The Substitution Principle within the REACH Regulation: Nuclear Receptor-Bound Endocrine Disruptors

Within the REACH Regulation (EC/1907/2006), the substitution principle for chemicals classified as Substances of Very High Concern (SVHC) for either human health or environmental risks has been implemented in order to support their replacement by suitable alternatives. Considering the thousands of chemicals to be tested within the frame of REACH, animal testing by internationally-accepted guidelines sounds unreasonable in terms of the required time, costs as well ethical issues. Hence, REACH recommended also the use of alternative methods to animal experimentation although no validated in silico or in vitro tools were available when regulation entried into force. To search for suitable alternatives to SVHC having an Endocrine Disruptor (ED)-like Mode-of-Action (MoA) by means of an integrated, tiered in silico-in vitro approach, the EU-granted project LIFE-EDESIA (contract no. LIFE12 ENV/IT/000633) is combining computational-based tools and cell-based bioassays, in order to develop a no-animal testing procedure to screen for chemicals having less or no toxicity in terms of endocrine disruption-like activities. A general view of the no-animal testing approach implementing REACH and the substitution principle will be given, emphasising ligand-nuclear receptor (NR) assessment by molecular docking (one of the LIFE-EDESIA in silico approaches) and the use of clinical biomarkers in in vitro toxicology to detect ED-like adverse effects in cell-based bioassays

Mixtures of Commercial Lentil Cultivars Show Inconsistent Results on Agronomic Parameters but Positive Effects on Yield Stability

Cultivar mixtures are a useful tool to enhance cultivated biodiversity to buffer crop biotic and abiotic stresses. There are multiple pieces of evidence of mixture advantages in terms of pathogen control and increase in yield amount, stability and quality. Lentil represents a founder crop in the Mediterranean, yet it experiences strong yield fluctuations in the face of abiotic stresses. The present study aims to assess the mixing ability of four Italian commercial lentil lines in terms of yield amount and stability, nodule number, total lentil biomass and sensitivity to weeds. Since there is very limited information on lentil genotype traits, two-, three- and four-cultivar mixtures were designed with a trait-blind approach and compared to sole cultivars. The nodule number was mainly influenced by cultivar and weather; no interaction between cultivars was observed. Treatments were differently sensitive to weeds, but the effect of spatial heterogeneity prevailed over that of the cultivar. The average yield stability of all mixtures was significantly higher than pure stands, but in terms of yield amount, individual mixtures either outperformed or were outperformed by pure stands. Against our expectations, cultivar mixtures showed the most advantages in the most productive year: likely, the reason lies in the supposed low genetic diversity of commercial lentil lines in Italy. We encourage further research, taking into account the diversity of Italian lentil landraces, in order to gain a broader genetic base for the implementation of a trait-based approach, which may lead to better-performing mixtures

Antioxidative Molecules in Human Milk and Environmental Contaminants

Breastfeeding provides overall beneficial health to the mother-child dyad and is universally recognized as the preferred feeding mode for infants up to 6-months and beyond. Human milk provides immuno-protection and supplies nutrients and bioactive compounds whose concentrations vary with lactation stage. Environmental and dietary factors potentially lead to excessive chemical exposure in critical windows of development such as neonatal life, including lactation. This review discusses current knowledge on these environmental and dietary contaminants and summarizes the known effects of these chemicals in human milk, taking into account the protective presence of antioxidative molecules. Particular attention is given to short- and long-term effects of these contaminants, considering their role as endocrine disruptors and potential epigenetic modulators. Finally, we identify knowledge gaps and indicate potential future research directions

The Role of Human Semen as an Early and Reliable Tool of Environmental Impact Assessment on Human Health

Several studies have shown a dramatic reduction of semen quality in many industrialized countries and infertility is becoming a public health top priority, whose incidence is associated to late-onset adult diseases, especially cancer, shorter life expectancy and trans-generational effects. The male reproductive system is particularly sensitive to a broad variety of reproductive and developmental toxicants, including many environmental pollutants and recent studies suggest that human semen is an early and sensitive environmental and health marker. A set of semen biomarkers is described for reproductive health effects in relation to environmental exposure, where human semen seems to be an early and sensitive source of biomarkers than blood to monitor high environmental pressure on human health. Environmental health should consider reproductive health and development, from intrauterine life to childhood and puberty: these are both vulnerable targets and high-value protection goals, inasmuch as they represent the future of our societies. Hence, biomarkers of reproductive health should be exploited as early signals of environmental pressure and increased risk of adverse chronic health effects so that the use of “human seminal model” might be the main objective to be considered in the agenda of public prevention policies for early detection and innovative programs of health surveillance in environmental risk areas

Evaluation of potsherds features using hyperspectral maps generated by μ-LIBS scanner

The micro-laser induced breakdown spectroscopy (µ-LIBS) technique allows performing fast elemental analyses, without sample preparation and thus making it specifically useful in the analysis of the composition of ancient potsherd. The µ-LIBS instrument is equipped with a microscope and a scanning system allowing to realize small craters (about Ø = 25 µm) in order to obtain detailed hyperspectral surfaces maps (up to a maximum size of one square centimeter). The data are processed by Self-Organizing Maps (SOMs) method to visualize in 2D representations allowing significant information on the technological features of ceramic samples.The micro-laser induced breakdown spectroscopy (µ-LIBS) technique allows performing fast elemental analyses, without sample preparation and thus making it specifically useful in the analysis of the composition of ancient potsherd. The µ-LIBS instrument is equipped with a microscope and a scanning system allowing to realize small craters (about Ø = 25 µm) in order to obtain detailed hyperspectral surfaces maps (up to a maximum size of one square centimeter). The data are processed by Self-Organizing Maps (SOMs) method to visualize in 2D representations allowing significant information on the technological features of ceramic samples

Biomarkers of effect in endocrine disruption: how to link a functional assay to an adverse outcome pathway

The development of in vitro testing strategies may achieve a cost-effective generation of  comprehensive datasets on a large number of chemicals, according to the requirements of the European Regulation REACH. Much emphasis is placed on in vitro methods based on subcellular mechanisms (e.g., nuclear receptor interaction), but it is necessary to define the predictive value of molecular or biochemical changes within an adverse outcome pathway (AOP). AOP pivots on the description of the flow from a molecular initiating event through a cascade of intermediate events needed to produce a specific adverse effect at organism level: downstream esponses at cell level are, therefore, essential to define an AOP. Several in vitro assays are based on human cell lines representative of endocrine-targeted tissues (e.g., prostate) and on functional biomarkers of clinical relevance (e.g., PSA secretion in human prostate epithelial cells). We discuss the implementation of such functional biomarkers in the AOP contex

Mineralogical, petrographic and physical-mechanical study of Roman construction materials from the Maritime Theatre of Hadrian's Villa (Rome, Italy)

This paper presents the study of various Roman materials used in the construction of the Maritime Theatre, one of the main buildings in the Hadrian’s Villa complex, a designated UNESCO World Heritage Site located in Tivoli (Rome, Italy), dating to the first half of the II century A.D. The plaster layers (arriccio and intonachino) and overlying original Roman paintings that form the concave wall of the portico as well as some bedding mortars of the pyramidal stone elements (i.e. cubilia) of the circular masonry have been studied in particular. In addition, the acid volcanic rocks of the cubilia have been investigated, aiming to understand their state of alteration and geological origin. By mineralogical-petrographic microscopy (OM), diffractometry (XRPD), Raman spectroscopy, Point Load Tests (PLT), helium pycnometry, and particle size analysis, the composition and granulometric distribution of the aggregate, type and characteristics of the binder, and various physical-mechanical properties (density, porosity, water absorption, imbibition and saturation indices, mechanical resistance) of mortars and stones were defined. In addition, through digital image analysis of thin sections, the binder/aggregate ratio and some geometric characteristics of the aggregates (e.g. circularity) were determined. The research aims to improve the knowledge of the constructive technologies of the Maritime Theatre through the analysis of its materials

Micro-chemical evaluation of ancient potsherds by μ-LIBS scanning on thin section negatives

In the study of ancient pottery, thin section analysis represents the basic approach to study mineralogical and petrografic features in order to obtain preliminary information about the production technology and origin of archaeological ceramics. However, even if thin section analysis allows investigating the textural and structural characteristics of potteries, peculiar features related to clay paste and temper composition, as well as provenance issues, can be detailed addressed only by quantitative mineralogical and chemical studies. In the realization of thin sections, a negative face is always produced, similar to the thin section itself; these remains can be used for additional analyses, such as high spatial resolution micro-chemical studies using, for example, a micro-laser induced breakdown spectroscopy (LIBS) scanner. LIBS is a spectroscopic technique that, exploiting the laser radiation, is able to bring into the plasma state micrometric portions of the sample, and to analyse its content through the study of the optical emission of the plasma itself. Unlike other techniques, LIBS can detect and quantify also light elements such as aluminium and magnesium. Images produced by the micro-LIBS instrument show the spatial distribution of the chemical elements within a portion of the sample, which may have dimensions from a few hundred microns up to several centimeters. The combination of these images with algorithms derived from image processing techniques may return interesting information and supporting data to in-depth investigate pottery components detected by optical microscopy observations. In this work, we present the results of an experimental study performed on thin-section negatives with different grain size, surface treatments and aggregates, coming from some Neolithic Italian sites, exploring the potential of the LIBS method in micro-chemical studies of ancient potsherds

Multivariate calibration in Laser-Induced Breakdown Spectroscopy quantitative analysis: The dangers of a ‘black box’ approach and how to avoid them

The introduction of multivariate calibration curve approach in Laser-Induced Breakdown Spectroscopy (LIBS) quantitative analysis has led to a general improvement of the LIBS analytical performances, since a multivariate approach allows to exploit the redundancy of elemental information that are typically present in a LIBS spectrum. Software packages implementing multivariate methods are available in the most diffused commercial and open source analytical programs; in most of the cases, the multivariate algorithms are robust against noise and operate in unsupervised mode. The reverse of the coin of the availability and ease of use of such packages is the (perceived) difficulty in assessing the reliability of the results obtained which often leads to the consideration of the multivariate algorithms as ‘black boxes’ whose inner mechanism is supposed to remain hidden to the user. In this paper, we will discuss the dangers of a ‘black box’ approach in LIBS multivariate analysis, and will discuss how to overcome them using the chemical-physical knowledge that is at the base of any LIBS quantitative analysis

Captação e aproveitamento de água das chuvas: atitude sustentável em uma indústria de artefatos de concreto

Atualmente, diversas organizações encontram-se empenhadas em ganhar mais espaço num mundo que valoriza cada vez mais atitudes sustentáveis. Dessa forma, o problema de pesquisa deste trabalho envolveu: quais seriam os possíveis benefícios econômicos e financeiros obtidos com a implantação de um sistema de utilização da água da chuva em uma indústria de artefatos de concreto? O objetivo principal foi analisar a administração e o consumo de água, comparando o consumo anterior e posterior a 2008 – ano do início da implantação do sistema de captação e aproveitamento de água das chuvas – e também o prazo para compensação dos investimentos realizados pela indústria. E, ainda, buscou-se demonstrar os benefícios alcançados pela organização, nos quesitos financeiros, ambiental e social, perante seus colaboradores e para a sociedade. O estudo de caso apresenta uma abordagem qualitativa e descritiva das práticas organizacionais e ações sustentáveis e análise SWOT. Com a implantação do sistema, tanto os funcionários quanto a administração da empresa contribuem para a utilização sustentável da água da chuva. Os ganhos não são apenas financeiros, pois implicam atitudes responsáveis ambiental e socialmente