DNA demethylating antineoplastic strategies: a comparative point of view.

Despite the involvement of genetic alterations in neoplastic cell transformation, it is increasingly evident that abnormal epigenetic patterns, such as those affecting DNA methylation and histone posttranslational modifications (PTMs), play an essential role in the early stages of tumor development. This finding, together with the evidence that epigenetic changes are reversible, enabled the development of new antineoplastic therapeutic approaches known as epigenetic therapies. Epigenetic modifications are involved in the control of gene expression, and their aberrant distribution is thought to participate in neoplastic transformation by causing the deregulation of crucial cellular pathways. Epigenetic drugs are able to revert the defective gene expression profile of cancer cells and, consequently, reestablish normal molecular pathways. Considering the emerging interest in epigenetic therapeutics, this review focuses on the approaches affecting DNA methylation, evaluates novel strategies and those already approved for clinical use, and compares their therapeutic potential

INTEGRATED KNOWLEDGE ON A THREE-DIMENSIONAL BASIS: THE FOUNDATION OF THE CONSERVATION PROJECT AND POSSIBLE IMPLICATIONS IN THE BIM AREA THE CASE OF THE PORTRAIT ROOM OF PALAZZO BOREA D'OLMO IN SANREMO

This involved a sort of preventive diagnostics based on the historical knowledge of the building, on the construction techniques and on the compositional style, on the other hand on diagnostic imaging using photographic techniques developed from the early 1800s.The preliminary cognitive approach, the stylistic identification, the classification of method, the historical reading, the critical analysis of the historical and compositional genesis of the building, the careful reading of the pathogenesis in progress, mark, from the mid-800 in then, the evolution of the concept of restoration that will take shape from time to time from what was "before" and only secondarily from what "is now". This perhaps unconscious diagnostic reading of the artifact (first artistic, then monumental and finally as a cultural testimony) has, in fact, involved generations of restorers painting the landscape of the restoration sometimes in different way, with results that well we know.In fact, almost never the fathers of restoration (or anti-restoration) explicitly touch the issues of preventive diagnosis, but certainly base their theories and their "restoration" on the observation of the factory, on the knowledge of construction techniques and historical period of belonging.The things begin to change starting from the first Charter of Restoration of 1883 with the enlargement of the diagnostic project (which finds paternity starting from the 1972 Italian Charter, which embraces many disciplines: from photography to relief, from chemistry to physics, from the historical reading to the knowledge of the materials, from the identification of the material pathogenesis to the reading of the static and structural framework of the buildings.With the passing of time and the explosion of science and technology in the field of cultural heritage all aspects related to preliminary knowledge aimed at their conservation evolve exponentially especially with the advent of electronics. Starting from the 60s of the 1900s, equipment with great calculation capacity and small dimensions was born, a real springboard for the new techniques of surveying by means of a laser scanner, photographic socket and photogrammetric restitution.Tools and techniques of restitution settle and even more solidly base the knowledge base linked to the project of conservation and enhancement of cultural heritage.Base of support and extraction of selected data and / or dedicated to the project continuously interface with the disciplines of knowledge that are now more and more "forced" to the table of confrontation to reveal information hidden in the folds of time.A recent work on an eighteenth-century Sanremese building it gives us information on the complexity of the construction of a process of knowledge articulated, composed of several activities to be correlated and integrated continuously, one in the other to try to give answers on evident problems of degradation never fully documented.The study, performed in a rather limited time frame, focused on the Hall of family portraits seriously degraded by "accidents of various kinds". The work was based on the three-dimensional conception of the acquired data, allowing to investigate the hall as an articulated organism, supporting the technicians in the three-dimensional understanding of the asset, constituting a complete database of the actual state, becoming support of the results of the various surveys conducted for the knowledge of the asset.In this cognitive path, the BIM method is understood as the possibility of constructing the "as built" model, complete not only of the dimensional graphic data of the asset, but also of those that contribute to the determination of the actual state. The graphic aspect of the model must be connected with historical, material, degradation, contextual information, with the results of any specialized investigations conducted.Only then will the BIM model of a historic building be the complete database, the "medical record" of its state of health, complete with diagnosis, care, indications of maintenance that can be shared and questioned over time.</p

Physical activity, sedentary behaviour and screen time among youths with Down syndrome during the COVID-19 pandemic

Background: The COVID-19-related restrictions hampered habitual physical activity (PA), particularly affecting the more vulnerable, such as people with Down syndrome (DS). The study aimed to investigate changes in PA, sedentary behaviour (SB) and screen time (ST) of youths with DS, before, during and after the restrictions, also in relation to parental PA levels. Methods: A cross-sectional design with a retrospective assessment of variables for the before and during restrictions periods was adopted. Parents of youths with DS completed an online questionnaire. Sociodemographic aspects, weekly PA levels and youths' daily SB and ST were investigated, referring to three time-points: before the pandemic, during the restrictions and the restrictions-easing phase. Results: A total of 57 parents voluntarily participated in the study, proxy-reporting on their child (male = 41, female = 16, age = 21.4 ± 7.7 years). A repeated measures multivariate analysis of variance showed negative effects of restrictions (P < 0.05) on PA levels, SB and ST, independently from sociodemographic characteristics. In the restrictions-easing phase, PA levels did not return to before the pandemic values (P < 0.05). A positive correlation between parents and their child's PA was detected before the pandemic (r = 0.38; P < 0.01), no longer reported in the restrictions-easing phase. Conclusions: The findings showed the negative impact of restrictions on youths with DS lifestyle. Moreover, the importance of addressing the needs of the disabled community including the whole family is highlighted

Dietary habits and psychological states during covid-19 home isolation in italian college students: The role of physical exercise

Social isolation has adverse effects on mental health, physical exercise, and dietary habits. This longitudinal observational study aimed to investigate the effects of mood states and exercise on nutritional choices, on 176 college students (92 males, 84 females; 23 &plusmn; 4 years old) during the COVID-19 lockdown. During 21 days, nutrition and exercise were daily monitored, and the mood states assessed. A factor analysis was used to reduce the number of nutritional variables collected. The relationships between exercise, mood and nutrition were investigated using a multivariate general linear model and a mediation model. Seven factors were found, reflecting different nutritional choices. Exercise was positively associated with fruit, vegetables and fish consumption (p = 0.004). Depression and quality of life were, directly and inversely, associated with cereals, legumes (p = 0.005; p = 0.004) and low-fat meat intake (p = 0.040; p = 0.004). Exercise mediated the effect of mood states on fruit, vegetables and fish consumption, respectively, accounting for 4.2% and 1.8% of the total variance. Poorer mood states possibly led to unhealthy dietary habits, which can themselves be linked to negative mood levels. Exercise led to healthier nutritional choices, and mediating the effects of mood states, it might represent a key measure in uncommon situations, such as home-confinement

PAT-ChIP coupled with laser microdissection allows the study of chromatin in selected cell populations from paraffin-embedded patient samples

Background: The recent introduction of pathology tissue-chromatin immunoprecipitation (PAT-ChIP), a technique allowing chromatin immunoprecipitation from formalin-fixed and paraffin-embedded (FFPE) tissues, has expanded the application potential of epigenetic studies in tissue samples. However, FFPE tissue section analysis is strongly limited by tissue heterogeneity, which hinders linking the observed epigenetic events to the corresponding cellular population. Thus, ideally, to take full advantage of PAT-ChIP approaches, procedures able to increase the purity and homogeneity of cell populations from FFPE tissues are required. Results: In this study, we tested the use of both core needle biopsies (CNBs) and laser microdissection (LMD), evaluating the compatibility of these methods with the PAT-ChIP procedure. Modifications of the original protocols were introduced in order to increase reproducibility and reduce experimental time. We first demonstrated that chromatin can be prepared and effectively immunoprecipitated starting from 0.6-mm-diameter CNBs. Subsequently, in order to assess the applicability of PAT-ChIP to LMD samples, we tested the effects of hematoxylin or eosin staining on chromatin extraction and immunoprecipitation, as well as the reproducibility of our technique when using particularly low quantities of starting material. Finally, we carried out the PAT-ChIP using chromatin extracted from either normal tissue or neoplastic lesions, the latter obtained by LMD from FFPE lung sections derived from mutant K-rasv12transgenic mice or from human adeno- or squamous lung carcinoma samples. Well characterized histone post-translational modifications (HPTMs), such as H3K4me3, H3K27me3, H3K27Ac, and H3K9me3, were specifically immunoselected, as well as the CTCF transcription factor and RNA polymerase II (Pol II). Conclusions: Epigenetic profiling can be performed on enriched cell populations obtained from FFPE tissue sections. The improved PAT-ChIP protocol will be used for the discovery and/or validation of novel epigenetic biomarkers in FFPE human samples

Malten, a new synthetic molecule showing in vitro antiproliferative activity against tumour cells and induction of complex DNA structural alterations

Background: Hydroxypyrones represent several classes of molecules known for their high synthetic versatility. This family of molecules shows several interesting pharmaceutical activities and is considered as a promising source of new anti neoplastic compounds. Methods: In the quest to identify new potential anti cancer agents, a new maltol (3-hydroxy-2-methyl-4-pyrone)-derived molecule, named malten (N,N′-bis((3-hydroxy-4-pyron-2-yl)methyl)-N,N′-dimethylethylendiamine), has been synthesised and analysed at both biological and molecular levels for its antiproliferative activity in eight tumour cell lines. Results: Malten exposure led to a dose-dependent reduction in cell survival in all the neoplastic models studied. Sublethal concentrations of malten induce profound cell cycle changes, particularly affecting the S and/or G2-M phases, whereas exposure to lethal doses causes the induction of programmed cell death (apopotosis). The molecular response to malten was also investigated in two biological models: JURKAT and U937 cells. It showed the modulation of genes having key roles in cell cycle progression and apoptosis. Finally, as part of the effort to clarify the action mechanism, we showed that malten is able to impair DNA electrophoretic mobility and drastically reduce both PCR amplificability and fragmentation susceptibility of DNA. Conclusion: Taken together, these results show that malten may exert its antiproliferative activity through the induction of complex DNA structural modifications. This evidence, together with the high synthetic versatility of maltol-derived compounds, makes malten an interesting molecular scaffold for the future design of new potential anticancer agents

miR-494-3p is a novel tumor driver of lung carcinogenesis

Lung cancer is the leading cause of tumor-related death worldwide and more efforts are needed to elucidate lung carcinogenesis. Here we investigated the expression of 641 miRNAs in lung tumorigenesis in a K-Ras(+/LSLG12Vgeo);RERTn(ert/ert) mouse model and 113 human tumors. The conserved miRNA cluster on chromosome 12qF1 was significantly and progressively upregulated during murine lung carcinogenesis. In particular, miR-494-3p expression was correlated with lung cancer progression in mice and with worse survival in lung cancer patients. Mechanistically, ectopic expression of miR-494-3p in A549 lung cancer cells boosted the tumor-initiating population, enhanced cancer cell motility, and increased the expression of stem cell-related genes. Importantly, miR-494-3p improved the ability of A549 cells to grow and metastasize in vivo, modulating NOTCH1 and PTEN/PI3K/AKT signaling.Overall, these data identify miR-494-3p as a key factor in lung cancer onset and progression and possible therapeutic target

Predictors of survival and toxicity in patients on adjuvant therapy with 5-fluorouracil for colorectal cancer

The present study aimed at investigating whether the simultaneous evaluation of pharmacokinetic, pharmacogenetic and demographic factors could improve prediction on toxicity and survival in colorectal cancer patients treated with adjuvant 5-fluorouracil (5FU)/leucovorin therapy. One hundred and thirty consecutive, B2 and C Duke's stage colorectal cancer patients were prospectively enrolled. 5FU pharmacokinetics was evaluated at the first cycle. Thymidylate synthase (TYMS) 5′UTR and 3′UTR polymorphisms and methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms were assessed in peripheral leukocytes. Univariate and multivariate analyses were applied to evaluate which variables could predict chemotherapy-induced toxicity, disease-free survival (DFS) and overall survival (OS). Multivariate analysis showed that: (a) low 5FU clearance was an independent predictive factor for severe toxicity (OR=7.32; P<0.0001); (b) high-5FU clearance predicted poorer DFS (HR=1.96; P=0.041) and OS (HR=3.37; P=0.011); (c) advanced age was associated with shorter DFS (HR=3.34; P=0.0008) and OS (HR=2.66; P=0.024); (d) the C/C genotype of the MTHFR C677T polymorphism was protective against grade 3–4 toxicity (P=0.040); (e) none of the TYMS polymorphisms could explain 5FU toxicity or clinical outcome