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In the present study, we addressed the knowledge gaps regarding the agglomeration behavior and fate of food-grade titanium dioxide (E 171) in human gastrointestinal digestion (GID). After thorough multi-technique physicochemical characterization including TEM, single-particle ICP-MS (spICP-MS), CLS, VSSA determination and ELS, the GI fate of E 171 was studied by applying the in vitro GID approach established for the regulatory risk assessment of nanomaterials in Europe, using a standardized international protocol. GI fate was investigated in fasted conditions, relevant to E 171 use in food supplements and medicines, and in fed conditions, with both a model food and E 171-containing food samples. TiO2 constituent particles were resistant to GI dissolution, and thus, their stability in lysosomal fluid was investigated. The biopersistence of the material in lysosomal fluid highlighted its potential for bioaccumulation. For characterizing the agglomeration degree in the small intestinal phase, spICP-MS represented an ideal analytical tool to overcome the limitations of earlier studies. We demonstrated that, after simulated GID, in the small intestine, E 171 (at concentrations reflecting human exposure) is present with a dispersion degree similar to that obtained when dispersing the material in water by means of high-energy sonication (i.e., ≥70% of particles

Human induced pluripotent stem cell (iPSC)-derived neuronal and glial cell models are suitable to assess the effects of environmental chemicals on the developing brain. Such test systems can recapitulate several key neurodevelopmental features, such as neural stem cell formation and differentiation towards different neuronal subtypes and astrocytes, neurite outgrowth, synapse formation and neuronal network formation and function, which are crucial for brain development. While monolayer, two-dimensional (2D) cultures of human iPSC-neuronal or glial derivatives are generally suited for high-throughput testing, they also show some limitations. In particular, differentiation towards myelinating oligodendrocytes can only be achieved after extended periods in differentiation. In recent years, the implementation of three-dimensional (3D) neuronal and glial models obtained from human iPSCs has been shown to compensate for such limitations, enabling robust differentiation towards both neuronal and glial cell populations, myelination and formation of more mature neuronal network activity. Here we compared the differentiation capacity of human iPSC-derived neural stem cells cultured either as 2D monolayer or as 3D neurospheres, and assessed chlorpyrifos (CPF) effects. Data indicate that 3D neurospheres differentiate towards neurons and oligodendroglia more rapidly than 2D cultures; however, the 2D model is more suitable to assess neuronal functionality by analysis of spontaneous electrical activity using multielectrode array. Moreover, 2D and 3D test systems are diversely susceptible to CPF treatment. In conclusion, the selection of the most suitable in vitro test system (either 2D or 3D) should take into account the context of use and intended research goals ('fit for purpose' principle).

Microplastics (MPs) represent a worldwide emerging relevant concern toward human and environmental health due to their intentional or unintentional release. Human exposure to MPs by inhalation is predicted to be among the most hazardous. MPs include both engineered, or primary MPs, and secondary MPs, materials obtained by fragmentation from any plastic good. The major part of the environmental MPs is constituted by the second ones that are irregular in size, shape and composition. These features make the study of the biological impact of heterogenous MPs of extremely high relevance to better estimate the real toxicological hazards of these materials on human and environmental organisms. The smallest fractions of plastic granules, relying on the micron-sized scale, can be considered as the most abundant component of the environmental MPs, and for this reason, they are typically used to perform toxicity tests using in vitro systems representative of an inhalation exposure scenario. In the present work, MPs obtained from industrial treatment of waste plastics (wMPs < 50 μm) were investigated, and after the physico-chemical characterization, the cytotoxic, inflammatory and genotoxic responses, as well as the modality of wMPs interactions with alveolar lung cells, were determined. Obtained results indicated that, at high concentrations (100 μg/ml) and prolonged exposure time (48 h), wMPs affect biological responses by inducing inflammation and genotoxicity, as a result of the cell–wMP interactions, also including the uptake of the smaller particles.

Plastic debris accounts for the most substantial part of marine pollution and its degradation byproducts are omnipresent. Beyond representing a serious potential hazard for the marine ecosystem, mi...

Rare earth elements (REEs) are critical raw materials with a wide range of industrial applications. As a result, the recovery of REEs via adsorption from REE-rich matrices, such as water streams from processed electric and electronic waste, has gained increased attention for its simplicity, cost-effectiveness and high efficacy. In this work, the potential of nanometric cerium oxide-based materials as adsorbents for selected REEs is investigated. Ultra-small cerium oxide nanoparticles (CNPs, mean size diameter ≈ 3 nm) were produced via a precipitation-hydrothermal procedure and incorporated into woven–non-woven polyvinyl alcohol (PVA) nanofibres (d ≈ 280 nm) via electrospinning, to a final loading of ≈34 wt%. CNPs, CNP–PVA and the benchmark material CeO2 NM-212 (JRCNM02102, mean size diameter ≈ 28 nm) were tested as adsorbents for aqueous solutions of the REEs Eu3+, Gd3+ and Yb3+ at pH 5.8. Equilibrium adsorption data were interpreted by means of Langmuir and Freundlich data models. The maximum adsorption capacities ranged between 16 and 322 mgREE gCeO2−1, with the larger value found for the adsorption of Yb3+ by CNP. The trend of maximum adsorption capacity was CNPs > NM-212 > CNP–PVA, which was ascribed to different agglomeration and surface area available for adsorption. Langmuir equilibrium constants KL were substantially larger for CNP–PVA, suggesting a potential higher affinity of REEs for CNPs due to a synergistic effect of PVA on adsorption. CNP–PVA were effectively used in repeated adsorption cycles under static and dynamic configurations and retained the vast majority of adsorptive material (>98% of CeO2 retained after 10 adsorption cycles). The small loss was attributed to partial solubilisation of fibre components with change in membrane morphology. The findings of this study pave the way for the application of CNP–PVA nanocomposites in the recovery of strategically important REEs from electrical and electronic waste.

Nanoplastic particulates (pNP) are widely considered as being potentially harmful to the environment and living organisms while also being technically difficult to detect and identify in the presence of biological matrices. In this study, we describe a method for the extraction and subsequent Raman analysis of pNP present in the tissues of salt-water mussels. The process combines a step of enzymatic digestion/filtering to eliminate the biological matrix with a detection/identification procedure, which uses a micro-machined surface, composed of arrays of cavities with well-defined sub-micron depths and diameters. This sensor surface, exploits capillary forces in a drying droplet of analyte solution to drive the self-assembly of suspended nanoparticles into the cavities leaving the individual particles isolated from each other over the surface. The resulting array, when analysed using confocal Raman microscopy, permits the size selective analysis of the individual sub-micron pNP trapped in the cavities structure.

The European Union (EU) continuously takes ensuring the safe use of manufactured nanomaterials (MNMs) in consumer products into consideration. The application of a common approach for testing MNMs, including the use of optimized protocols and methods' selection, becomes increasingly important to obtain reliable and comparable results supporting the regulatory framework. In the present study, we tested four representative MNMs, two titanium dioxides (NM100 and NM101) and two silicon dioxides (NM200 and NM203), using the EU FP7-NANoREG approach, starting from suspension and dispersion preparations, through to their characterization and final evaluation of biological effects. MNM dispersions were prepared following a refined NANOGENOTOX protocol and characterized by dynamic light scattering (DLS) in water/bovine serum albumin and in media used for in vitro testing. Potential genotoxic effects were evaluated on human bronchial BEAS-2B cells using micronucleus and Comet assays, and pro-inflammatory effects by cytokines release. Murine macrophages RAW 264.7 were used to detect potential innate immune responses using two functional endpoints (pro-inflammatory cytokines and nitric oxide [NO] production). The interaction of MNMs with RAW 264.7 cells was studied by electron microscopy. No chromosomal damage and slight DNA damage and an oxidative effect, depending on MNMs, were observed in bronchial cells. In murine macrophages, the four MNMs directly induced tumor necrosis factor α or interleukin 6 secretion, although at very low levels; lipopolysaccharide-induced NO production was significantly decreased by the titania and one silica MNM. The application of this approach for the evaluation of MNM biological effects could be useful for both regulators and industries.

International audience; The way nanoplastics aggregate in the environment is one of the key properties that control their final fate and impact on the environment. In the present work, to better predict their transportation pathways, nanoplastic homoaggregation was studied in saltwater to predict the behavior in seawater. We designed nanoplastic models that are free of additives with a chemical control of the surface to model surface weathering. The samples present a wide distribution of relevant surface properties such as functionality (ionizable carboxylic group, 0.10 to 1.7 mmol g–1), hydrophobicity (surface energy, 2.20 to 37.5 mJ m–2), surface morphology (smooth or “raspberry-textured),” zeta potential (−31 to −21 mV), and anisotropy in shape. The critical coagulation concentration (CCC) measurements demonstrate that spherical nanoparticles are more stable in seawater (CCC > 600 mmol L–1) than anisotropic nanoplastics (CCC ∼ 100 mmol L–1). The results highlight the importance of considering the surface properties and shape when assessing the behavior of nanoplastics in the environment.

Titanium dioxide is a white colourant authorised as food additive E171 in the EU and is applied in a range of food products. Currently the EU specifications for E171 do not refer to the characterisation of particle size distribution; however, this may be requested in the near future. Only a few studies have been published to date reporting data on the size distribution of food grade titanium dioxide. The aim of this study was to characterise the size distribution of titanium dioxide particles contained in eight confectionery products and the pristine titanium dioxide samples used in each of the products. This allowed the direct comparison of the particle size distribution in both the pristine and the extracted materials. By using various analytical techniques, such as transmission electron microscopy, single particle inductively coupled plasma mass spectrometry (sp-ICPMS) and centrifuge liquid sedimentation (CLS) for the characterisation and quantification of the titanium dioxide particle sizes, the impact of the instrumentation on the results was systematically studied. The volume-specific surface area (VSSA) and crystalline structure were also determined for all additives.

The possible neurodevelopmental consequences of SARS-CoV-2 infection are presently unknown. In utero exposure to SARS-CoV-2 has been hypothesized to affect the developing brain, possibly disrupting neurodevelopment of children. Spike protein interactors, such as ACE2, have been found expressed in the fetal brain, and could play a role in potential SARS-CoV-2 fetal brain pathogenesis. Apart from the possible direct involvement of SARS-CoV-2 or its specific viral components in the occurrence of neurological and neurodevelopmental manifestations, we recently reported the presence of toxin-like peptides in plasma, urine and fecal samples specifically from COVID-19 patients. In this study, we investigated the possible neurotoxic effects elicited upon 72-hour exposure to human relevant levels of recombinant spike protein, toxin-like peptides found in COVID-19 patients, as well as a combination of both in 3D human iPSC-derived neural stem cells differentiated for either 2 weeks (short-term) or 8 weeks (long-term, 2 weeks in suspension + 6 weeks on MEA) towards neurons/glia. Whole transcriptome and qPCR analysis revealed that spike protein and toxin-like peptides at non-cytotoxic concentrations differentially perturb the expression of SPHK1, ELN, GASK1B, HEY1, UTS2, ACE2 and some neuronal-, glia- and NSC-related genes critical during brain development. Additionally, exposure to spike protein caused a decrease of spontaneous electrical activity after two days in long-term differentiated cultures. The perturbations of these neurodevelopmental endpoints are discussed in the context of recent knowledge about the key events described in Adverse Outcome Pathways relevant to COVID-19, gathered in the context of the CIAO project (https://www.ciao-covid.net/).

Nanoforms (NFs) of a substance may be distinguished from one another through differences in their physicochemical properties. When registering nanoforms of a substance for assessment under the EU REACH framework, five basic descriptors are required for their identification: composition, surface chemistry, size, specific surface area and shape. To make the risk assessment of similar NFs efficient, a number of grouping frameworks have been proposed, which often require assessment of similarity on individual physicochemical properties as part of the group justification. Similarity assessment requires an understanding of the achievable accuracy of the available methods. It must be demonstrated that measured differences between NFs are greater than the achievable accuracy of the method, to have confidence that the measured differences are indeed real. To estimate the achievable accuracy of a method, we assess the reproducibility of six analytical techniques routinely used to measure these five basic descriptors of nanoforms: inductively coupled plasma mass spectrometry (ICP-MS), Thermogravimetric analysis (TGA), Electrophoretic light scattering (ELS), Brunauer-Emmett-Teller (BET) specific surface area and transmission and scanning electron microscopy (TEM and SEM). Assessment was performed on representative test materials to evaluate the reproducibility of methods on single NFs of substances. The achievable accuracy was defined as the relative standard deviation of reproducibility (RSD

Prostate cancer (PCa), one of the leading causes of cancer-related deaths, currently lacks effective treatment for advanced-stage disease. Paclitaxel (PTX) is a highly active chemotherapeutic drug and the first-line treatment for PCa; however, conventional PTX formulation causes severe hypersensitivity reactions and limits PTX use at high concentrations. In the pursuit of high molecular weight, biodegradable, and pH-responsive polymeric carriers, we conjugated PTX to a polyacetal-based nanocarrier to yield a tert-Ser-PTX polyacetal conjugate. tert-Ser-PTX conjugate provides sustained release of PTX over two weeks in a pH-responsive manner while also obtaining a degree of epimerization of PTX to 7-epi-PTX. Serum proteins stabilize tert-Ser-PTX, with enhanced stability in human serum vs. PBS (pH 7.4). In vitro efficacy assessments in PCa cells demonstrated IC50 values above those for the free form of PTX due to the differential cell trafficking modes; however, in vivo tolerability assays demonstrated that tert-Ser-PTX significantly reduced the systemic toxicities associated with free PTX treatment. tert-Ser-PTX also effectively inhibited primary tumor growth and hematologic, lymphatic, and coelomic dissemination, as confirmed by in vivo and ex vivo bioluminescence imaging and histopathological evaluations in mice carrying orthotopic LNCaP tumors. Overall, our results suggest the application of tert-Ser-PTX as a robust anti-tumor/antimetastatic treatment for PCa. This article is protected by copyright. All rights reserved.

Microplastic pollution represents a global problem with negative impacts on aquatic environment and organisms' health. To date, most of the laboratory toxicological studies on microplastics (MPs) have made use of single commercial micro and nano-polymers, which do not reflect the heterogeneity of environmental MPs. To improve the relevance of the hazard assessment, micrometer-sized plastic particles of miscellaneous non-reusable waste plastics, with size

Adjuvants have been used for decades to enhance the immune response to vaccines, in particular for the subunit-based adjuvants. Physicochemical properties of the adjuvant-protein antigen complexes, such as size, morphology, protein structure and binding, influence the overall efficacy and safety of the vaccine. Here we show how to perform an accurate physicochemical characterization of the nanoaluminum–ovalbumin complex. Using a combination of existing techniques, we developed a multi-staged characterization strategy based on measurements of increased complexity. This characterization cascade has the advantage of being very flexible and easily adaptable to any adjuvant-protein antigen combinations. It will contribute to control the quality of antigen–adjuvant complexes and immunological outcomes, ultimately leading to improved vaccines.

Nanotechnology is considered to be a key enabling technology and in recent years there has been much growth in the use of nanostructured materials in industrial applications and in consumer products. It is, therefore, important that prior to being commercialized in consumer products, engineered nanomaterials are subjected to a thorough physico-chemical characterization as part of broader risk assessment to evaluate their possible effects on human health and the environment. The proper dispersion of nanomaterials sourced as powders becomes a first crucial step in the characterization process. This paper focuses on the dispersion of multiwall carbon nanotubes - often hydrophobic and tangled - since it may be challenging to re-disperse them effectively in aqueous media prior to characterization. A comparison has been made of non-contact vial sonication and immersion probe sonication using tannic acid as a dispersant. Transmission electron microscopy and UV-Vis spectroscopy were the techniques used to evaluate the dispersions. We used High Content Imaging and Colony Forming Efficiency to perform in vitro cytotoxicity studies on Human Alveolar Epithelial cells. It was found that both sonication treatments produce equivalent stable dispersions. No cytotoxic effects from MWCNTs were observed although some toxicity was observed and attributed to excess of the tannic acid dispersant.

Rare earth elements (REEs) are critical raw materials with a wide range of industrial applications. As a result, the recovery of REEs

Plastic particulates in the environment pose an increasing concern for regulatory bodies due to their potential risk to higher organisms (including humans) as they enter the food chain. Nanoplastics (defined here as smaller than 1 μm) are particularly challenging to detect and analyze at environmentally relevant concentrations and in biological matrices. The tunicate Ciona Robusta is an effective bioindicator for microplastics and nanoplastic contamination in the marine environment, due to its capacity to filter substantial volumes of water and to accumulate particulates. In this proof-of-principle study that demonstrates a complete methodology, following controlled exposure using spiked samples of a model nanoplastic (100 nm diameter polystyrene spheres) the nanoparticles were separated from an enzymatically digested biological matrix, purified and concentrated for analysis. The described method yields an approximate value for nanoplastic concentration in the organism (with a limit of detection of 106 particles/organism, corresponding to 1 ng/g) and provides the chemical composition by Raman spectroscopy. Furthermore, this method can be extended to other biological matrices and used to quantitatively monitor the accumulation of nanoplastics in the environment and food chain.

In realistic exposure scenarios, the detection and quantification of engineered nanoparticles in complex environmental or biological matrixes is a challenge since nanoparticle concentrations are frequently low and have to be discerned from a background that may contain the same elements in various chemical forms in much higher concentrations. The use of radiolabelled nanoparticles may overcome these difficulties offering high detection sensitivity without the necessity of complex sample preparation procedures. However, the labelling procedure must not alter the physicochemical and biological properties of the nanoparticles. In the present work, the radiolabelling of three different types of TiO2 nanoparticles with primary particle sizes between 5 nm and 26 nm with commercially available 44Ti has been investigated applying a simple diffusion heat treatment at 180 °C for 2.5 h on nanoparticles impregnated with a solution containing the 44Ti radiolabel. The same treatment has been investigated to radiolabel amorphous SiO2 nanoparticles with 44Ti. The radiolabels are stably integrated in the nanoparticle matrix, and the release is less than 0.1% in aqueous suspension at neutral pH for at least 4 weeks. The method appears to be fast and reliable. By transmission electron microscopy, dynamic light scattering and ζ-potential measurements, only minor alterations of the nanoparticle size could be detected in the range of 1 to 2 nm.

The large-scale production of plastic and the resulting release of waste is leading to a huge accumulation of micro-sized particles in the environment that could have an impact on not only aquatic organisms but also on humans. Despite the extensive literature on the subject, there is still an insufficient harmonization of methodologies for the collection and analysis of microplastics (MPs) in complex matrices, especially for high density polymers, such as polyvinyl chloride (PVC), which tend to sink and accumulate in sediments, becoming available to benthonic organisms. In this article, mussels have been chosen as model for microplastic accumulation due to their extensive filtering activity and their wide distribution in both fresh and salt water basins. To facilitate the identification and quantification of microplastics taken up by mussels, novel fluorescent and metal-doped PVC microplastics (PVC-Platinum octaethylporphyrin (PtOEP) MPs in the size range of 100 &micro, m) have been synthesized and characterized. For the analysis of the mussels following exposure, an enzymatic protocol using amylase, lipase, papain, and SDS for organic material digestion and a sucrose-ZnCl2 density gradient for the selective separation of ingested microplastics has been developed. The final identification of MPs was performed by fluorescence microscopy. This work can greatly benefit the scientific community by providing a means to study the behavior of PVC MPs, which represent an example of a very relevant yet poorly studied high density polymeric contaminant commonly found in complex environmental matrices.

International audience; Nanoplastics (NPTs) are defined as colloids that originated from the unintentional degradation of plastic debris. To understand the possible risks caused by NPTs, it is crucial to determine how they are transported and where they may finally accumulate. Unfortunately, although most sources of plastic are land-based, risk assessments concerning NPTs in the terrestrial environmental system (soils, aquifers, freshwater sediments, etc.) have been largely lacking compared to studies concerning NPTs in the marine system. Furthermore, an important limitation of environmental fate studies is that the NPT models used are questionable in terms of their environmental representativeness. This study describes the fate of different NPT models in a porous media under unfavorable (repulsive) conditions, according to their physical and chemical properties: average hydrodynamic diameters (200–460 nm), composition (polystyrene with additives or primary polystyrene) and shape (spherical or polymorphic). NPTs that more closely mimic environmental NPTs present an inhomogeneous shape (i.e., deviating from a sphere) and are more deposited in a sand column by an order of magnitude. This deposition was attributed in part to physical retention, as confirmed by the straining that occurred for the larger size fractions. Additionally, different Derjaguin-Landau-Verwey-Overbeek (DLVO) models -the extended DLVO (XDLVO) and a DLVO modified by surface element integration (SEI) method-suggest that the environmentally relevant NPT models may alter its orientation to diminish repulsion from the sand surface and may find enough kinetic energy to deposit in the primary energetic minimum. These results point to the importance of choosing environmentally relevant NPT models.

Many chronic diseases manifest in prolonged inflammation and often ignored dysregulated lipid metabolism. When associated with inhalation of nanomaterials, limited information is available on the relevant molecular events and their causal connections. This prevents reliable prediction of outcomes by efficient testing strategies. To unravel how acute nanomaterial exposure leads to chronic conditions, we employed advanced microscopy and omics in vitro and in vivo together with in silico modelling. For selected metal-oxide nanomaterials, we show that lung epithelial cells survive the exposure by excreting internalized nanomaterials and passivating them on the surface, employing elevated lipid synthesis. Macrophages, on the contrary, lose their integrity whilst degrading the passivized bio-nano agglomerates, releasing the nanomaterials, which are taken up again by the epithelial cells. Constant proinflammatory signalling recruits new phagocytes that feed the vicious cycle of events, resulting in a long-lasting response to a single exposure. The proposed mechanism explains the nanomaterial-associated in vivo chronic outcomes and allows its prediction based on in vitro measurements. Similar mechanisms may trigger other chronic diseases affecting millions of lives worldwide.

This review paper reports a detailed characterization of some combustion or incineration residues and by-products produced in a medium-sized city in Northern Italy. The municipal solid waste incineration (MSWI) generates fly ash, which is a toxic waste. Coal fly ash (CFA) and flue gas desulfurization (FGD) derive from the thermoelectric coal plant located in the same city. Along with these ashes, silica fume and rice husk ash are also considered for the stabilization of fly ash based on their amorphous silica content with the aim to convert them into an inert material. The characterization of all the investigated ashes was performed using different techniques: X-ray diffraction, total reflection X-ray fluorescence, scanning electron microscopy, and transmission electron microscopy. The aim of this work is to describe the reuse possibilities that were proposed for these ashes, which were determined also on the basis of their structural properties. Several possible applications of the investigated ashes are proposed, and the most suitable reuse of stabilized fly ash samples seems to be the production of sustainable plastic composites. This paper shows that the reuse of the by-product materials can allow natural resources to be preserved following the principles of a circular economy.

Air pollution accounts as a critical environmental issue in different populated cities. With the introduction of smart cities concept, suitable measures to reduce people exposition to pollutants must be part of the policies for a suitable urban development. Generally urban policy makers have proposed some restrictions (for example for traffic), that were resulted insufficient. Then probably, different approaches may be considered to face the air pollution problem. This paper proposes a new porous hybrid material (SUNSPACE) based on the use of industrial silicate minerals and alginate materials to promote PM sequestration in urban areas. It was designed to be applied as a coating on all available external cities surfaces. The synthesis conditions were slightly modified, with respect to the original material, to avoid thermal treatment, and make the material directly applicable. SUNSPACE was characterized by using different techniques: nitrogen physisorption, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Results identify the presence of ink-bottle pores shape, suitable to sequestrate nano and micro PM. Moreover, PM reduction in the atmosphere generally occurs as a consequence of wet or dry deposition conditions, monodisperse Fe3O4 nanoparticles and indoor/outdoor exposure were used to investigate the ability of SUNSPACE in ultrafine PM sequestration.

In this paper a simple method for all waste recovery through municipal solid waste incineration (MSWI) is presented. For the first time, MSWI fly ash (FA) is treated using MSWI bottom ash (BA), which is considered as a new urban mining source due to its pozzolanic characteristics. The proposed technology involves mixing these ashes with two other components: flue gas desulphurization residue and coal fly ash. Thermal annealing at a low temperature (120 °C for 4 h) can also be performed, to accelerate some reactions, but it is not necessary for stabilization. The efficacy of the process is demonstrated by the comparison of elemental concentrations in leaching solutions that are detected in the untreated ashes and the obtained stabilized materials, which show the reduced solubility of leachable Zn and Pb (with respect to the starting MSWI FA). The chemical, morphological, and structural characterization of the obtained materials makes it possible to propose and discuss a stabilization mechanism, which was attributed to carbonation and formation of calcium silicate hydrate (C–S–H). In particular, for the stabilized sample, X-ray Diffraction analysis confirmed the formation of calcite, whereas the sample morphology shows some fibrils that may be due to amorphous C–S–H with a Ca/Si ratio that is slightly smaller than 1.5. Because the proposed method employs wastes produced at the same location, it is reasonable to conclude that the method may be directly applied to incinerator plants due to the simplification of the MSWI FA waste management strategy, which generally involves its transport and stabilization treatment before landfilling. Finally the saved carbon dioxide emissions and the economic benefits that are a result of the new urban mining proposed approach have been evaluated for Europe. In particular the avoid of MSWI FA landfilling was evaluated to save approximately 1.6 M€/day, and the saved carbon dioxide emissions introduced into the atmosphere is approximately 960 kt/year.

Background Significant progress of nanotechnology, including in particular biomedical and pharmaceutical applications, has resulted in a high number of studies describing the biological effects of nanomaterials. Moreover, a determination of so-called “critical quality attributes”, that is specific physicochemical properties of nanomaterials triggering the observed biological response, has been recognised as crucial for the evaluation and design of novel safe and efficacious therapeutics. In the context of in vitro studies, a thorough physicochemical characterisation of nanoparticles (NPs), also in the biological medium, is necessary to allow a correlation with a cellular response. Following this concept, we examined whether the main and frequently reported characteristics of NPs such as size and the agglomeration state can influence the level and the mechanism of NP cellular internalization. Results We employed fluorescently-labelled 30 and 80 nm silicon dioxide NPs, both in agglomerated and non-agglomerated form. Using flow cytometry, transmission electron microscopy, the inhibitors of endocytosis and gene silencing we determined the most probable routes of cellular uptake for each form of tested silica NPs. We observed differences in cellular uptake depending on the size and the agglomeration state of NPs. Caveolae-mediated endocytosis was implicated particularly in the internalisation of well dispersed silica NPs but with an increase of the agglomeration state of NPs a combination of endocytic pathways with a predominant role of macropinocytosis was noted. Conclusions We demonstrated that the agglomeration state of NPs is an important factor influencing the level of cell uptake and the mechanism of endocytosis of silica NPs. Electronic supplementary material The online version of this article (doi:10.1186/s12951-017-0281-6) contains supplementary material, which is available to authorized users.

pH-sensitive nonionic surfactant vesicles (niosomes) by polysorbate-20 (Tween-20) or polysorbate-20 derivatized by glycine (added as pH sensitive agent), were developed to deliver Ibuprofen (IBU) and Lidocaine (LID). For the physical-chemical characterization of vesicles (mean size, size distribution, zeta potential, vesicle morphology, bilayer properties and stability) dynamic light scattering (DLS), small angle X-ray scattering and fluorescence studies were performed. Potential cytotoxicity was evaluated on immortalized human keratinocyte cells (HaCaT) and on immortalized mouse fibroblasts Balb/3T3. In vivo antinociceptive activity (formalin test) and anti-inflammatory activity tests (paw edema induced by zymosan) in murine models were performed on drug-loaded niosomes. pH-sensitive niosomes were stable in the presence of 0 and 10% fetal bovine serum, non-cytotoxic and able to modify IBU or LID pharmacological activity in vivo. The synthesis of stimuli responsive surfactant, as an alternative to add pH-sensitive molecules to niosomes, could represent a promising delivery strategy for anesthetic and anti-inflammatory drugs.

The majority of active agents do not readily permeate into brain due to the presence of the blood-brain barrier and blood-cerebrospinal fluid barrier. Currently, the most innovative and promising non-invasive strategy in brain delivery is the design and preparation of nanocarriers, which can move through the brain endothelium. Niosomes can perform brain delivery, in fact polysorbates, can act as an anchor for apolipoprotein E from blood plasma. The particles mimic LDL and interact with the LDL receptor leading to the endothelial cells uptake. The efficacy of niosomes for anticancer therapeutic applications was correlated to their physicochemical and drug delivery properties. Dimensions and ζ-potential were characterized using dynamic light scattering and asymmetric flow-field fractionation system. Lipid bilayer was characterized measuring the fluidity, polarity and microviscosity by fluorescent probe spectra evaluation. Morphology and homogeneity were characterized using atomic force microscopy. Physicochemical stability and serum stability (45% v/v fetal bovine and human serum) were evaluated as a function of time using dynamic light scattering. U87-MG human glioblastoma cells were used to evaluate vesicle cytotoxicity and internalisation efficiency. From the obtained data, the systems appear useful to perform a prolonged (modified) release of biological active substances to the central nervous system.

BACKGROUND Arrhythmogenic cardiomyopathy (ACM) is associated with arrhythmias and risk of sudden death. Mutations in genes encoding proteins of cardiac intercalated discs account for similar to 60% of ACM cases, but the remaining 40% is still genetically elusive.OBJECTIVE The purpose of this study was to identify the underlying genetic cause in probands with ACM.METHODS DNA samples from 40 probands with ACM, negative for mutations in the 3 major ACM genes-DSP, PKP2, and DSG2, were screened by using a targeted gene panel consisting of 15 known ACM genes and 53 candidate genes.RESULTS About half of patients were found to carry rare variant(s) predicted to be damaging; specifically, 9 (22.5%) showed >= 1 variants in genes associated with ACM and/or with other inherited heart diseases and 10 (25%) showed variants in candidate genes. Among the latter, we focused on 2 novel variants in TP63 and PPP1R13L candidate genes (c.796C>T, p.(R266*) and c.1858G>C, p.(A620P), respectively). The encoded proteins p63 and inhibitor of apoptosis stimulating p53 protein are known to be interacting partners. Inhibitor of apoptosis stimulating p53 protein is a shuttling multifunctional protein: in the nucleus it is critical for inhibiting p63 function, whereas in the cytoplasm it regulates desmosome integrity. According to the American College of Medical Genetics and Genomics guidelines, the variant in TP63 has been scored as likely pathogenic and the variant in PPP1R13L as a variant of uncertain significance. Importantly, the mutant TP63 allele leads to nonsense-mediated messenger RNA decay, causing haploinsufficiency.CONCLUSION Our findings identify TP63 as a putative novel disease gene for ACM, while the possible involvement of PPP1R13L remains to be determined.

In this study we used 5 nm gold nanoparticles as delivery platforms to target cancer cells expressing the immune receptor Tim-3 using single chain antibodies. Gold surfaces were also covered with the cytotoxic drug rapamycin which was immobilised using a glutathione linker. These nanoconjugates allowed highly specific and efficient delivery of cytotoxic rapamycin into human malignant blood cells.

The World Health Organization reports that every year several million people die prematurely due to air pollution. Poor air quality is a by-product of unsustainable policies in transportation, energy, industry, and waste management in the world's most crowded cities. Particulate matter (PM) is one of the major element of polluted air. PM can be composed by organic and inorganic species. In particular, heavy metals present in PM include, lead (Pb), mercury (Hg), cadmium, (Cd), zinc (Zn), nickel (Ni), arsenic (As), and molybdenum (Mo). Currently, vegetation is the only existing sustainable method to reduce anthropogenic PM concentrations in urban environments. In particular, the PM-retention ability of vegetation depends on the surface properties, related to the plant species, leaf and branch density, and leaf micromorphology. In this work, a new hybrid material called SUNSPACE (SUstaiNable materials Synthesized from by-Products and Alginates for Clean air and better Environment) is proposed for air PM entrapment. Candle burning tests are performed to compare SUNSPACE with Hedera Helix L. leafs with respect to their efficacy of reducing coarse and fine PM. The temporal variation of PM10 and PM2.5 in presence of the trapping materials, shows that Hedera Helix L. surface saturates more rapidly. In addition, the capability of SUNSPACE in ultrafine PM trapping is also demonstrated by using titanium dioxide nanoparticles with 25 nm diameter. Scanning electron microscope (SEM) and Transmission electron microscope (TEM) images of SUNSPACE after entrapment tests highlight the presence of collected nanoparticles until to about 0.04 mm in depth from the sample surface. N2 physisorption measurements allow to demonstrate the possibility to SUNSPACE regeneration by washing.

In this work we investigated the genotoxicity of zinc oxide and titanium dioxide nanoparticles (ZnO NPs; TiO2 NPs) induced by oxidative stress on human colon carcinoma cells (Caco-2 cells). We measured free radical production in acellular conditions by Electron Paramagnetic Resonance technique and genotoxicity by micronucleus and Comet assays. Oxidative DNA damage was assessed by modified Comet assay and by measuring 8-oxodG steady state levels. The repair kinetics of DNA oxidation as well as the expression levels of hOGG1 were also analyzed. Even if both NPs were able to produce ROS in acellular conditions and to increase 8-oxodG levels in Caco-2 cells, only ZnO NPs resulted genotoxic inducing micronuclei and DNA damage. Furthermore, Caco-2 cells exposed to ZnO NPs were not able to repair the oxidative DNA damage that was efficiently repaired after TiO2 NPs treatment, through OGG1 involvement. These results indicate that the high oxidant environment caused by ZnO NPs in our cellular model can induce DNA damage and affect the repair pathways.

Refraction and absorption properties of soft X-rays provide distinct advantages for ‘fast’ imaging of matter consisting of light elements, as biological samples, with sensitivity better than conventional light microscopy. This is very attractive for nanotoxicology, in particular for rapid screening of nanoparticle distribution and their effects on exposed cells at submicrometer length scales. In this paper, we report on the first trials demonstrating the potential of scanning transmission X-ray microscopy to monitor morphological and chemical changes induced in Balb 3T3 mouse fibroblast cells exposed to CoFe2O4 nanoparticles. This research uses the complementary imaging methods of an advanced synchrotron soft X-ray microscopy instrument implementing a novel approach based on high-resolution absorption and phase contrast imaging for the identification of important cellular changes induced by the presence of nanoparticles. Copyright © 2015 John Wiley & Sons, Ltd.

Higher efficacy and safety of nano gold therapeutics require examination of cellular responses to gold nanoparticles (AuNPs). In this work we compared cellular uptake, cytotoxicity and RNA expression patterns induced in Caco-2 cells exposed to AuNP (5 and 30nm). Cellular internalization was dose and time-dependent for both AuNPs. The toxicity was observed by colony forming efficiency (CFE) and not by Trypan blue assay, and exclusively for 5nm AuNPs, starting at the concentration of 200μM (24 and 72h of exposure). The most pronounced changes in gene expression (Agilent microarrays) were detected at 72h (300μM) of exposure to AuNPs (5nm). The biological processes affected by smaller AuNPs were: RNA/zinc ion/transition metal ion binding (decreased), cadmium/copper ion binding and glutathione metabolism (increased). Some Nrf2 responsive genes (several metallothioneins, HMOX, G6PD, OSGIN1 and GPX2) were highly up regulated. Members of the selenoproteins were also differentially expressed. Our findings indicate that exposure to high concentration of AuNPs (5nm) induces metal exposure, oxidative stress signaling pathways, and might influence selenium homeostasis. Some of detected cellular responses might be explored as potential enhancers of anti-cancer properties of AuNPs based nanomedicines.

Single particle-inductively coupled plasma mass spectrometry (SP-ICPMS) is a promising technique able to generate the number based-particle size distribution (PSD) of nanoparticles (NPs) in aqueous suspensions. However, SP-ICPMS analysis is not consolidated as routine-technique yet and is not typically applied to real test samples with unknown composition. This work presents a methodology to detect, quantify and characterise the number-based PSD of Ag-NPs in different environmental aqueous samples (drinking and lake waters), aqueous samples derived from migration tests and consumer products using SP-ICPMS. The procedure is built from a pragmatic view and involves the analysis of serial dilutions of the original sample until no variation in the measured size values is observed while keeping particle counts proportional to the dilution applied. After evaluation of the analytical figures of merit, the SP-ICPMS method exhibited excellent linearity (r2>0.999) in the range (1–25) × 104 particles mL−1 for 30, 50 and 80 nm nominal size Ag-NPs standards. The precision in terms of repeatability was studied according to the RSDs of the measured size and particle number concentration values and a t-test (p = 95%) at the two intermediate concentration levels was applied to determine the bias of SP-ICPMS size values compared to reference values. The method showed good repeatability and an overall acceptable bias in the studied concentration range. The experimental minimum detectable size for Ag-NPs ranged between 12 and 15 nm. Additionally, results derived from direct SP-ICPMS analysis were compared to the results conducted for fractions collected by asymmetric flow-field flow fractionation and supernatant fractions after centrifugal filtration. The method has been successfully applied to determine the presence of Ag-NPs in lake water; tap water; tap water filtered by a filter jar; seven different liquid silver-based consumer products; and migration solutions (pure water and sweat simulant) from plasters. Results obtained by SP-ICPMS were supported by transmission electron microscopy and energy dispersive spectroscopy characterisation, suggesting that the proposed methodology can be applied as a positive screening test in the simultaneous quantification and size characterisation of Ag-NPs in samples of environmental interest. © 2017