bloodstream and responds to elevated forces with abrupt elongation, thereby increasing its adhesiveness to platelets and collagen. Remarkably, forces on VWF are elevated at sites of vascular injury, where VWF’s hemostatic potential is important to mediate platelet aggregation and to recruit platelets to the subendothelial layer. Adversely, elevated forces in stenosed vessels lead to an increased risk of VWF-mediated thrombosis. To dissect the remarkable force-sensing ability of VWF, we have performed atomic force microscopy (AFM)-based single-molecule force measurements on dimers, the smallest repeating subunits of VWF multimers. We have identified a strong intermonomer interaction that involves the D4 domain and critically depends on the presence of divalent ions, consistent with results from small-angle X-ray scattering (SAXS). Dissociation of this strong interaction occurred at forces above ∼50 pN and provided ∼80 nm of additional length to the elongation of dimers. Corroborated by the static conformation of VWF, visualized by AFM imaging, we estimate that in VWF multimers approximately one-half of the constituent dimers are firmly closed via the strong intermonomer interaction. As firmly closed dimers markedly shorten VWF’s effective length contributing to force sensing, they can be expected to tune VWF’s sensitivity to hydrodynamic flow in the blood and to thereby significantly affect VWF’s function in hemostasis and thrombosis

Early human impact on soils and hydro-sedimentary systems: Multi-proxy geoarchaeological analyses from La Narse de la Sauvetat (France)

International audienceWe analysed the late-Holocene pedo-sedimentary archives of La Narse de la Sauvetat, a hydromorphic depression in the southern Limagne plain (central France), where chronologically accurate studies are scarce. The multi-proxy geoarchaeological and palaeoenvironmental analysis of two cores from different areas of the basin was carried out through sedimentological, geochemical, micromorphological and malacological investigations. Integration of these datasets supported by a robust radiocarbon-based chronology allowed discussion of socio-environmental interactions and anthropogenic impacts from Late Neolithic to Early Middle Ages. Until the Middle Bronze Age, there was no clear evidence of anthropogenic impact on soils and hydro-sedimentary dynamics of the catchment, but two peaks of high alluvial activity probably related to the 4.2 and 3.5 kyr. BP climate events were first recorded in Limagne. Significant anthropogenic impacts started in the Late Bronze Age with increased erosion of the surrounding volcanic slopes. However, a major threshold was reached c. 2600 cal BP with a sharp increase in the catchment erosion interpreted as resulting from strong anthropogenic environmental changes related to agricultural activities and drainage. This implies an anthropogenic forcing on soils and hydro-sedimentary systems much earlier than was usually considered in Limagne. These impacts then gradually increased during Late Iron Age and Roman periods, but environmental effects were certainly contained by progress in agricultural management. Late Antiquity environmental changes are consistent with regional trend to drainage deterioration in lowlands, but marked asynchrony in this landscape change suggests that societal factors implying differential land management were certainly predominant here