Claudio Canale

Colloidal Synthesis of Quantum Confined Single Crystal CsPbBr3 Nanosheets with Lateral Size Control up to the Micrometer Range

We report the nontemplated colloidal synthesis of single crystal CsPbBr3 perovskite nanosheets with lateral sizes up to a few micrometers and with thickness of just a few unit cells (i.e., below 5 nm), hence in the strong quantum confinement regime, by introducing short ligands (octanoic acid and octylamine) in the synthesis together with longer ones (oleic acid and oleylamine). The lateral size is tunable by varying the ratio of shorter ligands over longer ligands, while the thickness is mainly unaffected by this parameter and stays practically constant at 3 nm in all the syntheses conducted at short-to-long ligands volumetric ratio below 0.67. Beyond this ratio, control over the thickness is lost and a multimodal thickness distribution is observed.

Collagen Plays an Active Role in the Aggregation of β2-Microglobulin under Physiopathological Conditions of Dialysis-related Amyloidosis

Dialysis-related amyloidosis is characterized by the deposition of insoluble fibrils ofβ2-microglobulin (β2-m) in the musculoskeletal system. Atomic force microscopy inspection of ex vivo amyloid material reveals the presence of bundles of fibrils often associated to collagen fibrils. Aggregation experiments were undertaken in vitro with the aim of reproducing the physiopathological fibrillation process. To this purpose, atomic force microscopy, fluorescence techniques, and NMR were employed. We found that in temperature and pH conditions similar to those occurring in periarticular tissues in the presence of flogistic processes, β2-m fibrillogenesis takes place in the presence of fibrillar collagen, whereas no fibrils are obtained without collagen. Moreover, the morphology ofβ2-m fibrils obtained in vitro in the presence of collagen is extremely similar to that observed in the ex vivo sample. This result indicates that collagen plays a crucial role in β2-m amyloid deposition under physiopathological conditions and suggests an explanation for the strict specificity of dialysis-related amyloidosis for the tissues of the skeletal system. We hypothesize that positively charged regions along the collagen fiber could play a direct role inβ2-m fibrillogenesis. This hypothesis is sustained by aggregation experiments performed by replacing collagen with a poly-l-lysine-coated mica surface. As shown by NMR measurements, no similar process occurs when poly-l-lysine is dissolved in solution with β2-m. Overall, the findings are consistent with the estimates resulting from a simplified collagen model whereby electrostatic effects can lead to high local concentrations of oppositely charged species, such as β2-m, that decay on moving away from the fiber surface.

Colloidal Synthesis of Strongly Fluorescent CsPbBr3 Nanowires with Width Tunable down to the Quantum Confinement Regime

We report the colloidal synthesis of strongly fluorescent CsPbBr3 perovskite nanowires (NWs) with rectangular section and with tuneable width, from 20 nm (exhibiting no quantum confinement, hence emitting in the green) down to around 3 nm (in the strong quan-tum-confinement regime, emitting in the blue), by introducing in the synthesis a short acid (octanoic acid or hexanoic acid) together with alkyl amines (octylamine and oleylamine). Temperatures below 70 {\deg}C promoted the formation of monodisperse, few unit cell thick NWs that were free from byproducts. The photoluminescence quantum yield of the NW samples went from 12% for non-confined NWs emitting at 524 nm to a maximum of 77% for the 5 nm diameter NWs emitting at 497 nm, down to 30% for the thinnest NWs (diameter ~ 3nm), in the latter sample most likely due to aggregation occurring in solution.

A large genomic deletion leads to enhancer adoption by the lamin B1 gene: a second path to autosomal dominant adult-onset demyelinating leukodystrophy (ADLD).

Chromosomal rearrangements with duplication of the lamin B1 (LMNB1) gene underlie autosomal dominant adult-onset demyelinating leukodystrophy (ADLD), a rare neurological disorder in which overexpression of LMNB1 causes progressive central nervous system demyelination. However, we previously reported an ADLD family (ADLD-1-TO) without evidence of duplication or other mutation in LMNB1 despite linkage to the LMNB1 locus and lamin B1 overexpression. By custom array-CGH, we further investigated this family and report here that patients carry a large (∼660 kb) heterozygous deletion that begins 66 kb upstream of the LMNB1 promoter. Lamin B1 overexpression was confirmed in further ADLD-1-TO tissues and in a postmortem brain sample, where lamin B1 was increased in the frontal lobe. Through parallel studies, we investigated both loss of genetic material and chromosomal rearrangement as possible causes of LMNB1 overexpression, and found that ADLD-1-TO plausibly results from an enhancer adoption mechanism. The deletion eliminates a genome topological domain boundary, allowing normally forbidden interactions between at least three forebrain-directed enhancers and the LMNB1 promoter, in line with the observed mainly cerebral localization of lamin B1 overexpression and myelin degeneration. This second route to LMNB1 overexpression and ADLD is a new example of the relevance of regulatory landscape modifications in determining Mendelian phenotypes.

A novel nanoscopic tool by combining AFM with STED microscopy

We present a new instrument for nanoscopic investigations by coupling an atomic force microscope (AFM) with a super resolution stimulated emission depletion (STED) microscope. This nanoscopic tool allows high resolution fluorescence imaging, topographical imaging and nano-mechanical imaging, such as, stiffness. Results obtained from technical and biological samples are shown illustrating different functions and the versatility of the presented tool. We assert that, this highly precise tractable tool paves the way to a new set of comprehensive studies in medicine, biology and materials science.

Effects of barium titanate nanoparticles on proliferation and differentiation of rat mesenchymal stem cells

Nanomaterials hold great promise in the manipulation and treatments of mesenchymal stem cells, since they allow the modulation of their properties and differentiation. However, systematic studies have to be carried out in order to assess their potential toxicological effects. The present study reports on biocompatibility evaluation of glycol-chitosan coated barium titanate nanoparticles (BTNPs) on rat mesenchymal stem cells (MSCs). BTNPs are a class of ceramic systems which possess interesting features for biological applications thanks to their peculiar dielectric and piezoelectric properties. Viability was evaluated up to 5 days of incubation (concentrations in the range 0-100 μg/ml) both quantitatively and qualitatively with specific assays. Interactions cells/nanoparticles were further investigated with analysis of the cytoskeleton conformation, with SEM and TEM imaging, and with AFM analysis. Finally, differentiation in adipocytes and osteocytes was achieved in the presence of high doses of BTNPs, thus highlighting the safety of these nanostructures towards mesenchymal stem cells.

Nanoscale structural and mechanical effects of beta-amyloid (1-42) on polymer cushioned membranes: a combined study by neutron reflectometry and AFM Force Spectroscopy.

The interaction of beta-amyloid peptides with lipid membranes is widely studied as trigger agents in Alzheimers disease. Their mechanism of action at the molecular level is unknown and their interaction with the neural membrane is crucial to elucidate the onset of the disease. In this study we have investigated the interaction of water soluble forms of beta-amyloid Aβ(1-42) with lipid bilayers supported by polymer cushion. A reproducible protocol for the preparation of a supported phospholipid membrane with composition mimicking the neural membrane and in physiological condition (PBS buffer, pH=7.4) was refined by neutron reflectivity. The change in structure and local mechanical properties of the membrane in the presence of Aβ(1-42) was investigated by neutron reflectivity and Atomic Force Microscopy (AFM) Force Spectroscopy. Neutron reflectivity evidenced that Aβ(1-42) interacts strongly with the supported membrane, causing a change in the scattering length density profile of the lipid bilayer, and penetrates into the membrane. Concomitantly, the local mechanical properties of the bilayer are deeply modified by the interaction with the peptide as seen by AFM Force Spectroscopy. These results may be of great importance for the onset of the Alzheimers disease, since a simultaneous change in the structural and mechanical properties of the lipid matrix could influence all membrane based signal cascades.

Inhibiting effect of αs1-casein on Aβ1–40 fibrillogenesis

BACKGROUND α(s1)-Casein is one of the four types of caseins, the largest protein component of bovine milk. The lack of a compact folded conformation and the capability to form micelles suggest a relationship of α(s1)-casein with the class of the intrinsically disordered (or natively unfolded) proteins. These proteins are known to exert a stabilizing activity on biomolecules through specific interaction with hydrophobic surfaces. In the present work we focused on the effect of α(s1)-casein on the fibrillogenesis of 1-40 β-amyloid peptide, involved in Alzheimers disease. METHODS The aggregation kinetics of β-peptide in presence and absence of α(s1)-casein was followed under shear at 37°C by recording the Thioflavine fluorescence, usually taken as an indicator of fibers formation. Measurements of Static and Dynamic Light Scattering, Circular Dichroism, and AFM imaging were done to reveal the details of α(s1)-casein-Aβ(1-40) interaction. RESULTS AND DISCUSSIONS α(s1)-Casein addition sizably increases the lag-time of the nucleation phase and slows down the entire fibrillization process. α(s1)-Casein sequesters the amyloid peptide on its surface thus exerting a chaperone-like activity by means a colloidal inhibition mechanism. GENERAL SIGNIFICANCE Insights on the working mechanism of natural chaperones in preventing or controlling the amyloid aggregation.

Force spectroscopy as a tool to investigate the properties of supported lipid membranes.

Solid supported lipid bilayers (SLB) are extensively used as a model for the investigation of cell membranes in a variety of spectroscopic and biophysical methods. It is nevertheless well known that the interaction with the solid substrate, such as mica or silicon, influences the properties of the membranes. In this article we have employed atomic force microscopy (AFM) in force spectroscopy mode (FS) to investigate the local mechanical properties of lipid membranes supported on mica and on polymer cushion. The lipid double layers were obtained by fusion of unilamellar vesicle of phospholipids. The polymer support was created by self‐assembly of charged polyelectrolytes. Force spectroscopy provided information about the breakthrough force, the breakthrough depth, and the sample adhesion. A batch analysis algorithm to process high‐resolution force mapping was developed. The breakthrough force to indent the bilayers down to the support and the adhesion force were measured as a function of the membrane charge. The comparison of the data obtained from SLB on mica and from bilayers on polymer cushion provides direct evidence about the influence of the substrate on the local mechanic properties of the membrane. As a major result, the yield force distribution of membranes on polymer cushion was bimodal, compared to the unimodal distribution obtained on mica. Microsc. Res. Tech. 73:965–972, 2010.

Sub-diffraction nano manipulation using STED AFM.

In the last two decades, nano manipulation has been recognized as a potential tool of scientific interest especially in nanotechnology and nano-robotics. Contemporary optical microscopy (super resolution) techniques have also reached the nanometer scale resolution to visualize this and hence a combination of super resolution aided nano manipulation ineluctably gives a new perspective to the scenario. Here we demonstrate how specificity and rapid determination of structures provided by stimulated emission depletion (STED) microscope can aid another microscopic tool with capability of mechanical manoeuvring, like an atomic force microscope (AFM) to get topological information or to target nano scaled materials. We also give proof of principle on how high-resolution real time visualization can improve nano manipulation capability within a dense sample, and how STED-AFM is an optimal combination for this job. With these evidences, this article points to future precise nano dissections and maybe even to a nano-snooker game with an AFM tip and fluorospheres.