Study of DNA methylation dynamics in a model of differential developmental competence in the ovine species

DNA methylation is an important epigenetic mark that plays a role in gene regulation by the addition of a methyl group to CpGs in the DNA. Despite being relatively stable in somatic cells, DNA methylation is subject to reprogramming during embryo development and gametogenesis. Aim of this work was to evaluate different aspects of DNA methylation in relation to oocyte quality in the ovine species. A model of differential developmental competence consisting in ovine oocytes and in vitro produced (IVP) embryos derived from prepubertal (P) and adult (A) donors was used. The methylation dynamics were analysed in terms of gene expression of a panel of genes involved in DNA methylation reprogramming (DNA methyltransferases (DNMTs), ten-eleven translocation dioxygenases (TET) and methylbinding proteins (DMB) and immunolocalization of TET3 protein, 5-methylcytosine (5-mc) and 5-hydroxymethylcitosine (5-hmc). Gene relative quantification was performed by RNA reverse transcription followed by Real-Time PCR. Pools of growing oocytes at different diameter (70/90 µm, 90/110 µm, 110/130 µm), germinal vesicles (GV), IVM metaphase II (MII) oocytes, IVP embryos at two (2C) and four cell (4C) stage and blastocysts derived from P and A donors were analysed. The analysis in growing oocytes revealed that DNMT1, DNMT3B, TET1 and TET3 transcript abundance increases in relation to oocyte diameter, suggesting that synthesis and storage occur during the final stage of oocyte growth. P-GV oocytes present lower expression level of TET1, TET2 and TET3 (ANOVA; P< 0.05). DNMT1 expression is lower in 4C embryos from prepubertal oocytes and MBD1 and MBD3 expression decreases during the transition from 2C to 4C stage in embryos derived from P-animals. No significant differences were observed in embryos at the blastocyst stage (ANOVA; P>0.05). These observations confirm different gene expression patterns in our model. Immunolocalization was performed by direct immunofluorescence. TET3 immunostaining revealed the presence of the protein only in the cytoplasm of early embryos (2C and 4C) from both P and A animals. The analysis of global DNA methylation showed high levels in all analyzed samples. Interestingly, pronuclear (PN) stage zygotes displayed a decrease in 5-mC signal in male PN compared to maternal PN from both young and adult animals; conversely, hydroxymethylation signal was scattered evident throughout the chromatin of all analyzed samples; the analysis of zygotes revealed 5-hmc signal only in male PN. Observations demonstrate that male PN undergoes a decrease in methylation and an increase in DNA hydroxylation suggesting that also in sheep an active DNA demethylation occurs soon after fertilization. Our results are in accordance with the recent hypothesis that the acquisition of epigenetic modifications may be involved in the gaining of oocyte competence during its growth. They indicate that epigenetic mechanisms may affect oocyte quality and pave the way to better understanding methylation dynamics during sheep pre-implantation development

Performance estimation of photovoltaic energy production

This article deals with the production of energy through photovoltaic (PV) panels. The efficiency and quantity of energy produced by a PV panel depend on both deterministic factors, mainly related to the technical characteristics of the panels, and stochastic factors, essentially the amount of incident solar radiation and some climatic variables that modify the efficiency of solar panels such as temperature and wind speed. The main objective of this work is to estimate the energy production of a PV system with fixed technical characteristics through the modeling of the stochastic factors listed above. Besides, we estimate the economic profitability of the plant, net of taxation or subsidiary payment policies, considered taking into account the hourly spot price curve of electricity and its correlation with solar radiation, via vector autoregressive models. Our investigation ends with a Monte Carlo simulation of the models introduced. We also propose the pricing of some quanto options that allow hedging both the price risk and the volumetric risk

Gene-specific methylation profiles in BRCA-mutation positive and BRCA-mutation negative male breast cancers

Male breast cancer (MBC) is a rare disease. Due to its rarity, MBC research and clinical approach are mostly based upon data derived from female breast cancer (FBC). Increasing evidence indicate that on molecular level MBC may be an heterogeneous disease different from FBC. In order to investigate whether epigenetic signatures could define molecular subgroups of MBCs, we performed promoter methylation analysis of genes involved in signal transduction and hormone signalling in BRCA1/2 mutation-positive and -negative MBCs. We examined 69 MBCs, paired blood samples, and 15 normal tissues for promoter methylation of hTERT, ESR1, RASSF1, AR, MYC and WNT1 genes. MBCs showed higher gene promoter methylation levels compared to paired blood and normal breast samples. Significantly higher RASSF1 methylation levels were observed in association with BRCA1/2 mutations, HER2 expression and high tumor grade. Significantly higher AR methylation levels were observed in BRCA1/2 wild-type cases and higher WNT1 methylation levels in PR negative cases. Overall, our results indicate that alterations in gene methylation profiles are common in MBC and that methylation pattern of tumor-associated genes may allow for the identification of MBC molecular subgroups, that could have implications in clinical management of MBC patients

Smoking and FGFR2 rs2981582 variant independently modulate male breast cancer survival: A population-based study in Tuscany, Italy

Aim: Male breast cancer (MBC) is a rare disease and recommendations for its clinical management are often extrapolated from those for female breast cancer, even if breast cancer (BC) has different characteristics in the two sexes. The purpose of this study was to assess the influence of several individual characteristics including clinico-pathological, lifestyle and genetic factors on overall survival (OS) of a relatively large and well characterized population-based series of 166 MBCs enrolled in Tuscany. Methods: We genotyped MBC cases at BRCA1/2 genes and at 9 candidate BC susceptibility SNPs. Kaplan-Meier method and multivariate Cox regression, adjusted for several individual characteristics were used. To reduce a possible selection bias related to the interval between diagnosis and enrolment of MBC cases into the study, we used the date of blood donation as the date of the start of observation for survival analysis. Results: Only smoking habits had a significant effect on OS at 10 years (for current smokers, HR: 3.34; 95% CI 1.45–7.68; p = 0.004), while lymph node status fell short of reaching statistical significance (for pN positive, HR: 2.07; 95% CI 0.93–4.55; p = 0.07). In the same multivariate analysis we found a significantly higher OS in cases with FGFR2 rs2981582 variant in the dominant transmission model (HR: 0.29; 95% CI: 0.13–0.62; p = 0.028). A sensitivity analysis with left truncation showed similar results. Conclusions: Our results may contribute to shed light on factors influencing MBC survival suggesting an important role for cigarette smoking and FGFR2 rs2981582 variant, and provide clues for better patient management

PALB2 mutations in male breast cancer: a population-based study in Central Italy

[No abstract available

Frequency and Determinants of Olfactory Hallucinations in Parkinson’s Disease Patients

Background: Olfactory dysfunctions and hallucinations are considered common nonmotor symptoms in Parkinson’s disease (PD). Visual and auditory hallucinations are well-known; however, olfactory hallucinations (OHs) are not fully investigated. The aim of this study was to evaluate OHs in PD patients, and their correlation to motor impairment, cognitive abilities, visual and auditory hallucinations, and olfactory and gustatory function. Methods: A sample of 273 patients was enrolled: 141 PD patients (mean age SD: 70.1 9.5 years) and 132 healthy controls (mean age SD: 69.4 9.6 years). In all patients, the following parameters were evaluated: motor symptoms (UPDRSIII), olfactory function, cognitive abilities, and occurrence of OH, gustatory hallucinations (GHs), and visual/auditory hallucinations. Results: OHs were found only in PD patients with a percentage of 11.3%. Among PD patients with OHs, 2.8% also presented GHs. High significant frequencies of females, the presence of visual/auditory hallucinations, and a high mean UPDRS-III score were found in patients with OHs related to patients without them. Binary logistic regression evidenced the presence of visual/auditory hallucinations and sex as main variables predicting the presence of OHs. Conclusions: Our data indicated that OHs occur frequently in PD patients, especially in women, and often concomitant with visual and auditory hallucinations, without any association with olfactory impairment

A gene-environment interaction between occupation and BRCA1/BRCA2 mutations in male breast cancer?

The association of male breast cancer (MBC) with a positive breast cancer (BC) family history and with BRCA1/2 germ-line mutations points to a genetic component; a relationship with occupation has also been reported. Recently, we identified pathogenetic BRCA1/2 mutations in a population-based series of Italian MBC patients: here in, we investigated interactions between a carrier status for BRCA1/2 mutations and occupation using a case-case design and estimating case-only odds ratios (CORs). Truck-driving was the most frequent occupation (3/4 BRCA-related cases and 2/19 unrelated cases). An interaction between carrier status and working as a truck-driver emerged, when we classified MBC cases as "ever/never-held" this job title (COR 25.5; 95% Confidence Limits (CL): 1.1-1,412.5) or according to truck-driving as the "longest-held" work (COR 54.0; 95% CL: 1.6-2,997.5). The possible modifying effect on MBC risk in subjects carrying BRCA1/2 germ-line mutations of an occupation characterised by exposure to chemicals such as polycyclic aromatic hydrocarbons (PAH) that are capable of inducing DNA damage, may provide clues to the role of environmental exposures in modifying BC risk in mutation carriers in both genders. © 2004 Elsevier Ltd. All rights reserved

Contribution of MUTYH variants to male breast cancer risk: results from a multicenter study in Italy

Inherited mutations in BRCA1, and, mainly, BRCA2 genes are associated with increased risk of male breast cancer (MBC). Mutations in PALB2 and CHEK2 genes may also increase MBC risk. Overall, these genes are functionally linked to DNA repair pathways, highlighting the central role of genome maintenance in MBC genetic predisposition. MUTYH is a DNA repair gene whose biallelic germline variants cause MUTYH-associated polyposis (MAP) syndrome. Monoallelic MUTYH variants have been reported in families with both colorectal and breast cancer and there is some evidence on increased breast cancer risk in women with monoallelic variants. In this study, we aimed to investigate whether MUTYH germline variants may contribute to MBC susceptibility. To this aim, we screened the entire coding region of MUTYH in 503 BRCA1/2 mutation negative MBC cases by multigene panel analysis. Moreover, we genotyped selected variants, including p.Tyr179Cys, p.Gly396Asp, p.Arg245His, p.Gly264Trpfs*7, and p.Gln338His, in a total of 560 MBC cases and 1,540 male controls. Biallelic MUTYH pathogenic variants (p.Tyr179Cys/p.Arg241Trp) were identified in one MBC patient with phenotypic manifestation of adenomatous polyposis. Monoallelic pathogenic variants were identified in 14 (2.5%) MBC patients, in particular, p.Tyr179Cys was detected in seven cases, p.Gly396Asp in five cases, p.Arg245His and p.Gly264Trpfs*7 in one case each. The majority of MBC cases with MUTYH pathogenic variants had family history of cancer including breast, colorectal, and gastric cancers. In the case-control study, an association between the variant p.Tyr179Cys and increased MBC risk emerged by multivariate analysis [odds ratio (OR) = 4.54; 95% confidence interval (CI): 1.17-17.58; p = 0.028]. Overall, our study suggests that MUTYH pathogenic variants may have a role in MBC and, in particular, the p.Tyr179Cys variant may be a low/moderate penetrance risk allele for MBC. Moreover, our results suggest that MBC may be part of the tumor spectrum associated with MAP syndrome, with implication in the clinical management of patients and their relatives. Large-scale collaborative studies are needed to validate these findings

Melatonin and pro-hypnotic effectiveness of the antidepressant Trazodone: A preliminary evaluation in insomniac mood-disorder patients

Objective To preliminary investigate the link between the darkness hormone melatonin (MLT) and the pro-hypnotic effectiveness of the atypical antidepressant Trazodone (TRZ) in a group of mood disorder patients suffering of insomnia. Design and methods The study's design comprised: i) the enrolment of insomniac outpatients, ii) baseline (t0) psychiatric and biochemical examinations; iii) the subsequent patients' introduction into a treatment with TRZ for 3–4 weeks, followed by post-therapy re-evaluations (t1). The MLT function was investigated by t0/t1 ELISA determinations of 6-hydroxy-MLT sulfate (6-OH-MLTs) levels in early-morning urines and HPLC analysis of morning MLT serum amount. Concomitantly, TRZ and its metabolite m-chloro-phenylpiperazine (m-CPP) were measured by HPLC in serum to monitor patients' compliance/metabolism. Results Seventeen insomniac outpatients, displaying mild symptoms of depression/anxiety resistant to antidepressants, completed TRZ therapy (dose:10–20 mg/day, bedtime). Serum TRZ levels (127 ± 57 ng ml− 1, mean ± SD) confirmed patients' compliance, while the anxiogenic metabolite m-CPP resulting almost undetectable. Moreover, the 6-OH-MLTs output was found increased at t1 vs. baseline values (t1: 58.4 ± 45.02 ng ml− 1; t0: 28.6 ± 15.8 ng ml− 1; mean ± SD, P < 0.05) in 9 patients who recovered both insomnia and depression/anxiety (P < 0.01). Unresponsive subjects showed instead no post-therapy 6-OH-MLTs variation (t1: 48.53 ± 50.70 ng ml− 1; t0: 49.80 ± 66.53 ng ml− 1). Morning MLT in serum slightly diminished at t1 without reaching the statistical significance, not allowing therefore to define the patients' outcome. Conclusions This initial investigation encourages to explore MLT networks as possible correlates of TRZ pro-hypnotic responses