COL10A1 expression distinguishes a subset of cancer-associated fibroblasts present in the stroma of high-risk basal cell carcinoma.

BACKGROUND: Basal cell carcinoma (BCC) is the most frequently diagnosed skin cancer and the most common malignancy in humans. Different morphological subtypes of BCC are associated with low- or high-risk of recurrence and aggressiveness, but the underlying biology of how the individual subtypes arise remains largely unknown. Because the majority of BCCs appear to arise from mutations in the same pathway, we hypothesized that BCC development, growth and invasive potential is also influenced by the tumor microenvironment and in particular by cancer-associated fibroblasts (CAFs) and their secreted factors. OBJECTIVE: We aimed to characterize the stroma of the different BCC subtypes with a focus on CAF populations. METHODS: To investigate the stromal features of the different BCC subtypes, we applied laser-capture microdissection (LCM) followed by RNA sequencing. A cohort of 15 BCC samples from 5 different "pure" subtypes (superficial, nodular, micronodular, sclerosing and basosquamous; n=3 each) were selected and included in the analysis. Healthy skin was used as a control (n=6). We confirmed the results by immunohistochemistry. We validated our findings in two independent, public single-cell RNA sequencing (scRNAseq) datasets and by RNAscope. RESULTS: The stroma of the different BCC subtypes have distinct gene expression signatures. Nodular and micronodular seem to have the most similar signatures, while superficial and sclerosing the most different. By comparing low- and high-risk BCC subtypes, we observed that Collagen 10A1 (COL10A1) is overexpressed in the stroma of sclerosing/infiltrative and basosquamous but not micronodular high-risk subtypes. Those findings were confirmed by immunohistochemistry in a cohort of 89 different BCC and 13 healthy skin samples. Moreover, scRNAseq analysis of BCCs of two independent datasets showed that the COL10A1-expressing population of cells is associated with the stroma adjacent to invasive BCC and shows extracellular matrix remodeling features. CONCLUSION: We identified COL10A1 as a marker of high-risk BCC, in particular of the sclerosing/infiltrative and basosquamous subtypes. We demonstrated at the single cell level that COL10A1 is expressed by a specific CAF population associated with the stroma of invasive BCC. This opens up new tailored treatment options as well as a new prognostic biomarker for BCC progression.

Lightning deaths in three outdoor dogs: A case study.

This case report presents findings in three German Shepherd placed outdoor, dead after a night of thunderstorm.

Exposure to environmental pollutants selects for xenobiotic-degrading functions in the human gut microbiome.

Environmental pollutants from different chemical families may reach the gut microbiome, where they can be metabolized and transformed. However, how our gut symbionts respond to the exposure to environmental pollution is still underexplored. In this observational, cohort study, we aim to investigate the influence of environmental pollution on the gut microbiome composition and potential activity by shotgun metagenomics. We select as a case study a population living in a highly polluted area in Campania region (Southern Italy), proposed as an ideal field for exposomic studies and we compare the fecal microbiome of 359 subjects living in areas with high, medium and low environmental pollution. We highlight changes in gut microbiome composition and functionality that were driven by pollution exposure. Subjects from highly polluted areas show higher blood concentrations of dioxin and heavy metals, as well as an increase in microbial genes related to degradation and/or resistance to these molecules. Here we demonstrate the dramatic effect that environmental xenobiotics have on gut microbial communities, shaping their composition and boosting the selection of strains with degrading capacity. The gut microbiome can be considered as a pivotal player in the environment-health interaction that may contribute to detoxifying toxic compounds and should be taken into account when developing risk assessment models. The study was registered at ClinicalTrials.gov with the identifier NCT05976126.

Use of Honey Bees and Hive Products as Bioindicators to Assess Environmental Contamination in Targeted Areas of the Campania Region (Italy).

In recent years, biomonitoring has gained more attention, particularly when assessing the environmental health of significant areas, such as those near waste-to-energy facilities. These requirements coincide with the chance to detect environmental pollutants using sensitive organisms. Bees were shown to be quite effective in evaluating the presence of certain compounds by analyzing their associated matrices, such as pollen, honey, or wax. In our study, we employed the honey bee (Apis mellifera) as an indicator to initially monitor the vicinity of the waste-to-energy plant in Acerra, which is situated in the Campania region of Italy. The primary aim was to determine whether the facility was accountable for any environmental releases of dioxins or dioxin-like compounds. Then, we assessed the presence of additional pollutants in the same area, including trace elements, polycyclic aromatic hydrocarbons, and pesticides, released by human activities. To obtain further information about environmental quality, a second biomonitoring station was installed near the Caivano S.T.I.R. (Waste Shredding, Sifting, and Packaging Plant). The results showed the dioxin levels did not exceed predetermined limitations at the Acerra site, thus demonstrating the efficacy of the waste-to-energy facility and the bees' ability to detect the presence of other pollutants. Additionally, this biomonitoring system exhibited sensitivity to environmental variations, thereby enabling the evaluation of xenobiotic flux between two proximate zones and across temporal scales. This pioneering study suggests the advantages of utilizing bees to detect a wide range of contaminants, thereby providing valuable insights into environmental quality and potential health risks for both ecosystems and human populations.

Occurrence and pattern of legacy and emerging per- and Poly-FluoroAlkyl substances (PFAS) in eggs of loggerhead turtle Caretta caretta from western Mediterranean.

Per-and Poly-FluoroAlkyl Substances (PFAS) are a class of persistent, toxic, and mobile and chemicals both from industrial sources and from the use and disposal of Consumers products containing PFAS, whose concentration in marine food webs could pose a toxicological risk for biota and humans. In 2021, unhatched eggs were sampled from 41 loggerhead turtle Caretta caretta nests from the Italian shores of the Campania Region (Southern Italy). Whole eggs were analysed for the presence of 66 legacy and emerging PFAS with Liquid Chromatography coupled to Hybrid High Resolution Mass Spectrometry. A median Σ66 Per- and Poly-FluoroAlkyl Substances value of 3.34 ng/g egg fresh weight was found; perfluoroctane sulfonate (PFOS) represented the most contributing congener (47%), followed by perfluoro-n-undecanoic acid, perfluoro-n-tridecanoic acid, perfluoro-n-decanoic acid, perfluoro-n-decanoic acid, and perfluoro-n-tetradecanoic acid, respectively. Such compounds showed a log-norm distribution, suggesting found concentrations could represent the baseline levels in the considered sampling area. Emerging ChloroPolyFluoroPolyEthers Carboxylic Acids (ClPFECAs) were found in 20 out of 41 samples in the range 0.01-1.59 ng/g. Four samples had 20-100 fold higher concentration compared to that of other samples, suggesting the presence of hot spot areas possibly related to presence of fluoropolymer-based marine litter turtles may ingest. The analysis of two paired eggs/liver samples recovered from stranded animals revealed PFAS concentration in the same order of magnitude, supporting the role of vitellogenin in their selective transfer to yolk. Significant (P = 0.0155) Kendall negative correlation coefficient of -0.2705 among PFOS content in eggs and the recorded hatching success prompts for further investigation on associated exposure assessment and related eco-toxicity risk. This work reports for the first time PFAS presence in georeferenced loggerhead turtle eggs of the Mediterranean Sea and results represent a starting point to study PFAS time-trends in this vulnerable species.

Comparative study of polycyclic aromatic hydrocarbons (PAHs) in salt gland and liver of loggerhead turtle Caretta caretta (Linnaeus, Cheloniidae) stranded along the Mediterranean coast, Southern Italy.

The levels of polycyclic aromatic hydrocarbons, PAHs, were determined in the liver and salt gland of 19 loggerhead turtles (Caretta caretta Linnaeus, Cheloniidae) stranded along the coasts of the south Tyrrhenian Sea, Italy, from 2019 to 2021. The 16 EPA's priority PAHs were determined by gas chromatography coupled with mass spectrometry (GC-MS). The average values of ΣPAHs in liver, 139 ± 55.0, were exceptionally high and up to one hundred times those of literature and appeared even more worrying in salt gland, 320 ± 97.6 ng/g, w.w. Naphthalene was the predominant contributor to PAHs richness and accounted for 90.0% and 93.7% of ΣPAHs in the two matrices, highlighting the petrogenic source of exposure. An overall higher bioaccumulation of NAP, more than two-fold, was detected in salt gland and especially in female and adults. Data of PAHs richness highlighted a potential risk of neoplastic disease development and that anthropogenic activities may seriously impair healthy state conditions of C. caretta. populations in south Tyrrhenian Sea.

Trace Elements and Persistent Organic Pollutants in Unhatched Loggerhead Turtle Eggs from an Emerging Nesting Site along the Southwestern Coasts of Italy, Western Mediterranean Sea.

Marine pollution is one of the major threats affecting loggerhead turtles, which due to their long life span, highly migratory behavior, and carnivorous diet, may be exposed to elevated levels of toxic elements throughout their life. The transfer of chemicals from mothers to their offspring is of particular conservation concern because it may affect embryonic development and hatching success. In this study, the concentrations of 16 toxic and potentially toxic trace elements, 6 indicator polychlorinated biphenyls (PCBs), and organochlorine pesticide residues (OCPs) were determined in 138 eggs from 46 loggerhead turtle nests laid during the 2021 nesting season in Campania, Italy, western Mediterranean Sea. The possible impact of pollutant levels on hatching success and early embryonic death was also investigated. Trace element analysis was performed using an ICP-MS, except for mercury, which was determined using a Direct Mercury Analyzer® (DMA). PCBs and OCPs were analyzed with high-resolution gas chromatography coupled with high-resolution mass spectrometry (HRGC-HRMS) and gas chromatography tandem quadrupole mass spectrometry GC-MS /MS, respectively. The concentrations of essential elements in the eggs were higher than those of non-essential elements. In addition, the highly chlorinated PCBs (153, 138, and 180) contributed the most to the total PCBs, while OCPs were not detected. No correlations were found between contaminant concentrations and reproductive parameters (hatching success and no obvious embryos). The results obtained suggest that the levels of contaminants found in the eggs do not affect the reproductive success of the species in the study area.

Wildfires as a Source of Potentially Toxic Elements (PTEs) in Soil: A Case Study from Campania Region (Italy).

The worldwide increase in fire events has attracted global attention, as potentially toxic elements (PTEs) have been widely recognised within the produced ash. Ash is transported, dispersed by wind, and deposited into the soil and surficial waters even far from fires. Considering that their composition can be enriched in PTEs, they represent a potential hazard for humans and other animals exposed to airborne particles and, afterwards, to resuspended matter, even at a considerable distance from the source. This study aimed to assess the environmental impact of fire events that occurred during the 2017 summer season at two different sites in the Campania region (Southern Italy). One of the fires affected a waste disposal site west of Caserta, and one involved a forest on the slopes of Mt. Somma-Vesuvius, a few kilometres southeast of Naples, the regional capital. Changes to the PTE concentration in the topsoil in the surroundings of both sites, after the fire events, were investigated. Enrichment factors (EFs) of a selection of PTEs were determined by comparing geochemical data from two sampling campaigns, one completed before and one after the fire events. A combined application of multivariate statistics (based on robust principal component analysis; RPCA) and geospatial analysis was used to determine the materials affected by the fire on the slopes of Mt. Somma-Vesuvius, and roughly locate their place. Specifically, a statistically significant enrichment of Hg was identified in the topsoil of both study areas. In addition, in soil samples collected at Mt. Somma-Vesuvius, more PTEs showed significant changes in their concentration. For both areas, Hg enrichments were related to the deposition of ash proceeding from waste burning; furthermore, as regards the soil of the Vesuvian area, Cr and Cd enrichments were associated with the fallout of ash generated during biomass combustion, and the increase in Cu and Zn concentrations was linked to the burning of crops on cultivated lands. Apart from the specific results obtained, concerning the examined case studies, the methods applied can be seen as a reliable option to determine the compositional characteristics of materials burned during a fire event, even with the prospect of improving the eventual assessment process of the related environmental hazards.

Transposons Acting as Competitive Endogenous RNAs: In-Silico Evidence from Datasets Characterised by L1 Overexpression.

LINE L1 are transposable elements that can replicate within the genome by passing through RNA intermediates. The vast majority of these element copies in the human genome are inactive and just between 100 and 150 copies are still able to mobilize. During evolution, they could have been positively selected for beneficial cellular functions. Nonetheless, L1 deregulation can be detrimental to the cell, causing diseases such as cancer. The activity of miRNAs represents a fundamental mechanism for controlling transcript levels in somatic cells. These are a class of small non-coding RNAs that cause degradation or translational inhibition of their target transcripts. Beyond this, competitive endogenous RNAs (ceRNAs), mostly made by circular and non-coding RNAs, have been seen to compete for the binding of the same set of miRNAs targeting protein coding genes. In this study, we have investigated whether autonomously transcribed L1s may act as ceRNAs by analyzing public dataset in-silico. We observed that genes sharing miRNA target sites with L1 have a tendency to be upregulated when L1 are overexpressed, suggesting the possibility that L1 might act as ceRNAs. This finding will help in the interpretation of transcriptomic responses in contexts characterized by the specific activation of transposons.

Bioaccumulation of PCDD/Fs and PCBs in free-range hens: Congener fingerprints and biotransfer factors.

The bioaccumulation of lipophilic environmental contaminants in farm animals is an important issue to control and prevent human exposure to toxic pollutants. Free-range hens were used as a model to assess the transfer and the bioaccumulation of PCDD/Fs, DL-PCBs and NDL-PCBs from the soil to eggs in an area in the Campania region particularly affected by pollution, the "Land of Fires". We determined the following ranges of concentrations in eggs: 0.90-5.51 pg WHO-TEQ g-1 fat for PCDD/Fs; 1.63-4.24 pg WHO-TEQ g-1 fat for DL-PCBs; 2.77-9.75 pg WHO-TEQ g-1 fat for the sum of PCDD/Fs and DL-PCBs and 12.53-30.94 ng g-1 fat for NDL-PCBs. Higher contamination levels were observed in the eggs from free-range hens, compared to those of the control group, that is hens raised indoors; this indicates that soil remains a major source of contamination. Livers showed low levels of contamination for both outdoor and indoor reared hens. Biotransfer factors (BTFs) were calculated for each of the 35 PCDD/F and PCB congeners analysed. The BTFs highlighted that the transfer and bioaccumulation depend on chlorination degree and substitution pattern, in particular they increase with increasing chlorination degree up to the hexa-chlorinated congeners and then decrease. An exception to these finding was observed for 2,3,7,8 TCDD and 2,3,7,8 TCDF, which showed very high BTFs. These results demonstrate the importance of promoting studies on the environmental contaminants bioaccumulation as they provide an effective support for the risk assessment and management of exposure to toxic chemicals.

Levels of non-dioxin-like PCBs (NDL-PCBs) in liver of loggerhead turtles (Caretta caretta) from the Tyrrhenian Sea (Southern Italy).

The levels of six non-dioxin-like polychlorinated biphenyls (PCBs 28, 52, 101, 138, 153, and 180) were determined in the liver of 84 loggerhead turtles (Caretta caretta) stranded along the coasts of the Tyrrhenian Sea in Campania Region (Southern Italy), from 2017 to 2021. The average value of the sum (∑6PCBIND) was 28.0 ± 52.2 ng/g (w.w.). The hexachlorobiphenyls PCB 153 and PCB 138 and the heptachlorobiphenyl PCB 180 were the main contributors to the ∑6PCBIND. A weak positive correlation was found between CCL and highly chlorinated PCBs, with adult females having lower PCB concentrations than juvenile females and adult males. This study provides more comprehensive information on the levels of NDL-PCB in Mediterranean loggerhead turtles and sets the basis for assessing anthropogenic impacts on this species.

Essential and Non-Essential Elements in Razor Clams (Solen marginatus, Pulteney, 1799) from the Domitio Littoral in Campania (Southwestern Tyrrhenian Sea, Italy).

The levels of essential (Cu, Cr, Co, Mn, Se, Zn) and non-essential (As, Be, Bi, Cd, Cs, Ga, Ni, Pb, Sr, Tl, U, V) trace elements were studied in razor clams (Solen marginatus) collected from the Tyrrhenian coast of Southern Italy at five selected sites along the Domitio littoral in the Campania region. The main objectives of this study were to assess the contamination status of these bivalve mollusks and to evaluate the risks to the environment and consumers due to metal contamination. The concentrations of 18 trace elements were determined after microwave-assisted mineralization and by inductively coupled plasma mass spectrometry (ICP-MS). Concentrations of the toxic elements Pb and Cd were below the maximum levels established by Commission Regulation (EC) 1881/2006, while higher average concentrations of arsenic were found at each of the five sites studied. Regarding the other trace elements, contamination levels followed the order: Zn > Sr > Mn > Cu > Se > Cr > V > Ni > Co > Ga > Cs > Be > U > Bi > Tl. No significant differences among the sites were found with regard to any of the trace elements analyzed, and element levels in razor clams did not reflect sediment contamination. The results demonstrated the substantial food safety of the razor clams in this area with respect to heavy metals but revealed a potential health risk due to arsenic contamination in all the areas sampled.

TEspeX: consensus-specific quantification of transposable element expression preventing biases from exonized fragments.

SUMMARY: Transposable elements (TEs) play key roles in crucial biological pathways. Therefore, several tools enabling the quantification of their expression were recently developed. However, many of the existing tools lack the capability to distinguish between the transcription of autonomously expressed TEs and TE fragments embedded in canonical coding/non-coding non-TE transcripts. Consequently, an apparent change in the expression of a given TE may simply reflect the variation in the expression of the transcripts containing TE-derived sequences. To overcome this issue, we have developed TEspeX, a pipeline for the quantification of TE expression at the consensus level. TEspeX uses Illumina RNA-seq short reads to quantify TE expression avoiding counting reads deriving from inactive TE fragments embedded in canonical transcripts. AVAILABILITY AND IMPLEMENTATION: The tool is implemented in python3, distributed under the GNU General Public License (GPL) and available on Github at https://github.com/fansalon/TEspeX (Zenodo URL: https://doi.org/10.5281/zenodo.6800331). SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Meta-Analysis Suggests That Intron Retention Can Affect Quantification of Transposable Elements from RNA-Seq Data.

Transposable elements (TEs), also known as "jumping genes", are repetitive sequences with the capability of changing their location within the genome. They are key players in many different biological processes in health and disease. Therefore, a reliable quantification of their expression as transcriptional units is crucial to distinguish between their independent expression and the transcription of their sequences as part of canonical transcripts. TEs quantification faces difficulties of different types, the most important one being low reads mappability due to their repetitive nature preventing an unambiguous mapping of reads originating from their sequences. A large fraction of TEs fragments localizes within introns, which led to the hypothesis that intron retention (IR) can be an additional source of bias, potentially affecting accurate TEs quantification. IR occurs when introns, normally removed from the mature transcript by the splicing machinery, are maintained in mature transcripts. IR is a widespread mechanism affecting many different genes with cell type-specific patterns. We hypothesized that, in an RNA-seq experiment, reads derived from retained introns can introduce a bias in the detection of overlapping, independent TEs RNA expression. In this study we performed meta-analysis using public RNA-seq data from lymphoblastoid cell lines and show that IR can impact TEs quantification using established tools with default parameters. Reads mapped on intronic TEs were indeed associated to the expression of TEs and influence their correct quantification as independent transcriptional units. We confirmed these results using additional independent datasets, demonstrating that this bias does not appear in samples where IR is not present and that differential TEs expression does not impact on IR quantification. We concluded that IR causes the over-quantification of intronic TEs and differential IR might be confused with differential TEs expression. Our results should be taken into account for a correct quantification of TEs expression from RNA-seq data, especially in samples in which IR is abundant.

Exposure study on susceptible people - SPES: An integrative biomonitoring approach.

BACKGROUND: The evaluation of environmental exposure risk requires a global analysis of pollution phenomena, including biological effects and potentially correlated clinical outcomes in susceptible populations. Although human biomonitoring plays a fundamental role in assessing the degree of contamination, it is not effective alone in identifying a direct link between exposure, biomolecular effects and outcomes on target organisms. While toxicogenomics and epidemiology are mainly focused on the investigation of molecular reactions and clinical outcomes, the monitoring of environmental matrices works independently to characterize the territorial distribution of toxic compounds, without proving any correlated health risk for residents. OBJECTIVES: We propose a new biomonitoring model based on a whole systemic analytical evaluation of environmental context. The paradigm of the method consists of identifying the sources of pollution, the migration pathways of those pollutants and their effects on target organisms. By means of this innovative, holistic epidemiological approach, we included healthy human subjects in a cohort to identify potential risks of exposure and predict possible correlated clinical outcomes. 4205 residents of the Campania region were enrolled in the "SPES" biomonitoring study, which especially focused on the areas dubbed "Land of Fires" in the recent decades. DISCUSSION: The analysis of environmental exposure risk suffers the lack of data integration from various science fields, and this comes down to a limited point of view and a limited knowledge of phenomena. In implementing our model, we first constructed an analytical picture of the Real-world situation. We next conducted a comparative risk assessment, in order to identify possible correlations between pollution and health within a holistic view. CONCLUSION: This type of research activities aims to support the implementation of public health interventions and to become a reference model in the evaluation of the risk of exposure to environmental pollutants.

Oyster Crassostrea gigas, a good model for correlating viral and chemical contamination in the marine environment.

To establish a relationship between viruses and chemicals, they were analysed in oyster Crassostrea gigas from an Italian experimental station. The chemicals concentrations were: Σ6 NDL-PCBs 0.82-7.12 ng g-1; BaP LOQ (<0.2 µg kg-1) to 1.2 µg kg-1; PAH4 LOQ (<0.2 µg kg-1) to 9.8 µg kg-1; Cd 0.073-0.365 mg kg-1; Pb 0.010-0.487 mg kg-1; and Hg < LOQ (0.089 mg kg-1). The viruses identified included: noroviruses (NoVGI/GII), astrovirus (AsV), rotavirus (RV), adenovirus (AdV), and sapovirus (SaV), while hepatitis A, hepatitis E, and Aichi viruses were not detected. Significant correlations were observed for NDL-PCBs with NoVGI, NoVGII, and AdV; BaP and PAH4 with NoVGI and AsV; Cd with RV; Pb with NoVGI and AsV; PAHs with Pb; AsV with NoVGI; and AdV with NoVGII. The study indicated as C. gigas is a model for correlating pollutants and foodborne viruses, whose co-presence may represent an additional food safety risk.

Dioxins and dioxin-like PCBs in buffalo milk from the Campania region (Italy): Decreasing trend and baseline assessment over 10 years (2008-2018).

Polychlorodibenzo-p-dioxins, polychlorodibenzo-furans (PCDD/Fs) and dioxin-like polychlorobiphenyls (DL-PCBs) enter the food chain from the environment. In this study, we report the 2008-2018 time-trends in the PCDD/F and DL-PCB contamination of milk from buffaloes fed on local forage in rural areas of the Campania region. Validated according to QA/QC criteria, the dataset (N = 808 on a total of 2068 samples, after excluding follow-up results and outliers) was computed on the upper-bound value pg WHO-TEQ2005 g-1fat. We assessed time-trends and assayed baseline contamination levels, which displayed log-normal distribution. A significant decreasing trend (p < 0.01) was observed from 2008 to 2009 and 2010; the P50-P95 range fell from 2.37-8.48 pg WHO-TEQ2005 g-1fat (N = 393) in 2008 to 1.73-4.61 in 2009 (N = 86) and to 0.67-1.46 in 2010 (N = 42). From 2010 to 2018 (N = 329), no significant variation was found among years and the related dataset fitted a log-normal distribution (p < 0.05). Occurrence descriptors indicated that the baseline contamination of dairy products (mean = 0.54; P50-P95 = 0.47-1.24) in the Campania Region was well below the EU regulatory limit in force (5.5 pg WHO-TEQ2005 g-1fat). Given the Tolerable Weekly Intake of 2 pg WHO-TEQ kg-1 body weight (bw) proposed by the EFSA for PCDD/Fs and DL-PCBs, this level of baseline contamination is discussed with regard to the orientation of food safety and food security risk connected with buffalo mozzarella cheese production.

Contamination by Aflatoxins B/G in Food and Commodities Imported in Southern Italy from 2017 to 2020: A Risk-Based Evaluation.

This study reports the results of aflatoxins B/G monitoring in food of vegetal origin, imported in Southern Italy from extra-European Union countries. From 2017 to 2020, we analyzed 1675 samples using an accredited HPLC method with fluorescence detection. We found out 295 samples (17.6%) were contaminated by aflatoxin B1, 204 by aflatoxins B/G (12.2%), while 75 (4.5%) resulted non-compliant to maximum limits set by the European Union law. Most of the batches tested were unprocessed food; the distribution of contamination levels, incidence of non-compliant samples, inference for different kinds of food are reported. The study focuses on the food more susceptible to contamination by aflatoxins; nuts are the food more controlled, showing the higher number of non-compliant samples. Our study confirms that pistachio nuts, hazelnuts and almonds are the major sources of exposure for consumers. Still, other products, such as chili pepper and Brazil nuts, need to get more information about their contamination levels. The study's findings are discussed in the perspective of the last opinion by EFSA about chronic exposure to aflatoxins. A case study to evaluate not compliance of a composed food to the European Union law is reported.

Polycyclic aromatic hydrocarbons (PAHs), arsenic, chromium and lead in warty crab (Eriphia verrucosa): occurrence and risk assessment.

This study assesses the PAH and heavy metal levels in muscle of warty crabs (Eriphia verrucosa), from the northern coast of the Campania region improving the data on toxic contaminants in this crustacean. The results showed a minimal PAH contamination; the mean concentrations were as follows: 0.2, 1.6 and 1.7 µg kg-1 wet weight (ww) for BaP, PAH4 and PAH6, respectively. Regarding the levels of the two PAHs not included in the European regulations, the BkF mean concentration was 0.1 µg kg-1 ww, while DahA was detected only in 10.7% of samples. Pb and Cr were also detected at low levels with mean values of 0.068 and 0.468 mg kg-1 ww, respectively; instead, high As levels, with a mean value of 5.021 mg kg-1 ww, were found. Considering the EWIs and the ILCRs calculated in this study, the PAH, Pb and Cr contamination levels found in the edible part of the crabs resulted safe for human consumption. Contrariwise, the ILCR calculated for the As exceeded the acceptable level of cancer risk, although the calculation did not refer to the inorganic form which is the only one recognized as carcinogenic. Hence, this study shows that warty crabs can accumulate environmental contaminants in their muscle tissue representing an important route of exposure to these toxics for the local population that regularly consumes them. This finding highlights the importance of monitoring the presence of these pollutants in crabs and in general in all fish and seafood in order to ensure food safety for consumers.