Plastic and environmental safety: the effects of EDCs on metabolism, reproduction and epigenetic processes

Per inquinamento causato dalle materie plastiche si intende l’accumulo in ambiente di prodotti plastici in grado di indurre problemi sia all’ambiente che alle specie selvatiche. Il Bisfenolo A (BPA) è uno dei distruttori endocrini maggiormente prodotti a livello mondiale durante la lavorazione della plastica. Numerosi studi hanno mostrato la capacità di questo inquinante di creare effetti dannosi sia sull’uomo che sulle specie selvatiche, per cui nel tempo sono stati compiuti diversi tentativi volti a trovare delle valide alternative. Tra i possibili sostituti, in questo studio di dottorato, l’attenzione è stata focalizzata sul Dhietylene dibenzoato (DGB) ed il Diisononilftalato (DiNP). Lo scopo principale di questo progetto è stato quindi quello di studiare l’impatto di questi plastificanti, focalizzando l’attenzione sui loro effetti nella riproduzione e nel metabolismo lipidico. Gli effetti del nonilfenolo, dell’ottilfenolo (t-OP) e del BPA sul metabolismo lipidico, sono stati inoltre analizzati in giovanili di orata. I risultati ottenuti hanno dimostrato come, sia il BPA, che il DiNP, siano in grado di interferire con il processo di oogenesi e che, sia il BPA che il DGB, sono in grado di interferire con il metabolismo lipidico nello zebrafish. La somministrazione a giovanili di orata di mangimi contaminati con diversi inquinanti tra cui il BPA, ha mostrato la loro capacità di indurre disordini metabolici, mostrando come oltre all’esposizione ambientale, anche il consumo di cibi contaminati possa essere considerata una fonte di esposizione importante ai contaminanti. Concludendo, il presente progetto di dottorato, mostra la capacità di alcuni distruttori endocrini, utilizzati nella lavorazione della plastica e nei detergenti, di interferire con la riproduzione ed il metabolismo lipidico nei teleostei, agendo prevalentemente in maniera dose dipendente, secondo curve non-monotoniche e, nel caso del BPA, interferendo con i meccanismi epigenetici.Plastic pollution involves the accumulation of plastic products in the environment that adversely affects wildlife and wildlife habitat. Bisphenol A (BPA) is one of the highest volume endocrine disruptor produced worldwide during the manufacturing of plastic. Due to his well-documented detrimental effects, several substitute for BPA, but also for other plasticizer of common use, have been suggested, such as Diethylene glycol dibenzoate (DGB) and Diisononyl Phthalate (DiNP). The aim of this project is therefore to investigate the impact of several plasticizers, which are supposed to act as endocrine disruptor compounds (EDCs), focusing the attention on their effects on metabolic and reproductive system. Due to the necessity of testing a different range of concentrations in various plastic pollutants, an experimental model easy to reproduce, treat and analyse was needed; thus the choice to introduce zebrafish as experimental model for these studies. Concomitantly, the effects of nonylpnenol (NP), 4-tert-octylphenol (t-OP) and BPA on lipid metabolism were studied in sea bream juveniles. The results we obtained, using a multidisciplinary approach ranging from molecular to spectroscopic techniques, showed the ability of BPA and DiNP to interfere with female’s reproduction in a dose dependent manner. Concerning BPA its epigenetic effects were also demonstrated and eventually both BPA and DGB were found able to interfere with lipid metabolism in a dose dependent manner. Furthermore t-OP, NP and BPA were demonstrated to give rise to hepatic metabolic disorders in sea bream juveniles. In conclusion, the present PhD project, demonstrates that endocrine disruptors compounds, used in the manufacturing of plastics, are able to interfere with reproductive and metabolic system of teleost fish. Furthermore, we demonstrates the capacity of BPA to affect gene expression through the deregulation of epigenetic patterns

BPA-Induced Deregulation of Epigenetic Patterns: Effects on Female Zebrafish Reproduction

Bisphenol A (BPA) is one of the commonest Endocrine Disruptor Compounds worldwide. It interferes with vertebrate reproduction, possibly by inducing deregulation of epigenetic mechanisms. To determine its effects on female reproductive physiology and investigate whether changes in the expression levels of genes related to reproduction are caused by histone modifications, BPA concentrations consistent with environmental exposure were administered to zebrafish for three weeks. Effects on oocyte growth and maturation, autophagy and apoptosis processes, histone modifications, and DNA methylation were assessed by Real-Time PCR (qPCR), histology, and chromatin immunoprecipitation combined with qPCR analysis (ChIP-qPCR). The results showed that 5 μg/L BPA down-regulated oocyte maturation-promoting signals, likely through changes in the chromatin structure mediated by histone modifications, and promoted apoptosis in mature follicles. These data indicate that the negative effects of BPA on the female reproductive system may be due to its upstream ability to deregulate epigenetic mechanism

Modulation of LDL receptor expression and promoter methylation in HepG2 cells treated with a Corylus avellana L. extract

Abstract The aim of our study was to evaluate the impact of an ethanolic extract of C. avellana on the molecular pathway(s) regulating the low-density lipoprotein receptor (LDLR) in HepG2 cells, mainly in terms of epigenetics. We demonstrated that viability, proliferation and cell cycle distribution were not affected up to 72 h of treatment, whereas LDLR expression was stimulated as early as 24 h following administration (

Biomarkers of myocardial injury with different energy sources for atrial fibrillation catheter ablation

Background: Our study aims to compare acute myocardial injury biomarker rise after atrial fibrillation ablation performed with different technologies.Methods and Results: One hundred and ten patients were treated with pulmonary vein isolation with 4 different technologies: open-irrigated tip radiofrequency (RF) catheter in35 patients (Group A), cryoballoon in 35 patients (Group B), visually guided laser balloon in 20 patients (Group C), open-irrigated tip RF catheter with contact-force-sensing technology in 20 patients (Group D). Post-procedure samples of cardiac troponin I (cTnI) and creatinine kinase-MB (CK-MB) were collected at 19 ± 3 h and 43 ± 3 h after ablation. At the first postprocedural sample, cTnI and CK-MB levels were found elevated in all 110 patients with a median value of 2.11 ng/mL and 8.95 ng/mL, respectively. Group B showed cTnI levels increased (median 5.96 ng/mL) compared to other groups (median Group A: 1.72 ng/mL, Group C: 1.54 ng/mL, Group D: 2.0 ng/mL; p < 0.001). Also CK-MB levels resulted higher in cryoablation (median 26.4 ng/mL) compared to other groups (median Group A: 6.40 ng/mL, Group C: 7.15 ng/mL, Group D: 6.50 ng/mL; p < 0.001). No significant association was observed between biomarker levels and recurrences of atrial fibrillation after a mean follow-up of 369 ± 196 days.Conclusions: Highest markers for myocardial injury were observed in the cryoballoon group. It is possible that a longer delivery energy duration and other factors affecting lesion size resulted in higher amount of cardiac injury in cryoablation. The higher levels of cardiac biomarkers did not translate into a better outcome and its physiologic significance is unknown.

Effect of the probiotic Lactobacillus rhamnosus on the expression of genes involved in European eel spermatogenesis

[EN] Positive effects of probiotics on fish reproduction have been reported in several species. In the present study, 40 male European eels were weekly treated with recombinant hCG for 9 weeks and with three different concentrations (10(3), 10(5), and 10(6) CFU/mL) of probiotic Lactobacillus rhamnosus IMC 501 (Sinbyotec, Italy). The probiotics were daily added to the water from the sixth week of the hCG treatment. Males from the treated and control groups were sacrificed after 1, 2, and 3 weeks of probiotic treatment (seventh ninth weeks of hCG treatment); at this point, sperm and testis samples were also collected. Sperm volume was estimated, and motility was analyzed by computer-assisted sperm analysis software. Alternations in transcription of specific genes involved in reproductive process such as activin, androgen receptors alpha and beta (ar alpha and ar beta), progesterone receptor 1 (pr1), bone morphogenetic protein 15 (bmp15), and FSH receptor (fshr) were analyzed in the testis. After 2 weeks of probiotic treatment, sperm production and sperm motility parameters (percentage of motile cells and percentage of straight-swimming spermatozoa) were increased in the European eel treated with 105 CFU/mL compared to controls or to the other probiotic doses. These changes were associated with increases in messenger RNA expression of activin, ar alpha, ar beta, pr1, and fshr. Conversely, after 3 weeks, activin and pr1 expression decreased. No significant changes were observed on bmp15 expression throughout the duration of the treatment with 10(5) CFU/mL dose. The lowest and highest probiotic dose (10(3) and 10(6) CFU/mL, respectively) inhibited the transcription of all genes along all the experiment, except for ar alpha and ar beta after 1 week of probiotic treatment when compared to controls. The changes observed by transcriptomic analysis and the sperm parameters suggest that a treatment with L rhamnosus at 10(5) CFU/mL for 2 weeks could improve spermatogenesis process in Anguilla anguilla. (C) 2015 Elsevier Inc. All rights reserved.This study was funded by the European Community’s 7th FP (grant agreement no. 245257, PRO-EEL) and COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE) Victor Gallego and M. Carmen Vilchez have predoctoral grants from MINISTERIO DE ECONOMIA Y COMPETITIVIDAD (BES-2009-020310) and UNIVERSIDAD POLITECNICA DE VALENCIA PAID Program (2011-S2-02-6521), respectively. Fondo Ateneo 2012 to Oliana Carnevali.Vilchez Olivencia, MC.; Santangeli, S.; Maradonna, F.; Gioacchini, G.; Verdenelli, C.; Gallego Albiach, V.; Peñaranda, D.... (2015). Effect of the probiotic Lactobacillus rhamnosus on the expression of genes involved in European eel spermatogenesis. Theriogenology. 84(8):1321-1331. https://doi.org/10.1016/j.theriogenology.2015.07.011S1321133184

Plastic and environmental safety: the effects of EDCs on metabolism, reproduction and epigenetic processes

Per inquinamento causato dalle materie plastiche si intende l’accumulo in ambiente di prodotti plastici in grado di indurre problemi sia all’ambiente che alle specie selvatiche. Il Bisfenolo A (BPA) è uno dei distruttori endocrini maggiormente prodotti a livello mondiale durante la lavorazione della plastica. Numerosi studi hanno mostrato la capacità di questo inquinante di creare effetti dannosi sia sull’uomo che sulle specie selvatiche, per cui nel tempo sono stati compiuti diversi tentativi volti a trovare delle valide alternative. Tra i possibili sostituti, in questo studio di dottorato, l’attenzione è stata focalizzata sul Dhietylene dibenzoato (DGB) ed il Diisononilftalato (DiNP). Lo scopo principale di questo progetto è stato quindi quello di studiare l’impatto di questi plastificanti, focalizzando l’attenzione sui loro effetti nella riproduzione e nel metabolismo lipidico. Gli effetti del nonilfenolo, dell’ottilfenolo (t-OP) e del BPA sul metabolismo lipidico, sono stati inoltre analizzati in giovanili di orata. I risultati ottenuti hanno dimostrato come, sia il BPA, che il DiNP, siano in grado di interferire con il processo di oogenesi e che, sia il BPA che il DGB, sono in grado di interferire con il metabolismo lipidico nello zebrafish. La somministrazione a giovanili di orata di mangimi contaminati con diversi inquinanti tra cui il BPA, ha mostrato la loro capacità di indurre disordini metabolici, mostrando come oltre all’esposizione ambientale, anche il consumo di cibi contaminati possa essere considerata una fonte di esposizione importante ai contaminanti. Concludendo, il presente progetto di dottorato, mostra la capacità di alcuni distruttori endocrini, utilizzati nella lavorazione della plastica e nei detergenti, di interferire con la riproduzione ed il metabolismo lipidico nei teleostei, agendo prevalentemente in maniera dose dipendente, secondo curve non-monotoniche e, nel caso del BPA, interferendo con i meccanismi epigenetici.Plastic pollution involves the accumulation of plastic products in the environment that adversely affects wildlife and wildlife habitat. Bisphenol A (BPA) is one of the highest volume endocrine disruptor produced worldwide during the manufacturing of plastic. Due to his well-documented detrimental effects, several substitute for BPA, but also for other plasticizer of common use, have been suggested, such as Diethylene glycol dibenzoate (DGB) and Diisononyl Phthalate (DiNP). The aim of this project is therefore to investigate the impact of several plasticizers, which are supposed to act as endocrine disruptor compounds (EDCs), focusing the attention on their effects on metabolic and reproductive system. Due to the necessity of testing a different range of concentrations in various plastic pollutants, an experimental model easy to reproduce, treat and analyse was needed; thus the choice to introduce zebrafish as experimental model for these studies. Concomitantly, the effects of nonylpnenol (NP), 4-tert-octylphenol (t-OP) and BPA on lipid metabolism were studied in sea bream juveniles. The results we obtained, using a multidisciplinary approach ranging from molecular to spectroscopic techniques, showed the ability of BPA and DiNP to interfere with female’s reproduction in a dose dependent manner. Concerning BPA its epigenetic effects were also demonstrated and eventually both BPA and DGB were found able to interfere with lipid metabolism in a dose dependent manner. Furthermore t-OP, NP and BPA were demonstrated to give rise to hepatic metabolic disorders in sea bream juveniles. In conclusion, the present PhD project, demonstrates that endocrine disruptors compounds, used in the manufacturing of plastics, are able to interfere with reproductive and metabolic system of teleost fish. Furthermore, we demonstrates the capacity of BPA to affect gene expression through the deregulation of epigenetic patterns

Roma : tre contesti. 1. Via Nova-Clivo Palatino. 2. Crypta Balbi. 3. Via Sacra-Via Nova

Ciotola Alberto, Picciola Stefania, Santangeli Valenzani Riccardo, Volpe Rita. Roma : tre contesti. 1. Via Nova-Clivo Palatino. 2. Crypta Balbi. 3. Via Sacra-Via Nova. In: Amphores romaines et histoire économique. Dix ans de recherche. Actes du colloque de Sienne (22-24 mai 1986) Rome : École Française de Rome, 1989. pp. 604-609. (Publications de l'École française de Rome, 114

Endocrine-disrupting chemicals in aquatic environment: what are the risks for fish gametes?

Over the past 25 years, extensive research in vertebrate species has identified several genomic pathways altered by exposures to anthropogenic chemicals with hormone-like activity mediated by their interaction with nuclear receptors. In addition, many pollutants have been shown to interfere with non-genomic (non-classical) pathways, but this mechanism of endocrine disruption is still poorly understood. Recently, the number of publications describing the effects of Endocrine disrupting chemicals (EDCs) on fish reproduction, focusing on the deregulation of the hypothalamus-pituitary-gonadal axis as well as on gamete quality, significantly increased. Depending on their ability to mimic endogenous hormones, the may differently affect male or female reproductive physiology. Inhibition of gametogenesis, development of intersex gonads, alteration of the gonadosomatic index, and decreased fertility rate have been largely documented. In males, alterations of sperm density, motility, and fertility have been observed in several wild species. Similar detrimental effects were described in females, including negative outcomes on oocyte growth and maturation plus the occurrence of apoptotic/autophagic processes. These pathways may affect gamete viability considered as one of the major indicators of reproductive endocrine disruption. Pollutants act also at DNA level producing DNA mutations and changes in epigenetic pathways inducing specific mechanisms of toxicity and/or aberrant cellular responses that may affect subsequent generation(s) through the germline. In conclusion, this review summarizes the effects caused by EDC exposure on fish reproduction, focusing on gametogenesis, giving a general overview of the different aspects dealing with this issue, from morphological alteration, deregulation of steroidogenesis, hormonal synthesis, and occurrence of epigenetic process