Polypeptide Chain Growth Mechanisms and Secondary Structure Formation in Glycine Gas-Phase Deposition on Silica Surfaces.

Peptide formation by amino acids condensation represents a crucial reaction in the quest of the origins of life as well as in synthetic chemistry. However, it is still poorly understood in terms of efficiency and reaction mechanism. In the present work, peptide formation has been investigated through thermal condensation of gas-phase glycine in fluctuating silica environments as a model of prebiotic environments. In-situ IR spectroscopy measurements under a controlled atmosphere reveal that a humidity fluctuating system subjected to both temperature and water activity variations results in the formation of more abundant peptides compared to a dehydrated system subjected only to temperature fluctuations cycles. A model is proposed in which hydration steps result in the hydrolysis and redistribution of the oligomers formed during previous deposition in dry conditions. This results in the formation of self-assembled aggregates with well-defined secondary structures (especially ß-sheets). Upon further monomers feeding, structural elements are conserved in newly growing chains, with indications of templated polymerization. The structural dynamics of peptides were also evaluated. Rigid self-assembled structures with a high resistance to further wetting/drying cycles and inaccessibility to isotopic exchange were present in the humidity fluctuating system compared to more flexible structures in the dehydrated system. The resistance and growth of self-assembled structures were also investigated for an extended duration of Gly deposition using isotope labeling.

Emergence of Order in Origin-of-Life Scenarios on Mineral Surfaces: Polyglycine Chains on Silica.

The polymerization of amino acids (AAs) to peptides on oxide surfaces has attracted interest owing to its high importance in biotechnology, prebiotic chemistry, and origin of life theories. However, its mechanism is still poorly understood. We tried to elucidate the reactivity of glycine (Gly) from the vapor phase on the surface of amorphous silica under controlled atmosphere at 160 °C. Infrared (IR) spectroscopy reveals that Gly functionalizes the silica surface through the formation of ester species, which represent, together with the weakly interacting silanols, crucial elements for monomers activation and polymerization. Once activated, ß-turns start to form as initiators for the growth of long linear polypeptides (poly-Gly) chains, which elongate into ordered structures containing both ß-sheet and helical conformations. The work also points to the role of water vapor in the formation of further self-assembled ß-sheet structures that are highly resistant to hydrolysis.

Structural and surface studies of luminescent Ca/Eu phosphate nanomaterials: From the bulk to surface features.

Three luminescent Eu-containing phosphate materials (Ca-doped europium phosphate monohydrate, Eu-doped carbonated-apatite, and europium phosphate monohydrate) were prepared and analyzed on the level of bulk structure and surface properties and compared to the biomimetic non-luminescent counterpart hydroxyapatite. Europium-containing phosphate materials exhibited nanosized dimensions but different luminescence emissions and luminescence lifetimes depending on their crystalline structures (i.e., lanthanide phosphate or apatites) and chemical composition. The introduction of Eu in the crystal lattice leads to a notable decrease in the overall Lewis acidity of the surface cationic sites detected by CO probing. Further, the mixed Eu/Ca-containing materials surfaces were found to be very similar to the reference hydroxyapatite in terms of water adsorption energy, while the pure europium phosphate resulted to have the notably higher energy values of direct interaction of water molecules with the surface cations with no detected propagation of this effect towards water overlayers.

TiO2-Chitosan Hybrid Materials for Drug Delivery Applications: Conjugation Reaction with a Model Drug and Evaluation of the Functional Properties.

The combination of TiO2 and chitosan is known to allow the achievement of implantable devices which combines the mechanical properties of TiO2, with the presence of chitosan, which ensures antibacterial properties combined with an in-situ drug-delivery of biomolecules physisorbed and/or covalently linked to chitosan. In this study, 5-aminofluorescein (5-AF), a derivative of fluorescein containing a primary amino group, has been used as model molecule to simulate a drug. This dye is characterized by low cost and low toxicity, and due to its high molar absorptivity it can easily be detected by means of absorption and emission spectroscopies. The combination of 5-AF and maleic anhydride (MA) with TiO2-chitosan materials has generated a range of novel hybrid materials tailored to applications in localized stimuli-responsive drug delivery systems. Maleic anhydride has been used as pH sensitive spacer for the covalent functionalization of the TiO2-chitosan hybrid with MA as linker molecule. This functionalization allowed to obtain a pH-sensitive hybrid material. The efficiency of the functionalization has been verified by means of different physico-chemical characterization techniques. The behaviour of the functionalized materials is related to different parameters, among which the ratio between physisorbed/coordinated and chemisorbed 5-AF and the matrix degradation. Moreover, delivery tests in simulated body solutions at different pH have been performed showing a pH-sensitive drug delivery behaviour and indicating that the release of 5-AF is favoured at basic pH.

Machine learning approach for elucidating and predicting the role of synthesis parameters on the shape and size of TiO2 nanoparticles.

In the present work a series of design rules are developed in order to tune the morphology of TiO2 nanoparticles through hydrothermal process. Through a careful experimental design, the influence of relevant process parameters on the synthesis outcome are studied, reaching to the develop predictive models by using Machine Learning methods. The models, after the validation and training, are able to predict with high accuracy the synthesis outcome in terms of nanoparticle size, polydispersity and aspect ratio. Furthermore, they are implemented by reverse engineering approach to do the inverse process, i.e. obtain the optimal synthesis parameters given a specific product characteristic. For the first time, it is presented a synthesis method that allows continuous and precise control of NPs morphology with the possibility to tune the aspect ratio over a large range from 1.4 (perfect truncated bipyramids) to 6 (elongated nanoparticles) and the length from 20 to 140 nm.

Nearly free surface silanols are the critical molecular moieties that initiate the toxicity of silica particles.

Inhalation of silica particles can induce inflammatory lung reactions that lead to silicosis and/or lung cancer when the particles are biopersistent. This toxic activity of silica dusts is extremely variable depending on their source and preparation methods. The exact molecular moiety that explains and predicts this variable toxicity of silica remains elusive. Here, we have identified a unique subfamily of silanols as the major determinant of silica particle toxicity. This population of "nearly free silanols" (NFS) appears on the surface of quartz particles upon fracture and can be modulated by thermal treatments. Density functional theory calculations indicates that NFS locate at an intersilanol distance of 4.00 to 6.00 Å and form weak mutual interactions. Thus, NFS could act as an energetically favorable moiety at the surface of silica for establishing interactions with cell membrane components to initiate toxicity. With ad hoc prepared model quartz particles enriched or depleted in NFS, we demonstrate that NFS drive toxicity, including membranolysis, in vitro proinflammatory activity, and lung inflammation. The toxic activity of NFS is confirmed with pyrogenic and vitreous amorphous silica particles, and industrial quartz samples with noncontrolled surfaces. Our results identify the missing key molecular moieties of the silica surface that initiate interactions with cell membranes, leading to pathological outcomes. NFS may explain other important interfacial processes involving silica particles.

Morphology, Surface Structure and Water Adsorption Properties of TiO2 Nanoparticles: A Comparison of Different Commercial Samples.

Water is a molecule always present in the reaction environment in photocatalytic and biomedical applications of TiO2 and a better understanding of its interaction with the surface of TiO2 nanoparticles is crucial to develop materials with improved performance. In this contribution, we first studied the nature and the surface structure of the exposed facets of three commercial TiO2 samples (i.e., TiO2 P25, SX001, and PC105) by electron microscopy and IR spectroscopy of adsorbed CO. The morphological information was then correlated with the water adsorption properties, investigated at the molecular level, moving from multilayers of adsorbed H2O to the monolayer, combining medium- and near-IR spectroscopies. Finally, we assessed in a quantitative way the surface hydration state at different water equilibrium pressures by microgravimetric measurements.

Migration study of organotin compounds from food packaging by surface-enhanced Raman scattering.

The potential of surface-enhanced Raman scattering (SERS) has been investigated for the rapid analysis of two representative organotin compounds (OTCs): dibutyltin maleate (DTM) and tributyltin chloride (TBT), after migration tests from polyvinyl chloride (PVC), as a model food packaging material in aqueous food simulant (acetic acid 3% w/v). OTCs, often used as heat stabilizers additives for PVC, are classified as endocrine disrupting chemicals (EDCs) and their migration potential has to be controlled in compliance with the normative prescriptions for food contact materials. In this study, colloidal silver nanoparticles (AgNPs) were applied as liquid SERS substrate for direct-in-liquid analysis of food simulant after standardized migration tests of PVC samples spiked with OTCs. Promising results were obtained, reaching detection limits below the permitted limits for the considered OTCs (i.e. 0.15 mg/l): DTM and TBT were detected down to 0.01 mg/l and 0.08 mg/l, respectively. Calibration curves were calculated for standard solutions of DTM and TBT in the dynamic range between 0 and 1 mg/l (reduced χ2 = 0.8), and 0.5-5 mg/l (reduced χ2 = 0.2), respectively. Migrated TBT and DTM were detected in the food simulant, specifically identified and quantified by SERS, with a measurement uncertainty around 10% in all cases. In particular, it was found that TBT can migrate in higher amount compared to DTM when the PVC film is in contact with a slightly acidic matrix. These results were further confirmed by inductively coupled plasma-mass spectrometry and UV-Vis spectroscopy. In the present study, direct-in-liquid SERS approach showed to be very promising because it provides a fast response and it allows to overcome most of the common drawbacks of solid SERS substrates due to inhomogeneity problems and low repeatability.

Monitoring the Reactivity of Formamide on Amorphous SiO2 by In-Situ UV-Raman Spectroscopy and DFT Modeling.

Formamide has been recognized in the literature as a key species in the formation of the complex molecules of life, such as nucleobases. Furthermore, several studies reported the impact of mineral phases as catalysts for its decomposition/polymerization processes, increasing the conversion and also favoring the formation of specific products. Despite the progresses in the field, in situ studies on these mineral-catalyzed processes are missing. In this work, we present an in situ UV-Raman characterization of the chemical evolution of formamide over amorphous SiO2 samples, selected as a prototype of silicate minerals. The experiments were carried out after reaction of formamide at 160 °C on amorphous SiO2 (Aerosil OX50) either pristine or pre-calcined at 450 °C, to remove a large fraction of surface silanol groups. Our measurements, interpreted on the basis of density functional B3LYP-D3 calculations, allow to assign the spectra bands in terms of specific complex organic molecules, namely, diaminomaleonitrile (DAMN), 5-aminoimidazole (AI), and purine, showing the role of the mineral surface on the formation of relevant prebiotic molecules.

Towards the control of the biological identity of nanobiomaterials: Impact of the structure of 011¯0 surface terminations of nanohydroxyapatite on the conformation of adsorbed proteins.

High-resolution transmission electron microscopy, ζ-potential and in-situ IR spectroscopy of adsorbed CO were combined for elucidating the ratio between {011¯0}_ Ca-rich: {011¯0}_ P-rich terminations of {011¯0} facets, i.e. the surfaces with the highest morphological importance, in two nanohydroxyapatite samples. Bovine serum albumin was found to form at least a monolayer on the surface left accessible to protein molecules by the agglomeration of nanoparticles when suspended in the buffered incubation medium. Noticeably, the conformation of adsorbed proteins appeared sensitive to the ratio between the two types of {011¯0} terminations, also resulting in a difference in the surface exposed toward the exterior by the adsorbed protein layer(s).

Toward a novel framework for bloodstains dating by Raman spectroscopy: How to avoid sample photodamage and subsampling errors.

Answers to questions about the time of bloodstains formation are often essential to unravel the sequence of events behind criminal acts. Unfortunately, the relevance of preserved evidence to the committed offence usually cannot be verified, because forensic experts are still incapable of providing an accurate estimate of the bloodstains' age. An antidote to this impediment might be substituting the classical dating approach - founded on the application of calibration models - by the comparison problem addressed using likelihood ratio tests. The key aspect of this concept involves comparing the evidential data with results characterizing reference bloodstains, formed during the process of supervised ageing so as to reproduce the evidence. Since this comparison requires data that conveys information inherent to changes accompanying the process of blood decomposition, this study provided a Raman-based procedure, designated for probing into the chemistry of ageing bloodstains. To circumvent limitations experienced with single-point measurements - the risk of laser-induced degradation of hemoglobin and subsampling errors - the rotating mode of spectral acquisition was introduced. In order to verify the performance of this novel sampling method, obtained spectra were confronted with those acquired during conventional static measurements. The visual comparison was followed by analysis of data structure using regularized MANOVA, which boosted the variance between differently-aged samples while minimizing the variance observed for bloodstains deposited at the same time. Studies of relation between these variances demonstrated the superiority of novel procedure, as it provided Raman signatures that enabled a better distinction between differently-aged bloodstains.

Flexible and Transparent Substrates Based on Gold Nanoparticles and TiO2 for in Situ Bioanalysis by Surface-Enhanced Raman Spectroscopy.

Flexible and transparent substrates are emerging as low cost and easy-to-operate support for surface-enhanced Raman spectroscopy (SERS). In particular, in situ SERS detection approach for surface characterization in transmission modality can be efficiently employed for non-invasive analysis of non-planar surfaces. Here we propose a new methodology to fabricate a homogenous, transparent, and flexible SERS membrane by the assistance of a thin TiO2 porous layer deposited on the PDMS surface, which supports the uniform loading of gold nanoparticles over large area. The substrate was first characterized for homogeneity, sensitivity and repeatability using a model molecule for SERS, i.e., 7-mercapto-4-methylcoumarin. Satisfactory intra-substrate uniformity and inter-substrates repeatability was achieved, showing an RSD of 10%, and an analytical sensitivity down to 10 nM was determined with an EF of 3.4 × 105 ± 0.4 × 105. Furthermore, SERS detection of pyrimethanil (PMT), a commonly employed pesticide in crops for human consumption, was performed in situ, exploiting the optical transparency of the device, using both model surfaces and non-flat bio-samples. PMT contamination at the phytochemical concentration levels corresponding to commonly used infield doses was successfully detected on the surface of the yellow Ficus benjiamina leaves, supporting the use of this substrate for food safety in-field application.

The puzzling issue of silica toxicity: are silanols bridging the gaps between surface states and pathogenicity?

BACKGROUND: Silica continues to represent an intriguing topic of fundamental and applied research across various scientific fields, from geology to physics, chemistry, cell biology, and particle toxicology. The pathogenic activity of silica is variable, depending on the physico-chemical features of the particles. In the last 50 years, crystallinity and capacity to generate free radicals have been recognized as relevant features for silica toxicity. The 'surface' also plays an important role in silica toxicity, but this term has often been used in a very general way, without defining which properties of the surface are actually driving toxicity. How the chemical features (e.g., silanols and siloxanes) and configuration of the silica surface can trigger toxic responses remains incompletely understood. MAIN BODY: Recent developments in surface chemistry, cell biology and toxicology provide new avenues to improve our understanding of the molecular mechanisms of the adverse responses to silica particles. New physico-chemical methods can finely characterize and quantify silanols at the surface of silica particles. Advanced computational modelling and atomic force microscopy offer unique opportunities to explore the intimate interactions between silica surface and membrane models or cells. In recent years, interdisciplinary research, using these tools, has built increasing evidence that surface silanols are critical determinants of the interaction between silica particles and biomolecules, membranes, cell systems, or animal models. It also has become clear that silanol configuration, and eventually biological responses, can be affected by impurities within the crystal structure, or coatings covering the particle surface. The discovery of new molecular targets of crystalline as well as amorphous silica particles in the immune system and in epithelial lung cells represents new possible toxicity pathways. Cellular recognition systems that detect specific features of the surface of silica particles have been identified. CONCLUSIONS: Interdisciplinary research bridging surface chemistry to toxicology is progressively solving the puzzling issue of the variable toxicity of silica. Further interdisciplinary research is ongoing to elucidate the intimate mechanisms of silica pathogenicity, to possibly mitigate or reduce surface reactivity.

The Case of Formic Acid on Anatase TiO2 (101): Where is the Acid Proton?

Carboxylic-acid adsorption on anatase TiO2 is a relevant process in many technological applications. Yet, despite several decades of investigations, the acid-proton localization-either on the molecule or on the surface-is still an open issue. By modeling the adsorption of formic acid on top of anatase(101) surfaces, we highlight the formation of a short strong hydrogen bond. In the 0 K limit, the acid-proton behavior is ruled by quantum delocalization effects in a single potential well, while at ambient conditions, the proton undergoes a rapid classical shuttling in a shallow two-well free-energy profile. This picture, supported by agreement with available experiments, shows that the anatase surface acts like a protecting group for the carboxylic acid functionality. Such a new conceptual insight might help rationalize chemical processes involving carboxylic acids on oxide surfaces.

The interaction of SiO2 nanoparticles with the neuronal cell membrane: activation of ionic channels and calcium influx.

AIM: To clarify the mechanisms of interaction between SiO2 nanoparticles (NPs) and the plasma membrane of GT1-7 neuroendocrine cells, with focus on the activation of calcium-permeable channels, responsible for the long lasting calcium influx and modulation of the electrical activity in these cells. MATERIALS & METHODS: Nontoxic doses of SiO2 NPs were administered to the cells. Calcium imaging and patch clamp techniques were combined with a pharmacological approach. RESULTS: TRPV4, Cx and Panx-like channels are the major components of the NP-induced inward currents. Preincubation with the antioxidant N-acetyl-L-cysteine strongly reduced the [Ca2+]i increase. CONCLUSION: These findings suggest that SiO2 NPs directly activate a complex set of calcium-permeable channels, possibly by catalyzing free radical production.

Enzyme activation by alternating magnetic field: Importance of the bioconjugation methodology.

Iron oxide nanoparticles (NPs) are attractive materials for enzyme immobilization and, thanks to their superparamagnetism, can be accessed by remote stimuli. This can be exploited to activate molecules that are not remotely actuable. Here, we demonstrate that thermophilic enzymes chemically linked to NPs can be activated in a "wireless" fashion by an external alternate magnetic field (AMF). To this aim, we have conjugated, with different binding strategies, the thermophilic enzymes α-amylase and l-aspartate oxidase to iron oxide NPs obtaining NP-enzyme systems with activities depending on the different orientations and stretching of the enzymes. Since enzyme activation occurs without a significant rise of the "overall" temperature of the systems, we have speculated a local NP-enzyme heating that does not immediately interest the rest of the solution that remains at relatively low temperature, low enough to allow non-thermophilic enzymes to work together with the NP-conjugated thermophilic enzymes. Nanoactuation of thermophilic enzymes by AMF has potential applications in different fields. Indeed, multi-enzymatic processes with enzymes with different temperature optima could be carried out in the same reaction pot and thermolabile products could be efficiently produced by thermophilic enzymes without suffering for the high temperatures. Moreover, our findings represent a proof of concept of the possibility to achieve a fine-tuning of the enzyme-NP system with the aim to intervene in cell metabolism.

Formamide Adsorption at the Amorphous Silica Surface: A Combined Experimental and Computational Approach.

Mineral surfaces have been demonstrated to play a central role in prebiotic reactions, which are understood to be at the basis of the origin of life. Among the various molecules proposed as precursors for these reactions, one of the most interesting is formamide. Formamide has been shown to be a pluripotent molecule, generating a wide distribution of relevant prebiotic products. In particular, the outcomes of its reactivity are strongly related to the presence of mineral phases acting as catalysts toward specific reaction pathways. While the mineral⁻products relationship has been deeply studied for a large pool of materials, the fundamental description of formamide reactivity over mineral surfaces at a microscopic level is missing in the literature. In particular, a key step of formamide chemistry at surfaces is adsorption on available interaction sites. This report aims to investigate the adsorption of formamide over a well-defined amorphous silica, chosen as a model mineral surface. An experimental IR investigation of formamide adsorption was carried out and its outcomes were interpreted on the basis of first principles simulation of the process, adopting a realistic model of amorphous silica.

Silica nanoparticles actively engage with mesenchymal stem cells in improving acute functional cardiac integration.

AIM: To assess functional effects of silica nanoparticles (SiO2-NPs) on human mesenchymal stem cell (hMSC) cardiac integration potential. METHODS: SiO2-NPs were synthesized and their internalization effects on hMSCs analyzed with particular emphasis on interaction of hMSCs with the cardiac environment Results: SiO2-NP internalization affected the area and maturation level of hMSC focal adhesions, accounting for increased in vitro adhesion capacity and augmented engraftment in the myocardial tissue upon cell injection in infarcted isolated rat hearts. SiO2-NP treatment also enhanced hMSC expression of Connexin-43, favoring hMSC interaction with cocultured cardiac myoblasts in an ischemia-like environment. CONCLUSION: These findings provide strong evidence that SiO2-NPs actively engage in mediating biological effects, ultimately resulting in augmented hMSC acute cardiac integration potential.

Detection of insect's meal in compound feed by Near Infrared spectral imaging.

Insects have recently emerged as a new protein source for both food and feed. Some studies have already demonstrated that insects' meal can be successfully added to animal feed without threaten animals' growth indices. However, effective and validated tests to individuate insects' meal in feed are strongly needed to meet traceability and safety concerns and to support the European legislation under development. Spectroscopic techniques represent valuable rapid and non-destructive methods that can be applied for in-situ analysis in feed production plants or in farms. In this work a Fourier Transform Near Infrared spectroscopy imaging (FT NIR) as a potential screening method for the detection and quantification of insects' meal in feed is presented. Discriminant analysis was used for the automatic recognition of insects' meal fragments into the feed matrix. Moreover, the possibility to quantify insect's meal in feed sample was successfully tested. The proposed method is a rapid and green strategy for feed contamination screening analysis.

Does the Abiotic Formation of Oligopeptides on TiO2 Nanoparticles Require Special Catalytic Sites? Apparently Not.

The oligomerization of non-activated amino acids catalyzed by nanostrucrured mineral oxide surfaces holds promises as a sustainable route for the industrial production of polypeptides. To analyze the influence of the surface type on the catalytic process, we performed, via a mild Chemical Vapor Deposition approach, the oligomerization of Glycine on two samples of TiO2 nanoparticles characterized by different relative amounts of defective surface terminations. Based on infrared spectroscopy and mass spectrometry data, we show herein that the formation of peptide bonds on titania nanoparticles does not require highly energetic surface terminations, but can occur also on the most abundant and thermodynamically most stable {101} facets of nanosized anatase.