Lipoprotein particles exhibit distinct mechanical properties

Lipoproteins (LPs) are micelle-like structures with a similar size to extracellular vesicles (EVs) and are therefore often co-isolated, as intensively discussed within the EV community. LPs from human blood plasma are of particular interest as they are responsible for the deposition of cholesterol ester and other fats in the artery, causing lesions, and eventually atherosclerosis. Plasma lipoproteins can be divided according to their size, density and composition into chylomicrons (CM), very-low-density lipoproteins (VLDL), low-density lipoproteins (LDL) and high-density lipoproteins (HDL). Here, we use atomic force microscopy for mechanical characterization of LPs. We show that the nanoindentation approach used for EV analysis can also be used to characterize LPs, revealing specific differences between some of the particles. Comparing LPs with each other, LDL exhibit a higher bending modulus as compared to CM and VLDL, which is likely related to differences in cholesterol and apolipoproteins. Furthermore, CM typically collapse on the surface after indentation and HDL exhibit a very low height after surface adhesion both being indications for the presence of LPs in an EV sample. Our analysis provides new systematic insights into the mechanical characteristics of LPs

Active probing of the mechanical properties of biological and synthetic vesicles

BACKGROUND: The interest in mechanics of synthetic and biological vesicles has been continuously growing during the last decades. Liposomes serve as model systems for investigating fundamental membrane processes and properties. More recently, extracellular vesicles (EVs) have been investigated mechanically as well. EVs are widely studied in fundamental and applied sciences, but their material properties remained elusive until recently. Elucidating the mechanical properties of vesicles is essential to unveil the mechanisms behind a variety of biological processes, e.g. budding, vesiculation and cellular uptake mechanisms. SCOPE OF REVIEW: The importance of mechanobiology for studies of vesicles and membranes is discussed, as well as the different available techniques to probe their mechanical properties. In particular, the mechanics of vesicles and membranes as obtained by nanoindentation, micropipette aspiration, optical tweezers, electrodeformation and electroporation experiments is addressed. MAJOR CONCLUSIONS: EVs and liposomes possess an astonishing rich, diverse behavior. To better understand their properties, and for optimization of their applications in nanotechnology, an improved understanding of their mechanical properties is needed. Depending on the size of the vesicles and the specific scientific question, different techniques can be chosen for their mechanical characterization. GENERAL SIGNIFICANCE: Understanding the mechanical properties of vesicles is necessary to gain deeper insight in the fundamental biological mechanisms involved in vesicle generation and cellular uptake. This furthermore facilitates technological applications such as using vesicles as targeted drug delivery vehicles. Liposome studies provide insight into fundamental membrane processes and properties, whereas the role and functioning of EVs in biology and medicine is increasingly elucidated

Mechanical Characterization of Liposomes and Extracellular Vesicles, a Protocol

Both natural as well as artificial vesicles are of tremendous interest in biology and nanomedicine. Small vesicles (<200 nm) perform essential functions in cell biology and artificial vesicles (liposomes) are used as drug delivery vehicles. Atomic Force Microscopy (AFM) is a powerful technique to study the structural properties of these vesicles. AFM is a well-established technique for imaging at nanometer resolution and for mechanical measurements under physiological conditions. Here, we describe the procedure of AFM imaging and force spectroscopy on small vesicles. We discuss how to image vesicles with minimal structural disturbance, and how to analyze the data for accurate size and shape measurements. In addition, we describe the procedure for performing nanoindentations on vesicles and the subsequent data analysis including mechanical models used for data interpretation

Regulation of Star Formation Rates in Multiphase Galactic Disks: a Thermal/Dynamical Equilibrium Model

We develop a model for regulation of galactic star formation rates Sigma_SFR in disk galaxies, in which ISM heating by stellar UV plays a key role. By requiring simultaneous thermal and (vertical) dynamical equilibrium in the diffuse gas, and star formation at a rate proportional to the mass of the self-gravitating component, we obtain a prediction for Sigma_SFR as a function of the total gaseous surface density Sigma and the density of stars + dark matter, rho_sd. The physical basis of this relationship is that thermal pressure in the diffuse ISM, which is proportional to the UV heating rate and therefore to Sigma_SFR, must adjust to match the midplane pressure set by the vertical gravitational field. Our model applies to regions where Sigma < 100 Msun/pc^2. In low-Sigma_SFR (outer-galaxy) regions where diffuse gas dominates, the theory predicts Sigma_SFR \propto Sigma (rho_sd)^1/2. The decrease of thermal equilibrium pressure when Sigma_SFR is low implies, consistent with observations, that star formation can extend (with declining efficiency) to large radii in galaxies, rather than having a sharp cutoff. The main parameters entering our model are the ratio of thermal pressure to total pressure in the diffuse ISM, the fraction of diffuse gas that is in the warm phase, and the star formation timescale in self-gravitating clouds; all of these are (in principle) direct observables. At low surface density, our model depends on the ratio of the mean midplane FUV intensity (or thermal pressure in the diffuse gas) to the star formation rate, which we set based on Solar neighborhood values. We compare our results to recent observations, showing good agreement overall for azimuthally-averaged data in a set of spiral galaxies. For the large flocculent spiral galaxies NGC 7331 and NGC 5055, the correspondence between theory and observation is remarkably close.Comment: 49 pages, 7 figures; accepted by the Ap.

Acidification increases microbial polysaccharide degradation in the ocean

© The Authors, 2010. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Biogeosciences 7 (2010): 1615–1624, doi:10.5194/bg-7-1615-2010.With the accumulation of anthropogenic carbon dioxide (CO2), a proceeding decline in seawater pH has been induced that is referred to as ocean acidification. The ocean's capacity for CO2 storage is strongly affected by biological processes, whose feedback potential is difficult to evaluate. The main source of CO2 in the ocean is the decomposition and subsequent respiration of organic molecules by heterotrophic bacteria. However, very little is known about potential effects of ocean acidification on bacterial degradation activity. This study reveals that the degradation of polysaccharides, a major component of marine organic matter, by bacterial extracellular enzymes was significantly accelerated during experimental simulation of ocean acidification. Results were obtained from pH perturbation experiments, where rates of extracellular α- and β-glucosidase were measured and the loss of neutral and acidic sugars from phytoplankton-derived polysaccharides was determined. Our study suggests that a faster bacterial turnover of polysaccharides at lowered ocean pH has the potential to reduce carbon export and to enhance the respiratory CO2 production in the future ocean.This study was supported by the Helmholtz Association (HZ-NG-102) and the Belgian Science Policy (SD/CS/03)

Effect of CO2 enrichment on bacterial metabolism in an Arctic fjord

he anthropogenic increase of carbon dioxide (CO2) alters the seawater carbonate chemistry, with a decline of pH and an increase in the partial pressure of CO2 (pCO2). Although bacteria play a major role in carbon cycling, little is known about the impact of rising pCO2 on bacterial carbon metabolism, especially for natural bacterial communities. In this study, we investigated the effect of rising pCO2 on bacterial production (BP), bacterial respiration (BR) and bacterial carbon metabolism during a mesocosm experiment performed in Kongsfjorden (Svalbard) in 2010. Nine mesocosms with pCO2 levels ranging from ca. 180 to 1400 μatm were deployed in the fjord and monitored for 30 days. Generally BP gradually decreased in all mesocosms in an initial phase, showed a large (3.6-fold average) but temporary increase on day 10, and increased slightly after inorganic nutrient addition. Over the wide range of pCO2 investigated, the patterns in BP and growth rate of bulk and free-living communities were generally similar over time. However, BP of the bulk community significantly decreased with increasing pCO2 after nutrient addition (day 14). In addition, increasing pCO2 enhanced the leucine to thymidine (Leu : TdR) ratio at the end of experiment, suggesting that pCO2 may alter the growth balance of bacteria. Stepwise multiple regression analysis suggests that multiple factors, including pCO2, explained the changes of BP, growth rate and Leu : TdR ratio at the end of the experiment. In contrast to BP, no clear trend and effect of changes of pCO2 was observed for BR, bacterial carbon demand and bacterial growth efficiency. Overall, the results suggest that changes in pCO2 potentially influence bacterial production, growth rate and growth balance rather than the conversion of dissolved organic matter into CO2

On the effect of low oxygen concentrations on bacterial degradation of sinking particles

In marine oxygen (O2) minimum zones (OMZs), the transfer of particulate organic carbon (POC) to depth via the biological carbon pump might be enhanced as a result of slower remineralisation under lower dissolved O2 concentrations (DO). In parallel, nitrogen (N) loss to the atmosphere through microbial processes, such as denitrification and anammox, is directly linked to particulate nitrogen (PN) export. However it is unclear (1) whether DO is the only factor that potentially enhances POC transfer in OMZs, and (2) if particle fluxes are sufficient to support observed N loss rates. We performed a degradation experiment on sinking particles collected from the Baltic Sea, where anoxic zones are observed. Sinking material was harvested using surface-tethered sediment traps and subsequently incubated in darkness at different DO levels, including severe suboxia (<0.5 mg l−1 DO). Our results show that DO plays a role in regulating POC and PN degradation rates. POC(PN) degradation was reduced by approximately 100% from the high to low DO to the lowest DO. The amount of NH4+ produced from the pool of remineralising organic N matched estimations of NH4+ anammox requirements during our experiment. This anammox was likely fueled by DON degradation rather than PON degradation

Membrane vesicles: Examination of biophysical properties with atomic force microscopy

Extracellular vesicles (EVs) are not only intensively studied to increase our fundamental knowledge on their functioning, but also for diagnosis, therapeutics and drug delivery purposes. To improve the current and potential applications of EVs, a fundamental understanding of their stability, structure, and function is crucial. Such studies can be conducted at the single particle level to gain biological and physical information about the vesicles and the particle to particle variability. A suitable technique to investigate EVs under near- to physiological conditions is atomic force microscopy (AFM). Operated in liquid, it provides images of the EVs while mechanical properties of the particles can be obtained as well. Here we present our approach and the latest results in studying the structure and mechanics of these particles