Optimized pinhole geometry for small-angle scattering

In order to make the most of the low photon flux in laboratory small-angle X-ray scattering instruments, the experimental geometry has to be chosen carefully, with special stress on the beam-shaping system. The optimum collimation scheme should enable accurate measurements over the desired range of the scattering variable, while yielding the highest flux and the lowest possible instrumental background. In order to identify the best setting, a phase-space optimization of the collimating scheme is carried out in the present work, including constraints on the beam size at the sample position and on the detector surface. The resulting formulae are implemented in a Python script with a graphical user interface, to aid the planning, construction and daily operation of pinhole small-angle scattering cameras

Osmotic shrinkage of sterically stabilized liposomes as revealed by time-resolved small-angle X-ray scattering

Time-resolved synchrotron small-angle X-ray scattering (SAXS) was used to study the structural changes during the osmotic shrinkage of a pharmacologi- cally relevant liposomal drug delivery system. Sterically stabilized liposomes (SSLs) with a diameter of 100nm and composed of hydrogenated soy phosphocholine, cholesterol and distearoyl-phosphoethanolamine-PEG 2000 prepared in a salt-free buffer were mixed with a buffered 0.3 M NaCl solution using a stopped flow apparatus. The changes in the liposome size and the bilayer structure were followed by using SAXS with a time resolution of 20 ms. A linear decrease in liposome size is observed during the firs

Effects of ursolic acid on the structural and morphological behaviours of dipalmitoyl lecithin vesicles.

Effects of ursolic acid on the structural and morphological characteristics of dipalmitoyl lecithin(DPPC)-water system was studied by using differential scanning calorimetry (DSC), small- and wide-angle X-ray scattering (SWAXS), freeze-fracture method combined with transmission electron-microscopy (FF-TEM) and infrared spectroscopy (FT-IR). The surface of the uncorrelated lipid system is rippled or grained and a huge number of small, presumably unilamellar vesicles are present if the UA/DPPC molar ratio is 0.1mol/mol or higher. Besides the destroyed layer packing of regular multilamellar vesicles, non-bilayer (e.g. cubic or hexagonal) local structures are evidenced by SAXS and FF-TEM methods. The ability of UA to induce non-bilayer structures in hydrated DPPC system originates from the actual geometry form of associated lipid and UA molecules as concluded from the FT-IR measurements and theoretical calculations. Beside numerous beneficial e.g. chemopreventive and chemotherapeutic effect of ursolic acid against cancer, their impact to modify the lipid bilayers can be utilized in liposomal formulations

charmm2gmx: An Automated Method to Port the CHARMM Additive Force Field to GROMACS

CHARMM is one of the most widely used biomolecular force fields. Although developed in close connection with a dedicated molecular simulation engine of the same name, it is also usable with other codes. GROMACS is a well-established, highly optimized, and multipurpose software for molecular dynamics, versatile enough to accommodate many different force field potential functions and the associated algorithms. Due to conceptional differences related to software design and the large amount of numeric data inherent to residue topologies and parameter sets, conversion from one software format to another is not straightforward. Here, we present an automated and validated means to port the CHARMM force field to a format read by the GROMACS engine, harmonizing the different capabilities of the two codes in a self-documenting and reproducible way with a bare minimum of user interaction required. Being based entirely on the upstream data files, the presented approach does not involve any hard-coded data, in contrast with previous attempts to solve the same problem. The heuristic approach used for perceiving the local internal geometry is directly applicable for analogous transformations of other force fields

A mechanistic view of lipid membrane disrupting effect of PAMAM dendrimers

The effect of 5th generation polyamidoamine (PAMAM G5) dendrimers on multilamellar dipalmitoylphosphocholine (DPPC) vesicles was investigated. PAMAM was added in two different concentration to the lipids (10-3 and 10-2 dendrimer/lipid molar ratios). The thermal behavior of the evolved systems was characterized by DSC; while the structure and the morphology were investigated with small- and wide-angel X-ray scattering (SWAXS), freeze-fracture electron microscopy (FFTEM) and phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy, respectively. IR spectroscopy was used to study the molecular interactions between PAMAM and DPPC. The obtained results show that the dendrimers added in 10-3 molar ratio to the lipids generate minor perturbations in the multilamellar structure and thermal character of liposomes, while added in 10-2 molar ratio dendrimers cause major disturbance in the vesicular system. The terminal amino groups of the dendrimers are in strong interaction with the phosphate headgroups and through this binding dendrimers disrupt the regular multilamellar structure of DPPC. Besides highly swollen, fragmented bilayers, small vesicles are formed

Dispersion and stabilization of cochleate nanoparticles

Cochleates, calcium-stabilized membrane rolls of nanoscale diameter, promise a unique and efficient way of delivering lipid-soluble drugs, proteins or nucleic acids into biological systems because they protect the encapsulated material against enzymatic or chemical degradation. Self-aggregation, which typically arises during production and storage is a major obstacle that has so far precluded the development of an efficient cochleate-based drug-delivery system. Here we show that citric acid, added transiently in a narrow concentration range, effectively disperses cochleate aggregates, stabilizes the disperse state for long-term storage and preserves the canonical ultrastructure and topological characteristics of cochleate nanoparticles

Structure and Function of Trypsin-Loaded Fibrinolytic Liposomes

Protease encapsulation and its targeted release in thrombi may contribute to the reduction of haemorrhagic complications of thrombolysis. We aimed to prepare sterically stabilized trypsin-loaded liposomes () and characterize their structure and fibrinolytic efficiency. Hydrogenated soybean phosphatidylcholine-based were prepared and their structure was studied by transmission electron microscopy combined with freeze fracture (FF-TEM), Fourier transform infrared spectroscopy (FT-IR), and small-angle X-ray scattering (SAXS). Fibrinolytic activity was examined at 45, 37, or 24°C on fibrin or plasma clots with turbidimetric and permeation-driven lysis assays. Trypsin was shown to be attached to the inner surface of vesicles (SAXS and FF-TEM) close to the lipid hydrophilic/hydrophobic interface (FT-IR). The thermosensitivity of was evidenced by enhanced fibrinolysis at 45°C: time to reduce the maximal turbidity to 20% decreased by 8.6% compared to 37°C and fibrin degradation product concentration in the permeation lysis assay was 2-fold to 5-fold higher than that at 24°C. exerted its fibrinolytic action on fibrin clots under both static and dynamic conditions, whereas plasma clot dissolution was observed only in the permeation-driven assay. The improved fibrinolytic efficiency of under dynamic conditions suggests that they may serve as a novel therapeutic candidate for dissolution of intravascular thrombi, which are typically exposed to permeation forces