Lisa Ghezzi

Physico-chemical characterization of protein–pigment interactions in tempera paint reconstructions: casein/cinnabar and albumin/cinnabar

AbstractIn this work, we characterized paint reconstructions using ovalbumin and casein as binders, and cinnabar (HgS) as a pigment, before and after artificial ageing. Egg and casein are common paint binders that were used historically in the technique of tempera painting. Despite extensive research on the identification of proteinaceous binders in paintings, there is a substantial lack of knowledge regarding the ageing pathway of their protein content, and their chemical interaction with inorganic pigments. Thermogravimetric analysis, infrared spectroscopy and size-exclusion chromatography (SEC) were used to reveal the physico-chemical processes involved in the ageing of proteins in paintings. Taken together, the three techniques highlighted that proteins are subject to both cross-linking and hydrolysis upon ageing, and to a lesser extent, to oxidation of the side chains. Mercury–protein interactions were also revealed using a cold vapour generation atomic fluorescence spectrometer mercury-specific detector coupled to SEC. The study clearly showed that HgS forms stable complexes with proteins and acts as a sensitizer in cross-linking, hydrolysis and oxidation. FigureA multi-techinque approach to the study of protein/cinnabar tempera paint recontructions: thermogravimetric analysis, Fourier Transform Infrared Spectroscopy and size exclusion chromatography

A simple and reliable methodology to detect egg white in art samples

A protocol for a simple and reliable dot-blot immunoassay was developed and optimized to test work of art samples for the presence of specific proteinaceus material (i.e. ovalbumin-based). The analytical protocol has been extensively set up with respect, among the other, to protein extraction conditions, to densitometric analysis and to the colorimetric reaction conditions. Feasibility evaluation demonstrated that a commercial scanner and a free image analysis software can be used for the data acquisition and elaboration, thus facilitating the application of the proposed protocol to commonly equipped laboratories and to laboratories of museums and conservation centres. The introduction of method of standard additions in the analysis of fresh and artificially aged laboratory-prepared samples, containing egg white and various pigments, allowed us to evaluate the matrix effect and the effect of sample aging and to generate threshold density values useful for the detection of ovalbumin in samples from ancient works of art. The efficacy of the developed dot-blot immunoassay was proved testing microsamples from 13th–16th century mural paintings of Saint Francesco Church in Lodi (Italy). Despite the aging, the altered conditions of conservation, the complex matrix, and the micro-size of samples, the presence of ovalbumin was detected in all those mural painting samples where mass-spectrometry-based proteomic analysis unambiguously detected ovalbumin peptides.

Interactions between inorganic pigments and rabbit skin glue in reference paint reconstructions

The thermal degradation of rabbit skin glue, a collagen-based proteinaceous material used as a paint binder in paintings, was investigated in this paper. Paint reconstructions of the glue on its own or mixed with azurite (Cu3(CO3)2(OH)2), calcium carbonate (CaCO3), hematite (Fe2O3·nH2O) and red lead (Pb3O4) were analysed using a thermoanalytical approach. This method enabled us to investigate the interactions between the glue and pigments before and after artificial indoor light ageing. The study was carried out using differential scanning calorimetry, thermogravimetry and thermogravimetry/FTIR analysis already successfully employed to characterize the paint binders. The results highlighted that all the inorganic pigments interact with rabbit skin glue, thus decreasing the thermal stability of the binder. Light ageing further decreased the thermal stability of pigmented paint replicas, suggesting a moderate increase in the rate of the degradation.

Kinetics and equilibria of the interaction of 8-hydroxyquinoline with gallium(III) in water and sodium dodecyl sulfate solution.

The kinetics and equilibria of binding of gallium(III) to the 8-hydroxyquinoline (HQ) have been investigated in water and in the presence of sodium dodecyl sulfate (SDS) micelles. Moreover, the pKA1 and pKA2 of HQ and first hydrolysis constant of Ga3+ ion have been measured in water and SDS solution. The analysis of the kinetic and thermodynamic behavior reveals that the reactive form of Ga(III) is GaOH2+ in both cases. Although in water the only bound form of Ga(III) appears to be the deprotonated complex GaL, evidence for stabilization of the protonated form, GaHL on the micelle surface and stabilization of Ga3+ with respect to GaOH2+ is provided by the kinetic behavior in SDS. The addition of SDS at concentrations around the critical micellar concentration, results in a large enhancement of the rate of complex formation. The large catalytic effect produced by the SDS micellar solution provides a promising basis for the extraction of gallium from water using the HQ/SDS system. A procedure for gallium(III) extraction and recovery based on ligand modified-micellar enhanced ultrafiltration method, using the HQ/SDS system, is described.

Chemical-physical and in vivo evaluations of a self-assembling amphiphilic peptide as an injectable hydrogel scaffold for biomedical applications

The self-aggregation and gelation of an amphiphilic peptide (C17H35CONH–A4G3ERGD, peptide amphiphile) were studied by light scattering, viscometry, nuclear magnetic resonance diffusometry, and atomic force microscopy. The peptide amphiphile critical aggregation concentration was evaluated to be 16 and 60 µM by light scattering and viscometry, respectively. The observed difference was attributed to the larger sensitivity of the latter technique to the presence of long fibrils. The addition of one equivalent or more of divalent cations (Ca2+ and Mg2+) to peptide amphiphile formed dense incoherent hydrogels. Based on the atomic force microscopy and nanoindentation data, both the hydrogel morphology and stiffness were independent of the cation type and peptide amphiphile concentration. However, gel stiffness increased on increasing Ca2+/peptide amphiphile molar ratio while a parallel decrease in the apparent water diffusion rate was observed by nuclear magnetic resonance diffusometry. The dispersions of endothelial progenitor cells in the peptide amphiphile hydrogels were evaluated in vivo on a rat tissue hypoxia model. Significant capillary formation at the injection site was observed by tissue appearance and histological examination, which indicated endothelial progenitor cell/peptide amphiphile hydrogel-enhanced angiogenesis in ischemic tissue.

Influence of environmental and anthropogenic parameters on thallium oxidation state in natural waters

The abandoned mining area of Valdicastello Carducci (Tuscany, Italy) is characterized by the massive presence of thallium in the acid mine drainages and in the valley stream crossing the region. We previously found that Tl(III), generally considered the less stable oxidation state of thallium, is present both in the stream and in tap water distributed in the area, whereas acid mine drainages only contain Tl(I). These findings posed some concern related to the reactivity and dispersion of this toxic element in the environment. Since the valence state of thallium determines its toxicity, distribution and mobility, the study of thallium redox speciation appears crucial to understand its environmental behaviour. In this work, water samples collected from the mine drainages and the contaminated stream were adopted as model to study the distribution of aqueous Tl(I)/Tl(III) as a function of light exposure and solution properties and composition. The influence of three light sources and organic acids was evaluated. Thallium speciation was also assessed in tap water after treatment with common oxidizing agents, and in the rust crust collected from the public waterworks.

Detection of heterologous bovine pancreatic trypsin inhibitor by capillary zone electrophoresis

Abstract A method based on the use of capillary zone electrophoresis (CZE) is proposed for the determination of the heterologous protein bovine pancreatic trypsin inhibitor (BPTI) expressed in yeast. Saccharomyces cerevisiae cells were transformed with plasmid pCB235U and the amount of inhibitor expressed in yeast was measured both in the culture medium (supernatant) and the cell extract. Supernatant samples were investigated by the CZE method preceded by affinity chromatography (the BPTI yield was between 2 and 5 μg ml−1); direct determination of BPTI in cell extract was performed taking advantage of the complex formation of BPTI and trypsin (BPTI yield was 5 μg ml−1).