Potential Effects of Environmental and Occupational Exposure to Microplastics: An Overview of Air Contamination.

Microplastics (MPs) are now ubiquitous environmental contaminants that lead to unavoidable human exposure; they have received increasing attention in recent years and have become an emerging area of research. The greatest concern is the negative impacts of MPs on marine, fresh-water, and terrestrial ecosystems, as well as human health, to the extent that the World Health Organization (WHO) calls for increased research and standardized methods to assess exposure to MPs. Many countries and international organizations are implementing or proposing legislation in this regard. This review aims to summarize the current state of legislation, indoor and outdoor contamination, and potential human health risk due to exposure to airborne MPs, considering that occupational exposure to MPs is also becoming a growing area of concern. Even though research regarding MPs has continuously increased in the last twenty years, the effects of MPs on human health have been scarcely investigated, and toxicity studies are still limited and not directly comparable, due to the lack of standardized studies in this field.

MicroRNAs diagnostic and prognostic value as predictive markers for malignant mesothelioma.

Malignant mesothelioma is an aggressive tumor resistant to current therapies with a latency period ranging between 20 and 60 years, caused by inhalation of asbestos fibers, that continues to represent a social and healthcare issue. The high percentage of people exposed to asbestos for professional or environmental reasons is associated with the high biopersistence of its fibers and with its widespread use in the last century. Approximately 20-40% of men report an occupational history that might have caused the workplace exposure (criteria Helsinki, 1997). Some authors are evaluating the possible use of bioindicators as a screening and early diagnosis tool. In this regard, the use of microRNAs has been proposed as powerful diagnostic and prognostic biomarkers for many tumors and human diseases. This review focuses on the current state of knowledge on the key role of microRNAs expression as new malignant mesothelioma biomarkers, in early clinical diagnostic applications.

Mobility of trace metals in serpentinite-derived soils of the Pollino Massif (Southern Italy): insights on bioavailability and toxicity.

This paper deals with the evaluation of geo- and bioavailability of trace elements, including heavy metals, present in a typical serpentinite-derived soil of the Pollino Massif (Southern Italy). Precisely, the research was aimed to (1) assess processes and factors controlling the mobility of metals in the soil in order to identify the elements "potentially harmful" for the human health, (2) estimate possible metals accumulation in horticultural crops, and (3) evaluate phyto- and genotoxicity of Cr naturally present in soils and water. The studied profile has a homogeneous mineralogical composition consisting of metal-rich phases as prevailing minerals. The three-step sequential extraction was performed on soil samples from which four fractions (exchangeable, reducible, oxidizable, and residual) were obtained and analysed by ICP-MS. The lowest contents of metals are in the exchangeable and oxidizable fractions suggesting a metal contamination is unlikely for the studied soil. Conversely, the residual and reducible fractions are the more metal enriched fractions. Among heavy metals, mainly Pb and Cd and subordinately Ni, Cr, Cu, and Zn are associated with Fe- and Mn-oxi/hydroxides that, under acidic conditions, may release the adsorbed metals in circulating water. The high contents of heavy metals, including Cr and Ni, of analysed vegetables suggest a metal transfer from soil to plant. However, no significant effects on Vicia faba seeds were observed by in vivo phyto- and genotoxicity tests. Conversely in vitro genotoxicity tests, performed on HepG2 human cell line, showed that DNA damage and cytotoxic effect depending on Cr concentration may occur.

Molecular and structural changes induced by essential oil treatments in Vicia faba roots detected by FTIR and FTNIR spectroscopy.

Essential oils have recognized antimicrobial and antifungal properties which allow their utilization in agriculture like an alternative to pesticides, but their utilization requires the knowledge of all the potential structural changes and damages produced by the interaction with the vegetal organisms. In this paper, we investigated the effects of two essential oils, the tea tree oil (TTO) and the mixture of clove and rosemary oils (C + R), on the molecular structure of Vicia faba roots by Fourier transform infrared (FTIR) and Fourier near infrared transform (FTNIR) spectroscopy. FTIR spectroscopy showed structural modifications of the absorption bands related to DNA (1100 and 1050 cm-1, carbohydrate backbones, and nucleotide bands within 900 and 850 cm-1), proteins (1700 and 1600 cm-1 amide I band, 1580 and 1520 cm-1 amide II band), and lipids (methylene group of aliphatic chains between 2950 and 2800 cm-1). The changes in the secondary structures of proteins consisted of a denaturation depending on increased presence of random coil structures. In addition, in the samples treated with TTO oils, we observed the presence of protein oxidation, an effect negligible instead for the C + R-treated samples. The modified shapes of the infrared methyl bands of aliphatic chains suggested an increased lipid disorder which could also determine lipid peroxidation. The changes observed for the DNA structures at the highest concentration of the above essential oils can be related to the genotoxic effect of eugenol, an important constituent of both TTO and C + R mixture oils. FTNIR spectroscopy showed the modified shape of the second overtone bands belonging to methyl and methylene groups, between 8500 and 8000 cm-1. This confirmed the increased lipid disorders already observed by FTIR spectroscopy. The results obtained on the probe organism V. faba show that FTIR and FTNIR spectroscopy can become a useful support to the conventional cytogenetic tests used in the evaluation of the allelopathic uses of essential oils in agriculture.

Molecular and structural changes induced by essential oils treatments in Vicia faba roots detected by genotoxicity testing.

Over the last few years, there has been an increased interest in exploiting allelopathy in organic agriculture. The aim of this investigation was to examine the effects of essential oil mixtures in order to establish their allelopathic use in agriculture. Two mixtures of essential oils consisting respectively of tea tree oil (TTO) and clove plus rosemary (C + R) oils were tested. Phytotoxicity and genotoxicity tests on the root meristems of Vicia faba minor were performed. A phytotoxic influence was particularly relevant for C + R mixture, while genotoxicity tests revealed significant results with both C + R oil mixture and TTO. Phenotypic analysis on Vicia faba minor primary roots following C + R oil mixture treatment resulted in callose production, an early symptom attributed to lipid peroxidation. The approach described in this study, based on genotoxicity bioassays, might identify specific DNA damage induced by essential oil treatments. These tests may represent a powerful method to evaluate potential adverse effects of different mixtures of essential oils that might be useful in alternative agriculture. Future studies are focusing on the positive synergism of more complex mixtures of essential oils in order to reduce concentrations of potentially toxic components while at the same time maintaining efficacy in antimicrobial and antifungal management.

Effects of essential oil treatments on the secondary protein structure of Vicia faba: a mid-infrared spectroscopic study supported by two-dimensional correlation analysis.

In this study we investigated the effects of essential oil treatments on the secondary protein structure of the Vicia faba roots, a bioindicator plant, in order to obtain information for the potential allelopathic uses of these oils as alternative to the use of pesticides in agriculture. We tested two mixtures of essential oils consisting of Tween 20-emulsions of tea tree oil (TTO) and Tween 20-emulsion of Clove and Rosemary (GARROM) essential oils respectively at three different oil concentrations each. The molecular modifications caused in Vicia faba by exposure to oil emulsions were investigated by FTIR spectroscopy in diffuse reflectance (DRIFT) mode. We considered the specific Amide I, Amide II and Amide VI bands by ordinary and second derivative spectroscopy and the results showed that both Tween 20-emulsion of GARROM and Tween 20-emulsion of TTO oils cause transitions among the secondary (α-helix, ß-sheet and ß-turn) structures with in addition the appearance of random coil structures in exposed samples. The Amide VI bands, placed between 500 and 600 cm(-1), confirmed the structural transitions observed for the Amide I bands. In addition we observed the presence of a protein oxidation effect for TTO treated samples, oxidation which resulted negligible instead for the GARROM oil samples. At last, FTIR spectra were also submitted to two-dimensional correlation analysis (2DCORR) and double two-dimensional correlation analysis (D2DCORR); the results confirmed the different effects caused by the two typologies of essential oils on the secondary protein structures of Vicia faba roots.

Arsenic exposure triggers a shift in microRNA expression.

Exposure to inorganic Arsenic (iAs) through drinking water is a major public health problem affecting most countries. iAs has been classified by the International Agency for Research on Cancer as Group 1: "Carcinogenic to humans". Although numerous studies have shown the related adverse effects of iAs, sensitive appropriate biomarkers for studies of environmental epidemiology are still required. The present work aims at investigate the role of microRNAs (miRNAs), powerful negative regulators of gene expression, playing a key role in many physiological and pathological cellular processes, in iAs exposure. To this end, we analyzed miRNA changes in expression profile triggered by iAs exposure in Jurkat cell line. We used microarray technology to profile the expression of miRNAs following 2 µmol/L sodium arsenite treatment at different time points. Moreover, we performed phenotypic analysis of iAs treated cells. Real Time Polymerase Chain Reaction (RT-PCR) was used to validate miRNA microarray data and to assay expression modulation of selected relevant mRNAs. Finally, bioinformatics techniques were applied to reconstruct iAs-relevant molecular pathways and miRNA regulatory networks from the expression data. We report miRNAs modulated after iAs treatment in Jurkat cells. In particular, we highlight 36 miRNAs exhibiting consistent dysregulation and particularly a panel of 8 miRNAs which we also validated by RT-PCR analysis. Computational analysis of lists of putative target genes for these 8 miRNAs points to an involvement in arsenic-response pathways, for a subset of them, that were analyzed by RT-PCR. Furthermore, iAs exposure reveals induction of cell cycle progression and the failure of apoptosis, supporting the idea of iAs carcinogenic activity. Our study provides a list of miRNAs whose expression levels are affected by iAs treatment, corroborating the importance of proceeding with the hunt for specific subset of miRNAs, which can serve as potential biomarkers of iAs effects with useful diagnostic value.

Use of arsenic contaminated irrigation water for lettuce cropping: effects on soil, groundwater, and vegetal.

The present study investigated the effects of using arsenic (As) contaminated irrigation water in Lactuca sativa L. cropping. Two different arsenic concentrations, i.e., 25 and 85 µg L(-1) and two different soils, i.e., sandy and clay loam, were taken into account. We determined the arsenic mobility in the different soil fractions, its amount in groundwater, and the phytotoxicity and genotoxicity. Nuclear magnetic resonance (NMR) and inductively coupled plasma (ICP) were used to assess the lettuce metabolic profile changes and the arsenic uptake by the plant, respectively, as a function of the various conditions studied, i.e., As content and type of soil. Data indicated that at both concentrations in sandy soil, arsenic is in part quickly leached and thus present in groundwater and in part absorbed by the vegetable, being therefore readily available for assimilation by consumption. NMR results reported a large modification of the metabolic pattern, which was depending on the pollutant amount. In clay loam soil, the groundwater had a low As content with respect to sandy soil, and NMR and ICP performed on the lettuce did not reveal severe changes related to As, most likely because the metalloid is bound to the colloidal fraction.

[microRNAs as biological indicators of environmental and occupational exposure to asbestos]. / I microRNA come indicatori biologici di esposizione ambientale e occupazionale ad amianto.

Malignant mesothelioma is an aggressive cancer refractory to current therapies, the incidence of which is expected to rise in the next decades. Exposure to asbestos is a well known risk factor, as InternationalAgency for Research and Cancer (IARC) classified this compound as group I (carcinogenic to humans). The lack of tumor biomarkers for diagnosis and medical survey plays a fundamental role for the development of a universally accepted therapeutic approach. In this review we evaluated the mechanism of asbestos carcinogenesis by analyzing activated oncogenes, genetic predisposition, and SV40 infection as cofactors. Therefore, interest has focused on microRNAs, 19-25 nucleotide-long single-stranded RNAs that negatively regulate gene expression by modulating translational efficiency of target genes involved in numerous cellular processes including development, differentiation, proliferation, apoptosis, and stress response. The analysis revealed a differential expression of miRNAs between mesothelioma and mesothelial cells, suggesting their potential role as oncogenes or tumor suppressor genes in mesothelioma oncogenesis. We have also investigated the role of polymorphism in the etiology and pathogenesis of mesothelioma, in order to evaluate the association between disease linked to asbestos exposure andgenetic variability. The identification of dysregulated miRNAs or frequent genetic polymorphisms as potential diagnostic biomarkers or as prognostic factors for malignant mesothelioma could facilitate the surveillance procedure of subjects exposed to asbestos.

Development of a multiresidue method for analyzing herbicide and fungicide residues in bovine milk based on solid-phase extraction and liquid chromatography-tandem mass spectrometry.

This report describes a liquid chromatography (LC)-tandem mass spectrometry (MS) multiresidue method for determining traces of 30 base/neutral/acid herbicides and fungicides in bovine whole milk. Four milliliters of milk was spiked with the analytes and two surrogate analytes (SAs) and then diluted with 35 mL of a water/methanol solution (50:50, v/v). This mixture was passed through a 0.5-g Carbograph 4 cartridge. After washings, analytes were re-extracted by back-flushing the cartridge with 1.5 mL of methanol followed by 6 mL of methylene chloride/methanol (80:20, v/v), 50 mmol/L formic acid. After partial solvent removal down to about 0.1, 0.15 mL of 1 mmol/L formic acid aqueous solution and an internal standard (IS) were added. After filtration, 50 microL of the final extract was then introduced into the LC analytical column. During the chromatographic run, the MS system was operated in both positive and negative ion modes. MS data acquisition was performed in the multi-reaction monitoring mode, selecting two precursor ion>product ion transitions for each target compound, except for pentachlorophenol. On analyzing six milk samples from different sources, absolute recovery of the analytes and the two SAs ranged between 78% and 104% with RSDs not larger than 13%. The accuracy of the method at three different spike levels was assessed by adding the two SAs to analyte-containing milk samples and varied between 82% and 120% with RSDs not larger than 11%. Limits of quantification were estimated to range between 0.008 and 1.4 microg/L. Compared to the Carbograph 4 cartridge, one filled with a N-vinylpyrrolidone-m-divinylbenzene co-polymer (Oasis HLB) sorbent was much less efficient in recovering several of the acidic herbicides considered and, in addition, its relative final extract produced a severe negative matrix effect that drastically weakened ion signal intensities of several non acidic analytes.

Deoxyribonucleic acid (DNA) biosensors for environmental risk assessment and drug studies.

In the present work, electrochemical DNA biosensors are proposed as a screening device for the rapid bio-analysis of environmental pollution and DNA-drug interaction studies. The binding of small molecules to DNA immobilised on disposable screen-printed electrodes has been measured through the variation of the electrochemical signal of guanine by square wave voltammetric scans. These kinds of biosensors were used to evaluate the soil contamination level in an Italian polluted area and the results were compared with several methods for the DNA damage detection, as Comet genotoxicity effects, aberrant anatelophases and micronucleated cells frequency on plant roots, and with fixed wavelength fluorescence (FF) by using 2-aminoanthracene as standard compound. The results showed the ability of the biosensors to distinguish in 11 min low, medium and high contaminated soils with good correlation with well established techniques as well as FF, Comet and genotoxicity tests. The same kind of biosensors was also used to evaluate the interaction of DNA with some anti-proliferative metallo drugs, and the electrochemical responses reflected the kind of interaction. The reproducibility of the electrochemical measurements of DNA guanine peak was estimated as less than 10% of relative standard deviation (R.S.D.%).