Human Adenovirus come indicatore di contaminazione biologica in matrici idriche

In Italia la normativa vigente non prevede virus come indicatori di contaminazione biologica. Nello specifico per le acque reflue, in uscita dall’impianto di recupero, gli unici indicatori presi in considerazione sono E. Coli e Salmonella (Allegato D.M. 185/03). Mentre per le acque di balneazione sono E. Coli ed Enterococchi (2006/7/CE), così come per le acque destinate al consumo umano (D.lgs. n. 31/2001). Per aerosol e superfici non vi sono normative a riguardo. Numerosi studi hanno però fatto emergere che la presenza di questi indicatori non è esaustiva per predire la contaminazione biologica delle matrici idriche in esame. Se da un lato sono in grado di rivelare una contaminazione e quindi un rischio per la salute, dall’altro non sono parametri correlabili con la presenza di virus e protozoi. Infatti, laddove E. Coli ed Enterococchi sono assenti, possono essere presenti virus umani come Human Adenovirus, Rotavirus, Epatite A (HAV), Enterovirus, Norovirus GII. Tra questi Human Adenovirus risulta essere ritrovato con maggior frequenza nelle matrici. In studi effettuati su acqua di mare tale virus risulta presente nel 21% dei campioni in cui invece erano totalmente assenti E. Coli ed Enterococchi (patogeni indice suggeriti dalla normativa). Tale dato si è riscontrato anche nelle acque di fiume dove il 72% dei campioni, negativo per E. Coli ed Enterococchi, è per il 63% positivo per Human Adenovirus. Nelle acque trattate il 46% dei campioni che è negativo per E. Coli ed Enterococchi è invece positivo invece per il 57% a Human Adenovirus. Questo lavoro si è concentrato appunto su Human Adenovirus, un virus della famiglia di Adenoviridae, genere Mastadenovirus, che conta 51 diversi sierotipi in grado di infettare l’uomo, divisi in 6 sottogeneri ( indicati con le lettere da A a F) e 51 sierotipi ( indicati con i numeri). E’ un virus a doppio filamento di DNA, formato da un capside icosaedrico privo di envelope e con un diametro di circa 35-40 nm. La trasmissione del virus avviene per via oro-fecale e respiratoria, mediante secrezioni respiratorie e oculari. Si localizza a livello di del tratto respiratorio, intestinale e della congiuntiva provocando molte patologie come: gastroenteriti, congiuntiviti, patologie respiratorie e infezioni del tratto urinario. E’ eliminato all’esterno mediante feci, urina, secrezioni oro-faringee e liquido congiuntivale. Lo scopo di questo lavoro è proporre Human Adenovirus come patogeno indice di contaminazione biologica in differenti matrici idriche da affiancare agli indicatori tradizionali. Tale scelta deriva dal fatto che Human Adenovirus è ubiquitario, ha elevata resistenza ambientale e ai trattamenti di disinfezione chimico-fisici. Inoltre ha elevata specificità per l’ospite umano il quale rilascia nell’ambiente un’alta concentrazione di particelle virali anche dopo molto tempo dall’infezione. Sono stati analizzati campioni di liquame in entrata e uscita dal depuratore, acqua sperimentalmente trattata, acqua di mare e acqua di fiume. I campioni successivamente sono stati esaminati con procedure standardizzate in house per passare poi all’analisi molecolare mediante la qPCR (Real Time PCR) con protocollo specifico per Human Adenovirus. Nella maggior parte dei campioni analizzati si è riscontrata la presenza del genoma di Human Adenovirus a concentrazioni variabili (liquame in entrata(1,7 x 1010 ± 7,9) e uscita dal depuratore(4,7 x 108 ± 11), acqua sperimentalmente trattata(1,6 x 102 ± 31), acqua di mare(9,3 x 103 ± 2,4) e acqua di fiume(8,5 x 102 ± 27), confermando la sua diffusione ubiquitaria nell’ambiente. I risultati ottenuti con questo lavoro concordano con quelli di precedenti monitoraggi confermano che Human Adenovirus potrebbe essere un ottimo candidato come indicatore di contaminazione biologica da affiancare a quelli tradizionali, per diverse matrici idriche, grazie anche alla facilità con cui, tramite qPCR, si può rilevare e quantificare la sua presenza

Landbouwpraktijk en waterkwaliteit op landbouwbedrijven aangemeld voor de derogatie. Beschrijving van de meetnetopzet voor de periode 2006-2009 en de inhoud van de rapportages vanaf 2008

Translation of RIVM report 680717001. Alleen digitaal verschenenHet RIVM en het LEI hebben in 2006 in Nederland een monitoringnetwerk opgezet dat de gevolgen meet als landbouwbedrijven mogen afwijken (derogatie) van de Europese gebruiksnorm voor dierlijke mest. Het meetnet volgt driehonderd landbouwbedrijven die zich hebben aangemeld voor derogatie. Het legt de gevolgen vast voor de landbouwpraktijk en de waterkwaliteit. In dit rapport is de opzet van het monitoringnetwerk beschreven, evenals de wijze waarop vanaf 2008 over de resultaten zal worden gerapporteerd. Het rapport geeft onder andere aan wanneer welke cijfers beschikbaar zijn, en welke rekenmethoden gebruikt zullen worden om onder andere de bemesting en gewasopbrengst te berekenen. De Europese Nitraatrichtlijn verplicht lidstaten het stikstofgebruik via dierlijke mest te beperken tot maximaal 170 kg per hectare. Een lidstaat kan de Europese Commissie vragen hiervan onder voorwaarden af te wijken. Nederland heeft in december 2005 toestemming gekregen om vanaf 2006 tot en met 2009 onder voorwaarden af te mogen wijken van de gestelde norm. Dit betekent dat landbouwbedrijven 250 kilo stikstof per hectare mogen toedienen via dierlijke mest afkomstig van graasdieren (vooral koeien). Een van die voorwaarden is dat minimaal 70 procent van het totale areaal grasland is. Daarnaast is de Nederlandse overheid verplicht een monitoringnetwerk in te richten en de Commissie over de resultaten daarvan te rapporteren. De driehonderd deelnemers die worden gevolgd, zijn een steekproef van de circa 27.000 Nederlandse landbouwbedrijven die zich hebben aangemeld voor derogatie. Het netwerk is een onderdeel van het Landelijk Meetnet effecten Mestbeleid (LMM).In 2006 RIVM and LEI set up a monitoring network that measures the consequences of the option for farmers to deviate from the European use-standard for animal manure (derogation). The monitoring network follows three hundred farms benefiting from individual derogations and records the consequences for agricultural practice and water quality. This report describes the design and set-up of the monitoring network, as well as the content and layout of the annual reports from 2008 onwards containing results of the monitoring network. The report indicates, for example, what data will be available at a certain point in time, and what methods will be used to calculate such factors as fertilisation and yield. The Nitrates Directive obliges Member States to limit the use of animal manure to a maximum of 170 kg of nitrogen per hectare. A Member State may request the European Commission to deviate from this obligation under certain conditions. In December 2005, the European Commission granted the Netherlands the right to derogate from the obligation from 2006 till 2009 inclusive, implying that farmers could use up to 250 kg of nitrogen via manure of grazing livestock (particularly cows). One of the conditions is that 70% or more of the acreage available for manure application is grass. The Netherlands authorities are also obliged to set up a monitoring network and to report the results of this network to the European Commission. The 300 farms participating in the monitoring represent a sample taken from the approximately 27,000 Netherlands farms benefiting from individual derogations. The network is part of the National Programme for Monitoring the Effectiveness of the Minerals Policy (LMM).VROM-BWLLNV-D

Functional divergence in the role of N-linked glycosylation in smoothened signaling

The G protein-coupled receptor (GPCR) Smoothened (Smo) is the requisite signal transducer of the evolutionarily conserved Hedgehog (Hh) pathway. Although aspects of Smo signaling are conserved from Drosophila to vertebrates, significant differences have evolved. These include changes in its active sub-cellular localization, and the ability of vertebrate Smo to induce distinct G protein-dependent and independent signals in response to ligand. Whereas the canonical Smo signal to Gli transcriptional effectors occurs in a G protein-independent manner, its non-canonical signal employs Gαi. Whether vertebrate Smo can selectively bias its signal between these routes is not yet known. N-linked glycosylation is a post-translational modification that can influence GPCR trafficking, ligand responsiveness and signal output. Smo proteins in Drosophila and vertebrate systems harbor N-linked glycans, but their role in Smo signaling has not been established. Herein, we present a comprehensive analysis of Drosophila and murine Smo glycosylation that supports a functional divergence in the contribution of N-linked glycans to signaling. Of the seven predicted glycan acceptor sites in Drosophila Smo, one is essential. Loss of N-glycosylation at this site disrupted Smo trafficking and attenuated its signaling capability. In stark contrast, we found that all four predicted N-glycosylation sites on murine Smo were dispensable for proper trafficking, agonist binding and canonical signal induction. However, the under-glycosylated protein was compromised in its ability to induce a non-canonical signal through Gαi, providing for the first time evidence that Smo can bias its signal and that a post-translational modification can impact this process. As such, we postulate a profound shift in N-glycan function from affecting Smo ER exit in flies to influencing its signal output in mice

Patients' and urologists' preferences for prostate cancer treatment: A discrete choice experiment

__Abstract__ Background: Patients' preferences are important for shared decision making. Therefore, we investigated patients' and urologists' preferences for treatment alternatives for early prostate cancer (PC). Methods: A discrete choice experiment was conducted among 150 patients who were waiting for their biopsy results, and 150 urologists. Regression analysis was used to determine patients' and urologists' stated preferences using scenarios based on PC treatment modality (radiotherapy, surgery, and active surveillance (AS)), and risks of urinary incontinence and erectile dysfunction.Results:The response rate was 110 out of 150 (73%) for patients and 50 out of 150 (33%) for urologists. Risk of urinary incontinence was an important determinant of both patients' and urologists' stated preferences for PC treatment (P<0.05). Treatment modality also influenced patients' stated preferences (P<0.05), whereas the risk of erectile dysfunction due to radiotherapy was mainly important to urologists (P<0.05). Both patients and urologists preferred AS to radical treatment, with the exception of patients with anxious/depressed feelings who preferred radical treatment to AS. Conclusion: Although patients and urologists generally may prefer similar treatments for PC, they showed different trade-offs between various specific treatment aspects. This implies that urologists need to be aware of potential differences compared with the patient's perspective on treatment decisions in shared decision making on PC treatment

Longer telomere length in peripheral white blood cells is associated with risk of lung cancer and the rs2736100 (CLPTM1L-TERT) polymorphism in a prospective cohort study among women in China.

A recent genome-wide association study of lung cancer among never-smoking females in Asia demonstrated that the rs2736100 polymorphism in the TERT-CLPTM1L locus on chromosome 5p15.33 was strongly and significantly associated with risk of adenocarcinoma of the lung. The telomerase gene TERT is a reverse transcriptase that is critical for telomere replication and stabilization by controlling telomere length. We previously found that longer telomere length measured in peripheral white blood cell DNA was associated with increased risk of lung cancer in a prospective cohort study of smoking males in Finland. To follow up on this finding, we carried out a nested case-control study of 215 female lung cancer cases and 215 female controls, 94% of whom were never-smokers, in the prospective Shanghai Women's Health Study cohort. There was a dose-response relationship between tertiles of telomere length and risk of lung cancer (odds ratio (OR), 95% confidence interval [CI]: 1.0, 1.4 [0.8-2.5], and 2.2 [1.2-4.0], respectively; P trend = 0.003). Further, the association was unchanged by the length of time from blood collection to case diagnosis. In addition, the rs2736100 G allele, which we previously have shown to be associated with risk of lung cancer in this cohort, was significantly associated with longer telomere length in these same study subjects (P trend = 0.030). Our findings suggest that individuals with longer telomere length in peripheral white blood cells may have an increased risk of lung cancer, but require replication in additional prospective cohorts and populations

Partial complementation of Sinorhizobium meliloti bacA mutant phenotypes by the Mycobacterium tuberculosis BacA protein

The Sinorhizobium meliloti BacA ABC transporter protein plays an important role in its nodulating symbiosis with the legume alfalfa (Medicago sativa). The Mycobacterium tuberculosis BacA homolog was found to be important for the maintenance of chronic murine infections, yet its in vivo function is unknown. In the legume plant as well as in the mammalian host, bacteria encounter host antimicrobial peptides (AMPs). We found that the M. tuberculosis BacA protein was able to partially complement the symbiotic defect of an S. meliloti BacA-deficient mutant on alfalfa plants and to protect this mutant in vitro from the antimicrobial activity of a synthetic legume peptide, NCR247, and a recombinant human \u3b2-defensin 2 (HBD2). This finding was also confirmed using an M. tuberculosis insertion mutant. Furthermore, M. tuberculosis BacA-mediated protection of the legume symbiont S. meliloti against legume defensins as well as HBD2 is dependent on its attached ATPase domain. In addition, we show that M. tuberculosis BacA mediates peptide uptake of the truncated bovine AMP, Bac71-16. This process required a functional ATPase domain. We therefore suggest that M. tuberculosis BacA is important for the transport of peptides across the cytoplasmic membrane and is part of a complete ABC transporter. Hence, BacA-mediated protection against host AMPs might be important for the maintenance of latent infections

Parental transfer of the antimicrobial protein LBP/BPI protects Biomphalaria glabrata eggs against oomycete infections

Copyright: © 2013 Baron et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was funded by ANR (ANR-07-BLAN-0214 and ANR-12-EMMA-00O7-01), CNRS and INRA. PvW was financially supported by the BBSRC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

SerpinB2 regulates stromal remodelling and local invasion in pancreatic cancer

Pancreatic cancer has a devastating prognosis, with an overall 5-year survival rate of ~8%, restricted treatment options and characteristic molecular heterogeneity. SerpinB2 expression, particularly in the stromal compartment, is associated with reduced metastasis and prolonged survival in pancreatic ductal adenocarcinoma (PDAC) and our genomic analysis revealed that SERPINB2 is frequently deleted in PDAC. We show that SerpinB2 is required by stromal cells for normal collagen remodelling in vitro, regulating fibroblast interaction and engagement with collagen in the contracting matrix. In a pancreatic cancer allograft model, co-injection of PDAC cancer cells and SerpinB2(-/-) mouse embryonic fibroblasts (MEFs) resulted in increased tumour growth, aberrant remodelling of the extracellular matrix (ECM) and increased local invasion from the primary tumour. These tumours also displayed elevated proteolytic activity of the primary biochemical target of SerpinB2-urokinase plasminogen activator (uPA). In a large cohort of patients with resected PDAC, we show that increasing uPA mRNA expression was significantly associated with poorer survival following pancreatectomy. This study establishes a novel role for SerpinB2 in the stromal compartment in PDAC invasion through regulation of stromal remodelling and highlights the SerpinB2/uPA axis for further investigation as a potential therapeutic target in pancreatic cancer