Comparison of Automated Ribotyping,spa Typing,and MLST in 108 Clinical Isolates of Staphylococcus aureus from Orthopedic Infections

108 isolates of Staphylococcus aureus, belonging to six large ribogroups according to the automated Ribo-Printer^® system, were studied with two highly used molecular methods for epidemiological studies, namely multi-locus sequence typing (MLST) and spa typing, followed by BURP and eBURST v3 analysis for clustering spa types and sequence (ST) types. The aim was to evaluate whether automated ribotyping could be considered a useful screening tool for identifying S. aureus genetic lineages with respect to spa typing and MLST. Clarifying the relationship of riboprinting with these typing methods and establishing whether ribogroups fit single clonal complexes were two main objectives. Further information on the genetic profile of the isolates was obtained from agr typing and the search for the mecA, tst genes, and the IS256 insertion sequence. Automated ribotyping has been shown to predict spa clonal complexes and MLST clonal complexes. The high cost and lower discriminatory power of automated ribotyping compared to spa and MSLT typing could be an obstacle to fine genotyping analyzes, especially when high discriminatory power is required. On the other hand, numerous advantages such as automation, ease and speed of execution, stability, typeability and reproducibility make ribotyping a reliable method to be juxtaposed to gold standard methods.     

The Anxiolytic Drug Buspirone Prevents Rotenone-Induced Toxicity in a Mouse Model of Parkinson’s Disease

A pharmacological and genetic blockade of the dopamine D3 receptor (D3R) has shown to be neuroprotective in models of Parkinson’s disease (PD). The anxiolytic drug buspirone, a serotonin receptor 1A agonist, also functions as a potent D3R antagonist. To test if buspirone elicited neuroprotective activities, C57BL/6 mice were subjected to rotenone treatment (10mg/kg i.p for 21 days) to induce PD-like pathology and were co-treated with increasing dosages of buspirone (1, 3, or 10 mg/kg i.p.) to determine if the drug could prevent rotenone-induced damage to the central nervous system (CNS). We found that high dosages of buspirone prevented the behavioural deficits caused by rotenone in the open field test. Molecular and histological analyses confirmed that 10 mg/kg of buspirone prevented the degeneration of TH-positive neurons. Buspirone attenuated the induction of interleukin-1β and interleukin-6 expression by rotenone, and this was paralleled by the upregulation of arginase-1, brain-derived neurotrophic factor (BDNF), and activity-dependent neuroprotective protein (ADNP) in the midbrain, striatum, prefrontal cortex, amygdala, and hippocampus. Buspirone treatment also improved mitochondrial function and antioxidant activities. Lastly, the drug prevented the disruptions in the expression of two neuroprotective peptides, pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP). These results pinpoint the neuroprotective efficacy of buspirone against rotenone toxicity, suggesting its potential use as a therapeutic agent in neurodegenerative and neuroinflammatory diseases, such as PD.     

Synthesis,Computational Studies and Assessment of in Vitro Activity of Squalene Derivatives as Carbonic Anhydrase Inhibitors

We report novel molecules incorporating the nontoxic squalene scaffold and different carbonic anhydrase inhibitors (CAIs). Potent inhibitory action, in the low-nanomolar range, was detected against isoforms hCA II for sulfonamide derivatives, which proved to be selective against this isoform over the tumor-associate hCA IX and XII isoforms. On the other hand, coumarin derivatives showed weak potency but high selectivity against the tumor-associated isoform CA IX. These compounds are interesting candidates for preclinical evaluation in glaucoma or various tumors in which the two enzymes are involved. In addition, an in silico study of inhibitor-bound hCA II revealed extensive interactions with the hydrophobic pocket of the active site and provided molecular insights into the binding properties of these new inhibitors.     

Assessment of heavy metal levels in Almendares River sediments--Havana City, Cuba

The Almendares River watershed covers a large portion of Havana, Cuba and is centrally important to both recreational and other activities in the region. In order to assess current water quality conditions prior to planned remediation efforts, the spatial distribution of six heavy metals and other compounds were determined in river sediments at fifteen sampling stations in the watershed. Metal concentrations in sediments ranged from 86.1 to 708.8 for Zn, 39.3 to 189.0 for Pb, 71.6 to 420.8 for Cu, 84.4 to 209.7 Cr, 1.5 to 23.4 for Co, and 1.0 to 4.3 for Cd microg/g dry weight sediment. Calculated enrichment factors (EF; measured metal versus background mineral conditions) were almost always greater than 1.0, suggesting significant anthropogenic impact on metal levels in the river. The highest EF values were seen immediately below Cotorro (EF>10 for Pb, Cu, and Cd), a suburban town that has an active secondary smelter, and below the largest municipal landfill in Havana (EF>10 for Pb, Cu, Cd, and Zn). Further, three sampling stations had multiple metals at concentrations higher than probable effects concentrations (PEC), implying possible local ecotoxicological impacts. Finally, sequential extractions of the sediments indicated that heavy metals were largely associated with the organic fraction, and it was estimated that up to 62% of metals in the sediments would be susceptible to release back into the water column if hydraulic or other changes occurred in the river. These data are being used to prioritize decisions related to the remediation of the river system.     

Study of ITER equatorial port plug handling system and vacuum sealing interface

In the field of the ITER port plug engineering and integration task, CEA has contributed to define proposals concerning the port plugs vacuum sealing interface with the vessel flange and the equatorial plug handling.The 2001 baseline vacuum flange sealing consisted of TIG welding of a 316L strip plate on to U shapes. This arrangement presented some issues like welding access, implementation of tools, lip consumption, complex local leak test, continuous leak checking. Therefore, an alternate sealing solution based on the use of metallic gaskets is proposed. The different technical aspects are discussed to explain how this design can simplify the maintenance and deal with safety and vacuum requirements.The design of the mechanical attachment and vacuum sealing of the plug has constantly evolved, but the associated remote handling equipment was not systematically reviewed. An update of the cask and maintenance procedure was studied in order to design it in accordance with the last generic plug flange design. This includes a concept of a gripping system that uses the plug flange bolting area and, to help the remote handling process, a cantilever assisting system is suggested to increase the reliability of the transfer operation between vacuum vessel and cask.     

Exploitation of Polyphenolic Extracts from Grape Marc as Natural Antioxidants by Encapsulation in Lipid-Based Nanodelivery Systems

Phenolic compounds were extracted from grape marc by means of high pressure and temperature extraction. In order to increase their dispersability in the aqueous phase, the polyphenolic extracts were encapsulated at a final concentration of 0.1 % (w/w) in nanoemulsion-based delivery systems, which were formulated with natural ingredients, using either a liquid (sunflower oil) or a solid (palm oil) lipid phase, as well as the combination of a hydrophilic and hydrophobic emulsifier, and were produced by high-pressure homogenization. The delivery systems were characterized in terms of physicochemical stability under accelerated ageing (storage at 4 °C, 30 °C, and 55 °C for 14 days), by recording the evolution of the mean droplet size, the creaming index as well as the UV–vis absorption spectra of the encapsulated polyphenols. The antioxidant activity of the encapsulated extracts was measured with two different chemical assays (FRAP and ORAC) and a cellular antioxidant assay. Sunflower oil-based nanoemulsions resulted to be the most physically and chemically stable, with no significant variation of the mean droplet size and no degradation of the encapsulated compounds under the different conditions tested. The FRAP and ORAC assays showed that the antioxidant compounds, when encapsulated, are as available as unencapsulated polyphenols in scavenging the peroxyl radicals (ORAC), but are less available in reducing the ferric tripyridyltriazine complexes (FRAP). Remarkably, the cellular antioxidant activity was significantly higher for the encapsulated grape marc polyphenols than for the unencapsulated ones, suggesting the fundamental role of the nanoemulsions in favoring the delivery through the biological membranes.     

The Role of Cytoskeleton Revealed by Quartz Crystal Microbalance and Digital Holographic Microscopy

The connection between cytoskeleton alterations and diseases is well known and has stimulated research on cell mechanics, aiming to develop reliable biomarkers. In this study, we present results on rheological, adhesion, and morphological properties of primary rat cardiac fibroblasts, the cytoskeleton of which was altered by treatment with cytochalasin D (Cyt-D) and nocodazole (Noc), respectively. We used two complementary techniques: quartz crystal microbalance (QCM) and digital holographic microscopy (DHM). Qualitative data on cell viscoelasticity and adhesion changes at the cell–substrate near-interface layer were obtained with QCM, while DHM allowed the measurement of morphological changes due to the cytoskeletal alterations. A rapid effect of Cyt-D was observed, leading to a reduction in cell viscosity, loss of adhesion, and cell rounding, often followed by detachment from the surface. Noc treatment, instead, induced slower but continuous variations in the rheological behavior for four hours of treatment. The higher vibrational energy dissipation reflected the cell’s ability to maintain a stable attachment to the substrate, while a cytoskeletal rearrangement occurs. In fact, along with the complete disaggregation of microtubules at prolonged drug exposure, a compensatory effect of actin polymerization emerged, with increased stress fiber formation.     

A highly efficient receiver for satellite‐based automatic identification system signal detection

An innovative receiver architecture for the satellite‐based automatic identification system has been recently proposed. In this paper, we describe a few modifications that can be introduced on the algorithms for synchronization and detection, which provide an impressive performance improvement. The receiver architecture has been designed for an on‐board implementation, and a prototype has been implemented by the University of Parma and CGS S.p.A. Compagnia Generale per lo Spazio under the European Space Agency project FENICE (Flexible innovative AIS receiver prototype). A few modifications are also here described that could allow a further performance improvement in case of processing moved to ground‐based stations, based on a priori information there available. Copyright © 2014 John Wiley & Sons, Ltd.     

BETaaS: A Platform for Development and Execution of Machine-to-Machine Applications in the Internet of Things

Wireless Personal Communications - The integration of everyday objects into the Internet represents the foundation of the forthcoming Internet of Things (IoT). Smart objects will be the building...