Site-selective protein glycation and PEGylation

A two step protocol has been set up to selectively conjugate PEG to buried amino acids of proteins. The process involves site-specific glycation followed by PEGylation of the oxidized glycosides. Aimed at glycating the cysteine groups of proteins, two maleimide-glycosylic linkers have been synthesised: galactosyl-glucono-CO–NH–(CH₂)₁₂–NH–CO–(CH₂)₂-maleimide and maltosyl-glucono-CO–NH–(CH₂)₁₂–NH–CO–(CH₂)₂-maleimide. The linkers were extensively characterized by ¹H NMR, FT-IR, ESI–TOF mass spectrometry and colorimetric assays. Complete conjugation of the activated linkers to Cys³⁴ of human serum albumin was obtained in about 2 h. The selective oxidation of the galactosyl and maltosyl moieties by periodate treatment yielded two and three available aldehyde groups, respectively. The PEG-hydrazide conjugation to the aldehyde groups was found to be 100% in about 40 h, whereas less than 30% protein modification was obtained by direct conjugation of commercial PEG-maleimide to Cys³⁴. The pH dependent PEG-glycosyl hydrazone bond hydrolysis at various pH values was verified. PEG release was faster under mild acidic and basic conditions than at neutral pH. Furthermore, the maltosyl derivatives, by virtue of the higher number of coupled PEG chains, showed a slower protein release as compared to the galactosyl counterpart, indicating that the choice of the glycosylic linker allows for control of protein release kinetics.     

An advanced multivariate approach for processing X-ray fluorescence spectral and hyperspectral data from non-invasive in situ analyses on painted surfaces

In the last decades, in situ non-invasive analytical techniques have been widely used for the analysis of paintings. These techniques are useful to extensively map the surface in a non-invasive way, in order to identify the most representative areas to be sampled. When spectroscopic investigations, such as X ray fluorescence (XRF), are conducted, they usually imply the acquisition of a huge amount of measurements. Subsequently, all these data should be processed in situ, in order to immediately support the sampling strategies. To this aim, an appropriate and fast strategy for multivariate treatment of XRF spectral and hyperspectral data sets is presented, able to account for inter-correlation among variables, which is an issue of high importance for elemental analyses. The main advantage of the approach is that XRF spectral profiles are analysed directly, without computation of derived parameters, by means of principal component analysis (PCA). This procedure allows a fast interpretation of results that can be accomplished in situ. Particular attention was paid to the selection of proper spectral pre-treatments to be applied on data together with the use of several chemometric tools (peak alignment, spectra normalisation and exploratory analysis) aimed at improving the interpretation of XRF results. In addition, the application of multivariate exploratory analysis on XRF hyperspectral maps was studied by using an interactive brushing procedure. The multivariate approach was validated on data obtained from the analysis of the famous Renaissance panel painting “The Ideal City”, exhibited in Palazzo Ducale of Urbino, Italy.     

Structural and vibrational characterization of medieval like glass samples

Within the framework of a project aimed to develop protective coatings for antique glass windows, three glass varieties of medieval-like composition were prepared using recipes deduced from archaeometric studies and different amounts of potassium for fluxing (15–20–25 K₂O wt.%). Batches were melted in mullite crucibles using an electric furnace at 1350 °C, the glass was fast cooled in air and annealed at temperatures 20 °C higher than the sample glass transition temperatures (726, 702 and 683 °C for V1, V2 and V3, respectively). The chemical composition of the glass was determined by X-Ray Fluorescence Spectroscopy (XRF) and the glass transition temperature was checked by differential thermal analysis (DTA). X-Ray Powder Diffraction (XRPD) data and vibrational spectroscopic experiments (FT-IR and Raman) revealed a direct dependence of glass de-polymerization on potassium content. In particular, the Raman data were interpreted on the basis of SiO₄ structural units Qⁿ and the polymerization index I_p.     

Synthesis of DnaK and GroEL in Escherichia coli cells exposed to different magnetic field signals

The effects of extremely low frequency magnetic field (ELF-MF)(1 mT, 50 Hz) on the heat shock protein (HSP) synthesis in Escherichia coli were investigated. Two magnetic field signals were studied: sinusoidal (SMF) and pulsed square wave (PMF). It was found that bacteria exposed to SMF showed a significantly higher level of DnaK and GroEL proteins as compared to sham-exposed bacteria as revealed by Western blot, whereas a lower level was observed after PMF exposure. Similar results were obtained when bacterial cells were exposed to heat shock (HS) after ELF-MF exposure: again SMF and PMF resulted in an increase and in a reduction of HSP amount in comparison with sham control, respectively. In conclusion, the MF influences the synthesis of HSPs in E. coli in a way that critically depends on the signal characteristics.     

A Raman Scattering Study of PbTiO3 and TiO2 Obtained by Sol-Gel

The mechanisms that give rise to the broadening and the shifts of the Raman peaks of titanium dioxide and lead titanate nanocrystals prepared by sol-gel are discussed. Phonon confinement and oxygen deficiency are competitive mechanisms in TiO2 obtained by different sol-gel preparations whereas pressure effects on the nanocrystals predominate in ferroelectric PbTiO3.     

An integrated Raman and petrographic characterization of Italian mediaeval artifacts in pietra ollare (soapstone)

Ten fragments of pots made by pietra ollare (basic and ultrabasic metamorphic rocks belonging to greenschist facies) found in the archaeological site of Crocetta di Sant'Agata Bolognese (Bologna, Italy) have been studied to characterize them and to define their provenance. The fragments, dated between the 9^th and 10^th century AD, show traces of blackening from fire, indicating the use of these findings as pots by fire. The extensive use of a non‐destructive technique, Raman spectroscopy, in both laboratory and portable forms, for the provenance analysis of pietra ollare artifacts has been evaluated. Micro‐Raman spectroscopy was used for a detailed study of the main components, to identify the secondary minerals and to study the distribution of the mineral phases in the samples. The samples show schistose texture, are fine grained and consist mainly of talc and chlorite, and subordinate carbonates, oxides and serpentine. Olivine is rare. Based on the mineralogical composition, the material can be defined as chlorite and carbonate talc‐schist. Our samples of pietra ollare may be assigned to chlorite and magnesite‐bearing talc‐schists. The alpine areas of origin of this lithotype are Valtellina, Valchiavenna and Val Bregaglia. In particular, we can hypothesize that the origin of these findings is Valchiavenna, which had great commercial importance in the Middle Ages. Further measurements, obtained with a portable Raman spectrometer, directly on a quarry near Chiavenna, support our hypothesis. Copyright © 2013 John Wiley & Sons, Ltd.     

A comparison between ab initio calculated and measured Raman spectrum of triclinic albite (NaAlSi3O8)

Albite is one of the most common minerals in the Earth's crust, and its polymorphs can be found in rocks with different cooling histories. The characteristic spectrum of vibration of the albite mineral reflects its structural Si/Al ordering. In this study, we report on the comparison between the Raman spectra measured on a natural and fully ordered (as deduced on the basis of single‐crystal X‐ray diffraction data) ‘low albite’, NaAlSi₃O₈, and those calculated at the hybrid Hartree–Fock/density functional theory level by employing the WC1LYP Hamiltonian, which has proven to give excellent agreement between calculated and experimentally measured vibrational wavenumbers in silicate minerals. All the 39 expected A_g modes are identified in the Raman spectra, and their wavenumbers and intensities, in different scattering configurations, correspond well to the calculated ones. The average absolute discrepancy is ~3.4 cm⁻¹, being the maximum discrepancy |Δv|_max ~ 10.3 cm⁻¹. The very good quality of the WC1LYP results allows for reliable assignments of the Raman features to specific patterns of atomic vibrational motion. Copyright © 2015 John Wiley & Sons, Ltd.     

Characterization of colorants and opacifiers in roman glass mosaic tesserae through spectroscopic and spectrometric techniques

Several glass mosaic tesserae were recovered during the archeological excavation of the thermal baths at the ‘Villa dei Quintili’ in Rome and dated to the second century ad . This work reports the results of an archeometrical investigation performed, through a multi‐technique approach, on 19 colored opaque tesserae. The aims of the study were (1) the characterization of coloring and opacifying agents used for the production of the glass tesserae and (2) the definition of the technological processes involved. Colorimetric measurements allowed us to classify the tesserae in color groups, while the glassy matrix and the dispersed crystallites were characterized in detail through micro‐Raman spectroscopy, field emission scanning electron microscopy with energy dispersive X‐ray spectroscopy, laser ablation‐inductively coupled plasma‐mass spectrometry, and X‐ray powder diffraction analyses. Most of the glass shows the typical soda‐lime‐silicate composition (except for the orange and red tesserae). Raman results and elemental analysis prove the use of Sn–Pb antimonates to create yellow glass and of Ca‐antimonates for the white tesserae. A mixture of Sn–Pb antimonates and copper ions was used to obtain the emerald green color, while Ca‐antimonates were employed in both copper‐colored and cobalt‐colored blue glass to obtain different shades (blue‐green, dark, and light blue). X‐ray powder diffraction analyses reveal the presence of metallic copper (Cu⁰) and Cu₂O particles (cuprite) in red and orange tesserae, respectively. These results confirm the high technological level reached by the glassmakers of the Imperial Age. Copyright © 2014 John Wiley & Sons, Ltd.     

The issue of pseudoreplication when applying a statistical exploratory approach to extract relevant features from ToF‐SIMS spectra

The issue of automated peak selection in time‐of‐flight secondary ion mass spectrometry (ToF‐SIMS) spectra is examined in relation to the hierarchical nature of the experimental design and the related existence of observations which are not statistically independent. To avoid unreliable results, the presence of pseudoreplication should be taken into account correctly. A combination of the recommended univariate peak selection approach with some multivariate techniques, namely principal component analysis and heat map data representation, is proposed to highlight and analyze obtained results for a set of ToF‐SIMS spectra from copper phthalocyanine thin films grown on TiO₂ substrates by a supersonic beam deposition apparatus. New insight is obtained on the effectiveness of the deposition for different working parameters of the process. Copyright © 2013 John Wiley & Sons, Ltd.