Reflectance Spectroscopy as a Novel Tool for Thickness Measurements of Paint Layers.

A major challenge in heritage science is the non-invasive cross-sectional analysis of paintings. When low-energy probes are used, the presence of opaque media can significantly hinder the penetration of incident radiation, as well as the collection of the backscattered signal. Currently, no technique is capable of uniquely and noninvasively measuring the micrometric thickness of heterogeneous materials, such as pictorial layers, for any painting material. The aim of this work was to explore the possibility of extracting stratigraphic information from reflectance spectra obtained by diffuse reflectance spectroscopy (DRS). We tested the proposed approach on single layers of ten pure acrylic paints. The chemical composition of each paint was first characterised by micro-Raman and laser-induced breakdown spectroscopies. The spectral behaviour was analysed by both Fibre Optics Reflectance Spectroscopy (FORS) and Vis-NIR multispectral reflectance imaging. We showed that there is a clear correlation between the spectral response of acrylic paint layers and their micrometric thickness, which was previously measured by Optical Coherence Tomography (OCT). Based on significant spectral features, exponential functions of reflectance vs. thickness were obtained for each paint, which can be used as calibration curves for thickness measurements. To the best of our knowledge, similar approaches for cross-sectional measurements of paint layers have never been tested.

The Degradation of Daguerreotypes and the Relationship with Their Multi-Material Structure: A Multimodal Investigation.

Preserving and analytically examining daguerreotypes is particularly challenging because of their multi-material and multi-component structure. Various sensors have been exploited to examine mainly the image plates of the daguerreotypes even though the degradation goes beyond this component. Micro-analyses have been the preferred method due to the nanoscale structure of the image particles. In this work, we propose comprehensive multi-modal non-invasive sensing to investigate the corrosion products present in nine daguerreotypes from the Fondazione Alinari per la Fotografia (FAF, Florence, Italy). The methodology proposed includes chemical and morphological analyses: portable X-ray fluorescence spectrometry (pXRF), Raman microspectroscopy (µ-Raman), and micro-Fourier transform infrared spectroscopy in reflection mode (µ-rFTIR) for the chemical identification. For the first time, optical coherence tomography (OCT) was deployed to record the cross-sectional and morphological data of the relevant corrosion formations on daguerreotypes in a contactless way. The results allowed the characterization, in a non-invasive mode at a microscopic level, of a wide range of degradation products produced by the interaction of the different elements present in the structure of the daguerreotypes. The aim was to verify the performance of the proposed methodology and to link the chemical and physical complexity of the entire structure, disclosed by the state-of-art sensors, to the daguerreotype degradation. The results draw attention to the need to monitor not only the image condition but the whole object as a partially closed system in constant interaction internally and with the environment.

Nonlinear imaging and vibrational spectroscopic analysis of cellulosic fibres treated with COEX® flame-retardant for tapestry preservation.

Cellulose-based fabrics are widely used in the preservation and storage of historic tapestries. Their ease of flammability is a serious concern that greatly limits their applications and requires the development of effective and safe flame-retardant treatments. In this work, we analysed linen and cotton textile samples before and after COEX® treatment, a patented green technology imparting anti-flame properties by functionalizing the cellulose molecules with phosphorus and sulphur groups. Some of the samples were also exposed to photo-induced ageing after the treatment. The resulting structural and chemical changes in both fibres were characterized by nonlinear optical imaging modalities, namely Second Harmonic Generation (SHG) and Two-Photon Excited Fluorescence microscopies (TPEF), and Raman and Attenuated Total Reflection - Fourier Transform Infrared (ATR-FTIR) spectroscopies. Complementary results evidenced a reduction in microfibril crystallinity, attributed mainly to the reduction of hydrogen bonding among cellulose macromolecules, with a concomitant increase in fluorescence possibly due to the introduction of ester groups into cellulose chains and to decomposition of lignin into fluorescent by-products.

Non-invasive contactless analysis of an early drawing by Raffaello Sanzio by means of optical methods.

Studying highly valuable and fragile Renaissance drawings requires a non-invasive contactless analytical approach. In this work, we study an early drawing by Raffaello Sanzio, one of the most important artists of the Italian Renaissance, realized during his stay in Florence (1504-1508). Our analyses aimed to reveal the features of the paper support significant for its dating, identify and map the drawing media to understand the artist's technical approach, and document the drawing condition with imaging and single-sited optical methods. Reflectance Vis-NIR multispectral imaging spectroscopy elaborated with False-Colour and Principal Component Analysis provided information about the paper support and the drawing media. Laser scanning micro-profilometry and Optical Coherence Tomography allowed revealing the superficial micro-scale features of the support. The chemical composition of the different drawing materials was characterized by µ-Raman spectroscopy, which provided also some evidence of the conservation history of the drawing. Integration of spectroscopic and imaging data shows that Raffaello used different dry drawing media (carbon-based and Pb stylus) to sketch the figure and then refined the details and shadows with iron-gall ink. The paper presents a methodological approach for the analytical examination of fragile paper artworks.

Correction: Dal Fovo et al. Multi-Analytical Characterization and Radiocarbon Dating of a Roman Egyptian Mummy Portrait. Molecules 2021, 26, 5268.

Due to the fact that the policy regarding the publication of images from the collection of the Papyrological Institute, the owner of the object under study, changed when the article was already in publication, the authors would like to make the following corrections to this paper [...].

Portable Sequentially Shifted Excitation Raman Spectroscopy to Examine Historic Powders Enclosed in Glass Vials.

Raman spectroscopy (RS) is a powerful non-invasive tool for the characterization of materials. However, the fluorescence effect often hampers the detectability of the relatively weak vibrational Raman signal. Several approaches were exploited to overcome this limit. This work, in particular, evaluates the performance of an in situ portable sequentially shifted excitation (SSE™) Raman spectrometer applied to the examination of artistic historical pigment powders enclosed in glass vials. The explored handheld spectrometer employs a dual, temperature-shifted, 785 nm and 852 nm laser excitation to optimize both spectral coverage and fluorescence subtraction. The study demonstrates the feasibility of the SSE RS approach for non-invasive identification of art materials, and its applicability in complex situations where the examined material cannot be removed from its container. Laboratory measurements using benchtop dispersive micro-Raman spectroscopy at 785 nm are reported for comparison.

Automated Phasor Segmentation of Fluorescence Lifetime Imaging Data for Discriminating Pigments and Binders Used in Artworks.

The non-invasive analysis of fluorescence from binders and pigments employed in mixtures in artworks is a major challenge in cultural heritage science due to the broad overlapping emission of different fluorescent species causing difficulties in the data interpretation. To improve the specificity of fluorescence measurements, we went beyond steady-state fluorescence measurements by resolving the fluorescence decay dynamics of the emitting species through time-resolved fluorescence imaging (TRFI). In particular, we acquired the fluorescence decay features of different pigments and binders using a portable and compact fibre-based imaging setup. Fluorescence time-resolved data were analysed using the phasor method followed by a Gaussian mixture model (GMM) to automatically identify the populations of fluorescent species within the fluorescence decay maps. Our results demonstrate that this approach allows distinguishing different binders when mixed with the same pigment as well as discriminating different pigments dispersed in a common binder. The results obtained could establish a framework for the analysis of a broader range of pigments and binders to be then extended to several other materials used in art production. The obtained results, together with the compactness and portability of the instrument, pave the way for future in situ applications of the technology on paintings.

Revealing Underdrawings in Wall Paintings of Complex Stratigraphy with a Novel Reflectance Photoacoustic Imaging Prototype.

Revealing precious hidden features by a completely non-invasive approach is one of the crucial issues in the Heritage Science field. In this regard, concealed fresco paintings still represent an analytical challenge. This paper addresses the specific issue in wall painting diagnostics by the photoacoustic (PA) imaging technique, already proven to be efficient in revealing underdrawings and internal stratigraphy in movable paintings on paper and canvas. A newly set-up reflection PA prototype was applied here for the first time to probe the charcoal, graphite and sinopia hidden sketch drawings in concealed (gypsum, limewash, overpainted) wall paintings. The results presented here push forward the frontiers of the PA imaging technique and point to its potential effectiveness of revealing hidden underdrawings in historical wall paintings with complex stratigraphy.

Multi-Analytical Characterization and Radiocarbon Dating of a Roman Egyptian Mummy Portrait.

Fayum mummy portraits, painted around 2000 years ago, represent a fascinating fusion of Egyptian and Graeco-Roman funerary and artistic traditions. Examination of these artworks may provide insight into the Roman Empire's trade and economic and social structure during one of its most crucial yet still hazy times of transition. The lack of proper archaeological documentation of the numerous excavated portraits currently prevents their chronological dating, be it absolute or relative. So far, their production period has been defined essentially on the basis of the relevant differences in their pictorial style. Our study introduces the use of Accelerator Mass Spectrometry (AMS) to assess the age of a fragment of an encaustic painting belonging to the corpus of the Fayum portraits. The unexpected age resulting from 14C analysis suggests the need to reconsider previous assumptions regarding the period of production of the Fayum corpus. Furthermore, our multi-analytical, non-invasive approach yields further details regarding the fragment's pictorial technique and constituting materials, based on spectral and morphological analysis and cross-sectional examination.

Multisensorial Assessment of Laser Effects on Shellac Applied on Wall Paintings.

The assessment of five different laser treatments in the conservation of wall paintings was devised on the basis of the surface temperature monitoring by infrared thermography (IRT), ultraviolet-induced fluorescence-visible (UV-VIS) imaging, and optical coherence tomography (OCT). A series of yttrium-aluminum-garnet (YAG) lasers were tested for removal of shellac layers from wall painting mock-ups. The mock-ups were realized as buon fresco with different mineral based pigments (earths and iron oxide) on a lime- and sand-based mortar. After the carbonatation process, all the samples were treated with shellac (5% in ethanol). The effects of neodymium (Nd):YAG, holmium (Ho):YAG, and erbium (Er):YAG laser sources, in different operative modes, on average temperature of the surface, color, and morphology were inspected with complementary sensors. The results show the necessity to adopt a combined approach in establishing safe laser operating conditions to avoid any undesired effects induced on the artefacts by the laser treatments. We demonstrate, for the first time, the performance of the Ho:YAG laser in the removal of a conservation treatment.

Synchronized Hardware-Registered VIS-NIR Imaging Spectroscopy and 3D Sensing on a Fresco by Botticelli.

We discuss a synchronised sensing technique for the analysis of painted surfaces of frescos. Specifically, the performance of Visible-Near Infrared (VIS-NIR) Reflectance Imaging Spectroscopy (RIS) synchronized with three-dimensional (3D) acquisition is demonstrated in the study of a detached mural painting by Alessandro Botticelli. Synchronized sensing generates georeferenced data for simplified data treatment and interpretation. We show how such output data can provide key information to interpret important fresco surface and subsurface features (e.g., painting technique, material composition, pentimenti).

Behind the Scene of "The Holy Family with St. Anne and the Young St. John" by Bernardino Luini: A Computer-Assisted Method to Unveil the Underdrawings.

Uncovering the underdrawings (UDs), the preliminary sketch made by the painter on the grounded preparatory support, is a keystone for understanding the painting's history including the original project of the artist, the pentimenti (an underlying image in a painting providing evidence of revision by the artist) or the possible presence of co-workers' contributions. The application of infrared reflectography (IRR) has made the dream of discovering the UDs come true: since its introduction, there has been a growing interest in the technology, which therefore has evolved leading to advanced instruments. Most of the literature either report on the technological advances in IRR devices or present case studies, but a straightforward method to improve the visibility of the UDs has not been presented yet. Most of the data handling methods are devoted to a specific painting or they are not user-friendly enough to be applied by non-specialized users, hampering, thus, their widespread application in areas other than the scientific one, e.g., in the art history field. We developed a computer-assisted method, based on principal component analysis (PCA) and image processing, to enhance the visibility of UDs and to support the art-historians and curators' work. Based on ImageJ/Fiji, one of the most widespread image analysis software, the algorithm is very easy to use and, in principle, can be applied to any multi- or hyper-spectral image data set. In the present paper, after describing the method, we accurately present the extraction of the UD for the panel "The Holy Family with St. Anne and the Young St. John" and for other four paintings by Luini and his workshop paying particular attention to the painting known as "The Child with the Lamb".

Nuclear receptor ligands induce TREM-1 expression on dendritic cells: analysis of their role in tumors.

Dendritic cells (DCs) initiate adaptive immune responses after their migration to secondary lymphoid organs. The LXR ligands/oxysterols and the RXR ligand 9-cis Retinoic Acid (9-cis RA) were shown to dampen DC migration to lymphoid organs through the inhibition of CCR7 expression. We performed transcriptomics of DCs undergoing maturation in the presence of the LXR ligand 22R-Hydroxycholesterol (22R-HC). The analysis highlighted more than 1500 genes modulated by 22R-HC treatment, including the triggering receptor expressed on myeloid cells (TREM)-1, which was found markedly up-regulated. We tested the effect of other nuclear receptor ligands (NRL) and we reported the induction of TREM-1 following RXR, RAR and VDR activation. From a functional point of view, triggering of TREM-1 induced by retinoids increased TNFα and IL-1ß release, suggesting an active role of NRL-activated TREM-1+ DCs in inflammation-driven diseases, including cancer. Consistently with this hypothesis we detected DCs expressing TREM-1 in pleural effusions and ascites of cancer patients, an observation validated by the induction of TREM-1, LXR and RAR target genes when monocyte-DCs were activated in the presence of tumor-conditioned fluids. Finally, we observed a better control of LLC tumor growth in Trem-1-/- bone marrow chimera mice as compared to wild type chimera mice. Future studies will be necessary to shed light on the mechanism of TREM-1 induction by distinct NRL, and to characterize the role of TREM-1+ DCs in tumor growth.

Combined photoacoustic imaging to delineate the internal structure of paintings.

In this Letter, we present a combined photoacoustic imaging method, based on consecutive excitation using either the fundamental or the second-harmonic wavelength of a pulsed Nd:YAG laser for the stratigraphy of painted artworks. Near-infrared excitation was employed for the imaging of hidden underdrawings in mock-up samples, whereas visible light was used for the thickness mapping of the overlying paint through the detection of photoacoustic signal attenuation. The proposed methodology was proven effective in measuring thick and strongly absorbing layers, which would not be possible by means of other pure optical techniques, while also enabling the visualization of features underneath the painted surface. Such an implementation expands significantly the applicability of the previously presented photoacoustic technique, which was limited to point-measurements, and paves the way for novel application in historical and technical studies, as well as in documenting restoring operations.

Multianalytical non-invasive characterization of phthalocyanine acrylic paints through spectroscopic and non-linear optical techniques.

The documentation and monitoring of cleaning operations on paintings benefit from the identification and determination of thickness of the materials to be selectively removed. Since in artworks diagnosis the preservation of the object's integrity is a priority, the application of non-invasive techniques is commonly preferred. In this work, we present the results obtained with a set of non-invasive optical techniques for the chemical and physical characterization of six copper-phthalocyanine (Cu-Pc) acrylic paints. Cu-Pc pigments have been extensively used by artists over the past century, thanks to their properties and low cost of manufacture. They can also be found in historical paintings in the form of overpaints/retouchings, providing evidence of recent conservation treatments. The optical behaviour and the chemical composition of Cu-Pc paints were investigated through a multi-analytical approach involving micro-Raman spectroscopy, Fibre Optics Reflectance Spectroscopy (FORS) and Laser Induced Fluorescence (LIF), enabling the differentiation among pigments and highlighting discrepancies with the composition declared by the manufacturer. The applicability of Non Linear Optical Microscopy (NLOM) for the evaluation of paint layer thickness was assessed using the modality of Multi-photon Excitation Fluorescence (MPEF). Thickness values measured with MPEF were compared with those retrieved through Optical Coherence Tomography (OCT), showing significant consistency and paving the way for further non-linear stratigraphic investigations on painting materials.

Spectral Imaging and Archival Data in Analysing Madonna of the Rabbit Paintings by Manet and Titian.

A concise insight into the outputs provided by the latest prototype of visible-near infrared (Vis-NIR) multispectral scanner (National Research Council-National Institute of Optics, CNR-INO, Italy) is presented. The analytical data acquired on an oil painting Madonna of the Rabbit by É. Manet are described. In this work, the Vis-NIR was complemented with X-ray fluorescence (XRF) mapping for the chemical and spatial characterization of several pigments. The spatially registered Vis-NIR data facilitated their processing by spectral correlation mapping (SCM) and artificial neural network (ANN) algorithm, respectively, for pigment mapping and improved visibility of pentimenti and of underdrawing style. The data provided several key elements for the comparison with a homonymous original work by Titian studied within the ARCHive LABoratory (ARCHLAB) transnational access project.

Using Optical Coherence Tomography to Reveal the Hidden History of The Landsdowne Virgin of the Yarnwinder by Leonardo da Vinci and Studio.

Optical coherence tomography (OCT) was used for non-invasive examination of a well-known, yet complex, painting from the studio of Leonardo da Vinci in combination with routine imaging in various bands of electromagnetic radiation. In contrast with these techniques, OCT provides depth-resolved information. Three post-processing modalities were explored: cross-sectional views, maps of scattering from given depths, and their 3D models. Some hidden alterations of the painting owing to past restorations were traced: retouching and overpainting with their positioning within varnish layers as well as indications of a former transfer to canvas.

Prognostic role of liver X receptor-alpha in resected stage II and III non-small-cell lung cancer.

INTRODUCTION: In the search of new therapeutical strategies against non-small-cell lung cancer (NSCLC), the identification of new prognostic factors is crucial. OBJECTIVES: In this study, we analyzed the prognostic value of the liver X receptor-alpha (LXR-alpha), a nuclear receptor of a family of cholesterol derivatives called oxysterols, in patients with radically resected NSCLC. METHODS: We retrospectively reviewed 140 stage II and III surgically treated NSCLC patients that were grouped by percentage of LXR-alpha-positive cells value above or below its median value. Tumor-related survival was evaluated as primary end point. RESULTS: The 5-year overall and tumor-related survival rates were 40% and 46%, respectively. The median percentage of LXR-alpha-positive cells was 20%. Patients with stage II NSCLC had higher LXR-alpha values than those with stage III (P = .04). Univariate analysis demonstrated that both TNM stage and LXR-alpha were significantly related to tumor-related survival (P = .006 and P = .004, respectively). The 5-year tumor-related survival rates in stage II and III NSCLC were 56% and 34%, respectively. The 5-year tumor-related survival rates in high and low LXR-alpha value were 57% and 32%, respectively. The multivariate analysis showed that both TNM stage and LXR-alpha were independent prognostic factors (P = .01 and P = .007, respectively) with hazard ratio of 1.92 and 0.49, respectively. CONCLUSION: LXR-alpha seems to be an independent prognostic factor indicating a better survival in completely resected stage II and III NSCLC patients.

Close to the diffraction limit in high resolution ATR FTIR mapping: demonstration on micrometric multi-layered art systems.

This paper is aimed at demonstrating the potentiality of high resolution Attenuated Total Reflection Fourier Transform Infrared micro-mapping (micro-ATR-FTIR) to reconstruct the images of micrometric multi-layered systems. This method can be an effective analytical alternative when the layer thickness requires high lateral resolution, and fluorescence or thermal effects prevent the deployment of conventional analytical techniques such as micro-Raman spectroscopy. This study demonstrates the high micro-ATR-FTIR setup performances in terms of lateral resolution, spectral quality and chemical image contrast using a new laboratory instrument equipped with a single element detector. The method has been first validated on mock-ups and then successfully applied on cross-sectional samples from real artworks: Leonardo da Vinci's mural painting, characterised by a few micrometers thin sequence of organic and inorganic layers, and an outdoor marble statue, with a complex sequence of decay products on its surface. This study paves the way to a new investigation modality of micrometric systems, combining high lateral resolution with excellent spectral quality, essential in the field of Cultural Heritage as well as in the wider area of materials and forensic sciences.

Side-Chain Modified Ergosterol and Stigmasterol Derivatives as Liver X Receptor Agonists.

A series of stigmasterol and ergosterol derivatives, characterized by the presence of oxygenated functions at C-22 and/or C-23 positions, were designed as potential liver X receptor (LXR) agonists. The absolute configuration of the newly created chiral centers was definitively assigned for all the corresponding compounds. Among the 16 synthesized compounds, 21, 27, and 28 were found to be selective LXRα agonists, whereas 20, 22, and 25 showed good selectivity for the LXRß isoform. In particular, 25 showed the same degree of potency as 22R-HC (3) at LXRß, while it was virtually inactive at LXRα (EC50 = 14.51 µM). Interestingly, 13, 19, 20, and 25 showed to be LXR target gene-selective modulators, by strongly inducing the expression of ABCA1, while poorly or not activating the lipogenic genes SREBP1 and SCD1 or FASN, respectively.