Using solution X-ray scattering to determine the high-resolution structure and morphology of PEGylated liposomal doxorubicin nanodrugs.

BACKGROUND: Among nanodrugs, PEGylated nanoliposomes loaded with an active agent are of major importance. In this paper we studied the structures and morphology of PEGylated nanoliposomes before and after remote loading with doxorubicin. METHODS: High-resolution structures were obtained by solution X-ray scattering combined with our advanced analysis tools. We studied the PEGylated liposomal doxorubicin (PLD) product Doxil(®), and its generics, where remote doxorubicin loading is performed by a gradient of ammonium sulfate, and LC100, a novel PLD under development, where remote loading was done by a gradient of ammonium methanesulfonate. The PLD structures were compared with drug-free nanoliposomes having identical composition. RESULTS: We determined the membrane electron density profiles of the empty and loaded PLDs, the thickness and density of the PEG layers, and the structure of the drug inside the liposomes. CONCLUSIONS: The liposomal membranes had the same structure for both ammonium salts. We found that the drug formed crystals inside PLDs loaded by ammonium sulfate, whereas it had an amorphous morphology in the PLD loaded by ammonium methanesulfonate. The variations of the drug's structural parameters between the generics of Doxil(®) are similar to the variations between batches of the same product, suggesting that all these products were structurally similar. GENERAL SIGNIFICANCE: This paper demonstrates that solution X-ray scattering, when combined with our powerful analysis tools, can determine the high-resolution structure of complex non-crystallized nanoparticle dispersions used in nanomedicine, thereby providing useful physical insights into their functions.

Thermal doping by vacancy formation in copper sulfide nanocrystal arrays.

A new approach for doping of Cu2S nanocrystal arrays using thermal treatment at moderate temperatures (T < 400 K) is presented. This thermal doping process yields conductance enhancement by 6 orders of magnitude. Local probe measurements prove this doping is an intraparticle effect and, moreover, tunneling spectroscopy data signify p-type doping. The doping mechanism is attributed to Cu vacancy formation, resulting in free holes. Thermal-doping temperature dependence exhibits an Arrhenius-like behavior, providing the vacancy formation energy of 1.6 eV. The moderate temperature conditions for thermal doping unique to these nanocrystals allow patterned doping of nanocrystal films through local heating by a focused laser beam, toward fabrication of nanocrystal-based electronic devices.

Fine Structural Analysis of Degummed Fibroin Fibers Reveals Its Superior Mechanical Capabilities.

Bombyx mori silk fibroin fibers constitute a class of protein building blocks capable of functionalization and reprocessing into various material formats. The properties of these fibers are typically affected by the intense thermal treatments needed to remove the sericin gum coating layer. Additionally, their mechanical characteristics are often misinterpreted by assuming the asymmetrical cross-sectional area (CSA) as a perfect circle. The thermal treatments impact not only the mechanics of the degummed fibroin fibers, but also the structural configuration of the resolubilized protein, thereby limiting the performance of the resulting silk-based materials. To mitigate these limitations, we explored varying alkali conditions at low temperatures for surface treatment, effectively removing the sericin gum layer while preserving the molecular structure of the fibroin protein, thus, maintaining the hierarchical integrity of the exposed fibroin microfiber core. The precise determination of the initial CSA of the asymmetrical silk fibers led to a comprehensive analysis of their mechanical properties. Our findings indicate that the alkali surface treatment raised the Young's modulus and tensile strength, by increasing the extent of the fibers' crystallinity, by approximately 40 % and 50 %, respectively, without compromising their strain. Furthermore, we have shown that this treatment facilitated further production of high-purity soluble silk protein with rheological and self-assembly characteristics comparable to those of native silk feedstock, initially stored in the animal's silk gland. The developed approaches benefits both the development of silk-based materials with tailored properties and the proper mechanical characterization of asymmetrical fibrous biological materials made of natural building blocks.

Micro and nano-scale compartments guide the structural transition of silk protein monomers into silk fibers.

Silk is a unique, remarkably strong biomaterial made of simple protein building blocks. To date, no synthetic method has come close to reproducing the properties of natural silk, due to the complexity and insufficient understanding of the mechanism of the silk fiber formation. Here, we use a combination of bulk analytical techniques and nanoscale analytical methods, including nano-infrared spectroscopy coupled with atomic force microscopy, to probe the structural characteristics directly, transitions, and evolution of the associated mechanical properties of silk protein species corresponding to the supramolecular phase states inside the silkworm's silk gland. We found that the key step in silk-fiber production is the formation of nanoscale compartments that guide the structural transition of proteins from their native fold into crystalline ß-sheets. Remarkably, this process is reversible. Such reversibility enables the remodeling of the final mechanical characteristics of silk materials. These results open a new route for tailoring silk processing for a wide range of new material formats by controlling the structural transitions and self-assembly of the silk protein's supramolecular phases.

Human microtubule-associated-protein tau regulates the number of protofilaments in microtubules: a synchrotron x-ray scattering study.

Microtubules (MTs), a major component of the eukaryotic cytoskeleton, are 25 nm protein nanotubes with walls comprised of assembled protofilaments built from alphabeta heterodimeric tubulin. In neural cells, different isoforms of the microtubule-associated-protein (MAP) tau regulate tubulin assembly and MT stability. Using synchrotron small angle x-ray scattering (SAXS), we have examined the effects of all six naturally occurring central nervous system tau isoforms on the assembly structure of taxol-stabilized MTs. Most notably, we found that tau regulates the distribution of protofilament numbers in MTs as reflected in the observed increase in the average radius R(MT) of MTs with increasing Phi, the tau/tubulin-dimer molar ratio. Within experimental scatter, the change in R(MT) seems to be isoform independent. Significantly, R(MT) was observed to rapidly increase for 0 < Phi < 0.2 and saturate for Phi between 0.2-0.5. Thus, a local shape distortion of the tubulin dimer on tau binding, at coverages much less than a monolayer, is spread collectively over many dimers on the scale of protofilaments. This implies that tau regulates the shape of protofilaments and thus the spontaneous curvature C(o)(MT) of MTs leading to changes in the curvature C(MT) (=1/R(MT)). An important biological implication of these findings is a possible allosteric role for tau where the tau-induced shape changes of the MT surface may effect the MT binding activity of other MAPs present in neurons. Furthermore, the results, which provide insight into the regulation of the elastic properties of MTs by tau, may also impact biomaterials applications requiring radial size-controlled nanotubes.

The conservative management of vesicoureteric reflux: a review of 121 children.

A group of 121 children with vesicoureteric reflux (VUR) grades 1 to 3 managed conservatively were followed-up for a period of six to ten years. In the majority of patients VUR grades 1 and 2 disappeared spontaneously. Prognosis was less favorable in those who were seen initially with grade 3 VUR. A statistically significant higher incidence of reflux disappearance was observed in children who were seen before 4 years of age. In the majority of patients with paraureteric-vesicle diverticulum, reflux persisted. Reflux was more likely to disappear in children with lower incidence of urinary tract infection than in those with multiple infections.

Fluidity of water confined to subnanometre films.

The fluidity of water in confined geometries is relevant to processes ranging from tribology to protein folding, and its molecular mobility in pores and slits has been extensively studied using a variety of approaches. Studies in which liquid flow is measured directly suggest that the viscosity of aqueous electrolytes confined to films of thickness greater than about 2-3 nm remains close to that in the bulk; this behaviour is similar to that of non-associative organic liquids confined to films thicker than about 7-8 molecular layers. Here we observe that the effective viscosity of water remains within a factor of three of its bulk value, even when it is confined to films in the thickness range 3.5 +/- 1 to 0.0 +/- 0.4 nm. This contrasts markedly with the behaviour of organic solvents, whose viscosity diverges when confined to films thinner than about 5-8 molecular layers. We attribute this to the fundamentally different mechanisms of solidification in the two cases. For non-associative liquids, confinement promotes solidification by suppressing translational freedom of the molecules; however, in the case of water, confinement seems primarily to suppress the formation of the highly directional hydrogen-bonded networks associated with freezing.

The polymer mat: arrested rebound of a compressed polymer layer.

Compression of an adsorbed polymer layer distorts its relaxed structure. Surface force measurements from different laboratories show that the return to this relaxed structure after the compression is released can require tens of minutes and that the recovery time can grow rapidly with molecular weight. We argue that the arrested state of the free layer before relaxation can be described as a Guiselin brush structure (O. Guiselin, Europhys. Lett. 17, 225 (1992)), in which the monomer density falls off only weakly with distance from the surface. This brush structure predicts an exponential falloff of the force at large distance with a decay length that varies as the initial compression distance to the 6/5 power. This exponential falloff is consistent with surface force measurements. We propose a relaxation mechanism that accounts for the increase in relaxation time with chain length.

Hypothyroidism due to ectopy in siblings.

Two families each had two siblings with hypothyroidism due to ectopy and hypoplasia of the thyroid. A genetic factor controlling normal thyroid development and descent may be responsible, and the importance of plasma thyroid-stimulating hormone estimation in borderline hypothyroid cases is emphasized. We suggest screening of siblings of patients with ectopic thyroid for hypothyroidism.