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The emerging impact on the planet of 3D printing, at all levels, doesn’t seem to limit itself to the simple modification of the traditional means of production, indicating a deeper and far more diffuse transformation, referred to by some as the “third industrial revolution”. 3D printing has become a process accessible to everyone. Up until a few years ago, in fact, those who built a company and produced, for example, spare parts for the automotive supply sector or for electronic devices, were considered the pioneers of their respective fields; while today, the true pioneers of modern craftsmanship are common people that, to fulfill their needs, autonomously build what they need. Those are the so-called makers. Once he has designed a tridimensional model, a maker can take advantage of 3D printing in different ways, some of which will be listed right now, while the others will be treated in deeper detail in the following paragraphs: the purchase and/or the building of a personal printer; the employment of share printing by non-profit associations like Fablab, Maker-spaces, Hacker; the use of Internet for direct contact with companies that develop personalized printing also for private production. “The true revolution is not the development of technology, but its democratization: to make it accessible to an enormous group of people who can experience it and improve it”, as said by Chris Anderson, former director of Wired magazine, author of the book “The Long Tail” and editor of the essay “Makers: The New Industrial Revolution”. “Today we have instruments like the world wide web to help us: the inventor of today isn’t an isolated genius anymore, as he can make use of the joint effort of other individuals. In its workshop, instead of a plane, a buzz saw and a drill, we would see the inevitable computer, on whose screen the item takes shape, and a three-dimensional printer”. In this sense, 3D printing is revolutionizing not only the world of fashion and design, but also the universes of the construction industry and of architecture. In fact, one of the most respected “contemporary masters”, Norman Foster, won an international contest launched by the European Space Agency (ESA) to design, through the use of a 3D printer, a lunar outpost capable of hosting four people, employing regolith as a construction unit, that is, dust, soil, broken rock, and other related materials present on the Moon. In China, the creations of the multinational corporation WinSun are causing quite a sensation: after realizing in recent years “10 houses in 24 hours”, and printing an 1100 square meters villa. The incredible feat has been achieved with a giant 3D printer – 32 meters of length, 10 of width and 6,6 of height – in which, as a construction unit, has been employed a mix of recycled dumping materials, like fiberglass, steel, concrete and special additives. Allowing a saving of materials between 30% and 60%, and a saving of building time between 50% and 70%. Holland is today the main European innovator of 3D architecture. Dutch architect Janjaap Ruijssenaars has devised a system that will allow the production of base elements of approximately 9×13,5 meters that will constitute the model of residential buildings. For its manufacturing, a sand compound will be employed as construction unit, mixed to a bonding agent of new conception that will make it sturdier than concrete. In Italy, the company WASP (acronym of World's Advanced Saving Project), has realized printers that operate through a process of additive manufacturing. The aim of these devices, called BigDelta, is to realize echo-compatible houses with a limited power consumption, with available natural materials like clay. Besides the advantages of reduction of energetic consumption and emissions, of fast building times, contained costs and easy adaptability to different situations, the research is also moving in the elaboration of new materials to substitute concrete, that are more ecological and sustainable. The thesis project, carried out by Valentina Sapio, with the relation of the Prof. Arch. Claudio Gambardella (SUN “Luigi Vanvitelli” –Aversa), the correlation of the Prof. Arch. Fabio Naselli (UNI Kore – Enna) and the referee of the Prof. Mohamed Assem Hanafi (Dep. of Architectures, Faculty of Engineering, University of Alexandria), concentrates on the requalification of the industrial district of Minet El Bassal in Alexandria. The proposal of project focuses, besides the urban intervention project to redevelop the surrounding district, on a tower, placed inside one of the seven buildings that comprise the area that accommodates the Fablab. Realized entirely with the use of 3D printing, the tower, that recalls the ancient lighthouse of Alexandria, and in line with the already existing skyline formed by the minarets, is conceived with materials that are typical of the Egyptian buildings on the rivers of Nile (with fire-bricks in straw and clay), that is a natural paste made up of clay, straw and adhesives. Intervening both from an evocative volumetric point of view, and in the use of materials in the full respect of the pre-existent, melting tradition and innovation. Ideally, the same Fablab, recalling another important concept, that is, the concept of self-building, and another value intrinsic of Egyptian culture, realizes the tower. Examining the study, the works and the thought of Egyptian architect, Hassan Fathy: “The quality and the values inherent the traditional answer of the man to the atmosphere (on the environmental challenges), should be preserved without renouncing to scientific progress. Science can be applied to several aspects of our job, if, at the same time, it is subordinated to philosophy, faith and spirituality”..

La riflessione che di seguito si sviluppa vuole condurre verso una più estesa considerazione delle implicazioni sociali dell’attività agricola. Il dibattito nel campo economico agrario, infatti, si concentra sui contenuti che l’attività agricola trasmette attraverso la modellazione del paesaggio, la conservazione della storia, la tutela dell’ambiente e della salute. La notorietà della Dieta mediterranea e il suo riconoscimento globale sembrano riassumere in larga parte queste proposizioni.

La crisi finanziaria globale coinvolge e sconvolge le economie reali e quindi anche i sistemi valoriali alla base delle strumentazioni di governo delle trasformazioni urbane e territoriali stressando le sperimentazioni di governance dei modelli di sviluppo place based a seconda delle tipologie territoriali e dei loro livelli di declino. Il turismo sostenibile sembra inserirsi in modo utile nelle strategie di resilienza dei territori. Nella prima parte si sottolinea il rapporto tra pianificazione territoriale e agricoltura che torna ad essere centrale. Le ICT spingono la condizione sociale a mutare verso quella di cittadini del mondo che vivono lo spazio ed il tempo immergendosi dentro una vita in costante movimento. Il turismo sembra scomparire al crescere della possibilità di scoprire e intrattenere nel tempo nuove relazioni umane che collaborano a minimizzare gli impatti del turismo sia sul patrimonio costruito che su quello rurale. Nella seconda parte è messa in luce la potenzialità ed il ruolo che l’agricoltura e la produzione alimentare di qualità gioca nelle politiche di sviluppo locale. Nella terza parte è illustrato un caso di studio della Sicilia centrale in cui è necessario e possibile tracciare strategie di recupero del patrimonio culturale, colturale e di alimentazione su livelli di eccellenza, grazie a politiche e pratiche trans-settoriali integrate. Nelle conclusioni si sottolinea la convenienza dell’approccio di turismo relazionale integrato. La visione rur-urbana che ne deriva ha l’ambizione di divenire un quadro di riferimento per nuovi scenari di riorganizzazione degli equilibri fra area urbana e area rurale.

Inflammation is a complex physiological response that requires the activity of a sophisticated regulatory network involving the activation of specific genes for defense, tissue repair and remodeling. Although transcriptional activation has been shown to be critical in the regulation of inflammatory genes (1) the role of epigenetic phenomena in the modulation of the inflammatory response is now emerging (2). Specifically, it has been recently reported that proinflammatory stimuli induce DNA demethylation in the interleukin IL-1 promoter of human articular chondrocytes (3). IL-1 cytokine, among several proinflammatory agents, represents an essential player in the inflammatory conditions of the gut (4): functioning as the strongest signal transduction to NF-kB, IL-1 increases in intestinal paracellular permeability and over-expression of proinflammatory genes (5). In this tissue, moreover, inflammatory response is crucial to maintain its structural integrity and function, thus, alteration and deregulation of inflammatory pathways contribute to tissue damage and ulceration, and are pivotal factors in the pathogenesis of several inflammatory gut diseases. In the present study we evaluate both wide-ranging and gene-specific epigenetic changes in the inflammatory response of Caco-2 cells differentiated into intestinal epithelial cells and exposed to the inflammatory actions of IL-1β. Our results clearly show that IL-1induces changes in the DNA methylation either at genome and gene level and that the local methylation changes are induced in two pro-inflammatory genes that are IL-1-regulated. In particular, we show that a cell exposure to IL-1β for 24 h induces, in a dose-dependent manner, hypomethylation of genomic DNA in respect to untreated cells. We also observe a reduced DNA methyltransferases activity of cell lysates obtained from IL-1 treated cells. Finally our data show that IL-1 is able to induce hypomethylation of specific CpG sites in IL-6 and IL-8 genes. These preliminary results suggest that IL-1 in intestinal epithelial cells is able to act as an epigenetic modulator towards the entire genome and specific genes. Modulation of epigenetic changes in inflammation may provide a new “reading frame” of the inflammatory diseases molecular basis. Since epigenetic modifications are potentially reversible, a thorough understanding of these changes during inflammatory response opens opportunities to develop efficient agents for specific targets.

Itatour. Visioni territoriali e nuove mobilità. Progetti integrati per il turismo nella città e nell’ambiente

Background and Aims: Hepatic enzyme CYP2E1 is involved in the metabolism of a number of exogenous and endogenous substances (i.e. ethanol, drugs and chemical carcinogens). Being polymorphic, CYP2E1 gene can give different xeno-metabolic capabilities in a population and it is well known that inadequate or no enzymatic deactivation of xenobiotics could induce an increased susceptibility to disease and cancer. In particular, one of the 5 -flanking region polymorphisms, able to differentiate CYP2E1 gene transcriptional activity, is caused by the appearance/disappearance of RsaI and PstI restriction sites, which generates two different alleles, namely *C1(Rsa+/Pst−) and *C2(Rsa−/Pst+) respectively, reported to be in complete linkage disequilibrium. Methods: To confirm the existence of a correlation between some particular CYP2E1 genotypes/haplotypes and hepatocarcinoma, we determined CYP2E1 PstI/RsaI genotypes/haplotypes by RFLP-PCR in a cohort of central western Sicily hepatocarcinoma patients and in a population of healthy students from the same geographic area. Results: In hepatocarcinoma patients, modal genotype association was Rsa++/Pst−−, corresponding to CYP2E1 *C1/*C1 haplotype, whereas the Rsa+−/Pst−+ association, equivalent to CYP2E1 *C1/*C2 haplotype, resulted to have the lowest frequency both in patients and in controls. Moreover, both in patients and in controls, noncanonical genotype associations were frequent and arose from a no-linkage disequilibrium between the two polymorphic sites. Other authors reported this finding as a rare occurrence. Thus, from analysis of only one restriction site, Rsa++ genotype was approximately 1.5-fold more frequent in patients than in controls, and the non-canonical Rsa+− genotype was found relatively frequent in patients. Moreover, HuH7 and HA22T transformed hepatocarcinoma cell lines also showed the Rsa+− genotype. Conclusions: These results suggest that the presence in CYP2E1 genotype of at least one allele with an Rsa I restriction site is correlated with hepatocarcinoma. As this site is known a consensus sequence for some specific CYP gene transcription factors, like HNF-1, it may be supposed that a single nucleotide polymorphism can alter the possibility of HNF-1 to bind CYP2E1 promoter. This could determine a marked change in the transcriptional activity of the gene, incompetence in xenobiotic metabolism or in toxic substance deactivation and an increased susceptibility to neoplastic diseases, such as hepatocarcinoma.

Context: The genetic polymorphism is considered a major source of variability, influencing the levels of gene expression. Cytochrome P450 2E1 (CYP2E1) is a mixed-function oxidase involved in the metabolism of the many endogenous and exogenous substances (ethanol, chemical carcinogens) in the hepatic and pancreatic tissue. CYP2E1 gene polymorphisms can cause various abilities of metabolize xenobiotic substances within a population with consequent increased susceptibility to various diseases,including cancer. One of the polymorphisms of the CYP2E1 gene is a VNTR (Variable Number Tandem Repeat) of some sequences in its "5 '- flanking region. Method : VNTR genotype CYP2E1 was determined by RFLP-PCR performed on DNA extracted from peripheral blood lymphocytes and paraffin embedded tissue of central-western Sicilia population with PA, to confirm or exclude a correlation between certain genotypes and specific disease. A population of university students, without specific overt disease, was used as control . Results: The modal genotype, found in all subjects, was CYP2E1 VNTR A2/A2, while CYP2E1 VNTR A2/A4 was the least represented in both populations. The A2/A3 genotype was different between patients with pancreatic cancer and healthy subjects, suggesting a correlation between this genotype and pancreatic cancer. Recent studies indicated A2 allele might be associated with a negative regulation of the gene and nothing about it has been reported for the allele A3. Conclusion: Our preliminary data indicate an association between genotype A2/A3 and PA and allow us to hypothesize that the A3 allele is associated with low activity of the CYP2E1 gene. Consequently, individuals with this allelic association may have inadequate ability to metabolize toxic xenobiotic substances, condition that might increase the susceptibility to develop some cancer, including PA.

Il turismo, inteso come viaggio di conoscenza e come svago, esiste da sempre, almeno da quando l’uomo ha manifestato l’esigenza di scoprire quello che gli stava intorno, di allargare quindi la propria sfera di conoscenza e di influenza nel territorio circostante. Con il suo aprirsi “a tutti” e con l’introduzione dell’aspetto simbolico dello status sociale, il turismo ha assunto una connotazione sempre più di“moda”, avviandosi in una direzione gestita da logiche industriali di “massa”2. Logiche economiche centrate sulla ricerca del massimo profitto, come unico parametro valido per la misura della performance aziendale che mostrano oggi, quanto mai prima, i loro difetti e le loro profonde contraddizioni, soprattutto nel confronto con le logiche dello sviluppo sostenibile. Uno sviluppo che per essere realmente sostenibile deve farsi promotore della conservazione e della valorizzazione della tipicità e della diversità di un territorio e quindi allargare il campo all’entrata di nuovi valori che negano la sola applicazione della logica del profitto. Una connotazione, quella industrialista, che col passare del tempo è arrivata a sostituire gli originari scopi del viaggiatore, facendo assumere al termine turista, una palese accezione negativa, proprio ad indizio dell’indifferenza ai luoghi e alle culture indotta da quella forma di turismo. Per nostra fortuna la funzione e gli scopi originari del viaggio non saranno cancellati da questa ondata di turismo di massa, ma resteranno relegati per alcuni decenni in una nicchia riservata a pochi specialisti, studiosi e ricercatori. Con il diffondersi di una cultura orientata verso la salvaguardia e la promozione delle risorse locali e, parallelamente, con l’avvento delle tecnologie della comunicazione, in particolar modo della telematica e dell’informatica, quindi nel corso dell’ultimo ventennio, si è timidamente manifestata una nuova inversione di tendenza verso una riappropriazione dello spirito originario del turismo, ossia: conoscenza, descrizione, scambio. Il più delle volte percorrendo strade e modalità anche molto differenti fra loro.

Contemporary cities could be considered vibrant organisms of places and communities, of data and information, of sensors and actuators, and of actions and reactions generated by people and environment both. Cities must be more responsive to our behavioral changes, enabling devices for enhancing our contemporary life. We would be able to build amore efficient urban environment, able to sense, to understand and to act every day and for everyone. In the post-city age and beyond the smart city, Augmented City is a new paradigm that perceives the demands of more networked, knowledge-based, and creative society that answers to the global change by a new circular metabolism. The Augmented City is a spatial/cultural/social/economic platform for enhancing our contemporary life, individual and collective, informal, and institutional, expanding the urban space generated by the effects of innovation. The Augmented City redefines dogmas of urbanism that we often thought of being more static and rule-based, recovering its prospective, incremental, responsive, and creative approach. We need to think, design, and manage cities more sentient, open source and intelligent, again productive, creative and based on recycle, cities resilient, fluid and reticular, and truly strategic. The Augmented City is not the city of the future, but it brings us in a different present.

Objective and design: Epigenetic regulation is important in the activation of inflammatory cells. In the present study, we evaluated if DNA-methylation variations are involved in Interleukin-1β (IL-1β)-induced intestinal epithelial cells activation. Materials and methods: Differentiated Caco-2 cells were exposed to IL-1β or to 5-azadeoxycytidine (5-azadC) for 24 or 48 h. Genome-wide methylation status was evaluated, while DNA methylation status at the promoter region of the gene encoding interleukin-6, 8 and 10 (IL-6, 8 and 10) was estimated. The levels of the corresponding gene products as well as DNA methyltransferases (DNMTs) quantity were assessed. Results: IL-1β decreased genomic methylation of human intestinal epithelial cells and induced demethylation at cg-specific sites at the promoter of pro-inflammatory genes IL6 and IL8; conversely it did not change the methylation of the IL10 promoter. IL-1β also increased the release of IL-6 and IL-8 but did not change the IL-10 expression. Finally, cell exposure to IL-1β decreased the DNMT3b expression, increased DNMT3a and was not able to change DNMT1 expression. Conclusions: Our results suggest a potential role of IL-1β as modulator of DNA methylation in activated differentiated Caco-2 cell line.

A fundamental task in cancer research aims at the identification of new pharmacological therapies that can affect tumor growth. Differentiation therapy might exploit this function not only for hematological diseases, such as acute promyelocytic leukemia (APML) but also for epithelial tumors, including lung cancer. Here we show that Retinoic Acid (RA) arrests in vitro and in vivo the growth of Tyrosine Kinase Inhibitors (TKI) resistant Non Small Cell Lung Cancer (NSCLC). In particular, we found that RA induces G0/G1 cell cycle arrest in TKI resistant NSCLC cells and activates terminal differentiation programs by modulating the expression of GATA6, a key transcription factor involved in the physiological differentiation of the distal lung. In addition, our results demonstrate that RA inhibits EGFR and Wnt signaling activation, two pathways involved in NSCLC progression. Furthermore, we uncovered a novel mechanism in NSCLC that shows how RA exerts its function; we found that RA-mediated GATA6 activation is necessary for EGFR and Wnt inhibition, thus leading to 1) increased differentiation and 2) loss of proliferation. All together, these findings prove that differentiation therapy might be feasible in TKI resistant NSCLCs, and shed light on new targets to define new pharmacological therapies.

Human PTHrP gene displays a complex organization with nine exons producing diverse mRNA variants due to alternative splicing at 5' and 3' ends and the existence of three different transcriptional promoters (P1, P2 and P3), two of which (P2 and P3) contain CpG islands. It is known that the expression of PTHrP isoforms may be differentially regulated in a developmental stage- and tissue-specific manner. To search for novel molecular markers of stemness/differentiation, here we have examined isoform expression in fat-derived mesenchymal stem cells both maintained in stem conditions and induced toward adipo- and osteogenesis. In addition, the expression of the splicing isoforms derived from P2 and P3 promoters was correlated to the state of methylation of the latter. Moreover, we also performed a quantitative evaluation of intracellular and secreted PTHrP protein product in undifferentiated stem cells and in parallel cultures at various differentiation stages. The data obtained indicate that from the stemness condition to that of osteo- and adipo-genic differentiated cells, the expression of isoforms becomes increasingly selective, thereby being a potential gene signature for the monitoring of cell stem or committed/differentiating state and that the switching-off of PTHrP isoform expression is mostly promoter methylation-dependent. Moreover, PTHrP intracellular retention is down-regulated in osteo-differentiating cells whereas the secretion of the protein in the extracellular medium is up-regulated with respect to stem cells, thereby suggesting that these variations of the intracellular and extracellular levels of PTHrP could potentially be enclosed in the list of the available protein signature of osteogenic differentiation.

We have previously reported a molecular and cytogenetic characterization of three different 5S rDNA clusters in the sea urchin Paracentrotus lividus and recently, demonstrated the presence of high heterogeneity in functional 5S rRNA. In this paper, we show some important distinctive data on 5S rRNA transcription for this organism. Using single strand conformation polymorphism (SSCP) analysis, we demonstrate the existence of two classes of 5S rRNA, one which is embryo-specific and encoded by the smallest (700 bp) cluster and the other which is expressed at every stage and encoded by longer clusters (900 and 950 bp). We also demonstrate that the embryo-specific class of 5S rRNA is expressed in oocytes and embryonic stages and is silenced in adult tissue and that this phenomenon appears to be due exclusively to DNA methylation, as indicated by sensitivity to 5-azacytidine, unlike Xenopus where this mechanism is necessary but not sufficient to maintain the silenced status.

The present invention relates to the obtainment of vegetable products showing pharmacological activity and their therapeutic use. In particular the invention refers to vesicles of nanometric dimensions obtained from the juice of plants of the family Rutaceae.

Background: Recently, we have shown anti-proliferative and pro-apoptotic effects of indicaxanthin associated with epigenetic modulation of the onco-suppressor p16INK4a in the human colon cancer cell line CACO2. In the present study, the epigenetic activity of indicaxanthin and the mechanisms involved were further investigated in other colorectal cancer cell lines. Methods: LOVO1, CACO2, HT29, HCT116, and DLD1 cells were used to evaluate the potential influence of consistent dietary concentrations of indicaxanthin on DNA methylation, and the epigenetic mechanisms involved were researched. Results: Indicaxanthin exhibited anti-proliferative activity in all cell lines but HT29, induced demethylation in the promoters of some methylation-silenced onco-suppressor genes involved in colorectal carcinogenesis (p16INK4a, GATA4, and ESR1), and left unchanged others which were basally hypermethylated (SFRP1 and HPP1). In apparent contrast, cell exposure to indicaxanthin increased DNMT gene expression, although indicaxanthin appeared to be an inhibitor of DNMT activity. Indicaxanthin also increased the expression of genes involved in DNA demethylation. Finally, an in silico molecular modelling approach suggested stable binding of indicaxanthin at the DNMT1 catalytic site. Conclusions: Our findings contribute to new knowledge in the field of phytochemicals and specifically suggest dietary indicaxanthin as a potential epigenetic agent to protect colon cells against tumoral alterations.

Bone disease is the most frequent complication in multiple myeloma (MM) resulting in osteolytic lesions, bone pain, hypercalcemia and renal failure. In MM bone disease the perfect balance between bone-resorbing osteoclasts (OCs) and bone-forming osteoblasts (OBs) activity is lost in favour of OCs, thus resulting in skeletal disorders. Since exosomes have been described for their functional role in cancer progression, we here investigate whether MM cell-derived exosomes may be involved in OCs differentiation. We show that MM cells produce exosomes which are actively internalized by Raw264.7 cell line, a cellular model of osteoclast formation. MM cell-derived exosomes positively modulate pre-osteoclast migration, through the increasing of CXCR4 expression and trigger a survival pathway. MM cell-derived exosomes play a significant pro-differentiative role in murine Raw264.7 cells and human primary osteoclasts, inducing the expression of osteoclast markers such as Cathepsin K (CTSK), Matrix Metalloproteinases 9 (MMP9) and Tartrate-resistant Acid Phosphatase (TRAP). Pre-osteoclast treated with MM cell-derived exosomes differentiate in multinuclear OCs able to excavate authentic resorption lacunae. Similar results were obtained with exosomes derived from MM patient's sera. Our data indicate that MM-exosomes modulate OCs function and differentiation. Further studies are needed to identify the OCs activating factors transported by MM cell-derived exosomes.

We isolated and characterized nanovesicles from edible Citrus limon with size and composition similar to mammalian-derived exosomes. Furthermore we show an in vitro and in vivo anti-proliferative and pro-apoptotic effect of these vesicles. This study opens to the possibility of using this natural plant-derived nanovesicles as antineoplastic agents.

Autophagy is a highly conserved catabolic process that degrades and recycles intracellular components through the lysosomes [1]. The role of this process in tumorigenesis and tumor progression is controversial: in the early stages, it can block tumor growth and conversely it can promote its progression in the later stages [2]. The tumor suppressor BECN1 gene, encodes the protein Beclin 1, a marker of autophagy down-regulated in several types of cancer, such as colorectal cancer [3]. There are a lot of both genetic and environmental risk factors for colorectal cancer, including diet: for this reason, in accordance with epidemiological studies, consumption of foods rich in phytochemicals is widely promoted. The betalain indicaxantin (Ind) is a phytochemical from the Opuntia Ficus-Indica fruit having several biological activities, such as antioxidant, anti-inflammatory. It showed antiproliferative and proapoptotic effects in colorectal adenocarcinoma (Caco2) cells where was able to regulate gene expression through modulation of methylation state of DNA at CpG islands [4]. For the first time, using Methylation-Sensitive Restriction Endonuclease PCR (MSRE-PCR), we report that Ind (50 e 100 µM) decreases the methylation of BECN1 promoter in Caco2 cells, to the same extent as 5-azacytidine (Zcyd, positive control). Interestingly, colorimetric detection of DNA Methyltransferases activity, indicates that Ind reduced the activity of these enzymes, like Zcyd did. These preliminary data, indicating that Ind is able to decrease the methylation of BECN1 gene, allow us to propose an epigenetic hypothesis of autophagy regulation in Caco2 cells.

Intestinal inflammation is a natural process crucial to maintain gut integrity, but its deregulation is involved in the pathogenesis of severe intestinal disorders[1]. Intestinal epithelial cells play a crucial role in the inflammatory response, modulating the immune cell exposure to antigens and by their ability to secrete many inflammatory mediators. IL-1β represents a pivotal player: secreted by infiltrated leucocytes, it induces the expression of several pro-inflammatory genes. Also the anti-inflammatory IL-10, whose function is to terminate the inflammatory process, modulates the intestinal physiology[2]. Recent clinical reports showed that patients with ulcerative colitis in remission phase have significantly higher IL10 gene expression in mucosa compared with active patients and controls[3]. Moreover, in the latest years aberrant epigenetic mechanisms were put in binomial relationship with chronic inflammatory diseases[4]. Previously, we described a demethylation of pro-inflammatory IL6 and IL8 genes in human colonic Caco-2 cells differentiated into an enterocyte-like phenotype and exposed to the inflammatory action of IL-1β[5]. In the present study we evaluate whether the IL-1β treatment affected the methylation status of the anti-inflammatory IL10 gene, in the same in vitro model. Our results showed that IL-1β treatment did not change the hypermethylation status of the IL10 promoter. Moreover, in cell lysates from IL-1β-treated Caco-2 cells, we observed a dose-dependent increase of DNMTs activity and, surprisingly, a decrease of DNMT3b expression. These findings put in evidence the complexity of relationship between IL-1β and DNMTs, and may suggest a potential role of IL-1β as pleiotropic modulator of DNA methylation in Caco-2 cell line.

Different individuals possess slightly different genetic information and show genetically-determined differences in several enzyme activities due to genetic variability. Following an integrated approach, we studied the polymorphisms and methylation of sites contained in the 5' flanking region of the metabolizing enzyme CYP2E1 in correlation to its expression in both tumor and non-neoplastic liver cell lines, since to date little is known about the influence of these (epi)genetic elements in basal conditions and under induction by the specific inductor and a demethylating agent. In treated cells, reduced DNA methylation, assessed both at genomic and gene level, was not consistently associated with the increase of enzyme expression. Interestingly, the Rsa/Pst haplotype differentially influenced CYP2E1 enzyme expression. In addition, regarding the Variable Number of Tandem Repeats polymorphism, cells with A4/A4 genotype showed a greater expression inhibition (ranging from 20% to 30%) compared with others carrying the A2/A2 one, while those cells bringing A2/A3 genotype showed an increase of expression (of 25%, about). Finally, we demonstrated for the first time that the A2 and A3 CYP2E1 alleles play a more important role in the expression of the enzyme, compared with other (epi)genetic factors, since they are binding sites for trans-acting proteins.

Inflammation is a complex physiological response that requires the activity of a sophisticated regulatory network involving the activation of specific genes for defense, tissue repair and remodeling. Although transcriptional activation has been shown to be critical in the regulation of inflammatory genes (1) the role of epigenetic phenomena in the modulation of the inflammatory response is now emerging (2). Specifically, it has been recently reported that proinflammatory stimuli induce DNA demethylation in the interleukin IL-1 promoter of human articular chondrocytes (3). IL-1 cytokine, among several proinflammatory agents, represents an essential player in the inflammatory conditions of the gut (4): functioning as the strongest signal transduction to NF-kB, IL-1 increases in intestinal paracellular permeability and over-expression of proinflammatory genes (5). In this tissue, moreover, inflammatory response is crucial to maintain its structural integrity and function, thus, alteration and deregulation of inflammatory pathways contribute to tissue damage and ulceration, and are pivotal factors in the pathogenesis of several inflammatory gut diseases. In the present study we evaluate both wide-ranging and gene-specific epigenetic changes in the inflammatory response of Caco-2 cells differentiated into intestinal epithelial cells and exposed to the inflammatory actions of IL-1β. Our results clearly show that IL-1induces changes in the DNA methylation either at genome and gene level and that the local methylation changes are induced in two pro-inflammatory genes that are IL-1-regulated. In particular, we show that a cell exposure to IL-1β for 24 h induces, in a dose-dependent manner, hypomethylation of genomic DNA in respect to untreated cells. We also observe a reduced DNA methyltransferases activity of cell lysates obtained from IL-1 treated cells. Finally our data show that IL-1 is able to induce hypomethylation of specific CpG sites in IL-6 and IL-8 genes. These preliminary results suggest that IL-1 in intestinal epithelial cells is able to act as an epigenetic modulator towards the entire genome and specific genes. Modulation of epigenetic changes in inflammation may provide a new “reading frame” of the inflammatory diseases molecular basis. Since epigenetic modifications are potentially reversible, a thorough understanding of these changes during inflammatory response opens opportunities to develop efficient agents for specific targets.

Nel saggio curato da Ferdinando Trapani affronta aspetti che legano il problema della gestione turistica delle risorse locali dal punto di vista della programmazione che della pianificazione dello sviluppo territoriale. Il contributo di Ferdinando Trapani affronta una riflessione sulla problematica della gestione nel rapporto tra domanda e offerta turistica.

D-Limonene is found widely in citrus and many other plant species; it is a major constituent of many essential oils and is used as a solvent for commercial purposes. With the discovery of its chemotherapeutic properties against cancer, it is important to investigate the biological effects of the exposure to D-Limonene and elucidate its, as yet unknown, mechanism of action. We reported here that D-Limonene is toxic in V79 Chinese hamster cells in a dose-dependent manner. Moreover, to determine the cellular target of D-Limonene, we performed morphological observations and immunocytochemical analysis and we showed that this drug has a direct effect on dividing cells preventing assembly of mitotic spindle microtubules. This affects both chromosome segregation and cytokinesis, resulting in aneuploidy that in turn can lead to cell death or genomic instability.

Cytochrome P450 2E1 (CYP2E1) is one of the main enzymes involved in the oxidation of ethanol and in the transformation of a number of potentially dangerous compounds. It has various polymorphic sites, one of which is a variable number tandem repeat (VNTR) polymorphism previously described in the 5'-flanking region. The aim of this study was to investigate the genotype-phenotype association between CYP2E1 VNTR polymorphisms and risky health habits in healthy subjects and to analyze the associations between these polymorphisms with drinking- and/or smoking-related cancers. We analyzed 166 healthy subjects by genotyping for the CYP2E1 VNTR polymorphism associated with drinking and/or smoking habits by the more sensitive restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method, using the NlaIV restriction enzyme. Sixty cases of pancreatic adenocarcinoma (PA) and 66 with hepatocellular carcinoma (HCC), were also genotyped. Statistical analysis was carried out to investigate the genotype-phenotype associations and to compare certain genotypes and cancer. We found 7 genotypes both in the healthy subjects and patients. The A1/A1 genotype was observed to be mainly associated with non-drinkers and -smokers (87.5 and 75.0%, respectively); moreover it was never found in the PA or HCC patients. Conversely, a weak association between A2/A3 with smokers (45.8%) and A4/A4 with drinkers (53.9%) was detected. In addition, the A4/A4 genotype was found to be significantly associated to PA [odds ratio (OR)=3.25; 95% confidence interval (CI) 1.21-7.50]. Our data demonstrate that certain CYP2E1 VNTR genotypes are associated with drinking and/or smoking habits; consequently, they may contribute either to the decreased or increased risk of developing drinking- and/or smoking-related cancers. In particular, we hypothesize that the A1/A1 VNTR genotype may have a protective role against drinking- and/or smoking-related cancers, and that A4/A4 may be a high-risk genotype during the early stages of cancer.

Il tema centrale di questo scritto è ragionare sui requisiti di territorialità che possono essere rintracciati nei sistemi ad economia diffusa, con particolare attenzione alle mutazioni intervenute con la transizione ad una fase “postindustriale” che molti sistemi economici maturi stanno affrontando da alcuni anni. Proprio l’avere assunto come riferimento la nozione di sistema locale ci mette a disposizione un apparato concettuale che, a nostro avviso, ben si presta a svelare alcuni degli equivoci terminologici (e, poi, anche operativi) comparsi di recente nella cultura dello sviluppo locale. Si proverà a affrontare la questione facendo ricorso al concetto di “esternalità”, un termine che ha le sue più larghe applicazioni nelle discipline economiche ma che, a nostro avviso, si presta a rivelare il contributo che le risorse territoriali endogene possono esercitare nei processi di sviluppo locale.

Phytochemicals play prominent roles in human diet and nutrition as protective redo-active substances in prevention of several disorders and chronic diseases in humans. Today, their function as potent modulators of the mammalian epigenome-regulated gene expression is rapidly emerging. In the present study antiproliferative effects of Indicaxanthin (Ind) from the fruits of Opuntia ficus-indica (1), and potential influence on DNA methylation has been investigated on Caco-2 cells, a human cell line of colorectal carcinoma. Ind caused a clear dose- and time-dependent decrease of the cell proliferation (IC(50) 50 M) associated to apoptosis as demonstrated by phosphatidylserine externalization and depolarization of mithocondrial membrane. Ind decreased the Go-G1phase whereas increased S and G2-M phases of the cell cycle. The phytochemical did not altered the intracellular ROS levels but decreased the [Ca2+]i. Investigation on DNA methylation using MESAP-PCR (Methylation-Sensitive Arbitrarily-Primed Polymerase Chain Reaction) (2), showed that 100 M Ind induced a slight global demethylation after a 48 h treatment. Analysis of epigenetic changes in the DNA methylation pattern at CpG promoter of p16 (INK4a), using MSRE (Methylation-Sensitive Restriction Endonucleases Multiplex-Polymerase Chain Reaction), showed that Ind caused CpG demethylation. Western blotting analysis carried out with p16 monoclonal antibody, confirmed the reactivation of the protein expression. Present data, suggesting that a long-term exposure to indicaxanthin in diet might potentially affect epigenetic machines of the intestinal cells, preventing or repairing initial derangements/disorders, encourage studies on the mechanism involved.

Si illustrano le nuove forme di stanzialità del turismo

Il contributo contiene il resoconto dello stato di avanzamento di una ricerca e azione nel quadro di un accordo tra l'Università di Palermo e del Cairo per la realizzazione di un Master di secondo livello Italia Egitto e di ricerca sui temi del turismo relazionale integrato.

Il turismo relazionale integrato è una nuova forma di sviluppo locale che coniuga istanze ambientali con forme indirette di redistribuzione del reddito (la spesa dei turisti sul territorio ospitante). Attività economica inevitabilmente territorializzata, il turismo relazionale integrato propone in Sicilia, rispetto ai processi di globalizzazione, un paradigma di sviluppo auto-centrato volto al riequilibrio tra pressione antropica costiera e aree interne in declino economico. In aree marginali, il recupero del patrimonio storico e culturale esistente, la natura dei luoghi e l’interazione con le attività in abbandono, sono motivi di riflessione sulle capacità creative delle comunità locali di partecipare consapevolmente alla ricostruzione delle identità territoriali, ricombinando in termini innovativi i materiali della memoria. Il contributo affronta il tema della partecipazione creativa al turismo, attraverso la descrizione di due casi studio, differentemente marginali rispetto a consuete mete turistiche e offerte di servizi più tradizionali, mostrando le molteplici motivazioni relazionali che s’intessono tra i luoghi da visitare, le comunità ospitanti e i viaggiatori. The integrated relational tourism is a new form of local development, which combines environmental instances with those of indirect income redistribution (tourists’ spending on host territory). Economic activity inevitably rooted on territory, this kind of tourism in Sicily, with respect to globalization processes, proposes a local development paradigm, which is aimed at rebalancing between anthropic pressures of coastal areas and decline of internal areas. In marginal areas, some issues, such as the recovery of historical settlements and of cultural heritage, the nature of places and the interaction with activities in abandonment, are reasons for reflection on the creative abilities of local communities to participate knowingly in the reconstruction of territorial identity, through the recombining the materials of memory in innovative terms. The contribution deals with the theme of the creative participation to tourism, through the description of two case studies, which are characterized by condition of different marginality and differ from typical tourist destinations and from more traditional tourist offerings, showing the various motivations that put in relation places, host communities and travellers.

In the present study antiproliferative effects of Indicaxanthin (Ind), a highly bioavailable pigment from the fruits of Opuntia ficus-indica (1), were investigated on a number of human cancer cell lines including hepatocarcinoma cells (HepG2, Ha22T, HUH 7), breast cancer cells (MCF7), cervix epithelial carcinoma (HeLa), and colorectal carcinoma cells (Caco-2). Cytotoxicity of Ind, in a concentration range between 25 to 100 M, was evaluated by Trypan blue exclusion method and MTT assay. Ind caused a clear dose- and time-dependent decrease in the proliferation of Caco-2 and Ha 22T cells with an IC(50) of about 50 M, with minor effect on the other cell lines. Flow cytometric analysis after Annexin V-FITC and propidium iodure double staining, at 24, 48 and 72 h of treatment with 100 M Ind, showed a pro-apoptotic effect of the pigment at 48 and 72 h. Effect of Ind on DNA methylation investigated on DNA from Ha22T cells line and Caco2 cells line at 48 h of treatment with 10 M Ind, using MESAP-PCR (Methylation-Sensitive Arbitrarily-Primed Polymerase Chain Reaction) (3) showed that Ind induces a slight global demethylation. While antiproliferative effects of indicaxanthin add further value to the nutritional characteristics of the fruits of O. ficus-indica (2), our results also are consistent with the emerging role of dietary phytochemicals on the epigenetic regulation of gene expression.

Contributo scientifico sulla diversità dei luoghi in relazione alla definizione di “Progetto” quale connotazione imprescindibile della sua natura disciplinare nell’ambito della ricerca Turismo Relazionale Integrato, per il progetto regionale e internazionale MOTRIS II. Curato dal Collegio Universitario ARCES con il supporto scientifico del Centro Ricerche nuovo Umanesimo Edoardo Caracciolo (CRUEC).

Cytochrome P450 (CYP2E1) is often associate to susceptibility to alcohol-related diseases and various cancers, because of its role in the metabolism of multiple environmental xenobiotics. In the 5’- flanking region of the human CYP2E1 gene there are restriction fragment length polymorphism which are involved in the transcriptional regulation of the CYP2E1 gene. Recently a tandem repeat polymorphism (VNTR) in the 5’-flanking region of CYP2E1 was found. Because cytochrome P450 2E1 catalyzes the metabolic activation of pro-carcinogen and cytotoxic compound, we value the genetic distribution of this tandem repeat polymorphism in a healthy population, and in patients with hepatocellular carcinoma living in same country, in order to found a correlation between CYP2E1 VNTR genotype and neoplasia. DNA was isolated from spit sample of 108 control subject and from the peripheral lymphocytes of 35 HCC patients. The 5’ flanking region of the CYP2E1 gene was amplified by polymerase chain reaction and examined for tandem repeat polymorphism using Nla IV restriction map. This study reports that only four of the ten possible genotype were found in all subjects. The modal genotype, found in both analyzed populations, is A2/A2. Interestingly, in a gender-based analysis of data this genotype was found more frequent in woman with disease that in control ones. These preliminary findings represent a first report of a gender-specific correlation between CYP2E1 VNTR polymorphism and hepatocarcinoma.

Cosa possa intendersi per "cultura aziendale" e importanza delle caratteristiche con cui essa può manifestarsi, in relazione allo sviluppo di un territorio.

L'azione di ricerca ha lo scopo di verificare la presenza, nei tre contesti di studio (Val d’Anapo, Valle dell’Halaesa, S. Vito) di un’ offerta turistica che dispone di un sistema di relazioni formali ed informali al fine di ottenere un'idonea “mappatura” dei tre territori per determinare, con rigore scientifico, il fabbisogno turistico in chiave relazionale. Un secondo aspetto riguarda la verifica della dimensione relazionale con riferimento alla domanda turistica effettiva e potenziale delle località. Si vuol stabilire quali e quanti sono le relazioni presenti tra domanda turistica e sistema dell’offerta, sia attuali che potenziali ed in che termini questi contatti siano in grado di influenzare il livello di qualità dell’offerta complessiva della località. Questo aspetto è utile per verificare se i tre contesti territoriali possono poi costruire un vantaggio competitivo basato sulla relazionalità del territorio e dove si deve intervenire per rafforzarlo e per renderlo visibile nell’offerta turistica complessiva. La ricerca si offre fondamentalmente quale indagine di campo, quale studio che, partendo dalla rilevazione di determinate risorse reali e dall’analisi del fabbisogno ampiamente turistico, miri a verificare l’opportunità di raccordare gli operatori privati e pubblici, direttamente e indirettamente impegnati nel settore turistico, al fine di progettare e quindi comporre, un'offerta turistica relazionale articolata e/o di promuoverla e/o di gestirla nella fase erogativa. La ricerca ha quindi come scopo quello di dare fondamento analitico e contenuti strategici a questa opzione di sviluppo, e vuole rappresentare un valido supporto funzionale per quei territori che intendono programmare e proporre processi di sviluppo turistico con connotazione relazionale.

Il contributo contenuto in questo capitolo, vuole indagare il sistema di relazioni esistenti tra turista-imprese, turista-enti pubblici, turista-comunità locale, nei tre ambiti territoriali. L’analisi accerta la presenza di relazionalità tra gli attori, verificandone l’intensità e la direzione, adottando una metodologia statistica, che fa uso di indicatori sintetici, sulla base dei dati raccolti in fase di rilevazione. I paragrafi seguenti, riportano per ciascuna relazione (turista verso la comunità e viceversa; turista verso gli enti, e viceversa; turista verso imprese e viceversa), la misura della relazionalità. Ciò consente di tracciare la mappa delle relazioni presenti all’interno di ciascun sistema territoririale (Anapo, S. Vito e Halesa), evidenziazione le difformità e le principali somiglianze. Dall’analisi del sistema relazionale interno ed esterno, si è provceduto all’analisi dei principali fabbisogni formativi presenti nei tre sistemi territoriali, evidenziandome, contestualmente i gaps esistenti e da colmare per il raggiungimento di su sistema d’offerta turistica locale particolarmente fondato sulla relazionalità degli attori locali.