Background Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as “Large Oncosomes” (LO), have been identified in highly migratory and invasive PCa cells. We recently developed and characterized the DU145R80 subline, selected from parental DU145 cells as resistant to inhibitors of mevalonate pathway. DU145R80 showed different proteomic profile compared to parental DU145 cells, along with altered cytoskeleton dynamics and a more aggressive phenotype. Methods Immunofluorescence staining and western blotting were used to identify blebbing and EVs protein cargo. EVs, purified by gradient ultra-centrifugations, were analyzed by tunable resistive pulse sensing and multi-parametric flow cytometry approach coupled with high-resolution imaging technologies. LO functional effects were tested in vitro by adhesion and invasion assays and in vivo xenograft model in nude mice. Xenograft and patient tumor tissues were analyzed by immunohistochemistry. Results We found spontaneous blebbing and increased shedding of LO from DU145R80 compared to DU145 cells. LO from DU145R80, compared to those from DU145, carried increased amounts of key-molecules involved in PCa progression including integrin alpha V (αV-integrin). By incubating DU145 cells with DU145R80-derived LO we demonstrated that αV-integrin on LO surface was functionally involved in the increased adhesion and invasion of recipient cells, via AKT. Indeed either the pre-incubation of LO with an αV-integrin blocking antibody, or a specific AKT inhibition in recipient cells are able to revert the LO-induced functional effects. Moreover, DU145R80-derived LO also increased DU145 tumor engraftment in a mice model. Finally, we identified αV-integrin positive LO-like structures in tumor xenografts as well as in PCa patient tissues. Increased αV-integrin tumor expression correlated with high Gleason score and lymph node status. Conclusions Overall, this study is the first to demonstrate the critical role of αV-integrin positive LO in PCa aggressive features, adding new insights in biological function of these large EVs and suggesting their potential use as PCa prognostic markers. Electronic supplementary material The online version of this article (10.1186/s13046-019-1317-6) contains supplementary material, which is available to authorized users.
In this work, biodegradable films based on poly (lactic acid) (PLA) and corn thermoplastic starch (TPS), additivated with epoxidized cardoon oil plasticizer (ECO) at 3% by weight with respect to PLA mass fraction, were prepared by melt extrusion process and compression molding. The effect of ECO on structural, thermal, mechanical, barrier, and spectral optical properties of the films was investigated. Spectroscopic analysis evidenced the development of physical interaction between oil and polymers, mainly PLA. In addition, no oil migration occurrence was detected after six months of film preparation, as evidenced by oil mass evaluation by precipitation as well as by 1H-NMR methods, thus highlighting the good inclusion of oil inside the polymeric network. The plasticizing action of the oil induced a lean improvement of the interfacial adhesion between hydrophobic PLA and hydrophilic TPS, particularly accentuated in PLA80_ECO composition, as evidenced by morphological analysis of blend fracture surfaces. TGA data underlined that, differently from TPS-based films, PLA-based systems followed one degradative thermal profile suggesting a slight compatibilization effect of epoxidized oil in these films. The shifting of Tg values, by differential scanning calorimetry (DSC) analysis, indicated a weak miscibility at molecular level. Generally, in the investigated blends, the phase separation between PLA and TPS polymers was responsible for the mechanical properties failing; in particular, the tensile strength evidenced a negative deviation from the rule of mixtures, particularly marked in TPS-based blends, where no physical entanglements occurred between the polymers since their immiscibility even in presence of ECO. The epoxidized oil strongly improved the barrier properties (water vapor permeability (WVP) and oxygen permeability (O2P)) of all the films, likely developing a physical barrier to water and oxygen diffusion and solubilization. With respect to neat PLA, PL80 and PL80_ECO films evidenced the improvement of surface wettability, due to the presence of polar groups both in TPS (hydroxyl residues) and in epoxidized oil (oxirane rings). Finally, following to the conditioning in climatic chamber at T = 25 °C and RH = 50%, PLA80 film became opaque due to TPS water absorption, causing a light transmittance decreasing, as evidenced by spectral optical analysis.
An inverted pyramidal metasurface was designed, fabricated, and studied at the nanoscale level for the development of a label-free pathogen detection on a chip platform that merges nanotechnology and surface-enhanced Raman scattering (SERS). Based on the integration and synergy of these ingredients, a virus immunoassay was proposed as a relevant proof of concept for very sensitive detection of hepatitis A virus, for the first time to our best knowledge, in a very small volume (2 μL), without complex signal amplification, allowing to detect a minimal virus concentration of 13 pg/mL. The proposed work aims to develop a high-flux and high-accuracy surface-enhanced Raman spectroscopy (SERS) nanobiosensor for the detection of pathogens to provide an effective method for early and easy water monitoring, which can be fast and convenient.
Nonlinear chiroptical effects of precisely designed chiral plasmonic nanomaterials can be much stronger than such effects observed in the linear regime. We take advantage of this property to demonstrate the use of circularly polarized second-harmonic generation microscopy towards the efficient read-out of a microscopic pattern encoded by an array of triangular gold nanoprisms forming arrangements of adequate chiral symmetry. Strong chiroptical effects are observed in the backscattered second-harmonic generation intensity, enabling clear distinction of the laterally arranged enantiomers, down to nearly 1 µm resolution with an overall intensity contrast of about 40 % (second-harmonic generation circular dichroism of 20 %). Numerical simulations show a noticeable change in the spatial distribution of plasmonic hot spots within the individual nanostructures under excitation by circularly polarized light of different handedness. This leads to rather weak chiroptical effect in the linear backscattering (theoretical circular dichroism not exceeding 3 %), in contrast with the much more significant change of the second-harmonic generation in the far-field (secondharmonic generation circular dichroism from 16 up to 37%). These results open the possibility of designing deeply subwavelength chiral nanostructures for encoding microscopic "watermarks", which cannot be easily accessed by linear optical methods, moreover requiring a nonlinear microscopy set-up for reading out the encoded pattern. Graphic Over the past two decades, research devoted to the second-harmonic generation (SHG) from plasmonic nanostructures evolved from the investigation of hyper-Rayleigh scattering in colloidal suspensions of metal nanoparticles [1-3] towards the measurements of SHG from individual plasmonic nano-objects [4][5][6]. Far-field SHG radiation pattern has been demonstrated as a highly sensitive probe of the nanoscale optical field distribution [7], allowing for the investigation of multipolar plasmon modes [8][9][10], the analysis of surface plasmon polariton (SPP)-mediated interactions [11,12], refractive index sensing [13] and precise characterization of the nanostructure geometry [14][15][16]. In parallel, advanced theoretical tools for SHG modelling were developed, based on numerical methods such as finite elements to solve Maxwell equations and yield the scattered SHG radiation [17][18][19]. Among the geometrical properties that can be addressed by SHG microscopy, chirality stands-out as one of the most interesting probes of light-matter interactions, especially in the context of chiral plasmonic meta-materials which have been reported to exhibit giant optical activity in the linear regime [20,21]. Valev et al. have thus reported extensive work on chirality in SHG from specially patterned superchiral meta-surfaces [7,[22][23][24]. The location of plasmonic hot spots and their influence on the chiroptical response have been thoroughly addressed [22,23], leading to reports of nonlinear chiroptical effects reaching magnitudes as high as 5...
The development of fast and ultrasensitive methods to detect bacterial pathogens at low concentrations is of high relevance for human and animal health care and diagnostics. In this context, surface-enhanced Raman scattering (SERS) offers the promise of a simplified, rapid, and high-sensitive detection of biomolecular interactions with several advantages over previous assay methodologies. In this work, we have conceived reproducible SERS nanosensors based on tailored multilayer octupolar nanostructures which can combine high enhancement factor and remarkable molecular selectivity. We show that coating novel multilayer octupolar metastructures with proper self-assembled monolayer (SAM) and immobilized phages can provide label-free analysis of pathogenic bacteria via SERS leading to a giant increase in SERS enhancement. The strong relative intensity changes of about 2100% at the maximum scattered SERS wavelength, induced by the Brucella bacterium captured, demonstrate the performance advantages of the bacteriophage sensing scheme. We performed measurements at the single-cell level thus allowing fast identification in less than an hour without any demanding sample preparation process. Our results based on designing well-controlled octupolar coupling platforms open up new opportunities toward the use of bacteriophages as recognition elements for the creation of SERS-based multifunctional biochips for rapid culture and label-free detection of bacteria.
Photomobile polymer (Pmp) films are fabricated by using a cheap and fast process. The working mechanism of the Pmp-film motion under illumination is explained. Details concerning the film structure and formation are given. Two related applications regarding light-induced caterpillar-miming motion and photocontrolled electrical switches are proposed.
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