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

Genetically engineered mesenchymal stromal cells produce IL-3 and TPO to further improve human scaffold-based xenograft models

This work was supported by grants from the EU (FP7-PEOPLE-2010-ITN EuroCancer StemCell Training network) and European Research Council (ERC-2011-StG 281474-huLSCtargeting)

Modeling BCR-ABL and MLL-AF9 leukemia in a human bone marrow-like scaffold based xenograft model

While NOD-SCID IL2Rγ(-/-) (NSG) xenograft mice are currently the most frequently used model to study human leukemia in vivo, the absence of a human niche severely hampers faithful recapitulation of the disease. We used NSG mice in which ceramic scaffolds seeded with human mesenchymal stromal cells were implanted to generate a human bone marrow (huBM-sc)-like niche. We observed that, in contrast to the murine bone marrow (mBM) niche, expression of BCR-ABL or MLL-AF9 was sufficient to induce both primary AML and ALL. Stemness was preserved within the human niches as demonstrated by serial transplantation assays. Efficient engraftment of AML MLL-AF9 and blast-crisis CML patient cells was also observed, whereby the immature blast-like phenotype was maintained in the huBM-sc niche, but to a much lesser extent in mBM niches. We compared transcriptomes of leukemias derived from mBM niches versus leukemias from huBM-like scaffold-based niches, which revealed striking differences in expression of genes associated with hypoxia, mitochondria and metabolism. Finally, we utilized the huBM-sc MLL-AF9 B-ALL model to evaluate the efficacy of the I-BET151 inhibitor in vivo. In conclusion, we have established human niche models in which the myeloid and lymphoid features of BCR-ABL(+) and MLL-AF9(+) leukemias can be studied in detail. Accepted article preview online 29 April 2016; Advance online publication 17 May 2016This work was supported by grants from the Dutch Cancer Society (2009-4411; VU2011-5127) and by the EU (ITN EuroCSC). I-BET151 was kindly provided by Nicholas Smithers (GSK R&D, UK)

Thoracic costotransverse joint pain patterns: a study in normal volunteers

<p>Abstract</p> <p>Background</p> <p>Pain referral patterns of asymptomatic costotransverse joints have not been established. The objective of this study was to determine the pain referral patterns of asymptomatic costotransverse joints via provocative intra-articular injection.</p> <p>Methods</p> <p>Eight asymptomatic male volunteers received a combined total of 21 intra-articular costotransverse joint injections. Fluoroscopic imaging was used to identify and isolate each costotransverse joint and guide placement of a 25 gauge, 2.5 inch spinal needle into the costotransverse joint. Following contrast medium injection, the quality, intensity, and distribution of the resultant pain produced were recorded.</p> <p>Results</p> <p>Of the 21 costotransverse joint injections, 16 (76%) were classified as being intra-articular via arthrograms taken at the time of injection, and 14 of these injections produced a pain sensation distinctly different from that of needle placement. Average pain produced was 3.3/10 on a 0–10 verbal pain scale. Pain was described generally as a deep, dull ache, and pressure sensation. Pain patterns were located superficial to the injected joint, with only the right T2 injections showing referred pain 2 segments cranially and caudally. No chest wall, upper extremity or pseudovisceral pains were reported.</p> <p>Conclusion</p> <p>This study provides preliminary data of the pain referral patterns of costotransverse joints. Further research is needed to compare these findings with those elicited from symptomatic subjects.</p

CT angiography; useful in non-selected outpatients?

Dance has been a part of the physical education (PE) curriculum in several countries for a longtime. In spite of this, studies demonstrate that the position of dance in the subject of PE iscontested and that little time is devoted to dance. The overall aim of this article is to examine theposition of dance as a pedagogical discourse in Swedish steering documents over time. Theempirical material consists of five Swedish curricula for PE over a period of 50 years (1962–2011).Discourse analysis is used to identify organised systems of meaning, including privileged andprioritised values. Our theoretical frame of reference draws on Bernstein’s concept of codes. Threedifferent knowledge areas within dance are found in the text material: ‘dance as cultural preserver’,‘dance as bodily exercise’ and ‘dance as expression’. Three pedagogical discourses emerge fromthese knowledge areas: an identity formation discourse, a public health discourse and an aestheticdiscourse. The identity formation discourse in earlier curricula focuses on the perpetuation ofSwedish and Nordic cultural traditions, while in later curricula, it emphasises the construction of abroader multicultural identity formation related to the understanding of different cultures. Thepublic health discourse constitutes a prioritised understanding of dance as physical training relatedto a healthy lifestyle. The aesthetic discourse, which has the weakest position over time, representsthe valuing of embodied experiences and feelings expressed through movements. This discourse isclosely linked to the construction of gender. Over time, a new performance code came to surpassthe former competence code in the steering documents. The performance code positions dance inPE as mainly a physical activity with little artistic or aesthetic value. The pedagogical discourse ofdance remains within a highly disciplinary framework of social control

Cardiac resynchronization therapy; the importance of evaluating cardiac metabolism

Cardiac Dysfunction and Arrhythmia

Evidence of scar tissue: contra-indication to cardiac resynchronization therapy?

Ventricular Dysfunction & Heart Failur

Transcriptional Silencing of the Wnt-Antagonist DKK1 by Promoter Methylation Is Associated with Enhanced Wnt Signaling in Advanced Multiple Myeloma

The Wnt/β-catenin pathway plays a crucial role in the pathogenesis of various human cancers. In multiple myeloma (MM), aberrant auto-and/or paracrine activation of canonical Wnt signaling promotes proliferation and dissemination, while overexpression of the Wnt inhibitor Dickkopf1 (DKK1) by MM cells contributes to osteolytic bone disease by inhibiting osteoblast differentiation. Since DKK1 itself is a target of TCF/β-catenin mediated transcription, these findings suggest that DKK1 is part of a negative feedback loop in MM and may act as a tumor suppressor. In line with this hypothesis, we show here that DKK1 expression is low or undetectable in a subset of patients with advanced MM as well as in MM cell lines. This absence of DKK1 is correlated with enhanced Wnt pathway activation, evidenced by nuclear accumulation of β-catenin, which in turn can be antagonized by restoring DKK1 expression. Analysis of the DKK1 promoter revealed CpG island methylation in several MM cell lines as well as in MM cells from patients with advanced MM. Moreover, demethylation of the DKK1 promoter restores DKK1 expression, which results in inhibition of β-catenin/TCF-mediated gene transcription in MM lines. Taken together, our data identify aberrant methylation of the DKK1 promoter as a cause of DKK1 silencing in advanced stage MM, which may play an important role in the progression of MM by unleashing Wnt signaling

Bioenergetic status modulates motor neuron vulnerability and pathogenesis in a zebrafish model of spinal muscular atrophy

Degeneration and loss of lower motor neurons is the major pathological hallmark of spinal muscular atrophy (SMA), resulting from low levels of ubiquitously-expressed survival motor neuron (SMN) protein. One remarkable, yet unresolved, feature of SMA is that not all motor neurons are equally affected, with some populations displaying a robust resistance to the disease. Here, we demonstrate that selective vulnerability of distinct motor neuron pools arises from fundamental modifications to their basal molecular profiles. Comparative gene expression profiling of motor neurons innervating the extensor digitorum longus (disease-resistant), gastrocnemius (intermediate vulnerability), and tibialis anterior (vulnerable) muscles in mice revealed that disease susceptibility correlates strongly with a modified bioenergetic profile. Targeting of identified bioenergetic pathways by enhancing mitochondrial biogenesis rescued motor axon defects in SMA zebrafish. Moreover, targeting of a single bioenergetic protein, phosphoglycerate kinase 1 (Pgk1), was found to modulate motor neuron vulnerability in vivo. Knockdown of pgk1 alone was sufficient to partially mimic the SMA phenotype in wild-type zebrafish. Conversely, Pgk1 overexpression, or treatment with terazosin (an FDA-approved small molecule that binds and activates Pgk1), rescued motor axon phenotypes in SMA zebrafish. We conclude that global bioenergetics pathways can be therapeutically manipulated to ameliorate SMA motor neuron phenotypes in vivo

Effectiveness of routine third trimester ultrasonography to reduce adverse perinatal outcomes in low risk pregnancy (the IRIS study): nationwide, pragmatic, multicentre, stepped wedge cluster randomised trial

Objectives To investigate the effectiveness of routine ultrasonography in the third trimester in reducing adverse perinatal outcomes in low risk pregnancies compared with usual care and the effect of this policy on maternal outcomes and obstetric interventions. Design Pragmatic, multicentre, stepped wedge cluster randomised trial. Setting 60 midwifery practices in the Netherlands. Participants 13 046 women aged 16 years or older with a low risk singleton pregnancy. Interventions 60 midwifery practices offered usual care (serial fundal height measurements with clinically indicated ultrasonography). After 3, 7, and 10 months, a third of the practices were randomised to the intervention strategy. As well as receiving usual care, women in the intervention strategy were offered two routine biometry scans at 28-30 and 34-36 weeks’ gestation. The same multidisciplinary protocol for detecting and managing fetal growth restriction was used in both strategies. Main outcome measures The primary outcome measure was a composite of severe adverse perinatal outcomes: perinatal death, Apgar score <4, impaired consciousness, asphyxia, seizures, assisted ventilation, septicaemia, meningitis, bronchopulmonary dysplasia, intraventricular haemorrhage, periventricular leucomalacia, or necrotising enterocolitis. Secondary outcomes were two composite measures of severe maternal morbidity, and spontaneous labour and birth. Results Between 1 February 2015 and 29 February 2016, 60 midwifery practices enrolled 13 520 women in mid-pregnancy (mean 22.8 (SD 2.4) weeks’ gestation). 13 046 women (intervention n=7067, usual care n=5979) with data based on the national Dutch perinatal registry or hospital records were included in the analyses. Small for gestational age at birth was significantly more often detected in the intervention group than in the usual care group (179 of 556 (32%) v 78 of 407 (19%), P<0.001). The incidence of severe adverse perinatal outcomes was 1.7% (n=118) for the intervention strategy and 1.8% (n=106) for usual care. After adjustment for confounders, the difference between the groups was not significant (odds ratio 0.88, 95% confidence interval 0.70 to 1.20). The intervention strategy showed a higher incidence of induction of labour (1.16, 1.04 to 1.30) and a lower incidence of augmentation of labour (0.78, 0.71 to 0.85). Maternal outcomes and other obstetric interventions did not differ between the strategies. Conclusion In low risk pregnancies, routine ultrasonography in the third trimester along with clinically indicated ultrasonography was associated with higher antenatal detection of small for gestational age fetuses but not with a reduced incidence of severe adverse perinatal outcomes compared with usual care alone. The findings do not support routine ultrasonography in the third trimester for low risk pregnancies. Trial registration Netherlands Trial Register NTR4367