Aggregation, dissolution, and stability of quantum dots in marine environments: importance of extracellular polymeric substances

There is an increasing concern that a considerable fraction of engineered nanoparticles (ENs), including quantum dots (QDs), will eventually find their way into the marine environment and have negative impacts on plankton. As ENs enter the ocean, they will encounter extracellular polymeric substances (EPS) from microbial sources before directly interacting with plankton cells. In this study, EPS harvested from four phytoplankton species, Amphora sp., Dunaliella tertiolecta, Phaeocystis globosa, and Thalassiosira pseudonana, were examined for potential interactions with CdSe nonfunctionalized and functionalized (carboxyl- and amine-) QDs in artificial seawater. Our results show that EPS do not reduce the solubility of QDs but rather decrease their stability. The degradation rate of QDs was positively correlated to the protein composition of EPS (defined by the ratio of protein/carbohydrate). Two approaches showed significant inhibition to the degradation of carboxyl-functionalized QDs: (1) the presence of an antioxidant, such as N-acetyl cysteine, and (2) absence of light. Owing to the complexity in evaluating integrated effects of QDs intrinsic properties and the external environmental factors that control the stability of QDs, conclusions must be based on a careful consideration of all these factors when attempting to evaluate the bioavailability of QDs and other ENs in the marine environments.     

The Effect of Ageing on Exciton Dynamics,Charge Separation,and Recombination in P3HT/PCBM Photovoltaic Blends

A study of how light‐induced degradation influences the fundamental photophysical processes in the active layer of poly(3‐hexylthiophene)/[6,6]‐phenyl C₆₁‐butyric acid methyl ester (P3HT/PCBM) solar cells is presented. Non‐encapsulated samples are systematically aged by exposure to AM 1.5 illumination in the presence of dry air for different periods of time. The extent of degradation is quantified by the relative loss in the absorption maximum of the P3HT, which is varied in the range 0% to 20%. For degraded samples an increasing loss in the number of excitons within the P3HT domains is observed with longer ageing periods. This loss occurs rapidly, within the first 15 ps after photoexcitation. A more pronounced decrease in the population of polarons than excitons is observed, which also occurs on a timescale of a few picoseconds. These observations, complemented by a quantitative analysis of the polaron and exciton population dynamics, unravel two primary loss mechanisms for the performances of aged P3HT/PCBM solar cells. One is an initial ultrafast decrease in the polaron generation, apparently not related to the exciton diffusion to the polymer/fullerene interface; the second, less significant, is a loss in the exciton population within the photoexcited P3HT domains. The steady‐state photoinduced absorption spectra of degraded samples exhibits the appearance of a signal ascribed to triplet excitons, which is absent for non‐degraded samples. This latter observation is interpreted considering the formation of degraded sites where intersystem crossing and triplet exciton formation is more effective. The photovoltaic characteristics of same blends are also studied and discussed by comparing the decrease in the overall power conversion efficiency of solar cells.     

Tuning morphology of mesoporous titanium oxides through fluorinated surfactants-based systems

Four different fluorinated surfactant-based systems were used as template for the synthesis of highly ordered TiO₂ mesoporous materials with large-pore wormhole structures. The calcined materials exhibit large pore diameters (up to 5.9 nm), high surface areas (900–1,180 m² g^?1), pore volumes (0.90–1.25 cm³ g^?1) and thick pore walls (4.6–7.7 nm) depending of the synthesis route. It is established that the self-aggregation behavior of fluorinated amphiphile systems can be manipulate and provides a rich phase behavior to obtain well-organized titania sieves with adjustable pore size and surface topography. By comparison to titania and silica sieves obtained by the same procedure, it can be established that there is a deep interaction between head groups of fluorinated surfactants and Ti(IV)(iPrO)₄ showing that the inorganic precursor has a great influence on the properties of the final materials.     

Coating-Activation and Antimicrobial Efficacy of Different Polyethylene Films with a Nisin-Based Solution

Antimicrobial packaging can be considered an extremely challenging technology that could have a significant impact on shelf-life extension and food safety of fresh meat and meat products. In this study, different commercial polyethylene films differing in vinyl acetate ethylene, erucamide contents, and oxygen permeability were used for the coating treatment with a nisin-based antimicrobial solution (NS). Detection and measurement of the activity of the NS was determined against different food spoilage bacteria. NS was then spread manually on food contact layer of different plastic films using coating rods providing thickness of 6, 40, 60, and 100 μm. The polyethylene films before and after treatment were analysed by atomic force microscopy (AFM). NS was active against Gram-positive bacteria and the best activity was obtained against Brochothrix thermosphacta. Viable staining and epifluorescence microscopy analysis of indicator strains in contact with activated plastic films showed that the effect of the film on the various indicator strains changed very much on the basis of both type of film and indicator strain. The highest numbers of lysed cells were shown by two polyethylene films that, according to the AFM and roughness parameters analyses, were characterized by significant increase or decrease of roughness after the coating treatment. AFM analysis showed that the homogeneity of the coating was much influenced by the type of plastic films used. In order to test the efficacy in food, portions of beef chuck tender slices were prepared and covered with the antimicrobial plastic films on both sides. After 1 h and 1, 7, and 12 days of storage at 4 °C the meat samples were analyzed by standard plate counting targeting spoilage associated microbial populations. The antimicrobial plastic films after 1 h of contact with the meat caused a significant reduction of lactic acid bacteria and B. thermosphacta. The most effective antimicrobial activity of films was shown against the same populations after 24 h of storage.     

Microencapsulation of Lactobacillus reuteri DSM 17938 Cells Coated in Alginate Beads with Chitosan by Spray Drying to Use as a Probiotic Cell in a Chocolate Soufflé

The main objective of this work was to obtain microencapsulated probiotic cells in order to improve their resistance to heat stress and gastrointestinal conditions. A further aim was to obtain a potentially probiotic chocolate soufflé. Lactobacillus reuteri DSM 17938 cells were microencapsulated by spray drying in alginate matrix and further coated with chitosan. Bacterial survival after exposure to different heat treatments and simulated gastrointestinal conditions were measured to test the microcapsules. They were also dyed by using a LIVE/DEAD^® BacLight? Bacterial Viability Kit and characterized by epifluorescence microscope observation. Furthermore, a potentially chocolate soufflé was prepared using microencapsulated cells. The results indicated that alginate microcapsules did not improve acid tolerance or heat resistance in “in vitro” experiments, while they were able to protect 7% of the Lactobacillus reuteri population during the baking of a chocolate soufflé, compared to a survival rate of 1% of free cells. By contrast, the cells microencapsulated with alginate coated with chitosan showed, compared to free cells, improved acid tolerance, allowing the cell population to remain constant after 3 h in simulated gastric conditions. Moreover, the heat resistance of cells in co-cross-linked microcapsules significantly improved compared to free cells, both in “in vitro” and “in food” experiments. Microencapsulation led to a survival rate of 10% after baking a chocolate soufflé. However, the final level of bacterial cells in the product was too low to consider the chocolate soufflé as a probiotic product.     

FRP Confinement of Square Masonry Columns

The problem of masonry columns subjected to structural deficiency under axial load was studied and reported in this paper. The results of an extensive experimental campaign are presented in order to show the behavior of columns built with clay or with calcareous blocks, commonly found in southern Italy, especially in historical buildings. Rectangular masonry columns were tested for a total of 33 specimens; uniaxial compression tests were conducted on columns taking into account the influence of several variables: different strengthening schemes (internal and/or external confinement), curvature radius of the corners, amount of fiber-reinforced polymer (FRP) reinforcement, cross-section aspect ratio, and material of masonry blocks. Materials characterization was preliminarily carried out including a mechanical test on plain masonry. For all cases the experimental results evidenced a significant increase in load carrying capacity and ductility after FRP strengthening, which identified the columns as ductile elements despite the brittle nature of the unconfined masonry. Differences in mechanical behavior, due to the geometry of the columns, to the nature of different materials, to different strengthening schemes, and to the amount of reinforcement, are presented and discussed in the paper. The calibration of design equations recently developed by Italian National Research Council, CNR was conducted to compare analytical prediction and experimental results. The same procedure was applied to calibrate an analytical model recently published, in which the existing coefficients are related only to clay. Here the model is applied to limestone for the first time. Thus, new important information is furnished to researchers and practitioners involved in structural assessment and strengthening of compressed elements in historical buildings.     

Tumor-Derived Small Extracellular Vesicles Induce Pro-Inflammatory Cytokine Expression and PD-L1 Regulation in M0 Macrophages via IL-6/STAT3 and TLR4 Signaling Pathways

Tumor-associated macrophages play a key role in promoting tumor progression by exerting an immunosuppressive phenotype associated with the expression of programmed cell death ligand 1 (PD-L1). It is well known that tumor-derived small extracellular vesicles (SEVs) affect the tumor microenvironment, influencing TAM behavior. The present study aimed to examine the effect of SEVs derived from colon cancer and multiple myeloma cells on macrophage functions. Non-polarized macrophages (M0) differentiated from THP-1 cells were co-cultured with SEVs derived from a colorectal cancer (CRC) cell line, SW480, and a multiple myeloma (MM) cell line, MM1.S. The expression of PD-L1, interleukin-6 (IL-6), and other inflammatory cytokines as well as of the underlying molecular mechanisms were evaluated. Our results indicate that SEVs can significantly upregulate the expressions of PD-L1 and IL-6 at both the mRNA and protein levels and can activate the STAT3 signaling pathway. Furthermore, we identified the TLR4/NF-kB pathway as a convergent mechanism for SEV-mediated PD-L1 expression. Overall, these preliminary data suggest that SEVs contribute to the formation of an immunosuppressive microenvironment.     

VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

VDAC (voltage-dependent anion selective channel) proteins, also known as mitochondrial porins, are the most abundant proteins of the outer mitochondrial membrane (OMM), where they play a vital role in various cellular processes, in the regulation of metabolism, and in survival pathways. There is increasing consensus about their function as a cellular hub, connecting bioenergetics functions to the rest of the cell. The structural characterization of VDACs presents challenging issues due to their very high hydrophobicity, low solubility, the difficulty to separate them from other mitochondrial proteins of similar hydrophobicity and the practical impossibility to isolate each single isoform. Consequently, it is necessary to analyze them as components of a relatively complex mixture. Due to the experimental difficulties in their structural characterization, post-translational modifications (PTMs) of VDAC proteins represent a little explored field. Only in recent years, the increasing number of tools aimed at identifying and quantifying PTMs has allowed to increase our knowledge in this field and in the mechanisms that regulate functions and interactions of mitochondrial porins. In particular, the development of nano-reversed phase ultra-high performance liquid chromatography (nanoRP-UHPLC) and ultra-sensitive high-resolution mass spectrometry (HRMS) methods has played a key role in this field. The findings obtained on VDAC PTMs using such methodologies, which permitted an in-depth characterization of these very hydrophobic trans-membrane pore proteins, are summarized in this review.     

Nusinersen Modulates Proteomics Profiles of Cerebrospinal Fluid in Spinal Muscular Atrophy Type 1 Patients

Spinal muscular atrophy (SMA) type 1 is a severe infantile autosomal-recessive neuromuscular disorder caused by a survival motor neuron 1 gene (SMN1) mutation and characterized by progressive muscle weakness. Without supportive care, SMA type 1 is rapidly fatal. The antisense oligonucleotide nusinersen has recently improved the natural course of this disease. Here, we investigated, with a functional proteomic approach, cerebrospinal fluid (CSF) protein profiles from SMA type 1 patients who underwent nusinersen administration to clarify the biochemical response to the treatment and to monitor disease progression based on therapy. Six months after starting treatment (12 mg/5 mL × four doses of loading regimen administered at days 0, 14, 28, and 63), we observed a generalized reversion trend of the CSF protein pattern from our patient cohort to that of control donors. Notably, a marked up-regulation of apolipoprotein A1 and apolipoprotein E and a consistent variation in transthyretin proteoform occurrence were detected. Since these multifunctional proteins are critically active in biomolecular processes aberrant in SMA, i.e., synaptogenesis and neurite growth, neuronal survival and plasticity, inflammation, and oxidative stress control, their nusinersen induced modulation may support SMN improved-expression effects. Hence, these lipoproteins and transthyretin could represent valuable biomarkers to assess patient responsiveness and disease progression.