Evaluation of Pure PFAS Decrease in Controlled Settings.

Since 1940, poly- or perfluorinated alkyl substances (PFAS) have been largely used in many applications, including paints, fire foaming, household items, product packaging, and fabrics. Because of their extremely high persistency, they have been defined as "forever chemicals". Although the EU is taking action to reduce their use, their widespread occurrence in environmental matrices and their harmful effects on human health require the use of highly performing analytical methods for efficient monitoring. Furthermore, novel PFAS are constantly revealed by both EU and National environmental agencies. The objective of this work is to investigate the cause of the signal decrease during the analysis of a standard PFAS mixture in water-based matrices, by proposing an efficient technical procedure for laboratory specialists. The analyses were carried out on a mixture of 30 PFAS, including both regulated and unknown substances (which are expected to be introduced in the guidelines), characterized by different chemical features, using LC-vials of two different materials, namely, glass and polypropylene, and dissolved in two solvents, namely, water and water-methanol. The temperature of analysis and the concentration of PFAS were also considered through LC-MS analyses at different times, in the 0-15 h range. Depending on the chemical structure and length of the PFAS, sampling and treatment procedures may be adopted to tackle the decrease and the release from the containers, reducing the risk of underestimating PFAS also in real water matrices.

A step forward on site-specific environmental risk assessment and insight into the main influencing factors of CECs removal from wastewater.

The presence of Contaminants of Emerging Concern (CECs) in water systems has been recognized as a potential source of risk for human health and the ecosystem. The present paper aims at evaluating the effects of different characteristics of full-scale Wastewater Treatment Plants (WWTPs) on the removal of 14 selected CECs belonging to the classes of caffeine, illicit drugs and pharmaceuticals. Particularly, the investigated plants differed because of the treatment lay-out, the type of biological process, the value of the operating parameters, the fate of the treated effluent (i.e. release into surface water or reuse), and the treatment capacity. The activity consisted of measuring concentrations of the selected CECs and also traditional water quality parameters (i.e. COD, phosphorous, nitrogen species and TSS) in the influent and effluent of 8 plants. The study highlights that biodegradable CECs (cocaine, methamphetamine, amphetamine, benzoylecgonine, 11-nor-9carboxy-Δ9-THC, lincomycin, trimethoprim, sulfamethoxazole, sulfadiazine, sulfadimethoxine, carbamazepine, ketoprofen, warfarin and caffeine) were well removed by all the WWTPs, with the best performance achieved by the MBR for antibiotics. Carbamazepine was removed at the lowest extent by all the WWTPs. The environmental risk assessed by using the site-specific value of the dilution factor resulted to be high in 3 out of 8 WWTPs for carbamazepine and less frequently for caffeine. However, the risk was reduced when the dilution factor was assumed equal to the default value of 10 as proposed by EU guidelines. Therefore, a specific determination of this factor is needed taking into account the hydraulic characteristics of the receiving water body.

Impact of COVID19 restrictions on organic micropollutants in wastewater treatment plants and human consumption rates.

COVID19 pandemic and the consequent restrictions to constrain SARS-CoV-2 spreading produced several impacts on the worldwide population. The present study focused on 10 Organic Micropollutants (illicit drugs, pharmaceuticals including some antibiotics and caffeine) and aimed to assess: (1) if COVID19 pandemic restrictions affected the load of those contaminants released into the sewage network and consequently the removal achieved by the Wastewater Treatment Plants; (2) if pursuant to the COVID19 pandemic, there was a change in population consumption rates of the same compounds through the wastewater-based epidemiology (WBE) approach. Two full-scale wastewater treatment plants (WWTPs) located in Central Italy were chosen as case studies, which are distinguished by different characteristics of the catchment area and water treatment layouts. The study was based on a 2-years monitoring activity of the concentration of the above organic micropollutants, traditional water quality parameters (COD, TSS, nitrogen compounds, total phosphorous) and flow rate in the influent and effluent. The statistical analysis of the monitoring data showed an increase of the influent load of most of the organic micropollutants. A decrease from 22% to -18% of the median removal efficiency was observed for carbamazepine in the WWTP with the lower treatment capacity only. The other compounds were removed roughly at the same rate. The application of the WBE approach demonstrated an increase in the consumption rate of cocaine, trimethoprim, sulfamethoxazole, sulfadiazine, carbamazepine and above all caffeine during the COVID19 restrictions period. These results highlight that COVID19 pandemic affected people's lifestyle and habits also as far as drugs consumption is concerned, which in turn might have an impact on the treatment efficacy of plants and finally on the receiving water body quality. Therefore, it is mandatory to keep monitoring to improve knowledge and eventually to implement the required measures to address this new problem.

Office Paper-Based Electrochemical Strips for Organophosphorus Pesticide Monitoring in Agricultural Soil.

Although the use of pesticides has highlighted obvious advantages on agricultural yields, intensive and widespread pesticide use raises serious environmental and health concerns. In particular, organophosphate pesticides represent >40% of the totality used in the field of agriculture, and developing countries face the issue of agricultural poisoning, also due to scarce monitoring programs. In this work, a decentralized, miniaturized, sustainable, and portable paper-based electrochemical biosensor for the quantification of organophosphorus pesticides' level has been realized. The proposed approach highlights the use of a very common paper-based substrate, namely, office paper. Office paper offers several advantages due to its nature: it allows one to print conductive strips for electrochemical connection, loading bio-hybrid nanosized probes (Prussian blue, carbon black, and butyrylcholinesterase), evaluating pesticides and reducing waste disposal compared to plastic-based strips. The portable system has been characterized by a low detection limit of 1.3 ng/mL, and accordingly to total discovered pesticide contents in EU agricultural soils, up to ca. 3 µg/mL, it can offer a valuable tool for fast monitoring. To demonstrate its effectiveness, soil and fruit vegetables have been used to perform in situ quantification. Good recovery percentages between 90 and 110% have been achieved in different matrices, highlighting to be suitable for field measurements, and a good correlation has been obtained in comparison with LC-MS analysis.

A functional role for interleukin-21 in promoting the synthesis of the T-cell chemoattractant, MIP-3alpha, by gut epithelial cells.

BACKGROUND & AIMS: Interleukin (IL)-21, a T-cell-derived cytokine, is produced in excess in inflammatory bowel diseases (IBD). The IL-21 receptor (IL-21R) is expressed by immune and nonimmune cells, raising the possibility that IL-21 has broad effects in gut inflammation. In this study we examined whether intestinal epithelial cells express IL-21R and respond to IL-21 in IBD. METHODS: IL-21R was evaluated in intestinal samples of IBD patients and controls by immunohistochemistry and Western blotting. Intestinal epithelial cells were stimulated with IL-21, and cell-free supernatants were evaluated by a protein array and enzyme-linked immunosorbent assay. The effect of IL-21-treated epithelial cell supernatants on blood lymphocyte migration was assessed using a chemotaxis assay. Finally, we evaluated the effect of a neutralizing IL-21 antibody on MIP-3alpha synthesis in ex vivo organ cultures of IBD mucosal explants. RESULTS: Constitutive expression of IL-21R was seen in intestinal epithelial cells, but was higher in IBD patients than in controls. Stimulation of intestinal epithelial cells with IL-21 resulted in enhanced phosphorylation of ERK1/2 and p38 and increased synthesis of macrophage inflammatory protein-3 alpha (MIP-3alpha), a T-cell chemoattractant. Inhibition of ERK1/2 but not p38 suppressed IL-21-induced MIP-3alpha production. IL-21-treated cell culture supernatants enhanced in vitro lymphocyte migration, and this effect was inhibited by anti-MIP-3alpha antibody. Treatment of IBD explants with anti-IL-21 reduced MIP-3alpha production. CONCLUSIONS: These data show that intestinal epithelial cells are a target of IL-21 and that IL-21 is involved in the cross-talk between epithelial and immune cells in the gut.

Protease-activated receptor-2 activation in gastric cancer cells promotes epidermal growth factor receptor trans-activation and proliferation.

Dysregulated epidermal growth factor receptor (EGFR) signaling is involved in gastric cancer (GC) cell growth. However, the mechanism that sustains EGFR signaling in GC remains unknown. Since protease-activated receptor-2 (PAR-2), a G protein-coupled receptor, has been shown to trans-activate EGFR in several cell types, we examined the role of PAR-2 in GC. We show here that in vitro activation of PAR-2 enhances the growth of two GC cell lines, AGS and MKN28. In both these cell lines, PAR-2 trans-activated EGFR and inhibition of EGFR tyrosine kinase activity by AG1478 or specific EGFR siRNA completely prevented PAR-2-driven proliferation. Antibody blockade of EGF-like ligands to EGFR did not modify EGFR signaling or cell growth induced by PAR-2 activation. In contrast, PAR-2 promoted Src activation and interaction of this kinase with EGFR. In support of this, inhibition of Src kinase activity by PP1 or siRNA blocked PAR-2-induced EGFR signaling cascade and cell growth. Finally, PAR-2 was detectable in both normal and GC specimens, but its expression was more pronounced in GC than controls and correlated with activated EGFR. These data show that PAR-2 is overexpressed in GC and suggest a role of PAR-2 in EGFR trans-activation and cell growth.

Post-transcriptional regulation of Smad7 in the gut of patients with inflammatory bowel disease.

BACKGROUND & AIMS: Transforming growth factor (TGF)-beta1 is one of the most powerful endogenous negative regulators of inflammation. In patients with inflammatory bowel disease, despite abundant local TGF-beta1, there is a failure of TGF-beta-mediated negative regulation of nuclear factor kappaB activation and proinflammatory cytokine production because of increased intracellular expression of the endogenous inhibitor of TGF-beta1 signaling, Smad7. In this study, we examined the molecular mechanism underlying the induction of Smad7 in the human gut. METHODS: Whole intestinal mucosal and lamina propria mononuclear cell samples were analyzed for Smad7 by real-time polymerase chain reaction and Western blotting. Smad7 ubiquitination and acetylation, and interaction of Smad7 with the intrinsic histone acetyltransferase, p300, were examined by immunoprecipitation and Western blotting. The effect of p300 silencing on Smad7 expression was determined in Crohn's disease lamina propria mononuclear cells. RESULTS: We showed that Smad7 is not transcriptionally regulated in human gut but that its increase in patients with inflammatory bowel disease is due to posttranscriptional acetylation and stabilization by p300, which prevents Smad7 ubiquitination and degradation in the proteasome. Hence, Smad7 protein in cells from normal gut is ubiquitinated and rapidly degraded. In contrast, in inflamed gut, Smad7 is acetylated and not ubiquitinated, is not degraded, and can be decreased by short interfering RNA to p300. CONCLUSIONS: These results identify posttranslational protein modification as of importance in chronic gut inflammation in humans.