Non-invasive detection of lead carboxylates in oil paintings by in situ infrared spectroscopy: How far can we go?

This paper demonstrates the efficacy of portable mid-infrared spectroscopy as a diagnostic tool for revealing the presence of lead carboxylates on artworks, in situ, without sampling. Samples of cerussite and hydrocerussite - the main components of lead white - were separately mixed with linseed oil, and artificially aged in two steps. Their compositional changes have been monitored over time by infrared spectroscopy in two investigation modes: absorption (benchtop instrument) and reflection (portable instrument), and by XRD spectroscopy. Each component of lead white showed different behavior depending on aging conditions, giving important information about the degradation products which are found in real cases. The accordance of results obtained in both modalities demonstrates that portable FT-MIR is a reliable technique for individuating and identifying lead carboxylates directly on paintings. Examples of the efficacy of this application are provided, by studying paintings from the 17th and 18th centuries.

One drop only. Easy and rapid Raman evaluation of ß-carotene in olive oil and its relevance as an index of olive fly attack.

This paper presents, for the first time, a method for the rapid quantification of ß-carotene in olive oil by Raman spectroscopy. Using a 532 nm Raman laser source, our procedure requires only one drop (100 µL) of oil, for ß-carotene content to be determined. Results show that ß-carotene content is associated with the lutein/ß-carotene ratio, a parameter whose value describes how healthy the olives were before processing, specifically whether an olive fly attack occurred. Since olive fly attacks are not always visible to the oil producers, this method gives them the means to control the validity of the prevention strategies they adopted.

Pulmonary carcinoma associated with cystic airspaces in two dogs.

Malignant pulmonary neoplasia associated with cystic airspaces is a well-recognised disease entity in humans. Two elderly dogs, previously diagnosed with a solitary emphysematous bulla, presented with non-specific clinical signs. At presentation, pulmonary auscultation was unremarkable. In both cases, thoracic CT demonstrated the transformation of the cystic airspace lesions characterised by a progressive increase of the solid component and reduction of the air component. Cytological evaluation and subsequent surgical excision followed by histopathology confirmed pulmonary carcinoma in both cases. These two cases represent the first demonstration of possible malignant transformation of pulmonary cystic airspace in dogs. Veterinarians should consider neoplastic transformation as a differential diagnosis in cases of cystic airspaces, particularly cases with features including thickening or irregularity of the wall, associated soft-tissue nodules or solid and non-solid tissue intermixed within clusters of multiple cystic airspaces. Ongoing monitoring of cystic airspace lesions through diagnostic imaging is recommended.

Mediastinal histoplasmosis with cardiac involvement in a cat.

Histoplasmosis is the second most common fungal infection reported among domestic felines in the United States. Dissemination of the organism after inoculation is common and affected organ systems include the respiratory tract, gastrointestinal tract, reticuloendothelial organs, skeletal system, integument, and ocular system. However, histoplasmosis presenting as a discrete granulomatous mass identified on echocardiogram has never been reported in the veterinary literature. Here, we describe the first case of feline histoplasmosis presenting as a granuloma with cardiac involvement. The patient, a 6-year-old male neutered domestic longhair feline, was referred for tachypnea and dyspnea. A mass in the cranial mediastinum abutting the heart was diagnosed via two-dimensional echocardiography. Cytology of fine needle aspirates from the mass revealed round yeast structures consistent with Histoplasma spp. The patient was treated with oral fluconazole therapy, and subsequent rechecks have shown marked improvement in clinical parameters, lesion size, and antigen concentrations.

Thermal mud maturation: organic matter and biological activity.

OBJECTIVE: Many of the therapeutic and cosmetic treatments offered in spas are centred on mud therapy, to moisturize the skin and prevent skin ageing and rheumatic diseases. Thermal mud is a complex matrix composed of organic and inorganic elements which contribute to its functions. It is a natural product derived from the long mixing of clay and thermal water. During its maturation, organic substances are provided by the microalgae, which develop characteristic of the composition of thermal water. METHODS: The aim of this study was to identify methods for introducing objective parameters as a basis for characterizing thermal mud and assessing its efficacy. Samples of thermal mud were collected at the Saturnia spa, where there are several sulphureous pools. The maturation of the mud was evaluated by organic component determination using extractive methods and chromatographic analysis (HPLC, GC-MS, SPME). We also studied the radical scavenging activity of mud samples at different stages of maturation, in a homogeneous phase, using several tests (DPPH, ORAC, ABTS). RESULTS: We identified several classes of compounds: saturated and unsaturated fatty acids, hydroxyl acids, dicarboxylic acids, ketoacids, alcohols and others. SPME analysis showed the presence of various hydrocarbons compounds (C(11) -C(17)) and long-chain alcohols (C(12) -C(16)). Six or seven months seemed appropriate to complete the process of maturation, and the main effect of maturation time was the increase of lipids. Six-month mud showed the highest activity. The hydrophilic extract was more active than the lipophilic extract. CONCLUSION: The results indicate that maturation of thermal mud can be followed on the basis of the changes in its organic composition and antioxidant properties along the time. They also highlight the need to develop reference standards for thermal muds in relation to assess their use for therapeutic and cosmetic purposes.

4-Methylbenzylidene camphor microspheres: reconstituted epidermis (Skinethic®) permeation and distribution.

OBJECTIVE: The UV filter 3(4-methylbenzylidene) camphor (4-MBC) is a common ingredient in sunscreen cosmetic products. However, different 'in vitro' and 'in vivo' studies suggest that 4-MBC can cause endocrine disrupting effects. Therefore, there is a need for new systems able to minimize the skin penetration of this UV filter. The aim of this study was to evaluate cutaneous permeation and distribution, through and into EPISKIN reconstituted epidermis (RE) from an O/W emulsion containing 4-MBC free or encapsulated in polymeric substantive microspheres. METHODS: Microspheres containing 4-MBC were prepared using the emulsification-solvent evaporation method and characterized for shape and surface morphology and encapsulation efficiency. O/A emulsions containing sunscreen free or encapsulated in microspheres were undergone to permeation tests through RE using vertical diffusion cells. At the end of the in vitro permeation experiments, the skin was subjected to tape stripping procedure to separate stratum corneum from viable epidermis. Each part was properly treated to extract the sunscreen retained and subject to quantitative analysis. RESULTS: The encapsulation of the sunscreen in the microspheres remarkably reduced the permeation of 4-MBC and increased its retention on the skin surface where its action is more desirable. CONCLUSIONS: The results of this study confirm the validity of substantive microspheres as an ideal formulation candidate to use in sunscreen preparation as they appear minimizing its systemic uptake and the potential associate toxicological risks. Therefore, more of the active sunscreen remains on the surface of the skin where it is intended to act and a higher activity it will explicate.

A non-invasive NMR relaxometric characterization of the cyclododecane-solvent system inside porous substrates.

With the aim of deepening the knowledge on the behavior of cyclododecane (CDD) as a temporary consolidant agent for weathered stones, NMR longitudinal and transverse relaxation decays have been exploited to follow the distribution of cyclododecane solutions into porous matrices. By measuring as function of time the relaxation decay constants of CDD solutions dropped onto porous supports, it has been possible to differentiate the step encompassing the solvent evaporation, which determines the consolidant migration within the matrix, from that governing the consolidant sublimation, which is related to the consolidation effectiveness over time.

Skin photoprotection improvement: synergistic interaction between lipid nanoparticles and organic UV filters.

A photoprotective formulation was developed with an increased sunprotection factor (SPF), compared to a conventional nanoemulsion, but having the same concentration of three molecular sunscreens, namely ethylhexyl triazone, bis-ethylhexyloxyphenol methoxyphenyl triazine, and ethylhexyl methoxycinnamate. The sunscreen mixture was incorporated into nanostructured lipid carriers (NLCs). The ability of nine different solid lipids to yield stable aqueous NLC suspensions was assessed. After the production by hot high pressure homogenization, the NLC were analyzed in terms of particle size, physical state, particle shape, ultraviolet absorbance and stability. The particle size for all NLC was around 200 nm after production. The NLC suspension with carnauba wax had superior UV absorbance, NLC from bees wax showed similar efficiency as the reference emulsion. The NLC formulations were incorporated into hydrogel formulations and the in vitro SPF was measured. This study demonstrated that approximately 45% higher SPF values could be obtained when the organic UV filters were incorporated into carnauba wax NLC, in comparison to the reference nanoemulsion and bees wax NLC. The data showed that the synergistic effect of NLC and incorporated sunscreens depends not only on the solid state of the lipid but also on its type.

The interaction of humic substances with the human prion protein fragment 90-231 affects its protease K resistance and cell internalization.

In this paper we analyzed the determinants and the structural effects of the interaction of human prion protein fragment 90-231 (HuPrP) with humic substances, (HS) including humic (HA) and fulvic (FA) acids, natural refractory organic polyanions widely diffused in soils and waters. We show that this interaction is mainly driven by non-specific electrostatic attraction involving regions situated within alpha-helix A and beta-sheet S1 of human PrP. FA binding to HuPrP altered its ability to acquire some PrPSc-like characteristics induced by the mild thermal denaturation of the peptide (1 h at 53 degrees C). In particular, in the presence of FA, HuPrP shows a reduced amount of beta-sheet content (as demonstrated by the reduced binding of thioflavin T), an increased sensitivity to protease K and an inhibition of the entering in the fibrillogenic pathway. FA/HuPrP interaction caused the aggregation of the peptide in unstructured macrocomplexes, as demonstrated by the altered electrophoretic migration in semi-denaturing detergent-agarose gel assay. Importantly, in the presence of FA the rate of internalization of HuPrP in human neuroblastoma cells was significantly reduced as compared to that of the beta-structured peptide. Therefore, HS inhibited the acquisition of PrP(Sc)-like structural properties that, in turn, are responsible for HuPrP intracellular accumulation and lead to neuronal death. Important implications of these data are that HuPrP-HS complexes, being unable to be internalized in living cells may represent a molecular mechanism for the reduced transmission of prion transmission from HS-rich soil also in the presence of contamination from infected animals.

Association of rs2200733 at 4q25 with atrial flutter/fibrillation diseases in an Italian population.

BACKGROUND: Atrial fibrillation (AF) and atrial flutter (AFL) are common cardiac conduction disorders affecting many people. Recent studies on sporadic cases of AF/AFL showed a significant association of the single nucleotide polymorphism rs2200733T with the disease, suggesting a genetic factor in the development of the disease. OBJECTIVES: To determine the association of rs2200733 with AF/AFL derived from an Italian population sample. SUBJECTS: 78 patients with AF/AFL and 348 controls took part in the study. DESIGN: Genetic case-control study. RESULTS: The results indicate that there is a positive, significant association between the rs2200733 T allele and patients with AF/AFL of Italian origin (allelic p<0.001 with OR = 2.17). CONCLUSION: These results derived from a sample of the Italian population agree with previously reported findings from an Icelandic study, which also found that the minor allele rs2200733 was associated with AF/AFL disease.

Preparation and characterization of n-dodecyl-ferulate-loaded solid lipid nanoparticles (SLN).

Solid lipid nanoparticles (SLN) containing a novel potential sunscreen n-dodecyl-ferulate (ester of ferulic acid) were developed. The preparation and stability parameters of n-dodecyl-ferulate-loaded SLN have been investigated concerning particle size, surface electrical charge (zeta potential) and matrix crystallinity. The chemical stability of n-dodecyl-ferulate at high temperatures was also assessed by thermal gravimetry analysis. For the selection of the appropriated lipid matrix, chemically different lipids were melted with 4% (m/m) of active and lipid nanoparticles were prepared by the so-called high pressure homogenization technique. n-Dodecyl-ferulate-loaded SLN prepared with cetyl palmitate showed the lowest mean particle size and polydispersity index, as well as the highest physical stability during storage time of 21 days at 4, 20 and 40 degrees C. These colloidal dispersions containing the sunscreen also exhibited the common melting behaviour of aqueous SLN dispersions.

New microencapsulated sunscreens: technology and comparative evaluation.

The aim of this work is to obtain new technologically improved microencapsulated sunscreens characterised by UV-radiation stability, good substantivity, low toxicity, a better tolerability and easiness to formulation. For this purpose we prepared two different systems using semisynthetic Hyaluronic Acid (HA) benzyl ester and a synthetic polymer (patent pending). We obtained these systems using two different methodologies: emulsification/solvent evaporation and emulsification/solvent extraction. The comparison between the two formulated systems was carried out in terms of their chemical-physical and biological properties.

From the sequence to the superstructural properties of DNAs.

A theoretical model for predicting intrinsic and induced DNA superstructures as well as their thermodynamic properties is presented. Intrinsic sequence-dependent superstructures are evaluated by integrating local deviations from the canonical B-DNA of the different dinucleotide steps. Induced superstructures are obtained by adopting the principle of minimum deformation free energy, evaluated in the Fourier space, in the framework of first-order elasticity. Finally dinucleotide stacking energies and melting temperatures are considered to account for local flexibility. In fact the two scales are strongly correlated. The model works very satisfactorily in predicting the sequence-dependent effects on the DNA experimental behavior, such as the gel electrophoresis retardation, the writhe transitions in topologically constrained domains, the thermodynamic constants of circularization reactions as well as the nucleosome thermodynamic stability constants.

Identification of protein domains on topological basis.

A theoretical method is proposed to identify structural domains in proteins of known structures. It is based on the distribution of the local axes of the polypeptide chain. In particular, a statistical analysis is applied to the contributions of the local axes to the absolute writhing number, a topological property of a space curve resulting from the number of self-crossings in the curve projections onto a unit sphere. This finding supports the hypothesis that topological requirements should be satisfied in the process of protein folding and in the final organization of the tertiary structures.

A theoretical model for the prediction of sequence-dependent nucleosome thermodynamic stability.

A theoretical model for predicting nucleosome thermodynamic stability in terms of DNA sequence is advanced. The model is based on a statistical mechanical approach, which allows the calculation of the canonical ensemble free energy involved in the competitive nucleosome reconstitution. It is based on the hypothesis that nucleosome stability mainly depends on the bending and twisting elastic energy to transform the DNA intrinsic superstructure into the nucleosomal structure. The ensemble average free energy is calculated starting from the intrinsic curvature, obtained by integrating the dinucleotide step deviations from the canonical B-DNA and expressed in terms of a Fourier series, in the framework of first-order elasticity. The sequence-dependent DNA flexibility is evaluated from the differential double helix thermodynamic stability. A large number of free-energy experimental data, obtained in different laboratories by competitive nucleosome reconstitution assays, are successfully compared to the theoretical results. They support the hypothesis that the stacking energies are the major factor in DNA rigidity and could be a measure of DNA stiffness. A dual role of DNA intrinsic curvature and flexibility emerges in the determination of nucleosome stability. The difference between the experimental and theoretical (elastic) nucleosome-reconstitution free energy for the whole pool of investigated DNAs suggests a significant role for the curvature-dependent DNA hydration and counterion interactions, which appear to destabilize nucleosomes in highly curved DNAs. This model represents an attempt to clarify the main features of the nucleosome thermodynamic stability in terms of physical-chemical parameters and suggests that in molecular systems with a large degree of complexity, the average molecular properties dominate over the local features, as in a statistical ensemble.

Unsaturated hydrocarbons with fruity and floral odors.

Hydrocarbons usually do not exhibit odors of interest or well-defined character. However, certain cyclic alkenes have been associated with typical and pleasant notes, such as fruity, green, and floral. One of the best known examples is represented by the isomeric megastigmatrienes, endowed with a pleasant smell of tropical fruits. From the structures of these odorants, 24 analogues and homologues, most of them cyclic alkenes, but including also some open-chain alkenes, have been synthesized to define structural parameters related to the characteristic odors of these compounds. The number and position of double bonds, the substitution on the ring, and the size of the ring are the variables taken into account. Most of the new compounds present a mainly fruity character, associated in several cases with floral and green notes, producing an overall sensation described as "tropical fruit".

Neurologically active plant compounds and peptide hormones: a chirality connection.

The most dramatic, but seldom mentioned, difference between alkaloid and peptide opioids is the change of chirality of the alpha carbon of the tyramine moiety. We propose that the presence of Gly2 or D-Ala2 in the two most common message domains compensates this change by allowing the attainment of unusual conformations. A thorough conformational search of Tyr-D-Ala-Phe-NH-CH3 and of its isomer Tyr-L-Ala-Phe-NH-CH3 backs this view and establishes a solid link between alkaloid and peptide opioids. This finding supports the notion that morphine, like other neurologically active plant compounds, may bind to endogenous receptors in plants to regulate cell-to-cell signaling systems.

Dual role of DNA intrinsic curvature and flexibility in determining nucleosome stability.

A statistical mechanistic approach to evaluate the sequence-dependent thermodynamic stability of nucleosomes is proposed. The model is based on the calculation of the DNA intrinsic curvature, obtained by integrating the nucleotide step deviations from the canonical B-DNA structure, and on the evaluation of the first order elastic distortion energy to reach the nucleosomal superstructure. Literature data on the free energy of nucleosome formation as obtained by competitive nucleosome reconstitution of a significant pool of different DNA sequences were compared with the theoretical results, and a satisfactorily good correlation was found. A striking result of the comparison is the emergence of two opposite roles of the DNA intrinsic curvature and flexibility in determining nucleosome stability. Finally, the obtained results suggest that the curvature-dependent DNA hydration should play a relevant role in the sequence-dependent nucleosome stability.

A non-parametric method for the analysis of experimental tumour growth data.

Analysis of tumour growth is required to investigate the biology of tumours and to determine the effects of new anti-tumour therapies. A non-parametric mathematical method for the analysis of a set of experimental tumour growth data is described. The method is based on the similarity between time series of tumour size measurements (e.g. tumour volume), similarity being defined as the Euclidean distance between data measured for each tumour at the same time. Subsets of similar time series are found for a given population of tumours. A biologically meaningful parameter H has been derived which is a measure of the scattering of experimental volume samples. The method has been applied to the analysis of the growth of (i) untreated multicellular tumour spheroids obtained with different cell lines and (ii) spheroids treated with cytotoxic drugs (immunotoxins). Results are compared with those previously obtained by applying the classical Gompertz growth model to the analysis of treated and untreated spheroids.