Polymer Encapsulated Liposomes for Oral Co-Delivery of Curcumin and Hydroxytyrosol

Curcumin (Cur) is a hydrophobic polyphenol from the rhizome of Curcuma spp., while hydroxytyrosol (HT) is a water-soluble polyphenol from Olea europaea. Both show outstanding antioxidant properties but suffer from scarce bioavailability and low stability in biological fluids. In this work, the co-encapsulation of Cur and HT into liposomes was realized, and the liposomal formulation was improved using polymers to increase their survival in the gastrointestinal tract. Liposomes with different compositions were formulated: Type 1, composed of phospholipids and cholesterol; Type 2, also with a PEG coating; and Type 3 providing an additional shell of Eudragit® S100, a gastro-resistant polymer. Samples were characterized in terms of size, morphology, ζ-potential, encapsulation efficiency, and loading capacity. All samples were subjected to a simulated in vitro digestion and their stability was investigated. The Eudragit®S100 coating demonstrated prevention of early releases of HT in the mouth and gastric phases, while the PEG shell reduced bile salts and pancreatin effects during the intestinal digestion. In vitro antioxidant activity showed a cumulative effect for Cur and HT loaded in vesicles. Finally, liposomes with HT concentrations up to 40 μM and Cur up to 4.7 μM, alone or in combination, did not show cytotoxicity against Caco-2 cells

Nanostructured Photoelectrochemical Biosensing Platform for Cancer Biomarker Detection

The innovative photoelectrochemical properties of multifunctional nanomaterials are here investigated for the development of biosensing platforms for rapid and sensitive detection of a class of cancer biomarker candidates, known as microRNAs. Many different transducers have been proposed, so far, for microRNA detection. Recently, with the emergence of novel photoelectrochemically active species and new detection schemes, photoelectrochemistry has received increasing attention. Gold nanostructures have been, here, used to modify TiO2 electrodes. The surface of the nanostructured platform has been modified by nucleic acid capture probes (CPs). Biotinylated target miRNAs have been recognized by the specific CPs. The biosensing platform has been incubated with streptavidin alkaline phosphatase and exposed to a proper substrate. The product of the enzymatic reaction has been photoelectrochemically monitored. A compact and hand-held analytical device has been developed in order to have a final prototype in line with the concept of point of care testing. (C) 2017 The Authors. Published by Elsevier Ltd

Characterization of anti-proliferative and anti-oxidant effects of nano-sized vesicles from Brassica oleracea L. (Broccoli)

In this in vitro study, we test our hypothesis that Broccoli-derived vesicles (BDVs), combining the anti-oxidant properties of their components and the advantages of their structure, can influence the metabolic activity of different cancer cell lines. BDVs were isolated from homogenized fresh broccoli (Brassica oleracea L.) using a sucrose gradient ultracentrifugation method and were characterized in terms of physical properties, such as particle size, morphology, and surface charge by transmission electron microscopy (TEM) and laser doppler electrophoresis (LDE). Glucosinolates content was assessed by RPLC–ESI–MS analysis. Three different human cancer cell lines (colorectal adenocarcinoma Caco-2, lung adenocarcinoma NCI-H441 and neuroblastoma SHSY5Y) were evaluated for metabolic activity by the MTT assay, uptake by fluorescence and confocal microscopy, and anti-oxidant activity by a fluorimetric assay detecting intracellular reactive oxygen species (ROS). Three bands were obtained with average size measured by TEM based size distribution analysis of 52 nm (Band 1), 70 nm (Band 2), and 82 nm (Band 3). Glucobrassicin, glucoraphanin and neoglucobrassicin were found mostly concentrated in Band 1. BDVs affected the metabolic activity of different cancer cell lines in a dose dependent manner compared with untreated cells. Overall, Band 2 and 3 were more toxic than Band 1 irrespective of the cell lines. BDVs were taken up by cells in a dose- and time-dependent manner. Pre-incubation of cells with BDVs resulted in a significant decrease in ROS production in Caco-2 and NCI-H441 stimulated with hydrogen peroxide and SHSY5Y treated with 6-hydroxydopamine, with all three Bands. Our findings open to the possibility to find a novel “green” approach for cancer treatment, focused on using vesicles from broccoli, although a more in-depth characterization of bioactive molecules is warranted

Rapid Atlantification along the Fram Strait at the beginning of the 20th century.

The recent expansion of Atlantic waters into the Arctic Ocean represents undisputable evidence of the rapid changes occurring in this region. Understanding the past variability of this “Atlantification” is thus crucial in providing a longer perspective on the modern Arctic changes. Here, we reconstruct the history of Atlantification along the eastern Fram Strait during the past 800 years using precisely dated paleoceanographic records based on organic biomarkers and benthic foraminiferal data. Our results show rapid changes in water mass properties that commenced in the early 20th century—several decades before the documented Atlantification by instrumental records. Comparison with regional records suggests a poleward expansion of subtropical waters since the end of the Little Ice Age in response to a rapid hydrographic reorganization in the North Atlantic. Understanding of this mechanism will require further investigations using climate model simulations

First insight into extracellular vesicle-miRNA characterization in a sheep in vitro model of inflammation

Extracellular vesicles (EVs) and their microRNA (miRNA) cargoes have garnered attention in the veterinary field for their regulatory role in various biological processes. This study aimed to (i) evaluate two techniques of EV isolation from sheep peripheral blood mononuclear cell (PBMC) supernatants using the ultracentrifugation (UC) and reagent (REA) methods and (ii) characterize the EV-miRNA profiles after an in vitro inflammatory environment mediated by lipopolysaccharides (LPS). Sheep peripheral blood was collected, and PBMCs were separated using a density gradient reagent. Subsequently, PBMCs were cultured at 37°C for 24 h (5% CO2), and the supernatants were collected to perform the EV isolation. The presence of CD81+ extracellular vesicle marker was determined, and the purity of isolated EVs was calculated as a ratio between the number of isolated EVs and the protein concentration. Moreover, the morphological characterization revealed mainly round-shaped structures with average sizes of 211 nm for EVs isolated by the UC method and 99 nm for EVs isolated by the REA method. Illumina NextSeq sequencing in a single-end mode was used to characterize the miRNA profile, and the differentially expressed (DE) miRNAs were analyzed using a combination of bioinformatics tools. The results revealed that the REA method is reliable for EV isolation from sheep supernatants. It was considered an improvement of the recovery rate and purity of EVs with the enhancement of the number and the expression levels of characterized miRNAs. The EVs isolated by the UC method after an LPS challenge showed 11 DE miRNAs, among which eight miRNAs were upregulated and three were downregulated. On the other hand, the REA method revealed an EV cargo in which eight DE miRNAs were upregulated and 21 DE miRNAs were downregulated. The master miRNA regulators of the biological process were identified by performing the MIRNA-mRNA network analysis, showing that, among the higher representative miRNAs based on the centrality and betweenness, the miR-26a-5p could have a crucial role in the resolution of inflammation. Moreover, the identification of the let-7 miRNA family in all the EVs showed potential targeted genes that regulate the inflammation and immune responses

A population genomics insight by 2b‐RAD reveals populations' uniqueness along the Italian coastline in Leptopsammia pruvoti (Scleractinia, Dendrophylliidae)

Aim Marine bioconstructions such as coralligenous formations are hotspot of biodiversity and play a relevant ecological role in the preservation of biodiversity by providing carbon regulation, protection and nursery areas for several marine species. For this reason, the European Union Habitat Directive included them among priority habitats to be preserved. Although their ecological role is well established, connectivity patterns are still poorly investigated, representing a limit in conservation planning. The present study pioneers a novel approach for the analysis of connectivity in marine bioconstructor species, which often lack suitable genetic markers, by taking advantage of next‐generation sequencing techniques. We assess the geographical patterns of genomic variation of the sunset cup coral Leptopsammia pruvoti Lacaze‐Duthiers, 1897, an ahermatypic, non‐zooxanthellate and solitary scleractinian coral species common in coralligenous habitats and distributed across the Mediterranean Sea. Location The Italian coastline (Western and Central Mediterranean). Methods We applied the restriction site‐associated 2b‐RAD approach to genotype over 1,000 high‐quality and filtered single nucleotide polymorphisms in 10 population samples. Results The results revealed the existence of a strongly supported genetic structure, with highly significant pairwise FST values between all the population samples, including those collected about 5 km apart from each other. Moreover, genomic data indicate that the strongest barriers to gene flow are between the western (Ligurian–Tyrrhenian Sea) and the eastern side (Adriatic Sea) of the Italian peninsula. Main conclusions The strong differentiation found in L. pruvoti is similar to that found in other species of marine bioconstructors investigated in this area, but it strongly contrasts with the small differences found in many fish and invertebrates at the same geographical scale. All in one, our results highlight the importance of assessing connectivity in species belonging to coralligenous habitats as, due to their limited dispersal ability, they might require specific spatial conservation measures

Magnetic nanocrystal modified epoxy photoresist for microfabrication of AFM probes

Nanocomposites based on an organic polymer and inorganic nanocrystals (NCs) represent a class of high impact functional materials able to convey the unique size and shape dependent properties of nano-objects to highly processable resists.[1] In this work, a novel magnetic nanocomposite based on a negative tone epoxy photoresist and magnetic colloidal Fe2O3 NCs has been manufactured for fabricating AFM probes. Epoxy-type photoresist grant superior lithographic performances when patterned by standard near- ultraviolet (UV) optical lithography, providing structures with high aspect-ratio and nearly vertical sidewalls. Such resists are at present employed in optical and micromechanical applications for the fabrication of optical waveguides, microfluidic systems and scanning probes.[2] However, these materials lack of any inherent functionality, e.g. electrical conductivity, luminescence, magnetism, piezoresistivity and dielectricity. Hence, the incorporation of NCs can confer them new properties maintaining the patterning resolutions required for the manufacturing of highly miniaturized devices. These added properties can be interesting for the fabrication of novel micro/nanoelectromechanical systems (MEMS/NEMS)

Tolerability of vortioxetine compared to selective serotonin reuptake inhibitors in older adults with major depressive disorder (VESPA): a randomised, assessor-blinded and statistician-blinded, multicentre, superiority trial.

BACKGROUND Major depressive disorder (MDD) is prevalent and disabling among older adults. Standing on its tolerability profile, vortioxetine might be a promising alternative to selective serotonin reuptake inhibitors (SSRIs) in such a vulnerable population. METHODS We conducted a randomised, assessor- and statistician-blinded, superiority trial including older adults with MDD. The study was conducted between 02/02/2019 and 02/22/2023 in 11 Italian Psychiatric Services. Participants were randomised to vortioxetine or one of the SSRIs, selected according to common practice. Treatment discontinuation due to adverse events after six months was the primary outcome, for which we aimed to detect a 12% difference in favour of vortioxetine. The study was registered in the online repository clinicaltrials.gov (NCT03779789). FINDINGS The intention-to-treat population included 179 individuals randomised to vortioxetine and 178 to SSRIs. Mean age was 73.7 years (standard deviation 6.1), and 264 participants (69%) were female. Of those on vortioxetine, 78 (44%) discontinued the treatment due to adverse events at six months, compared to 59 (33%) of those on SSRIs (odds ratio 1.56; 95% confidence interval 1.01-2.39). Adjusted and per-protocol analyses confirmed point estimates in favour of SSRIs, but without a significant difference. With the exception of the unadjusted survival analysis showing SSRIs to outperform vortioxetine, secondary outcomes provided results consistent with a lack of substantial safety and tolerability differences between the two arms. Overall, no significant differences emerged in terms of response rates, depressive symptoms and quality of life, while SSRIs outperformed vortioxetine in terms of cognitive performance. INTERPRETATION As opposed to what was previously hypothesised, vortioxetine did not show a better tolerability profile compared to SSRIs in older adults with MDD in this study. Additionally, hypothetical advantages of vortioxetine on depression-related cognitive symptoms might be questioned. The study's statistical power and highly pragmatic design allow for generalisability to real-world practice. FUNDING The study was funded by the Italian Medicines Agency within the "2016 Call for Independent Drug Research"

Copper-Triggered Aggregation of Ubiquitin

Neurodegenerative disorders share common features comprising aggregation of misfolded proteins, failure of the ubiquitin-proteasome system, and increased levels of metal ions in the brain. Protein aggregates within affected cells often contain ubiquitin, however no report has focused on the aggregation propensity of this protein. Recently it was shown that copper, differently from zinc, nickel, aluminum, or cadmium, compromises ubiquitin stability and binds to the N-terminus with 0.1 micromolar affinity. This paper addresses the role of copper upon ubiquitin aggregation. In water, incubation with Cu(II) leads to formation of spherical particles that can progress from dimers to larger conglomerates. These spherical oligomers are SDS-resistant and are destroyed upon Cu(II) chelation or reduction to Cu(I). In water/trifluoroethanol (80∶20, v/v), a mimic of the local decrease in dielectric constant experienced in proximity to a membrane surface, ubiquitin incubation with Cu(II) causes time-dependent changes in circular dichroism and Fourier-transform infrared spectra, indicative of increasing β-sheet content. Analysis by atomic force and transmission electron microscopy reveals, in the given order, formation of spherical particles consistent with the size of early oligomers detected by gel electrophoresis, clustering of these particles in straight and curved chains, formation of ring structures, growth of trigonal branches from the rings, coalescence of the trigonal branched structures in a network. Notably, none of these ubiquitin aggregates was positive to tests for amyloid and Cu(II) chelation or reduction produced aggregate disassembly. The early formed Cu(II)-stabilized spherical oligomers, when reconstituted in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes and in POPC planar bilayers, form annular and pore-like structures, respectively, which are common to several neurodegenerative disorders including Parkinson's, Alzheimer's, amyotrophic lateral sclerosis, and prion diseases, and have been proposed to be the primary toxic species. Susceptibility to aggregation of ubiquitin, as it emerges from the present study, may represent a potential risk factor for disease onset or progression while cells attempt to tag and process toxic substrates

Elena Mendia and women’s professionalism in Neapolitan post-war architecture

The XVIII edition of the Forum titled ‘World Heritage and Contamination' aims to create a cross-critical dialogue, open to cultural contamination and ‘without limits', in a logic of integration between competences which extends, and is not limited to, the following disciplines: Architecture, Culture, Environment, Agriculture, Health, Landscape, Design, Territorial Governance, Archeology, Economy, e-Learning. The activities of protection and promotion of World Heritage, understood as a asset shared by all Humanity, are particularly relevant in Italy, responsible towards the world of custody of the largest number of assets protected by the UNESCO. To the World Heritage Properties is added a landscape heritage of enormous variety and beauty to be protected also through the regeneration of degradation and of the ‘minor heritage', in line with the provisions of the UNESCO Conventions on material and intangible assets as well as the European Landscape Convention