First evaluation of pesticides occurrence in groundwater of Tidone Valley, an area with intensive viticulture.

Abstract Agricultural practice often involves an intensive and incorrect use of pesticides and fertilizers. These chemicals can leach through the soil profile and contaminate groundwater, including drinking water. For this reason, an effective groundwater monitoring is strongly advisable. The aim of this study was to investigate the groundwater contamination by plant protection products (PPPs) on a hilly area situated in the Tidone Valley, North-West of Italy, a region characterized by an intensive viticulture production. This area is not included in the national groundwater monitoring plan and therefore scarce information is available regarding the quality of groundwater, even though the local Environmental Agency previously revealed the occurrence of PPPs at values higher than the Environmental Quality Standard downstream this area. Hence, a monitoring wells network was developed following an upstream-downstream criterion, a list of pesticides to be monitored, based on a multi-actor approach, and an analytical method for PPPs detection and quantification. The analytical approach involved solid phase extraction followed by High-Performance Liquid Chromatography tandem mass spectrometry. The results of three monitoring campaigns revealed the occurrence of seven PPPs at a level higher than EQS for groundwater (0.1 μg/L) in 30% of the wells. The main pesticides detected were Chlorantraniliprole, Dimethomorph, Fluopicolide, Metalaxyl-M, Penconazole, and Tetraconazole, all commonly used in viticulture, together with S-metolachlor, authorized for cereal cropping. Statistical analysis revealed a significant influence of the sampling time, slope of the soil surrounding the wells, wells depth and wells location on the concentration of five PPPs. Therefore, the results obtained show that the improper use of PPPs for grapevine cultivation may cause groundwater contamination and suggest the need for a deeper analysis of territorial reality, including hydrology studies and farmer behavior and for an urgent introduction of best management practices and mitigation measures to promote a sustainable use of PPPs in viticulture

Multi-actor approach and engagement strategy to promote the adoption of best management practices and a sustainable use of pesticides for groundwater quality improvement in hilly vineyards.

The adoption of pesticide mitigation measures and innovation at farm level, are seen as a drivers to reach the sustainable water policy objectives. With the aim to prevent the pesticide pressure of hilly vineyards on groundwater contamination, a stepwise approach in Tidone Valley was applied using different consultation mechanisms and involvement strategies throughout the overall process. Face to face meetings, direct surveys, participatory monitoring and planning of several activities aiming to inform, educate, improve skills, change of individual behaviour or raise awareness, or even initiatives to build institutional trust or support for new investment in innovation are some examples. These activities allowed us to involve key actors of water use and governance (such as farmers, advisors, representatives of drinking water management, farmer's associations, Winemaking cooperatives, local Health Authority), and to have a deeper knowledge of the context agricultural practices, of farmer's knowledge and skills concerning factors influencing water contamination and also to promote the most suitable Best Management Practices aimed at limiting the pesticide occurrence in groundwater. Indeed, the surveys results highlighted that the majority of the farms are small (64% of vineyards10 ha), that most of the farmers (62%) are not aware of the current legislation on water, even if 64% of them declare to participate regularly to training courses for the prevention of water contamination and that there is a low to moderate level of adoption of Best Practices able to prevent contamination by pesticides. At the end of the overall process, it can be stated that the multi-actor approach and engagement strategy adopted were successful in improving attitudes to more sustainable practices. This is supported also by the monitoring data that show in 2019 a decrease by 44% of pesticides occurrences and a fall by 68% of values above EQ

Evaluation of groundwater contamination sources by plant protection products in hilly vineyards of Northern Italy

Abstract In Europe, 25% of groundwater has poor chemical status. One of the main stressors is agriculture, with nitrates and plant protection products (PPPs) causing failure in 18% and 6.5%, respectively, of groundwater bodies (by area). EU legislation for the placement of the PPPs on the market is one of the most stringent in the world. However, recent monitoring studies in hilly vineyards of Tidone Valley, north-west of Italy, show presence of PPPs used for grapevine cultivation in 15 out of 26 groundwater wells monitored, at values above the Environment Quality Standard (EQS) for groundwater (0.1 μg/L). However, no information about the contamination sources are available. Therefore, the objective of the present work is to evaluate the groundwater contamination sources by PPPs, in a small catchment with intensive viticulture, by collecting and integrating monitoring data, sub-surface water movement data and territorial characteristics. The results show that in wells used for PPP's mixture preparation and sprayer washing located at the top of hilly vineyards, with low slope and no water movement in the surrounding soil, the contamination is most likely from point sources. On the contrary, for wells located in a fenced area at the bottom of the hill, far away from vineyards and being used for drinking water production, the contamination is most likely from diffuse sources. Our results were used to raise awareness on groundwater contamination from PPPs among farmers in the study area; moreover a waterproof platform for sprayers washing, equipped with wastewater recovery and disposal system, able to avoid point-source contamination, was implemented in a local demonstration farm. Several demonstration activities were then organised with the farmers of the entire Valley in order to show its functionality and promote its diffuse use

Pro-inflammatory gene expression in solid glioblastoma microenvironment and in hypoxic stem cells from human glioblastoma

<p>Abstract</p> <p>Background</p> <p>Adaptation to hypoxia and consequent pro-inflammatory gene expression of prostate and breast carcinomas have been implicated in the progression toward cancer malignant phenotype. Only partial data are available for the human tumor glioblastoma multiforme (GBM). The aim of our study was to analyze the hypoxic and pro-inflammatory microenvironment in GBMs and to demonstrate that in a stem/progenitor cell line derived from human glioblastoma (GBM-SCs), hypoxia activates a coordinated inflammatory response, evidencing an invasive and migratory phenotype.</p> <p>Methods</p> <p>From each of 10 human solid glioblastomas, clinically and histopathologically characterized, we obtained three surgical samples taken from the center and the periphery of the tumor, and from adjacent host normal tissue. Molecular and morphological analyses were carried out using quantitative real-time PCR and western blot (WB). GBM stem and differentiated cells were incubated under hypoxic conditions and analyzed for pro-inflammatory gene expression and for invasive/migratory behavior.</p> <p>Results</p> <p>A panel of selected representative pro-inflammatory genes (RAGE and P2X7R, COX2, NOS2 and, PTX3) were analyzed, comparing tumor, peritumor and host normal tissues. Tumors containing leukocyte infiltrates (as assessed using CD45 immunohistochemistry) were excluded. Selected genes were overexpressed in the central regions of the tumors (i.e. in the more hypoxic areas), less expressed in peripheral regions, and poorly expressed or absent in adjacent normal host tissues. Western blot analysis confirmed that the corresponding pro-inflammatory proteins were also differently expressed. Hypoxic stem cell lines showed a clear time-dependent activation of the entire panel of pro-inflammatory genes as compared to differentiated tumor cells. Biological assays showed that invasive and migratory behavior was strengthened by hypoxia only in GBM stem cells.</p> <p>Conclusions</p> <p>In human solid glioblastoma we have observed a coordinated overexpression of a panel of pro-inflammatory genes as compared to host normal tissue. We have also evidenced a similar pattern of overexpressed genes in GBM-SCs after hypoxic treatment, showing also a gain of invasive and migratory function that was lost when these stem cells differentiated. We suggest that, as has been previously described for prostatic and mammary carcinoma, in human glioblastoma acquisition of a proinflammatory phenotype may be relevant for malignant progression.</p

Direct routes to functional RAFT agents from substituted N-alkyl maleimides

N-substituted maleimides have become an indispensable tool for the synthesis of bioconjugates and functional materials. Herein, we present three strategies for the incorporation of N-alkyl substituted maleimides into RAFT agents and show that these maleimide-derived CTAs can be used to easily introduce a range of chemical functionality at the β-position of polymer chains, resulting in α,β,ω-functional RAFT polymers. With both functional maleimides and RAFT agents that are increasingly available on the market, the approach presented in this study could facilitate the synthesis of end-functional macromolecules and will complement well the range of existing synthetic routes, including those utilising N-substituted maleimides, to functional polymeric materials

The Italian Network for Tumor Biotherapy (NIBIT): Getting together to push the field forward

As for a consolidated tradition, the 5th annual meeting of the Italian Network for Cancer Biotherapy took place in the Certosa of Pontignano, a Tuscan monastery, on September 20–22, 2007. The congress gathered more than 40 Italian leading groups representing academia, biotechnology and pharmaceutical industry. Aim of the meeting was to share new advances in cancer bio-immunotherapy and to promote their swift translation from pre-clinical research to clinical applications. Several topics were covered including: a) molecular and cellular mechanisms of tumor escape; b) therapeutic antibodies and recombinant constructs; c) clinical trials up-date and new programs; d) National Cooperative Networks and their potential interactions; e) old and new times in cancer immunology, an "amarcord". Here, we report the main issues discussed during the meeting

Insight into contact force local impedance technology for predicting effective pulmonary vein isolation

BackgroundHighly localized impedance (LI) measurements during atrial fibrillation (AF) ablation have the potential to act as a reliable predictor of the durability of the lesions created.ObjectiveWe aimed to collect data on the procedural parameters affecting LI-guided ablation in a large multicenter registry.MethodsA total of 212 consecutive patients enrolled in the CHARISMA registry and undergoing their first pulmonary vein (PV) isolation for paroxysmal and persistent AF were included.ResultsIn all, 13,891 radiofrequency (RF) applications of ≥3 s duration were assessed. The first-pass PV isolation rate was 93.3%. A total of 80 PV gaps were detected. At successful ablation spots, baseline LI and absolute LI drop were larger than at PV gap spots (161.4 ± 19 Ω vs. 153.0 ± 13 Ω, p &lt; 0.0001 for baseline LI; 22.1 ± 9 Ω vs. 14.4 ± 5 Ω, p &lt; 0.0001 for LI drop). On the basis of Receiver operating characteristic curve analysis, the ideal LI drop, which predicted successful ablation, was &gt;21 Ω at anterior sites and &gt;18 Ω at posterior sites. There was a non-linear association between the magnitude of LI drop and contact-force (CF) (r = 0.14, 95% CI: 0.13–0.16, p &lt; 0.0001) whereas both CF and LI drop were inversely related with delivery time (DT) (−0.22, −0.23 to −0.20, p &lt; 0.0001 for CF; −0.27, −0.29 to −0.26, p &lt; 0.0001 for LI drop).ConclusionAn LI drop &gt;21 Ω at anterior sites and &gt;18 Ω at posterior sites predicts successful ablation. A higher CF was associated with an increased likelihood of ideal LI drop. The combination of good CF and adequate LI drop allows a significant reduction in RF DT.Clinical trial registrationhttp://clinicaltrials.gov/, identifier: NCT03793998

Good practices for the development of budget impact models at regional level

Introduction: The present work aims to discuss the current scenario of procedures and regulations regarding budget impact analysis/models (BIA/BIM) at regional level in Italy and to provide a standardized approach and detailed recommendations for developing these analyses. Method: A systematic review of the literature was conducted in order to collect existing guidelines or specific regional procedures for budget impact analysis in Italy. All the records were analysed in qualitative terms according to a pre-specified analytical framework, based on the ISPOR BIA guidelines. At the end of the analysis, a consensus questionnaire was developed to establish agreed approaches and to provide possible solutions to any critical issues. A list of 39 statements was developed. The survey was distributed to 69 experts who rated their level of agreement with each statement on a 5-point Likert scale. Consensus was predefined as more than 66% of the panel agreeing/disagreeing with any given statement. Results: Sisty-nine experts answered the questionnaire; a total of 30/39 statements achieved consensus. There was agreement on most of the statements. Time horizon to consider and costs were the issues on which no agreement was found. The results allowed the working group to define a list of good practices. Conclusion: While the structure and development of BIM are now well-known and well-applied at national level, there remains a great diversity of management of BIM tools at regional level. Consensus was reached among participating experts, as to the main characteristics, determinants and features of regional BIA/BIM in the perspective of the Italian payer

Hsp90 Interacts Specifically with Viral RNA and Differentially Regulates Replication Initiation of Bamboo mosaic virus and Associated Satellite RNA

Host factors play crucial roles in the replication of plus-strand RNA viruses. In this report, a heat shock protein 90 homologue of Nicotiana benthamiana, NbHsp90, was identified in association with partially purified replicase complexes from BaMV-infected tissue, and shown to specifically interact with the 3′ untranslated region (3′ UTR) of BaMV genomic RNA, but not with the 3′ UTR of BaMV-associated satellite RNA (satBaMV RNA) or that of genomic RNA of other viruses, such as Potato virus X (PVX) or Cucumber mosaic virus (CMV). Mutational analyses revealed that the interaction occurs between the middle domain of NbHsp90 and domain E of the BaMV 3′ UTR. The knockdown or inhibition of NbHsp90 suppressed BaMV infectivity, but not that of satBaMV RNA, PVX, or CMV in N. benthamiana. Time-course analysis further revealed that the inhibitory effect of 17-AAG is significant only during the immediate early stages of BaMV replication. Moreover, yeast two-hybrid and GST pull-down assays demonstrated the existence of an interaction between NbHsp90 and the BaMV RNA-dependent RNA polymerase. These results reveal a novel role for NbHsp90 in the selective enhancement of BaMV replication, most likely through direct interaction with the 3′ UTR of BaMV RNA during the initiation of BaMV RNA replication

CHK1-targeted therapy to deplete DNA replication- stressed, p53-deficient, hyperdiploid colorectal cancer stem cells

Objective Cancer stem cells (CSCs) are responsible for tumour formation and spreading, and their targeting is required for tumour eradication. There are limited therapeutic options for advanced colorectal cancer (CRC), particularly for tumours carrying RAS-activating mutations. The aim of this study was to identify novel CSC-targeting strategies. Design To discover potential therapeutics to be clinically investigated as single agent, we performed a screening with a panel of FDA-approved or investigational drugs on primary CRC cells enriched for CSCs (CRC-SCs) isolated from 27 patients. Candidate predictive biomarkers of efficacy were identified by integrating genomic, reverse-phase protein microarray (RPPA) and cytogenetic analyses, and validated by immunostainings. DNA replication stress (RS) was increased by employing DNA replication-perturbing or polyploidising agents. Results The drug-library screening led to the identification of LY2606368 as a potent anti-CSC agent acting in vitro and in vivo in tumour cells from a considerable number of patients ( 3c36%). By inhibiting checkpoint kinase (CHK)1, LY2606368 affected DNA replication in most CRC-SCs, including RAS-mutated ones, forcing them into premature, lethal mitoses. Parallel genomic, RPPA and cytogenetic analyses indicated that CRC-SCs sensitive to LY2606368 displayed signs of ongoing RS response, including the phosphorylation of RPA32 and ataxia telangiectasia mutated serine/threonine kinase (ATM). This was associated with mutation(s) in TP53 and hyperdiploidy, and made these CRC-SCs exquisitely dependent on CHK1 function. Accordingly, experimental increase of RS sensitised resistant CRC-SCs to LY2606368. Conclusions LY2606368 selectively eliminates replication-stressed, p53-deficient and hyperdiploid CRC-SCs independently of RAS mutational status. These results provide a strong rationale for biomarker-driven clinical trials with LY2606368 in patients with CRC