Micro-RNA and Proteomic Profiles of Plasma-Derived Exosomes from Irradiated Mice Reveal Molecular Changes Preventing Apoptosis in Neonatal Cerebellum

Cell communication via exosomes is capable of influencing cell fate in stress situations such as exposure to ionizing radiation. In vitro and in vivo studies have shown that exosomes might play a role in out-of-target radiation effects by carrying molecular signaling mediators of radiation damage, as well as opposite protective functions resulting in resistance to radiotherapy. However, a global understanding of exosomes and their radiation-induced regulation, especially within the context of an intact mammalian organism, has been lacking. In this in vivo study, we demonstrate that, compared to sham-irradiated (SI) mice, a distinct pattern of proteins and miRNAs is found packaged into circulating plasma exosomes after whole-body and partial-body irradiation (WBI and PBI) with 2 Gy X-rays. A high number of deregulated proteins (59% of WBI and 67% of PBI) was found in the exosomes of irradiated mice. In total, 57 and 13 miRNAs were deregulated in WBI and PBI groups, respectively, suggesting that the miRNA cargo is influenced by the tissue volume exposed to radiation. In addition, five miRNAs (miR-99b-3p, miR-200a-3p, miR-200a, miR-182-5p, miR-182) were commonly overexpressed in the exosomes from the WBI and PBI groups. In this study, particular emphasis was also given to the determination of the in vivo effect of exosome transfer by intracranial injection in the highly radiosensitive neonatal cerebellum at postnatal day 3. In accordance with a major overall anti-apoptotic function of the commonly deregulated miRNAs, here, we report that exosomes from the plasma of irradiated mice, especially in the case of WBI, prevent radiation-induced apoptosis, thus holding promise for exosome-based future therapeutic applications against radiation injury.     

Microgravity Modifies the Phenotype of Fibroblast and Promotes Remodeling of the Fibroblast–Keratinocyte Interaction in a 3D Co-Culture Model

Microgravity impairs tissue organization and critical pathways involved in the cell–microenvironment interplay, where fibroblasts have a critical role. We exposed dermal fibroblasts to simulated microgravity by means of a Random Positioning Machine (RPM), a device that reproduces conditions of weightlessness. Molecular and structural changes were analyzed and compared to control samples growing in a normal gravity field. Simulated microgravity impairs fibroblast conversion into myofibroblast and inhibits their migratory properties. Consequently, the normal interplay between fibroblasts and keratinocytes were remarkably altered in 3D co-culture experiments, giving rise to several ultra-structural abnormalities. Such phenotypic changes are associated with down-regulation of α-SMA that translocate in the nucleoplasm, altogether with the concomitant modification of the actin-vinculin apparatus. Noticeably, the stress associated with weightlessness induced oxidative damage, which seemed to concur with such modifications. These findings disclose new opportunities to establish antioxidant strategies that counteract the microgravity-induced disruptive effects on fibroblasts and tissue organization.     

Secretome from Human Mesenchymal Stem Cells-Derived Endothelial Cells Promotes Wound Healing in a Type-2 Diabetes Mouse Model

Tissue regeneration is often impaired in patients with metabolic disorders such as diabetes mellitus and obesity, exhibiting reduced wound repair and limited regeneration capacity. We and others have demonstrated that wound healing under normal metabolic conditions is potentiated by the secretome of human endothelial cell-differentiated mesenchymal stem cells (hMSC-EC). However, it is unknown whether this effect is sustained under hyperglycemic conditions. In this study, the wound healing effect of secretomes from undifferentiated human mesenchymal stem cells (hMSC) and hMSC-EC in a type-2 diabetes mouse model was analyzed. hMSC were isolated from human Wharton’s jelly and differentiated into hMSC-EC. hMSC and hMSC-EC secretomes were analyzed and their wound healing capacity in C57Bl/6J mice fed with control (CD) or high fat diet (HFD) was evaluated. Our results showed that hMSC-EC secretome enhanced endothelial cell proliferation and wound healing in vivo when compared with hMSC secretome. Five soluble proteins (angiopoietin-1, angiopoietin-2, Factor de crecimiento fibroblástico, Matrix metallopeptidase 9, and Vascular Endothelial Growth Factor) were enriched in hMSC-EC secretome in comparison to hMSC secretome. Thus, the five recombinant proteins were mixed, and their pro-healing property was evaluated in vitro and in vivo. Functional analysis demonstrated that a cocktail of these proteins enhanced the wound healing process similar to hMSC-EC secretome in HFD mice. Overall, our results show that hMSC-EC secretome or a combination of specific proteins enriched in the hMSC-EC secretome enhanced wound healing process under hyperglycemic conditions.     

Immunotherapy for Biliary Tract Cancer in the Era of Precision Medicine: Current Knowledge and Future Perspectives

Biliary tract cancers (BTC) represent a heterogeneous and aggressive group of tumors with dismal prognosis. For a long time, BTC has been considered an orphan disease with very limited therapeutic options. In recent years a better understanding of the complex molecular landscape of biology is rapidly changing the therapeutic armamentarium. However, while 40–50% of patients there are molecular drivers susceptible to target therapy, for the remaining population new therapeutic options represent an unsatisfied clinical need. The role of immunotherapy in the continuum of treatment of patients with BTC is still debated. Despite initial signs of antitumor-activity, single-agent immune checkpoint inhibitors (ICIs) demonstrated limited efficacy in an unselected population. Therefore, identifying the best partner to combine ICIs and predictive biomarkers represents a key challenge to optimize the efficacy of immunotherapy. This review provides a critical analysis of completed trials, with an eye on future perspectives and possible biomarkers of response.     

Synthesis,Computational Studies and Assessment of in Vitro Activity of Squalene Derivatives as Carbonic Anhydrase Inhibitors

We report novel molecules incorporating the nontoxic squalene scaffold and different carbonic anhydrase inhibitors (CAIs). Potent inhibitory action, in the low-nanomolar range, was detected against isoforms hCA II for sulfonamide derivatives, which proved to be selective against this isoform over the tumor-associate hCA IX and XII isoforms. On the other hand, coumarin derivatives showed weak potency but high selectivity against the tumor-associated isoform CA IX. These compounds are interesting candidates for preclinical evaluation in glaucoma or various tumors in which the two enzymes are involved. In addition, an in silico study of inhibitor-bound hCA II revealed extensive interactions with the hydrophobic pocket of the active site and provided molecular insights into the binding properties of these new inhibitors.     

Semicontinuous heterophase copolymerization of vinyl acetate and butyl acrylate

Vinyl acetate (VAc) and butyl acrylate (BuA) were copolymerized in heterophase by a semicontinuous process (unseeded) and compared with the seeded semicontinuous microemulsion polymerization of the same monomers. A mixture of sodium dodecyl sulfate (SDS) and poly(ethylene oxide) dodecyl ether (Brij‐35®) were used as surfactants. The effects of monomer addition rate (R_a) and surfactants concentrations (4 or 1 wt % with respect to the initial mixture of reaction) on polymer and latex properties were studied. High copolymer content latexes (24–36 wt %) with average particle diameters (D_p) from 38 to 55 nm and relatively narrow particle size distributions, high polymerization rates, weight ratios of polymer to surfactant (P/S) from 13.3 to 32.8 were obtained. The number‐average molecular weights (M_n) were between 96,000 and 188,000 g/mol. Homogeneous copolymer compositions were obtained throughout the reaction for both, seeded and unseeded processes, which is not possible by the usual batch microemulsion process. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Azacrown Ethers with Naphthyl Branches. Fluorescence Properties,Protonation and Metal Coordination

Starting from 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane, we have prepared two compounds by replacing the amine hydrogens with naphthyl or 3,5-bis(2′-oxymethylnaphthyl)benzyl units. The absorption and emission spectra of compounds 2 (N,N′-bis(2-naphthylmethyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane) and 3 (N,N′-bis[3,5-bis(2′-oxymethylnaphthyl)benzyl]-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane) have been studied in CH₃CN:CH₂Cl₂ 1:1 (v/v) solution. For comparison purposes, the spectroscopic properties of N-methyl(2-methylnaphthalene)ethylamine (1) have also been investigated. For each compound, the absorption spectra are qualitatively very similar to that of naphthalene, with molar absorption coefficients as expected for the presence of one (1), two (2), and four (3) naphthyl chromophoric groups. The fluorescence spectra, however, are quite different from that of naphthalene. The naphthalene-type fluorescence (λ_max = 337 nm) is strongly quenched, particularly for compounds 1 and 2 which also exhibit a broad emission band in the visible region (λ_max ≈ 480 nm) assigned to a low lying charge-transfer excited state. In the case of compound 3, a quenched naphthalene-type band is accompanied by weak exciplex and excimer emissions. Upon titration with CF₃SO₃H, the charge transfer bands of 1 and 2 and the exciplex emission of 3 disappear and the naphthalene-type bands regain intensity. Titration plots show that in compounds 2 and 3 the protonation of the two nitrogens occurs stoichiometrically in two distinct steps. Titration with Zn²⁺ gives rise to 2.Zn²⁺ and 3.Zn²⁺ complexes. This article is dedicated to Professor Dedier Astruc.     

Use of ionic liquids based on phosphonium salts for preparing biocomposites by in situ polymerization

A sodium montmorillonite, Dellite HPS, was modified with ionic liquids based on phosphonium salts, such as octadecyltriphenylphosphonium tetrafluoroborate and octadecyltriphenylphosphonium bromide. Thanks to their high thermal stability, these salts can be used during in situ polymerization, a method that favors the achievement of a good dispersion of the clay. Poly(1,4‐dimethylcyclohexane adipate) (PCHA), was chosen as an example of aliphatic polyester which can be a suitable matrix for new biocomposites with organo‐clays. The organo modified clays prepared were characterized by X‐ray diffraction (XRD), ATR‐FTIR spectroscopy, and thermal gravimetric analysis (TGA) while biocomposites were analyzed in terms of molecular structure, thermal and thermomechanical properties. The degree of dispersion of the clays in the polymer matrix was studied by XRD. The results show that the clays are well dispersed in the biocomposites, despite an intercalated structure highlighted by XRD analysis. Moreover, the clays confer a certain improvement in mechanical properties to the final materials. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42467.     

Cold groundwater temperatures and conductive heat flow in the Mt. Amiata geothermal area, Tuscany, Italy

The hydrogeochemical study of the cold springs present in the Mt. Amiata geothermal area, where the Bagnore and Piancastagnaio geothermal fields are situated, has defined the different shallow groundwater systems.The cold groundwater temperature of the volcanic phreatic aquifer is largely correlated with the geothermal heat flow. Through the analysis of the temperature of cold groundwaters, a possible method for geothermal prospection has been developed, supported by the results obtained in the studied area. Through the enthalpic balance of the aquifer and in agreement with available data, a geothermal flow of about 200 mW m^-2 has been calculated.