Maria Paola Bracciale

Graphite Nanoplatelets and Caenorhabditis elegans: Insights from an in Vivo Model

We evaluated the toxicity of graphite nanoplatelets (GNPs) in the model organism Caenorhabditis elegans. The GNPs resulted nontoxic by measuring longevity as well as reproductive capability end points. An imaging technique based on Fourier transform infrared spectroscopy (FT-IR) mapping was also developed to analyze the GNPs spatial distribution inside the nematodes. Conflicting reports on the in vitro antimicrobial properties of graphene-based nanomaterials prompted us to challenge the host-pathogen system C. elegans-Pseudomonas aeruginosa to assess these findings through an in vivo model.

Zinc oxide microrods and nanorods: Different antibacterial activity and their mode of action against Gram-positive bacteria

The development of antibiotic resistance among pathogenic bacteria combined with increased implant-associated infections have determined a great interest towards new bactericidal materials containing various organic and inorganic substances. Among them, zinc oxide (ZnO) derived materials have received considerable attention due to their unique antibacterial, antifungal, and UV filtering properties as well as high catalytic and photochemical activities. In the present work, we investigate the antimicrobial properties against two Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) of ZnO microrods (MRs) and nanorods (NRs), produced in bulk quantities through simple and inexpensive methods. We demonstrate that the antimicrobial effect is strongly dependent on the rod size and dose. Scanning electron microscopy analysis revealed that the two investigated microbial types interact differently with the ZnO-MRs and NRs due to their different morphology. This resulted in different outcomes as reported by their respective Colony Forming Unit (CFU) capabilities. Moreover, Fourier Transform Infrared (FT-IR) spectroscopy revealed that changes in cell outer structures, i.e. membrane and exopolysaccharides (EPS), produced by the interaction with the ZnO structures, are responsible for the antimicrobial mechanism without the accumulation of reactive oxygen species. This was further strengthened by the increased survival observed in the case of bacterial cells treated in the presence of an osmotic support, like glycerol. In addition, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis showed that reduced cell viability is not strictly correlated to increased zinc ion release in the suspension. We then concluded that ZnO-NRs have a superior antimicrobial effect against both S. aureus and B. subtilis at much lower doses when compared to ZnO-MRs. This is mainly due to the smaller diameter of the NRs, which promotes surface damaging and protein alteration of the cell wall. Finally, the lack of toxicity and the antimicrobial properties of ZnO-NRs versus S. aureus, validated in vivo using the nematode Caenorhabditis elegans as host infection model, confirm the promising exploitation of ZnO-NRs in biomedical applications.

Effect of an enzymatic treatment with cellulase and mannanase on the structural properties of Nannochloropsis microalgae

The effects of an enzymatic treatment with cellulase and mannanase on the properties of marine microalgae Nannochloropsis sp. were investigated. The combined use of these enzymes synergistically promoted the recovery of lipids from the microalgae, increasing the extraction yield from 40.8 to over 73%. Untreated and enzymatically treated microalgae were characterized by chemical analysis and by TGA/DTG, FTIR, XRD and SEM. Significant changes were observed in the chemical composition and thermal behavior of the microalgae. The enzymatic treatment also resulted in an increase of the crystalline-to-amorphous cellulose ratio. SEM images revealed dramatic changes in cell morphology, extensive cell damage and release of intracellular material. Overall, the results obtained indicate that the enzymes used are capable of disrupting the microalgal cell wall and that a combination of common analytical techniques can be used to assess the enzyme-induced damage.

Aqueous polythiophene electrosynthesis: A new route to an efficient electrode coupling of PQQ-dependent glucose dehydrogenase for sensing and bioenergetic applications

In this study, polythiophene copolymers have been used as modifier for electrode surfaces in order to allow the immobilization of active pyrroloquinoline quinone dependent glucose dehydrogenase (PQQ-GDH) and to simultaneously improve the direct electrical connection of the enzyme with the electrode. Polymer films are electrosynthesized in aqueous solution without the need of surfactants onto carbon nanotubes modified gold electrodes from mixtures of 3-thiopheneacetic acid (ThCH2CO2H) and 3-methoxythiophene (ThOCH3) using a potentiostatic pulse method. Polythiophene deposition significantly improves the bioelectrocatalysis of PQQ-GDH: the process starts at - 200 mV vs. Ag/AgCl and allows well-defined glucose detection at 0 V vs. Ag/AgCl with high current density. Several parameters of the electro-polymerization method have been evaluated to maximize the anodic current output after enzyme coupling. The polymer deposited by this new procedure has been morphologically and chemically characterized by different methods (SEM, EDX, FT-IR, UV-Vis, XPS and Raman spectroscopy). The bioelectrocatalytic response towards increasing glucose concentrations exhibits a dynamic range extending from 1 μM to 2 mM. The low applied potential allows to avoid interferences from easily oxidizable substances such as uric acid and ascorbic acid. Short and long-term stability has been evaluated. Finally, the PQQ-GDH electrode has been coupled to a bilirubin oxidase (BOD)- and carbon nanotube-based cathode in order to test its performance as anode of a biofuel cell. The promising results suggest a further investigation of this kind of polymers and, in particular, the study of the interaction with other enzymes in order to employ them in building up biosensors and biofuel cells.

Effectiveness of Phosphocitrate as Salt Crystallization Inhibitor in Porous Materials: Case Study of the Roman Mosaic of Orpheus and the Beasts (Perugia, Italy)

Salt crystallization in porous materials constitutes one of the major causes of decay of buildings/archaeological sites in a wide range of environments. Desalination is among the most common methodologies of conservative treatment for salt decay. However, classic desalination techniques might be not suitable for long-term conservation. During the past decade interest has been an increasing towards crystallization inhibitors as a new means of controlling crystallization damage. This study deals with the first in-field application of an environment-friendly inhibitor system— phosphocitrate (PC). In particular, the case study of the Roman mosaic of Orpheus and the Beasts in Perugia (Italy) is presented. The inhibitor is completely soluble in water or alcohol, non-toxic, and easy to apply, thus enabling its use in accordance with the volatile organic compounds emission control and safety during the conservation works. Relevant samples from control and treated mosaic areas were collected and analyzed comparatively by means of Fourier transform-infrared spectroscopy, scanning electron microscopy, and energy dispersive spectrometry to study the potential of the inhibitor system in preventing/controlling salt damage in such archaeological site.

Characterization and Digital Restauration of XIV-XV Centuries Written Parchments by Means of Nondestructive Techniques: Three Case Studies

Parchment is the primary writing medium of the majority of documents with cultural importance. Unfortunately, this material suffers of several mechanisms of degradation that affect its chemical-physical structure and the readability of text. Due to the unique and delicate character of these objects, the use of nondestructive techniques is mandatory. In this work, three partially degraded handwritten parchments dating back to the XIV-XV centuries were analyzed by means of X-ray fluorescence spectroscopy, µ-ATR Fourier transform infrared spectroscopy, and reflectance and UV-induced fluorescence spectroscopy. The elemental and molecular results provided the identification of the inks, pigments, and superficial treatments. In particular, all manuscripts have been written with iron gall inks, while the capital letters have been realized with cinnabar and azurite. Furthermore, multispectral UV fluorescence imaging and multispectral VIS-NIR imaging proved to be a good approach for the digital restoration of manuscripts that suffer from the loss of inked areas or from the presence of brown spotting. Indeed, using ultraviolet radiation and collecting the images at different spectral ranges is possible to enhance the readability of the text, while by illuminating with visible light and by collecting the images at longer wavelengths, the hiding effect of brown spots can be attenuated.

Multi-technique characterisation of commercial alizarin-based lakes

The characterization of ancient and modern alizarin-based lakes is a largely studied topic in the literature. Analytical data on contemporary alizarin-based lakes, however, are still poor, though of primary importance, since these lakes might be indeed present in contemporary and fake paintings as well as in retouchings. In this work we systematically investigate the chemical composition and the optical features of fifteen alizarin-based lakes, by a multi-analytical technique approach combining spectroscopic methods (i.e. Energy Dispersive X-ray Fluorescence Spectroscopy, EDXRF; Attenuated Total Reflectance Fourier-Transform Infrared Spectroscopy, ATR-FTIR; X-ray Powder Diffraction, XRD; UV induced fluorescence and reflectance spectroscopies) and chromatography (i.e. High-performance Liquid Chromatography coupled with a Photodiode Array Detector, HPLC-PDA). Most of the samples contain typical compounds from the natural roots of madder, as occurring in ancient and modern lakes, but in two samples (23600-Kremer-Pigmente and alizarin crimson-Zecchi) any anthraquinonic structures were identified, thus leading to hypothesize the presence of synthetic dyes. The detection of lucidin primeveroside and ruberythrique acid in some lakes suggest the use of Rubia tinctorum. One sample (23610-Kremer-Pigmente) presents alizarin as the sole compound, thereby revealing to be a synthetic dye. Moreover, gibbsite, alunite and kaolinite were found to be used as substrates and/or mordants. Visible absorption spectra of the anthraquinonic lakes show two main absorption bands at about 494-511nm and 537-564nm, along with a shoulder at about 473-479nm in presence of high amounts of purpurin. Finally, from the results obtained by UV induced fluorescence spectroscopy it is possible to figure out that, although it is commonly assumed that the madder lake presents an orange-pink fluorescence, the inorganic compounds, added to the recipe, could induce a quenching phenomenon or an inhibition of the fluorescence, as occurring in some commercial alizarin-based lakes.

Boosting biomass valorisation. Synergistic design of continuous flow reactors and water-tolerant polystyrene acid catalysts for a non-stop production of esters

An effective flow protocol exploiting the features of a specifically designed solid catalytic system has been defined for the direct esterification of biomass-derived carboxylic acids with structurally diverse alcohols on grams per hour scale and with minimal cost in terms of waste disposal. The potential of the catalyst to work indefinitely under pump-free continuous flow conditions has been explored.

Piezo-resistive properties of graphene based PVDF composite films for strain sensing

Graphene based polyvinylidene fluoride (PVDF) composite films have been produced and characterized in order to investigate their piezo-resistive properties. To this purpose, a new production process has been developed, with the aim of fabricating at low cost PVDF films filled with graphene nanoplatelets (GNPs). The produced films, having a GNP content of 1.5%wt and 2% wt, have been characterized from morpholocial and chemical points of view. Moreover, their piezo-resistive properties have been investigated both in static and cyclic conditions. The produced films show a stable and repeatable response to the applied strain. A sensitivity of 15 has been measured for the PVDF/GNP film filled at 1.5% wt, corresponding to an applied strain of 1.5%.

Fluorescence lidar measurements at the archaeological site House of Augustus at Palatino, Rome

Early diagnostics and documentation fulfill an essential role for an effective planning of conservation and restoration of cultural heritage assets. In particular, remote sensing techniques that do not require the use of scaffolds or lifts, such as fluoresence lidar, can provide useful information to obtain an overall assessment of the status of the investigated surfaces and can be exploited to address analytical studies in selected areas. Here we present the results of a joint Italian-Swedish project focused on documenting and recording the status of some sections of the part closed to the public by using fluorescence hyperspectral imaging lidar. The lidar used a tripled-frequency Nd:YAG laser emitting at 355 nm as excitation source and an intensified, gated 512x512-pixel CCD as detector. The lidar had imaging capabilities thanks to a computer-controlled scanning mirror. The fluorescence characteristics of fresco wall paintings were compared to those of fresco fragments found at the same archaeological site and separately examined in the lab using FT-IR and Raman techniques for the identification of pigments. The fluorescence lidar was also used to remotely detect the growth of phototrophic biodeteriogens on the walls. The fluorescence lidar data were compared with results from biological sampling, cultivation and laboratory analysis by molecular techniques.