DNA-methylation dependent regulation of embryo-specific 5S ribosomal DNA cluster transcription in adult tissues of sea urchin Paracentrotus lividus

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 (700bp) cluster and the other which is expressed at every stage and encoded by longer clusters (900 and 950bp). 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

TIRANA-NEXT: UNA RICERCA PILOTA SUGLI STRATI URBANI DELLA CITTÀ

Tirana-Next is pilot research that aims to provide an updated analysis-based diagnosis of the Capital and the greatest City of Albania: Tirana. How it was in the past, how it became nowadays, what are its actual conditions and what is going on through the in-force Master Plan “Tirana 030” (2018). Across diverse pilot investigations, within the Tirana neighbourhoods (super-blocks), researchers want to explore the embodied experience of inner spaces and the changing relations between them and the surrounding humanity and spatiality. The pilot cases have been studied and analysed by considering the physical space (hard-scape), the social life on the streets (soft-scape), the human interactions (human scape), the virtual spaces in addition to the conventional physical ones (digital-scape), and the processes framework (process-scape). By investigating and analysing the links between these elements it was expected that this research work might provide a set of practical recommendations and suggestions, pilot-based, for a more suitable systemic-punctual urban regeneration in the favour of Tirana neighbourhoods’ quality of life, even “beyond” the in-force city planning formal process

The Diaptomidae (Copepoda, Calanoida) of Tunisia and the role of spatial and environmental factors as drivers of their distribution patterns

Even in a biodiversity hotspot such as the Mediterranean basin, aspects relating to the distribution of some groups of microcrustaceans still need clarification. In this paper, we critically analyse the available information on diaptomid copepods in Tunisian inland waters and, based on the largest sampling campaign to date carried out in the country, report new data on their distribution. In the frame of this study, 248 crustacean samples were collected from 190 sites, and 10 diaptomid species belonging to 7 genera and 2 subfamilies were found in the samples. Their distribution follows a climate gradient determined by precipitation, and ecological variables mainly affect diaptomid distribution in the study area, while pure spatial factors are of negligible importance. Diaptomid biodiversity is therefore strongly linked to the diversity of inland waters that characterise the country. This is especially true in the wetlands of the Medjerda alluvial plain, where six diaptomid species co-occurred sympatrically, with up to four or five species coexisting syntopically and synchronically in some sites. Unfortunately, the natural water bodies of this plain are currently strongly threatened by urban expansion and agricultural reclamation, and the risk of local extinction for the species they host is increasingly high

Abnormal mitotic spindle assembly and cytokinesis induced by D-Limonene in cultured mammalian cells

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

Role and importance of polymorphisms with respect to DNA methylation for the expression of CYP2E1 enzyme

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 andmethylation of sites contained in the 5′ flanking region of themetabolizing 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 associatedwith 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

Cytochrome P450 2E1 variable number tandem repeat polymorphisms and health risks: A genotype-phenotype study in cancers associated with drinking and/or smoking

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

Osteoarthritis in the Elderly Population: Preclinical Evidence of Nutrigenomic Activities of Flavonoids

Osteoarthritis (OA) is a degenerative joint disease that is age-related and progressive. It causes the destruction of articular cartilage and underlying bone, often aggravated by inflammatory processes and oxidative stresses. This pathology impairs the quality of life of the elderly, causing pain, reduced mobility, and functional disabilities, especially in obese patients. Phytochemicals with anti-inflammatory and antioxidant activities may be used for long-term treatment of OA, either in combination with current anti-inflammatories and painkillers, or as an alternative to other products such as glucosamine and chondroitin, which improve cartilage structure and elasticity. The current systematic review provides a comprehensive understanding of the use of flavonoids. It highlights chondrocyte, cartilage, and subchondral bone activities, with a particular focus on their nutrigenomic effects. The molecular mechanisms of these molecules demonstrate how they can be used for the prevention and treatment of OA in the elderly population. However, clinical trials are still needed for effective use in clinical practice

Pro-apoptotic activity of the phytochemical Indicaxanthin on colorectal carcinoma cells (Caco-2) and epigenetic CpG demethylation of the promoter and reactivation of the expression of p16

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