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Nell’ambito dell’elaborazione della Cartografia di Rischio dei Beni Culturali in Sicilia, sono stati presi accuratamente in considerazione le caratteristiche di alcuni siti archeologici, mediante la sperimentazione di metodologie e tecniche già applicate positivamente per l’investigazione di rocce e formazioni geologiche porose. Il Patrimonio Culturale è relativamente diffuso nell’Isola, grazie alle passate civiltà, fornendo numerose opportunità di studio sia in campagna, sia in laboratorio, ponendo le basi di successive integrazioni ed elaborazioni dei dati raccolti. I materiali dei monumenti storici appartengono a diversi terreni sedimentari, che presentano una relativa complessità geologica. Queste rocce generalmente presentano una porosità elevata, la quale può portare ad un notevole deterioramento dei luoghi che ospitano le testimonianze del passato, come anche dei monumenti medesimi. S’intendono qui investigare alcune proprietà di siti e monumenti inseriti nel Patrimonio Culturale della Sicilia e appartenenti, tra gli altri, alla Magna Grecia ed all’Età del Bronzo. Questi, verosimilmente sottoposti ad elevata vulnerabilità nei confronti dell’ingressione di acque più o meno contaminate, sono positivamente stati oggetto di studio per la caratterizzazione di quei fenomeni che sono, direttamente o indirettamente, connessi con l’assorbimento di liquidi. Fra i metodi più adatti all’esplorazione dei siti e dei materiali, sono state - in prima istanza - considerate tecniche fisiche non distruttive, quali XRD, XRF, NMR e TAC, confrontando i dati su campioni con misure in situ. Nel dettaglio, vengono qui considerati alcuni campioni di roccia provenienti dai siti archeologici di Agrigento e Noto. Inoltre, in considerazione dell’alta vulnerabilità nei confronti dell’inquinamento presentata dalle formazioni porose degli stessi siti archeologici e dalle antiche cave di estrazione, si accenna, ai risultati di recenti studi di rischio idrogeologico condotti in aree archeologiche ed alla possibilità offerta da indagini di questo tipo ai fini della definizione del rischio archeologico complessivo.

In questo lavoro, la tecnica di Imaging a Risonanza magnetica Nucleare (MRI) è stata impiegata per studiare in modo quantitativo, non invasivo, non distruttivo, la cinetica di assorbimento capillare di acqua in campioni di Pietra di Lecce, una biocalcarenite omogenea nella quale il fronte di bagnamento avanza pressoché piatto. MRI permette di misurare in funzione del tempo l’altezza raggiunta dal fronte in qualunque sezione interna del campione. Al fine di studiare l’anisotropia del fenomeno dovuta ad eventuali strati sedimentari, alcuni esperimenti sono stati effettuati variando l’asse principale di assorbimento (ortogonale al piano dove avviene l’assorbimento) attraverso la semplice rotazione del campione cubico. Le differenze osservate nella cinetica di assorbimento a seguito di trattamenti con prodotti idrorepellenti permettono di studiare l’efficacia dei trattamenti stessi. Le misure sono state ripetute dopo il trattamento con una resina acrilica (Paraloid B72) comunemente usata per tali trattamenti. I dati ottenuti sono stati interpretati mediante il modello di Washburn per la risalita capillare. Nell’intervallo di tempo esaminato è stata osservata la prevista dipendenza lineare dell’altezza di risalita dalla radice quadrata del tempo, sia per assorbimento dalla faccia trattata, sia per assorbimento da quella non trattata. I valori del coefficiente angolare (sorptivity, S) ottenuti nelle diverse condizioni permettono di trarre conclusioni circa l’efficacia del trattamento. Quando il litotipo è molto eterogeneo, come nel caso di una biocalcarenite proveniente da una cava nei pressi dell’area archeologica di Agrigento, MRI evidenzia un avanzamento stocastico del fluido, come confermato da immagini ottenute mediante tomografia computerizzata.

Aurora-A is a centrosome-associated serine/threonine kinase that is overexpressed in multiple types of human tumors. Primarily, Aurora-A functions in centrosome maturation and mitotic spindle assembly. Overexpression of Aurora-A induces centrosome amplification and G 2 /M cell cycle progression. Recently, it was observed that overexpression of Aurora-A renders cells resistant to cisplatin (CDDP)-, etoposide-, and paclitaxel-induced apoptosis.Our results indicate that already in initial stages of cancer progression Aurora-A overexpression could have a major role in inducing supernumerary centrosomes and aneuploidy, as shown by immunohistochemistry on tissue sections from various stages of human colon cancer. Aneuploidy was also observed after Aurora-A ectopic overexpression in colon cancer cells with MIN phenotype. Silencing of Aurora-A by RNA interference in tumor cell lines triggered arrest of the cell cycle associated to apoptosis/ mitotic catastrophe. Finally, Aurora-A transcriptional silencing seems to confer cancer cells a greater sensitivity to chemotherapy by vincristine, indicating Aurora-A as a possible gene target in cancer therapy.

Background Aneuploidy has been acknowledged as a major source of genomic instability in cancer, and it is often considered the result of chromosome segregation errors including those caused by defects in genes controlling the mitotic spindle assembly, centrosome duplication and cell-cycle checkpoints. Aneuploidy and chromosomal instability has been also correlated with epigenetic alteration, however the molecular basis of this correlation is poorly understood. Results To address the functional connection existing between epigenetic changes and aneuploidy, we used RNA-interference to silence the DNMT1 gene, encoding for a highly conserved member of the DNA methyl-transferases. DNMT1 depletion slowed down proliferation of near-diploid human tumor cells (HCT116) and triggered G1 arrest in primary human fibroblasts (IMR90), by inducing p53 stabilization and, in turn, p21waf1 transactivation. Remarkably, p53 increase was not caused by DNA damage and was not observed after p14-ARF post-transcriptional silencing. Interestingly, DNMT1 silenced cells with p53 or p14-ARF depleted did not arrest in G1 but, instead, underwent DNA hypomethylation and became aneuploid. Conclusion Our results suggest that DNMT1 depletion triggers a p14ARF/p53 dependent cell cycle arrest to counteract the aneuploidy induced by changes in DNA methylation.

The investigations on structure and micro-chemical composition of archaeological metal alloys are needed in archaeometry. The aim of this study is devoted both to acquire information about their provenance and production technology, and to improve our understanding about the corrosion processes. In this paper we present the study of the corrosion phenomena of bronze samples, laboratory-made according to binary, ternary and quaternary alloys typical of Roman archaeometallurgical production through an integrated methodology based on the use of non or micro invasive physical techniques. Among the analysed samples, two were artificially aged through burial in the archaeological site of Tharros, along the west coast of Sardinia (Italy). The corrosion products, typical of the bronzes in archaeological sites near the sea, have been characterized by non invasive and micro-destructive measurements. In particular, the corrosion patinas were examined through optical microscopy, scanning electron microscopy and microanalysis, X-ray fluorescence and laser ablation spectroscopy. The use of integrated technologies allowed us to determine both the elemental composition and surface morphology of the patina, highlighting the correlation between patina nature and chemical composition of the burial context. Moreover, data obtained by the laser-induced breakdown spectroscopy along the depth profile on the samples, have yielded information about the stratigraphic layers of corrosion products and their growth. Finally, the depth profiles allowed us to verify both the chemical elements constituting the patina, the metal ions constituting the alloy and the occurrence of migration phenomena from bulk to the surface.

The laser-induced breakdown spectroscopy (LIBS) technique is often used as atomic spectroscopic technique for elemental analysis of materials. However, it presents some drawbacks that make an accurate quantitative analysis difficult. Since the plasma properties, such as spatial inhomogeneity and plume stoichiometry strongly depend on the experimental conditions, the measurements are less reproducible. In order to evaluate the measurement fluctuations, we propose to use the more established micro X-Ray fluorescence (μ-XRF) technique for validating LIBS data. In particular, the quantitative data, obtained by varying the laser fluence, the shot numbers and the temporal acquisition parameters, were compared with those obtained by μ-XRF on laboratory made samples of binary, ternary and quaternary bronze alloys. For LIBS measurements a mobile double pulse laser instrument equipped with an high resolution Echelle type monochromator coupled to an intensified CCD camera was used. μ-XRF analyses were performed with a portable instrument that uses a micro collimated X-Ray beam and it is equipped with an high resolution detector. The LIBS results show a strong dependence both on the instrumental set up and the chemical-physical properties of the sample. With our findings we could identify the most suitable parameters to be used in the investigation of the different bronze alloys. The possibility to carry out a quantitative analysis by using the LIBS technique was checked through the comparison with related μ-XRF data. In particular in this paper we identified a set of reliable LIBS parameters for the quantitative analysis of copper, tin and zinc. Further analyses will be necessary to reach this goal also for the minor constituents as lead.

The present study confirms how the integrated use of non-destructive or micro-destructive spectroscopic techniques are capable to provide a clear response to the investigative needs in the conservation science. In particular, the X-ray fluorescence analysis in association with laser-induced breakdown spectroscopy can be useful for collecting information about chemical composition and elemental stratigraphic distribution of the external layers. The complementarity of these techniques has been tested by analysing bronze samples, made in the laboratory according to binary, ternary and quaternary alloys, typical of Roman archaeometallurgical production and buried for one year. The findings have been useful for acquiring information about the thickness and the composition of the corroded surfaces.

Lo studio dei materiali pittorici è uno degli obiettivi primari per la caratterizzazione storico- artistica delle opere d’arte poiché fornisce informazioni utili circa i materiali originali e di degrado, la tecnica di esecuzione ed eventuali interventi di restauro pregressi. Tuttavia, la complessità delle indagini archeometriche è legata alla necessità di garantire la salvaguardia del bene culturale che rende essenziale l’uso di tecniche di indagini non o micro invasive. Tra le metodologie d’indagine integrata, l’uso della fluorescenza a raggi X (XRF) e della spettroscopia mediante ablazione laser (LIBS) risponde a tale esigenza e risulta particolarmente utile per la complementarietà delle informazioni fornite sia in termini composizionali che di spessori indagati. Infatti, a differenza dell’analisi XRF, la LIBS non solo permette l’identificazione degli elementi chimici con basso numero atomico ma, attraverso impulsi laser consecutivi sullo stesso punto campione, consente anche di realizzare profili di profondità. In tal modo è possibile ottenere informazioni sulle differenze composizionali tra la superficie ed il materiale di bulk e, dunque, ricostruire la successione stratigrafica caratteristica del campione. Seguendo tale approccio metodologico, sono state condotte analisi XRF e LIBS, sia su frammenti di pitture murali provenienti da diversi siti, sia su stesure campione realizzate in laboratorio con differenti tecniche pittoriche. In particolare, mediante l’analisi LIBS è stato possibile analizzare la stratigrafia dalla superficie degli strati pittorici fino allo strato di preparazione. I risultati hanno permesso di valutare le potenzialità, in campo archeometrico, della tecnica LIBS, rispetto alle tecniche spettroscopiche convenzionali.

Ad approfondimento ed integrazione dei risultati analitici già presentati, riguardanti la caratterizzazione in situ tramite fluorescenza a raggi X (XRF) dei materiali pittorici presenti su opere marmoree della collezione statuaria della Galleria Regionale di Palazzo Abatellis (Palermo), sono state effettuate ulteriori indagini su un campione di cinabro (HgS) caratterizzato da una colorazione nera. Tali analisi, condotte tramite misure XRF e XRD presso la beam line ID18 dell’European Syncroton Radiation Facility (Grenoble, Francia), hanno fornito informazioni complementari fondamentali per la corretta interpretazione dello stato di conservazione dello strato pittorico studiato.

The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures faithful chromosome segregation during mitosis and its failure can result in aneuploidy. Previously, it was suggested that reduction of the MAD2 gene, encoding a major component of the SAC, induced aneuploidy in human tumor cells. However, tumor cell lines contain multiple mutations that might affect or exacerbate the cellular response to Mad2 depletion. Thus, the scenario resulting by Mad2 depletion in primary human cells could be different and more complex that the one depicted so far. We used primary human fibroblasts (IMR90) and epithelial breast cells (MCF10A) to gain further insight on the effects of genomic instability caused by transient Mad2 depletion. To this aim we depleted Mad2 by RNAi to a level shown by Mad2 haplo-insufficient cells and found that induced aneuploidy caused premature cellular senescence in IMR90 cells. IMR90 cells showed typical features of senescent cells, like senescence-associated (SA) beta galactosidase expression, including up-regulation of p53 and p14ARF proteins and of p21(waf1) as well, but not of p16(INK4A) cyclin-dependent kinase (Cdk) inhibitor. In contrast, after MAD2 post-transcriptional silencing MCF10A cells in which the INK4A/ARF locus is deleted, showed both aneuploidy and a small increase of p53 and p21(waf1) proteins, but not premature cellular senescence. Finally, our results provides an explanation of how a p53 controlled pathway, involving initially p21(waf1) and then p14ARF, could minimize the occurrence of genomic alterations derived from chromosome instability induced by low amounts of MAD2 protein.

In the present work results on the degradation phenomena of stone materials in the Ancient Theatre of Taormina, one of the most important Greek–Roman monuments of Sicily, are reported. Artificial stone materials in different conservation conditions were investigated. Samples of salt efflorescences from brick walls and degraded setting mortars were taken from the open gallery in “summa cavea”. The chemical, physical and structural characterization was performed by means of X-Ray Photoelectron Spectroscopy (XPS), X-Ray Diffraction (XRD) and X-Ray Fluorescence (XRF), both in situ and ex situ. Results showed that the deterioration of stone materials is due to the aggressive action of the environment agents. Soluble salts, such as chlorides, sulphates and nitrates, were found in efflorescence samples. Mortars were affected by a decomposition process induced by sulphates’ attack.

Highly Expressed in Cancer protein 1 (Hec1) is a subunit of the Ndc80 complex, a constituent of the mitotic kinetochore. HEC1 has been shown to be overexpressed in many cancers, suggesting that HEC1 upregulation is involved in the generation and/or maintenance of the tumour phenotype. However, the regulation of Hec1 expression in normal and tumour cells and the molecular alterations promoting accumulation of this protein in cancer cells are still unknown. Here we show that elevated Hec1 protein levels are characteristic of transformed cell lines of different origins and that kinetochore recruitment of this protein is also increased in cancer cell lines in comparison with normal human cells. Using different cell synchronization strategies, Hec1 expression was found to be tightly regulated during the cell cycle in both normal and cancer cells. A limited proteasome-dependent degradation of Hec1 cellular content was observed at mitotic exit, with no evident differences between normal and cancer cells. Interestingly, increased expression of HEC1 mRNA and Hec1 protein was observed after transient silencing of the retinoblastoma gene by siRNA or following microRNA-mediated permanent depletion of the retinoblastoma protein in HCT116 cells. Our data provide evidence for a functional link between Hec1 expression and the pRb pathway. These observations suggest that disruption of pRb function may lead to chromosome segregation errors and mitotic defects through Hec1 overexpression. This may importantly contribute to aneuploidy and chromosomal instability in RB-defective cancer cells.

Background Aneuploidy is a hallmark of most human cancers that arises as a consequence of chromosomal instability and it is frequently associated with centrosome amplification. Functional inactivation of the Retinoblastoma protein (pRb) has been indicated as a cause promoting chromosomal instability as well centrosome amplification. However, the underlying molecular mechanism still remains to be clarified. Results Here we show that pRb depletion both in wild type and p53 knockout HCT116 cells was associated with the presence of multipolar spindles, anaphase bridges, lagging chromosomes and micronuclei harbouring whole chromosomes. In addition aneuploidy caused by pRb acute loss was not affected by p53 loss. Quantitative real-time RT-PCR showed that pRB depletion altered expression of genes involved in centrosome duplication, kinetochore assembly and in the Spindle Assembly Checkpoint (SAC). However, despite MAD2 up-regulation pRb-depleted cells seemed to have a functional SAC since they arrested in mitosis after treatments with mitotic poisons. Moreover pRb-depleted HCT116 cells showed BRCA1 overexpression that seemed responsible for MAD2 up-regulation. Post-transcriptional silencing of CENPA by RNA interference, resulting in CENP-A protein levels similar to those present in control cells greatly reduced aneuploid cell numbers in pRb-depleted cells. Conclusion Altogether our findings indicate a novel aspect of pRb acute loss that promotes aneuploidy mainly by inducing CENPA overexpression that in turn might induce micronuclei by affecting the correct attachment of spindle microtubules to kinetochores.

This paper proposes the X-Ray CT imaging as appropriate tool for investigating the capillary water absorption in sedimentary rocks. This technique, in fact, provides information useful for deeping the knowledge about of the porosity and the kinetics of the water capillary absorption in porous materials. The possibility to improve in non invasive manner, the understanding of this phenomenon, constitutes a fundamental aspect to take actions in the restoration and conservation of lapideous artifact and monuments from cultural heritages. The investigated sedimentary rocks come from different Sicilian quarries and were used for the building of the Greek temples in the archaeological areas of Agrigento, Selinunte and in the baroque monuments of the Val di Noto area. The image acquisition was carried out at different times after contact with water. The wetting front progression along the height of the sample was evaluated on the basis of the estimated CT attenuation value maps. An average of the CT attenuation values measured by the ROI (Region of Interest) was used to evaluate the mean height of the wetting front, with different CT scans at fixed time intervals after sample wetting, in order to describe the dynamic behaviour of the imbibition process. CT imaging results have shown that the water absorption is highly subject to the sample porous morphology.