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This study reports the data on the contamination caused by heavy metals in the groundwater of the Campania Plain (CP) in Southern Italy. A total of 1093 groundwater samples were obtained from the following aquifers: coastal plains (GAR, VCP, VES, SAR, and SEL), volcanic districts (PHLE and VES), and carbonate massifs (MAS and LAT). In this study, the investigation depth ranged from 5 m (GAR) to 200 m (PHLE). The sequence of heavy metal content in groundwater samples was B > Fe > Al > Mn > Zn > Ba > Ni > As > Cu > V > Se > Pb > Cd. The heavy metal pollution index (HPI) and heavy metal evaluation (HEI) demonstrated that the study areas in which groundwater samples were sampled are not risk zones. Moreover, health risk assessment shows that hazard index (HI) values for heavy metals were found to be significantly low in groundwater samples. In non-carcinogenic risk evaluation for the adult group, the risk was low, whereas for children and infants, the risk was >1 for arsenic alone. Carcinogenic risk assessment (CR) was found lower for adults, children, and infants. The Jenks optimization method was used to evaluate the distribution of heavy metals in the groundwater of CP, and the principal component analysis technique (PCA) was employed to determine the source of heavy metals, and it was found that mixed sources (natural and anthropogenic) may be responsible for heavy metals presence.

Among the therapies against the pandemic SARS-CoV-2 virus, monoclonal Antibodies (mAbs) targeting the Spike glycoprotein represent good candidates to interfere in the Spike/ACE2 interaction, preventing virus cell entry. Since anti-spike mAbs, used individually, might be unable to block the virus entry in the case of resistant mutations, we designed an innovative strategy for the isolation of multiple novel human scFvs specific for the binding domain (RBD) of Spike. By panning a large phage display antibody library on immobilized RBD, we obtained specific binders by eluting with ACE2 in order to identify those scFvs recognizing the epitope of Spike interacting with its receptor. We converted the novel scFvs into full size IgG4, differently from the previously isolated IgG1 mAbs, to avoid unwanted potential side effects of IgG1 potent effector functions on immune system. The novel antibodies specifically bind to RBD in a nanomolar range and interfere in the interaction of Spike with ACE2 receptor, either used as purified protein or when expressed on cells in its native conformation. Furthermore, some of them have neutralizing activity for virus infection in cell cultures by using two different SARS-CoV-2 isolates including the highly contagious VOC 202012/01 variant and could become useful therapeutic tools to fight against the SARS-CoV-2 virus.

ATP2B1 is a known regulator of calcium (Ca2+) cellular export and homeostasis. Diminished levels of extra- or intra-cellular Ca2+ content have been suggested to block SARS-CoV-2 replication. Here, we demonstrate that a newly nontoxic caloxin-derivative compound (PI-7) inhibits ATP2B1, reduces the extra- and intra-cellular Ca2+ levels and impairs SARS-CoV-2 replication and propagation (VOCs: Delta and Omicron 2), as also measured by inhibition of syncytia in vitro. Furthermore, a FOXO3 transcriptional site of regulation of expression at the 5’ end of the ATP2B1 locus, together with a rare homozygous intronic variant in the ATP2B1 locus (rs11337717; chr12:89643729, T>C), are shown to be associated with severity of COVID19 (symptomatic versus asymptomatic patients). Here, we identify the mechanism of action during SARS-CoV-2 infection, which involves the PI3K/Akt signaling pathway, inactivation of FOXO3 (i.e., phosphorylation), and inhibition of transcriptional control of both membrane and reticulum Ca2+ pumps (ATP2B1 and ATP2A1 [i.e., SERCA1], respectively). The pharmacological action of compound PI-7 on sustaining both ATP2B1 and ATP2A1 expression reduces the intracellular cytoplasmic Ca2+ pool and thus negatively influences SARS-CoV-2 replication and propagation. As compound PI-7 shows a lack of toxicity, its prophylactic use as a therapy against the COVID19 pandemic is here proposed.In briefDe Antonellis et al. shows the importance of the Ca2+ channel pump ATP2B1 in the regulation of extracellular and intracellular Ca2+ levels that positively influence SARS-CoV-2 replication in human cells. Our study identifies the mechanism of action of SARS-CoV-2 in the regulation of the expression of ATP2B1 and ATP2A1 loci during infection via FOXO3 transcriptional factor. Furthermore, a small caloxin-derivative molecule (compound PI-7) can inhibit ATP2B1 activity, thus resulting in SARS-CoV-2 impairment. In further support, we have identified a genetic variant within the noncoding upstream region of ATP2B1 in symtomatic patients affected by severe COVID19, thus indicating this polymorphism as a genetic predisposition factor to SARS-CoV-2 infection.HighlightsAn anti-viral model of network of action for ATP2B1 against SARS-CoV-2 at the intracellular level that involves the PI3K/Akt signaling pathway, inactivation (i.e., phosphorylation) of FOXO3 and its transcriptional control, and inhibition of both membrane and reticulum Ca2+ pumps (i.e., ATP2B1, ATP2A1, respectively).A new drug and its lack of toxicity “compound PI-7”, thus envisioning both preventive and therapeutic applications in patients with COVID-19.The specificity of action in the context of Ca2+ homeostasis is one of the strategies that coronaviruses (including SARS-CoV-2 and any new VOC, including Omicron 2) use to infect host cells and promote organ dysfunction.Therapeutic applications for compound PI-7 against all other viruses belonging to the Coronoviridae family (e.g., SARS-CoV, MERS-CoV), and against the main families of positive sense ssRNA viruses from other hosts (e.g., Nidovirales), as these are all Ca2+ dependent.Identification of a rare homozygous intronic variant in the ATP2B1 locus (rs11337717; chr12:89643729, T>C) that is associated with severity of COVID19 (i.e., symptomatic versus asymptomatic patients). This variant can be used as a marker to identify those patients that might show severe COVID19 following their SARS-COV-2 infection.

In 2015, the "Campania Trasparente" project (http://www.campaniatrasparente.it), a monitoring plan focused on assessing the environmental conditions of the territory of the Campania region, started thanks to the financial support of the regional government. The project's general management was in charge of the Experimental Zooprophylactic Institute of Southern Italy (IZSM).In the project framework, the collection and analysis of many environmental and biological samples (including soil and air and human blood specimen) were completed. The primary aim of the whole project was to explore the existence of a link between the presence of some illnesses in the local population and the status of the environment and generate a reliable database to assess local foodstuff healthiness.Six research units were active in the framework of the project. As for soil and air, the Environmental Geochemistry Working Group (EGWG) at the Department of Earth, Environment and Resources Sciences, University of Naples Federico II, was in charge of most of the research activities. Specifically, the EGWG completed the elaboration of the data on potentially toxic metals/metalloids (PTMs) and organic contaminants (PAHs, OCPs, Dioxins) in the regional soils and air.The monitoring of air contaminants lasted more than one year, and it was completed employing passive air samplers (PAS) and deposimeters spread across the whole region.Three volumes were published, including statistical elaborations and geochemical maps of all the contaminants analysed to provide both the regional government and local scientific and professional community with a reliable tool to approach local environmental problems starting from a sound base of knowledge.Geochemical distribution patterns of potentially toxic elements (PTEs), for example, were used to establish local geochemical background/baseline intervals for those metals (naturally enriched in regional soils) found to systematically overcome the national environmental guidelines (set by the Legislative Decree 152/2006).Data from the air, analysed in terms of concentration and time variation, were, instead, fundamental to discriminate the areas of the regional territory characterised by heavy contamination associated with the emission of organic compounds from anthropic sources.The integration of all the data generated within the "Campania Trasparente" framework, including the data proceeding from the Susceptible Population Exposure Study (SPES), focusing on human biomonitoring (based on blood), allowed the development of a regional-wide conceptual model to be used as a base to generate highly specialised risk assessments for regional population and local communities affected by specific environmental problems.

Campania Region, Southern Italy, in the last 10 years was facing potential environmental issues which needed to be addressed, with the proper scientific approach, to alleviate pressure from public opinion, based more on emotions than on scientific data. Such pressure indicated an increase of oncological incidence, not supported by scientific data on the presence of anomalous pollutants in different natural media (soil, water, air, agricultural products). To face environmental/health alarm, the Campania Regional Government in 2015 funded a large, multidisciplinary, environmental project known as Campania Trasparente, to Istituto Zooprofilattico del Mezzogiorno (IZSM), to get a deeper and scientific knowledge of the Campania territory carrying out geochemical investigations, to: 1) characterize the geochemical composition of agricultural soil, air and groundwater at regional and local scale; 2) define the level of bio-availability of the toxic elements; 3) try to demonstrate a direct relationship between the presence of contaminants in the environmental matrices, in agricultural products and finally in the human matrices (hair, urine, blood). Within this project we got data on the presence of the potentially toxic metals (PTMs) and hazardous persistent organic pollutants (POPs: OCPs, PCBs, PAHs, PAEs, PBDEs) in different media of the entire Region. The new large dataset complemented our research and monitoring activities, which before 2015, were focused mostly on PTMs in soils, both at regional and local scale. In Campania Trasparente project, samples (9,000) of top and bottom soils, air and bulk deposition (150 passive air samplers, over 7 seasons), waters (1,200), vegetation (2,500) and biological (4,200) media, were collected to characterize the status of PTMs and POPs. The results obtained showed that: a) most of these elements and compounds, in higher concentrations, occur predominantly in critical areas of Napoli Urban and Metropolitan Area (NMA) and in the Sarno river basin; b) the infamous area, in the Caserta and Napoli provincial territory, known as Terra dei Fuochi (Land of Fires), is only marginally interested by anomalous occurrence of PTMs and POPs in some spot areas, not justifying the emotional alarms calling for an increase of oncological cases due to diffuse illegal practice of wastes disposal in the area; c) the agricultural crops of the Terra dei Fuochi are not affected by anomalous PTM. Specifically, the ecological risk conditions for PAHs and some OCPs (Endosulfan) occur, mostly in NMA; PCBs are sourced mostly in urban areas, being dissipated in rural areas, whereas PAEs and PBDEs occur, in general, in concentrations similar to those in other Italian regions, with some higher hot spot values in NMA and south of Salerno town. The interactional complexity between metropolitan and the surrounding rural areas is also confirmed, as it is the role that urban areas play in the migration and transformation process of POPs. High urban-rural gradients for atmospheric PAHs, PCBs and OCPs are observed mostly in the NMA and the urban areas, identified as the main emission source of POPs. Only OCPs, originating from the nearby agricultural areas, experienced long-term soil re-emission, continuously influencing conterminous urban environment via atmospheric transport processes.

Il volume illustra i risultati del monitoraggio geochimico-ambientale finalizzato alla valutazione dell’inquinamento da Contaminanti Organici Persistenti (noti nella letteratura come POP - Persistent Organic Pollutants) e Elementi Potenzialmente Tossici (EPT) della matrice aria della Regione Campania. Tale lavoro è stato svolto nell’ambito del “Programma Campania Trasparente - Attività di Monitoraggio Integrato per la Regione Campania”. Progetto finanziato dalla Regione Campania, attraverso l’IZSM, nell’ambito del Del.G.R. n. 497/2013: Fondo per le Misure Anticicliche e la Salvaguardia dell’Occupazione - Azione B4 “Mappatura del Territorio” approvato con il Decreto Esecutivo DG “Sviluppo Economico” n. 585, 14/09/2015. Le mappe prodotte rivestono particolare importanza in quanto mostrano la distribuzione nell’aria delle concentrazioni dei vari composti ed elementi analizzati (IPA, PCB, OCP, PBDE e EPT). Negli ultimi anni, la Campania è stata oggetto di attenzione mediatica per il presunto degrado dei territori agricoli di un’area tristemente nota come “Terra dei Fuochi”. La mancanza di una copertura di dati geochimici ad alta densità spaziale attestanti la presenza effettiva di contaminanti nelle diverse matrici ambientali dell’area ha favorito l’insorgere e la diffusione di allarmi relativi ad emergenze ambientali non meglio definite in termini di estensione territoriale ed impatto sanitario. Le indagini effettuate, con il Progetto Campania Trasparente, sono servite ad implementare la conoscenza geochimica esaustiva dell’intero territorio regionale- che rende la Campania la Regione meglio monitorata in Italia dal punto di vista ambientale. L’auspicio è che il monitoraggio (e la relativa cartografia) prodotto in Campania venga preso a modello dalle altre Regioni italiane, in modo da coprire l’intero territorio nazionale.

I dati illustrati in questo volume sono il frutto del lavoro svolto nell’ambito del Programma Campania Trasparente e della decennale attività di ricerca nell’ambito del monitoraggio ambientale della Regione Campania del gruppo di ricerca del Prof. Benedetto De Vivo. In questo libro, sono sinteticamente illustrati, cercando di semplificare gli aspetti tecnici rendendoli di facile comprensione al pubblico non addetto ai lavori, i principali risultati sulla distribuzione delle concentrazioni dei Contaminanti Organici Persistenti (noti nella letteratura come POP - Persistent Organic Pollutants) e Elementi Potenzialmente Tossici (EPT). Per una consultazione esaustiva delle mappe prodotte, si rimanda alle 1679 pagine dei 3 Volumi De Vivo et al. (2021a, 2021b, 2021c). Tale lavoro è stato svolto nell’ambito del “Programma Campania Trasparente - Attività di Monitoraggio Integrato per la Regione Campania”. Progetto finanziato dalla Regione Campania, attraverso l’IZSM, nell’ambito del Del.G.R. n. 497/2013: Fondo per le Misure Anticicliche e la Salvaguardia dell’Occupazione - Azione B4 “Mappatura del Territorio” approvato con il Decreto Esecutivo DG “Sviluppo Economico” n. 585, 14/09/2015. Le mappe prodotte rivestono particolare importanza in quanto mostrano la distribuzione nell’aria e nei suoli delle concentrazioni dei vari composti ed elementi analizzati (IPA, PCB, OCP, PBDE e EPT). Negli ultimi anni, la Campania è stata oggetto di attenzione mediatica per il presunto degrado dei territori agricoli di un’area tristemente nota come “Terra dei Fuochi”. La mancanza di una copertura di dati geochimici ad alta densità spaziale attestanti la presenza effettiva di contaminanti nelle diverse matrici ambientali dell’area ha favorito l’insorgere e la diffusione di allarmi relativi ad emergenze ambientali non meglio definite in termini di estensione territoriale ed impatto sanitario. Le indagini effettuate, con il Progetto Campania Trasparente, sono servite ad implementare la conoscenza geochimica esaustiva dell’intero territorio regionale- che rende la Campania la Regione meglio monitorata in Italia dal punto di vista ambientale. L’auspicio è che il monitoraggio (e la relativa cartografia) prodotto in Campania venga preso a modello dalle altre Regioni italiane, in modo da coprire l’intero territorio nazionale.

The development of prophylactic agents against the SARS-CoV-2 virus is a public health priority in the search for new surrogate markers of active virus replication. Early detection markers are needed to follow disease progression and foresee patient negativization. Subgenomic RNA transcripts (with a focus on sgN) were evaluated in oro/nasopharyngeal swabs from COVID-19-affected patients with an analysis of 315 positive samples using qPCR technology. Cut-off Cq values for sgN (Cq < 33.15) and sgE (Cq < 34.06) showed correlations to high viral loads. The specific loss of sgN in home-isolated and hospitalized COVID-19-positive patients indicated negativization of patient condition, 3–7 days from the first swab, respectively. A new detection kit for sgN, gene E, gene ORF1ab, and gene RNAse P was developed recently. In addition, in vitro studies have shown that 2’-O-methyl antisense RNA (related to the sgN sequence) can impair SARS-CoV-2 N protein synthesis, viral replication, and syncytia formation in human cells (i.e., HEK-293T cells overexpressing ACE2) upon infection with VOC Alpha (B.1.1.7)-SARS-CoV-2 variant, defining the use that this procedure might have for future therapeutic actions against SARS-CoV-2.

Long-chain polyphosphates inhibit SARS-CoV-2 infection by targeting a host receptor and a viral RNA polymerase., Polyphosphates versus SARS-CoV-2 Long-chain, inorganic polyphosphates (polyPs), which are found in many cells in the blood, have cytoprotective and antiviral activities, particularly against HIV-1 infection. Ferrucci et al. tested the effects of polyPs of various lengths on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro. Molecular docking and binding analyses showed that polyPs bound to the host receptor ACE2, which facilitates viral entry, and a viral RNA polymerase required for replication. Both proteins underwent proteasomal degradation in cells incubated with polyP120, the optimal species tested, resulting in inhibition of SARS-CoV-2 replication and a reduced inflammatory response. Given that polyPs have low toxicity, these results suggest that their potential therapeutic use should be further explored., Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO43−) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano– LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2–infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro.

In response to the complex social, environmental and economic situation caused by the "Terra dei Fuochi" phenomenon, the IZSM collaborated closely with the Terra dei Fuochi working group, carrying out additional monitoring surveys both on food (QR Code Campania project[30]), and on the environment (Campania Trasparente project[31]). This model was developed in the context of the experience accumulated in the field of environmental and food monitoring, and represents an innovative tool aimed at increasing knowledge of the environmental context of the Campania Region through an objective, integrated and organic synthesis of complex environmental phe-nomena and territorial dynamics. The model proposed here is useful for the global and synthetic assessment of environmental pressure on a municipal basis. As shown, it can also be applied to aggregations of municipalities. Furthermore, it can be used in the context of institutional actions for the planning and monitoring of improvements on a local or regional scale. Finally, the pro-posed municipality-based environmental pressure index represents the basis for geo-stratification of the sample in the context of population biomonitoring studies on a regional scale, as in the described biomonitoring study design applicable to the Campania Region.

The term "hemp" refers to Cannabis sativa cultivars grown for industrial purposes that are characterized by lower levels of tetrahydrocannabinol (THC), the active principle responsible for Cannabis psychotropic effects. Hemp is an extraordinary crop, with enormous social and economic value, since it can be used to produce food, textiles, clothing, biodegradable plastics, paper, paint, biofuel, and animal feed, as well as lighting oil. Various parts of the hemp plant represent a valuable source of food and ingredients for nutritional supplements. While hemp inflorescence is rich in nonpsychoactive, yet biologically active cannabinoids, such as cannabidiol (CBD), which exerts potent anxiolytic, spasmolytic, as well as anticonvulsant effects, hempseed has a pleasant nutty taste and represents a valuable source of essential amino acids and fatty acids, minerals, vitamins, and fibers. In addition, hempseed oil is a source of healthy polyunsaturated fatty acids, and hemp sprouts are rich in antioxidants. This review article aims to provide a comprehensive outlook from a multidisciplinary perspective on the scientific evidence supporting hemp beneficial properties when consumed as food or supplement. Marketing of hemp-derived products is subjected to diversified and complex regulations worldwide for several reasons, including the fact that CBD is also the active principal of pharmaceutical agents and that regulatory bodies in some cases ban Cannabis inflorescence regardless of its THC content. Some key regulatory aspects of such a complex scenario are also analyzed and discussed in this review article.

Fermented foods have been proposed in limiting SARS-CoV-2 infection. Emerging evidence suggest the efficacy of cow's milk fermented with the probiotic L. paracasei CBAL74 (FM-CBAL74) in preventing infectious diseases. We evaluated the protective action of FM-CBAL74 against SARS-CoV-2 infection in human enterocytes. Relevant aspects of SARS-CoV-2 infection were assessed: infectivity, host functional receptor angiotensin-converting enzyme-2 (ACE2), transmembrane protease serine 2 (TMPRSS2), and pro-inflammatory cytokines expression (IL-6, IL-15, IL-1β, VEGFβ, TNF-α, MCP-1, CXCL1). Pre-incubation with FM-CBA L74 reduced the number of infected cells. The expression of ACE2 and the pro-inflammatory cytokines IL-6, VEGFβ, IL-15, IL-1β was downregulated by the pre-treatment with this fermented food. No effect on TMPRSS2, MCP-1, TNF-α and CXCL1 expression was observed. Modulating the crucial aspects of the infection, the fermented food FM-CBAL74 exerts a preventive action against SARS-CoV-2. These evidence could pave the way to innovative nutritional strategy to mitigate the COVID-19.

The Italian municipality of Ariano Irpino (Avellino, Campania, Italy) was locked down by the regional authorities from March until April 2020 after several citizens tested positive for SARS coronavirus 2 (SARS-CoV-2). A serological mass screening campaign targeting the Ariano Irpino population using the Roche Cobas Elecsys anti-SARS-CoV-2 assay was organized by the Zoo-Prophylactic Institute of Southern Italy (Portici, Italy) and conducted in cooperation with the Local Health Unit (Azienda Sanitaria Locale – ASL – Avellino, Avellino, Italy), the Department of Public Health of University Federico II (Naples, Italy) and Department of Health Services of Azienda Ospedaliera dei Colli-Cotugno and Monaldi Hospital (Naples, Italy) in May 2020. A total of 13,218 asymptomatic individuals were reviewed in this analysis. A total of 738 citizens tested positive for anti-SARS-CoV-2 antibodies (398 females, 340 males). The overall prevalence in the sample was 5.6% (95% CI: 5.2–6.0). Among seropositive citizens, 101 cases tested positive on RT-PCR (0.76% of the overall population). Among citizens aged 14–18, 18–65 and >65 years, the seroprevalence was equal to 6.1 (95% CI: 4.1–8.7), 5.6 (95% CI: 5.1–6.1) and 4% (95% CI: 3.3–4.8), respectively. In the pediatric cohort (, Lay abstract A mass screening campaign targeting the Ariano Irpino population in Italy, including a total of 13,218 asymptomatic individuals, showed that 5.6% of the tested individuals had antibodies against the virus responsible for COVID-19 in the blood. Among citizens aged 65 years, the proportion of positive individuals was equal to 13, 6.1, 5.6 and 4%, respectively. These asymptomatic individuals, who were identified by a simple blood draw using an inexpensive test (costing only a few euros), could inadvertently spread the infection. A serological-based screening strategy could therefore be a cost-effective public health intervention.

The COVID-19 disease, caused by the SARS-Cov-2, presents a heterogeneous clinical spectrum. The risk factors do not fully explain the wide spectrum of disease manifestations, so it is possible that genetic factors could account for novel insights into its pathogenesis. In our previous study, we hypothesized that common variants on chromosome 21, near TMPRSS2 and MX1 genes, may be genetic risk factors associated to the different clinical manifestations of COVID-19. Here, we performed an in-depth genetic analysis of chromosome 21 exploiting the genome-wide association study data including 6,406 individuals hospitalized for COVID-19 and 902,088 controls with European genetic ancestry from COVID-19 Host Genetics Initiative. We found that five single nucleotide polymorphisms (SNPs) within TMPRSS2 and near MX1 gene show suggestive associations (P≤1x10-5) with severe COVID-19. All five SNPs replicated the association in two independent cohorts of Asian subjects while two and one out of the 5 SNPs replicated in African and Italian populations, respectively (P≤0.05). The minor alleles of these five SNPs correlated with a reduced risk of developing severe COVID-19 and increased level of MX1 expression in blood. Our findings provide further evidence that host genetic factors can contribute to determine the different clinical presentations of COVID-19 and that MX1, an antiviral effector of type I and III interferon pathway, may be a potential therapeutic target.

Il volume illustra, anche attraverso immagini, grafici e tabelle, i risultati del monitoraggio geochimico–ambientale finalizzato alla valutazione dell’inquinamento da elementi potenzialmente tossici dei suoli superficiali e profondi della Regione Campania nell’ambito del “Programma Campania Trasparente – Attività di Monitoraggio Integrato per la Regione Campania”. Progetto finanziato dalla Regione Campania, attraverso l’IZSM, nell’ambito del Del.G.R. n. 497/2013: Fondo per le Misure Anticicliche e la Salvaguardia dell’Occupazione – Azione B4 “Mappatura del Territorio” approvato con il Decreto Esecutivo DG “Sviluppo Economico” n. 585, 14/09/2015. Le mappe prodotte rivestono particolare importanza in quanto, oltre a mostrare la distribuzione spaziale delle concentrazioni dei vari elementi inorganici (metalli e metalloidi potenzialmente tossici – MPT), servono a definire la biodisponibilità dei MPT, e i valori background (tenori di fondo naturale) di riferimento in ottemperanza al d.l. 152/2006 e al d.m. 46/2019, laddove viene stabilito che i limiti di tolleranza possono essere elevati in funzione appunto delle concentrazioni naturali esistenti nei suoli. Negli ultimi anni, la Campania è stata oggetto di attenzione mediatica per il presunto degrado dei territori agricoli di un’area tristemente nota come “Terra dei Fuochi”. La mancanza di una copertura di dati geochimici ad alta densità spaziale attestanti la presenza effettiva di contaminanti nelle diverse matrici ambientali dell’area ha favorito l’insorgere e la diffusione di allarmi relativi a emergenze ambientali non meglio definite in termini di estensione territoriale ed impatto sanitario. Le indagini effettuate sui suoli superficiali e profondi, con il Progetto Campania Trasparente, sono servite ad implementare la conoscenza geochimica esaustiva dell’intero territorio regionale, con una copertura di campioni ad alta densità, che rende la Campania la Regione meglio monitorata in Italia dal punto di vista ambientale. L’auspicio è che il monitoraggio (e la relativa cartografia) prodotto in Campania venga preso a modello dalle altre Regioni italiane, in modo da coprire l’intero territorio nazionale con una maglia di campioni di pari.

Ovine papillomaviruses (OaPVs) were detected and quantified, for the first time, using droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (qPCR) via blood samples of 165 clinically healthy sheep. OaPV DNA was detected in 126 blood samples (~76.4%). DdPCR detected OaPV DNA in 124 samples; in only two additional samples positive for real-time qPCR, ddPCR failed to detect the presence of any OaPVs. In 70 of the positive samples (~55.6%), a single OaPV infection was observed, 12 of which were caused by OaPV1 (~17.1%) and 14 by OaPV2 (20%). OaPV3 was responsible for 19 single infections (~27.1%), and OaPV4 for 25 single infections (~35.7%). Multiple OaPV coinfections were observed in 56 (~44.4%) positive samples. OaPV coinfections caused by two genotypes were observed in 31 positive samples (~55.4%), with dual OaPV3/OaPV4 infection being the most prevalent as seen in 11 blood samples. In addition, five OaPV1/OaPV4, four OaPV1/OaPV2, four OaPV2/OaPV3, four OaPV1/OaPV3, and three OaPV2/OaPV4 dual coinfections were also detected. OaPV coinfections by triple and quadruple genotypes were detected in 24 (~42.8%) and only one (~1.8%) of coinfected blood samples, respectively. Multiple infections caused by OaPV1/OaPV3/OaPV4 genotypes were the most prevalent, as observed in 12 (50%) blood samples harboring triple OaPV infections. This study showed that ddPCR is the most sensitive and accurate assay for OaPV detection and quantification thus outperforming real-time qPCR in terms of sensitivity and specificity. Therefore, ddPCR may represent the molecular diagnostic tool of choice, ultimately providing useful insights into OaPV molecular epidemiology and field surveillance.

Genome-wide association studies (GWAS) found locus 3p21.31 associated with severe COVID-19. CCR5 resides at the same locus and, given its known biological role in other infection diseases, we investigated if common noncoding and rare coding variants, affecting CCR5, can predispose to severe COVID-19. We combined single nucleotide polymorphisms (SNPs) that met the suggestive significance level (P ≤ 1 × 10−5) at the 3p21.31 locus in public GWAS datasets (6406 COVID-19 hospitalized patients and 902,088 controls) with gene expression data from 208 lung tissues, Hi-C, and Chip-seq data. Through whole exome sequencing (WES), we explored rare coding variants in 147 severe COVID-19 patients. We identified three SNPs (rs9845542, rs12639314, and rs35951367) associated with severe COVID-19 whose risk alleles correlated with low CCR5 expression in lung tissues. The rs35951367 resided in a CTFC binding site that interacts with CCR5 gene in lung tissues and was confirmed to be associated with severe COVID-19 in two independent datasets. We also identified a rare coding variant (rs34418657) associated with the risk of developing severe COVID-19. Our results suggest a biological role of CCR5 in the progression of COVID-19 as common and rare genetic variants can increase the risk of developing severe COVID-19 by affecting the functions of CCR5.

Simple Summary Several studies have explored the role of the inflammatory cells and cytokines involved in the protection or pathogenesis of coronavirus disease 2019. Unfortunately, the results have been controversial, and further studies are needed to better understand not only the roles but also the balance of these parameters, which are crucial data to improve prevention and treatment. As COVID-19 has a well-determined phasic progression and rapidly deteriorates approximately seven days after the onset of symptoms, it is extremely necessary to detect the clinical signs that are predictive of the outcome as early as possible. To this end, in this preliminary study, we evaluated the data relating to the monocyte intracellular TNF-α expression and lymphocyte subpopulations in peripheral blood collected from patients at admission and every day of hospitalization until day 7. Our findings point to a modulation of the different cellular mediators of the immune system, which probably play a key role in the outcome of the coronavirus disease 2019. Abstract In December 2019, a novel coronavirus, “SARS-CoV-2”, was recognized as the cause of coronavirus disease 2019 (COVID-19). Several studies have explored the changes and the role of inflammatory cells and cytokines in the immunopathogenesis of the disease, but until today, the results have been controversial. Based on these premises, we conducted a retrospective assessment of monocyte intracellular TNF-α expression (iTNF-α) and on the frequencies of lymphocyte sub-populations in twenty-five patients with moderate/severe COVID-19. We found lymphopenia in all COVID-19 infected subjects compared to healthy subjects. On initial observation, in patients with favorable outcomes, we detected a high absolute eosinophil count and a high CD4+/CD8+ T lymphocytes ratio, while in the Exitus Group, we observed high neutrophil and CD8+ T lymphocyte counts. During infection, in patients with favorable outcomes, we observed a rise in the lymphocyte count, in the monocyte and in Treg lymphocyte counts, and in the CD4+ and in CD8+ T lymphocytes count but a reduction in the CD4+/CD8+ T lymphocyte ratio. Instead, in the Exitus Group, we observed a reduction in the Treg lymphocyte counts and a decrease in iTNF-α expression. Our preliminary findings point to a modulation of the different cellular mediators of the immune system, which probably play a key role in the outcomes of COVID-19.

To identify host genetic determinants involved in humoral immunity and associated with the risk of developing severe COVID-19, we analyzed 500 SARS-CoV-2 positive subjects from Southern Italy. We examined the coding sequences of 10 common variable immunodeficiency-associated genes obtained by the whole-exome sequencing of 121 hospitalized patients. These 10 genes showed significant enrichment in predicted pathogenic point mutations in severe patients compared with the non-severe ones. Moreover, in the TNFRSF13C gene, the minor allele of the p.His159Tyr variant, which is known to increase NF-kB activation and B-cell production, was significantly more frequent in the 38 severe cases compared to both the 83 non-severe patients and the 375 asymptomatic subjects further genotyped. This finding identified a potential genetic risk factor of severe COVID-19 that not only may serve to unravel the mechanisms underlying the disease severity but, also, may contribute to build the rationale for individualized management based on B-cell therapy.

The COVID-19 disease, caused by the SARS-Cov-2, presents a heterogeneous clinical spectrum. The risk factors do not fully explain the wide spectrum of disease manifestations, so it is possible that genetic factors could account for novel insights into its pathogenesis.In our previous study, we hypothesized that common variants on chromosome 21, nearTMPRSS2andMX1genes, may be genetic risk factors associated to the different clinical manifestations of COVID-19. Here, we performed an in-depth genetic analysis of chromosome 21 exploiting the genome-wide association study data including 6,406 individuals hospitalized for COVID-19 and 902,088 controls with European genetic ancestry from COVID-19 Host Genetics Initiative. We found that five single nucleotide polymorphisms (SNPs) withinTMPRSS2and nearMX1gene show suggestive associations (P≤1×10−5) with severe COVID-19. All five SNPs replicated the association in two independent cohorts of Asian subjects while two and one out of the 5 SNPs replicated in African and Italian populations, respectively (P≤0.05). The minor alleles of these five SNPs correlated with a reduced risk of developing severe COVID-19 and increased level ofMX1expression in blood.Our findings provide further evidence that host genetic factors can contribute to determine the different clinical presentations of COVID-19 and that MX1, an antiviral effector of type I and III interferon pathway, may be a potential therapeutic target.

Because most relevant studies have used small sample sizes, to date, representative atmospheric monitoring of persistent organic pollutants (POPs) on a regional scale has been very limited, which makes it difficult to precisely identify “hotspots” and possible pollution sources. In this study, an ultrahigh resolution monitoring technique was used to measure the atmospheric spatial variations in POP concentrations on a regional scale, throughout Campania, Italy. The occurrence of specific POPs—including polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs) and phthalate esters (PAEs)—were investigated using polyurethane foam-based passive air samplers (PUF-PAS), which were deployed at 129 sites across the Campania Territory between April and July 2016. The results show that the highest POP levels occurred in the Naples metropolitan area (NMA), although there were other problematic spots throughout the Territory. More specifically, hotspot areas in the NMA that depict serious POP pollution were found in the Bagnoli brownfield site, Sarno River Basin, and parts of the northeastern NMA sector. The atmospheric POP contamination in Campania is jointly controlled by the contributions of local emissions and long-range atmospheric transport. Diffusion model was employed to identify the potential sources of various POPs. The simulation showed that all the POP sources are located in the NMA and are closely related to industrial sites. This study demonstrates the advantage of using large sample sizes to identify POP source locations and achieve geospatial visualization of POP concentration and risk assessment levels.

Immunohistochemistry (IHC) and polymerase chain reaction (PCR) and fragment separation by capillary electrophoresis represent the current clinical laboratory standard for the evaluation of microsatellite instability (MSI) status. The importance of reporting MSI status in colorectal cancer is based on its potential for guiding treatment and as a prognostic indicator. It is also used to identify patients for Lynch syndrome testing. Our aim was to evaluate pre-analytical factors, such as age of formalin-fixed and paraffin-embedded (FFPE) block, neoplastic cell percentage, mucinous component, and DNA integrity, that may influence the accuracy of MSI testing and assess the concordance between three different MSI evaluation approaches. We selected the mucinous colorectal cancer (CRC) histotype for this study as it may possibly represent an intrinsic diagnostic issue due to its low tumor cellularity. Seventy-five cases of mucinous CRC and corresponding normal colon tissue samples were retrospectively selected. MMR proteins were evaluated by IHC. After DNA quality and quantity evaluation, the Idylla&trade, and TapeStation 4200 platforms were adopted for the evaluation of MSI status. Seventy-three (97.3%) cases were successfully analyzed by the three methodologies. Overall, the Idylla&trade, platform showed a concordance rate with IHC of 98.0% for microsatellite stable (MSS)/proficient MMR (pMMR) cases and 81.8% for MSI/deficient MMR (dMMR) cases. The TapeStation 4200 system showed a concordance rate with IHC of 96.0% for MSS/pMMR cases and 45.4% for MSI/dMMR cases. The concordance rates of the TapeStation 4200 system with respect to the Idylla&trade, platform were 98.1% for MSS profile and 57.8% for MSI profile. Discordant cases were analyzed using the Titano MSI kit. Considering pre-analytical factors, no significant variation in concordance rate among IHC analyses and molecular systems was observed by considering the presence of an acellular mucus cut-off &gt, 50% of the tumor area, FFPE year preparation, and DNA concentration. Conversely, the Idylla&trade, platform showed a significant variation in concordance rate with the IHC approach by considering a neoplastic cell percentage &gt, 50% (p-value = 0.002), and the TapeStation 4200 system showed a significant variation in concordance rate with the IHC approach by considering a DNA integrity number (DIN) &ge, 4 as cut-off (p-value = 0.009). Our data pinpoint a central role of the pre-analytical phase in the diagnostic outcome of MSI testing in CRC.