Sicurezza alimentare: cinquant’anni di evoluzione normativa

In questo articolo si sono analizzati gli ultimi cinquanta anni di legislazione in campo alimentare per capire le premesse che hanno portato all’emanazione del pacchetto igiene, fornendo poi una analisi critic

Impact of Lentiviral Vector-Mediated Transduction on the Tightness of a Polarized Model of Airway Epithelium and Effect of Cationic Polymer Polyethylenimine

Lentiviral (LV) vectors are promising agents for efficient and long-lasting gene transfer into the lung and for gene therapy of genetically determined pulmonary diseases, such as cystic fibrosis, however, they have not been evaluated for cytotoxicity and impact on the tightness of the airway epithelium. In this study, we evaluated the transduction efficiency of a last-generation LV vector bearing Green Fluorescent Protein (GFP) gene as well as cytotoxicity and tight junction (TJ) integrity in a polarized model of airway epithelial cells. High multiplicities of infection (MOI) showed to be cytotoxic, as assessed by increase in propidium iodide staining and decrease in cell viability, and harmful for the epithelial tightness, as demonstrated by the decrease of transepithelial resistance (TER) and delocalization of occludin from the TJs. To increase LV efficiency at low LV:cell ratio, we employed noncovalent association with the polycation branched 25 kDa polyethylenimine (PEI). Transduction of cells with PEI/LV particles resulted in 2.5–3.6-fold increase of percentage of GFP-positive cells only at the highest PEI:LV ratios (1×107 PEI molecules/transducing units with 50 MOI LV) as compared to plain LV. At this dose PEI/LV transduction resulted in 6.5 ± 2.4% of propidium iodide-positive cells. On the other hand, PEI/LV particles did not determine any alteration of TER and occludin localization. We conclude that PEI may be useful for improving the efficiency of gene transfer mediated by LV vectors in airway epithelial cells, in the absence of high acute cytotoxicity and alteration in epithelial tightness

Assembly and functional analysis of an S/MAR based episome with the cystic fibrosis transmembrane conductance regulator gene

Improving the efficacy of gene therapy vectors is still an important goal toward the development of safe and efficient gene therapy treatments. S/MAR (scaffold/matrix attached region)-based vectors are maintained extra-chromosomally in numerous cell types, which is similar to viral-based vectors. Additionally, when established as an episome, they show a very high mitotic stability. In the present study we tested the idea that addition of an S/MAR element to a CFTR (cystic fibrosis transmembrane conductance regulator) expression vector, may allow the establishment of a CFTR episome in bronchial epithelial cells. Starting from the observation that the S/MAR vector pEPI-EGFP (enhanced green fluorescence protein) is maintained as an episome in human bronchial epithelial cells, we assembled the CFTR vector pBQ-S/MAR. This vector, transfected in bronchial epithelial cells with mutated CFTR, supported long term wt CFTR expression and activity, which in turn positively impacted on the assembly of tight junctions in polarized epithelial cells. Additionally, the recovery of intact pBQ-S/MAR, but not the parental vector lacking the S/MAR element, from transfected cells after extensive proliferation, strongly suggested that pBQ-S/MAR was established as an episome. These results add a new element, the S/MAR, that can be considered to improve the persistence and safety of gene therapy vectors for cystic fibrosis pulmonary disease

SGSI project at CNAF

The Italian Tier1 center is mainly focused on LHC and physics experiments in general. Recently we tried to widen our area of activity and established a collaboration with the University of Bologna to set-up an area inside our computing center for hosting experiments with high demands of security and privacy requirements on stored data. The first experiment we are going to host is Harmony, a project part of IMI's Big Data for Better Outcomes programme (IMI stands for Innovative Medicines Initiative). In order to be able to accept this kind of data we had to make a subset of our computing center compliant with the ISO 27001 regulation. In this article we will describe the SGSI project (Sistema Gestione Sicurezza Informazioni, Information Security Management System) with details of all the processes we have been through in order to become ISO 27001 compliant, with a particular focus on the separation of the project dedicated resources from all the others hosted in the center. We will also describe the software solutions adopted to allow this project to accept in the future any experiment or collaboration in need for this kind of security procedures

Gauge Identities and the Dirac Conjecture

The gauge symmetries of a general dynamical system can be systematically obtained following either a Hamiltonean or a Lagrangean approach. In the former case, these symmetries are generated, according to Dirac's conjecture, by the first class constraints. In the latter approach such local symmetries are reflected in the existence of so called gauge identities. The connection between the two becomes apparent, if one works with a first order Lagrangean formulation. Our analysis applies to purely first class systems. We show that Dirac's conjecture applies to first class constraints which are generated in a particular iterative way, regardless of the possible existence of bifurcations or multiple zeroes of these constraints. We illustrate these statements in terms of several examples.Comment: 21 page

Characterization of Pupillary Light Response Features for the Classification of Patients with Optic Neuritis

Pupillometry is a promising technique for the potential diagnosis of several neurological pathologies. However, its potential is not fully explored yet, especially for prediction purposes and results interpretation. In this work, we analyzed 100 pupillometric curves obtained by 12 subjects, applying both advanced signal processing techniques and physics methods to extract typically collected features and newly proposed ones. We used machine learning techniques for the classification of Optic Neuritis (ON) vs. Healthy subjects, controlling for overfitting and ranking the features by random permutation, following their importance in prediction. All the extracted features, except one, turned out to have significant importance for prediction, with an average accuracy of 76%, showing the complexity of the processes involved in the pupillary light response. Furthermore, we provided a possible neurological interpretation of this new set of pupillometry features in relation to ON vs. Healthy classification

Microrna-221 and Microrna-222 Modulate Differentiation and Maturation of Skeletal Muscle Cells

BACKGROUND:MicroRNAs (miRNAs) are a class of small non-coding RNAs that have recently emerged as important regulators of gene expression. They negatively regulate gene expression post-transcriptionally by translational repression and target mRNA degradation. miRNAs have been shown to play crucial roles in muscle development and in regulation of muscle cell proliferation and differentiation. METHODOLOGY/PRINCIPAL FINDINGS:By comparing miRNA expression profiling of proliferating myoblasts versus differentiated myotubes, a number of modulated miRNAs, not previously implicated in regulation of myogenic differentiation, were identified. Among these, miR-221 and miR-222 were strongly down-regulated upon differentiation of both primary and established myogenic cells. Conversely, miR-221 and miR-222 expression was restored in post-mitotic, terminally differentiated myotubes subjected to Src tyrosine kinase activation. By the use of specific inhibitors we provide evidence that expression of miR-221 and miR-222 is under the control of the Ras-MAPK pathway. Both in myoblasts and in myotubes, levels of the cell cycle inhibitor p27 inversely correlated with miR-221 and miR-222 expression, and indeed we show that p27 mRNA is a direct target of these miRNAs in myogenic cells. Ectopic expression of miR-221 and miR-222 in myoblasts undergoing differentiation induced a delay in withdrawal from the cell cycle and in myogenin expression, followed by inhibition of sarcomeric protein accumulation. When miR-221 and miR-222 were expressed in myotubes undergoing maturation, a profound alteration of myofibrillar organization was observed. CONCLUSIONS/SIGNIFICANCE:miR-221 and miR-222 have been found to be modulated during myogenesis and to play a role both in the progression from myoblasts to myocytes and in the achievement of the fully differentiated phenotype. Identification of miRNAs modulating muscle gene expression is crucial for the understanding of the circuits controlling skeletal muscle differentiation and maintenance

The immunological basis of tumor therapy by targeted delivery of TNFa to tumor vessels

ISSN:1742-469

Thromboxane inhibition improves renal perfusion and excretory function in severe congestive heart failure

AbstractObjectivesThe aim of this study was to evaluate whether thromboxane inhibition can favorably affect renal perfusion and clinical conditions in patients affected by severe heart failure.BackgroundThe renal formation of the vasoconstrictor thromboxane A2(TxA2) is increased during cardiac failure.MethodsBy oral administration of picotamide (a renal TxA2synthase and TxA2/prostaglandin H2receptor inhibitor), we blocked renal TxA2. Fourteen patients in New York Heart Association functional class IV were studied according to a randomized, double-blinded, cross-over design. Each of the two eight-day periods of testing was preceded by a three-day period during which certain vasoactive medications were stopped.ResultsDaily 24-h total urinary thromboxane B2(TxB2), the stable metabolite of TxA2, dropped at the end of picotamide treatment (p < 0.01 vs. baseline). Compared with placebo, effective renal plasma flow and the glomerular filtration rate increased (p < 0.01 and p < 0.05, respectively), thus leading to a significant decrease in the filtration fraction (p < 0.01). Renal vascular resistance decreased consistently (p < 0.01). In all patients, picotamide treatment was associated with an increase in diuresis and natriuresis (p < 0.001 vs. baseline). Plasma creatinine decreased (p < 0.05 vs. baseline). Patients also showed improvement in several clinical parameters, including a significant decrease in both pulmonary and venous pressure (p < 0.01 vs. baseline).ConclusionsThese results indicate that renal thromboxane formation plays an important role in renal vascular resistance in patients with severe heart failure, such as those described in the present study. Inhibition of TxA2improves renal hemodynamics and kidney function and favorably affects indexes of cardiac performance