Differential Action of Connexin Hemichannel and Pannexin Channel Therapeutics for Potential Treatment of Retinal Diseases

Dysregulation of retinal function in the early stages of light-induced retinal degeneration involves pannexins and connexins. These two types of proteins may contribute to channels that release ATP, leading to activation of the inflammasome pathway, spread of inflammation and retinal dysfunction. However, the effect of pannexin channel block alone or block of both pannexin channels and connexin hemichannels in parallel on retinal activity in vivo is unknown. In this study, the pannexin channel blocker probenecid and the connexin hemichannel blocker tonabersat were used in the light-damaged rat retina. Retinal function was evaluated using electroretinography (ERG), retinal structure was analyzed using optical coherence tomography (OCT) imaging and the tissue response to light-induced injury was assessed immunohistochemically with antibodies against glial fibrillary acidic protein (GFAP), Ionized calcium binding adaptor molecule 1 (Iba-1) and Connexin43 (Cx43). Probenecid did not further enhance the therapeutic effect of connexin hemichannel block in this model, but on its own improved activity of certain inner retina neurons. The therapeutic benefit of blocking connexin hemichannels was further evaluated by comparing these data against results from our previously published studies that also used the light-damaged rat retina model. The analysis showed that treatment with tonabersat alone was better than probenecid alone at restoring retinal function in the light-damaged retina model. The results assist in the interpretation of the differential action of connexin hemichannel and pannexin channel therapeutics for potential treatment of retinal diseases.     

Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents

We report here the synthesis and structural characterization of novel cationic (phenothiazinyl)vinyl-pyridinium (PVP) dyes, together with optical (absorption/emission) properties and their potential applicability as fluorescent labels. Convective heating, ultrasound irradiation and mechanochemical synthesis were considered as alternative synthetic methodologies proficient for overcoming drawbacks such as long reaction time, nonsatisfactory yields or solvent requirements in the synthesis of novel dye (E)-1-(3-chloropropyl)-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium bromide 3d and its N-alkyl-2-methylpyridinium precursor 1c. The trans geometry of the newly synthesized (E)-4-(2-(7-bromo-10-ethyl-10H-phenothiazin-3-yl)vinyl)-1-methylpyridin-1-ium iodide 3b and (E)-1-methyl-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium tetrafluoroborate 3a′ was confirmed by single crystal X-ray diffraction. A negative solvatochromism of the dyes in polar solvents was highlighted by UV-Vis spectroscopy and explanatory insights were supported by molecular modeling which suggested a better stabilization of the lowest unoccupied molecular orbitals (LUMO). The photostability of the dye 3b was investigated by irradiation at 365 nm in different solvents, while the steady-state and time-resolved fluorescence properties of dye 3b and 3a′ in solid state were evaluated under one-photon excitation at 485 nm. The in vitro cytotoxicity of the new PVP dyes on B16-F10 melanoma cells was evaluated by WST-1 assay, while their intracellular localization was assessed by epi-fluorescence conventional microscopy imaging as well as one- and two-photon excited confocal fluorescence lifetime imaging microscopy (FLIM). PVP dyes displayed low cytotoxicity, good internalization inside melanoma cells and intense fluorescence emission inside the B16-F10 murine melanoma cells, making them suitable staining agents for imaging applications.     

Reliability Analysis Applied on Land Subsidence Effects of Groundwater Remediation: Probabilistic vs. Deterministic Approach

The quantification of soil variability is one of the most important aspects in the geo-engineering context. The uncertainty analysis is the main part of the reliability assessment for which a quantitative evaluation was performed in this study. The Reliability Index and the Probability of Failure using the First-Order Reliability Method (FORM) represents both, an effective method which is easy to implement at the same time. This work analyzes possible effects of compaction induced into the aquifer of the Scarlino Plain, caused by the extension of the hydraulic barrier for groundwater remediation. The currently implemented vertical barrier is composed of 12 wells which reach the depth of 10 m. The improvement of the project involves the construction of a further 40 clusters, each consisting of a doublet which intercepts different depths (10 and 18 m). The models of the subsoil stratigraphy and of the groundwater were built using a numerical model. The groundwater flow and the piezometric surface in the current configuration of the barrier were studied and the project configuration was evaluated. Using the Aquitard drainage model, the land subsidence was estimated to calculate the maximum admissible displacement related to exhibited goods, the so called territorial vulnerability. The evaluation analysis was performed using a traditional deterministic approach, followed by a reliability method based on probabilistic models. Finally, the respective results were reported in a soil mapping with overlapping layers.     

Evaluation of hydraulic excavator Human–Machine Interface concepts using NASA TLX

This study evaluated newly proposed Human–Machine Interface (HMI) design concepts for improving the ergonomics of hydraulic excavators. The design concepts were based on an augmented interaction technique which involved the use of heads-up display (HUD) and coordinated control as HMI elements. Two alternative HMI designs were elaborated in order to separately evaluate the ergonomic impacts of the head-up display and the coordinated control by comparing them to the standard HMI design. The effectiveness of these three HMI designs in terms of the reduction of the operators' mental and physical workload were assessed by conducting experiments utilizing human subjects, ages 23–35 years. The National Aeronautics and Space Administration's Task Load Index (NASA TLX) method was used for collecting subjective workload scores based on a weighted average of ratings of six factors: Mental Demand, Physical Demand, Temporal Demand, Own Performance, Effort, and Frustration Level. The results showed that the type of HMI design affects different aspects of the operator's workload. Indeed, it showed how the proposed augmented interaction is an effective solution for reducing the ergonomic gaps in terms of mental workload, and to a lesser extent the physical workload, subjected by the standard HMI design.     

Information Sharing Among Disaster Responders - An Interactive Spreadsheet-Based Collaboration Approach

Recent natural disasters have led crisis management organizations to revise their protocols so as to rely on the contribution of a wider range of actors, including simple citizens as well as expert operators, to support decision making activities. Reliable and timely information sharing among members of distributed teams of disaster responders has become paramount for the success of the overall crisis management process. In this paper we propose a crisis management system based on spreadsheet-mediated collaboration among on-site responders and decision makers. To share data a common spreadsheet artifact has been developed by using a participatory design approach which is accessed through mobile user interfaces. The evaluation results showed that the use of the spreadsheet artifact has resulted in more effective decision making relating to set of earthquake management scenarios in high-risk areas located in Italy.     

Methanol oxidation on Au/TiO2 catalysts

We have investigated the adsorption and reaction of methanol with Au/TiO₂ catalysts using a pulsed flow reactor, DRIFTS and TPD. The TiO₂ (P25) surface adsorbed a full monolayer of methanol, much of it in a dissociative manner, forming methoxy groups associated with the cationic sites, and hydroxyl groups at the anions. The methoxy is relatively stable until 250 °C, at which point decomposition occurs, producing mainly dimethyl ether by a bimolecular surface reaction. As the concentration of methoxy on the surface diminishes, so the mechanism reverts to a de-oxygenation pathway, producing mainly methane and water (at ~330 °C in TPD), but also with some coincident CO and hydrogen. Au catalysts were prepared by the deposition-precipitation method to give Au loadings between 0.5–3 wt %. The effect of low levels of Au on the reactivity is marked. The pathway which gives methane, which is characteristic of titania, remains, but a new feature of the reaction is the evolution of CO₂ and H₂ at lower temperature (a peak is seen in TPD at 220 °C), and the elimination of the DME-producing state. Clearly this is associated with the presence of Au and appears to be due to the production of a formate species on the surface of the Au component. This formate species is mainly involved in the reaction of methanol with the Au/TiO₂ catalysts which results in a combustion pathway being followed, with complete conversion occurring by ~130 °C.     

Digestion and absorption of free and esterified fish oil fatty acids in rats

This study was designed to test the hypotheses that digestibility and post-absorption metabolism of fish oil are influenced by impaired lipolysis and by the stereospecific composition of its triacylglycerols. Male Wistar rats were fed nonpurified diets containing one of the following fat sources: 9% native fish oil (NFO), 9% autorandomized fish oil (RFO), 8.1% fish oil-derived free fatty acids (FO-FFA) plus 0.9% glycerol, or 9% soybean oil (SO) as a reference fat. In a 24-day balance study, apparent digestibility of total dietary fat averaged 93.1% in the SO, NFO and RFO groups, and 90.9% in the FO-FFA group. Randomization of fish oil had no effect on apparent digestibility of individual fatty acids. In rats fed FO-FFA, apparent absorption of saturated and monounsaturated fatty acids was lower when compared to the NFO and RFO groups. Feeding the FO-FFA diet tended to increase plasma triglyceride content. The hypocholesterolemic effect of polyunsaturated n−3 fatty acids was not influenced by the dietary source. Similar effects on fatty acid profiles of plasma and liver phospholipids were caused by the NFO, RFO and the FO-FFA diets. We conclude that once polyunsaturated n−3 fatty acids are absorbed, their effect on lipid metabolism is not determined by the dietary source.     

Surface properties of fluorinated hybrid coatings

A commercial perfluoropolyether containing alkoxysilane functionalities was employed to prepare organic–inorganic hybrid coatings by using the sol‐gel process in the presence of tetraethoxysilane. Contact angle analysis revealed a strong hydrophobic and oleophobic character of the coatings almost independently from the molecular weight of the starting fluorinated oligomer. Surface tension values were in the range of 14–16 mN/m, suggesting a preferential segregation of fluorinated segments onto the surface of the coating. Atomic force microscopy showed the presence very smooth surfaces permitting to neglect the contribution of the surface roughness to wettability. Friction coefficient values were markedly lower with respect to the value of uncoated glass substrate. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102: 1483–1488, 2006     

A second-generation system for unbiased reading frame selection

Reading frame selection of nucleic acids has important implications for protein engineering and genomics. Current methods are limited because selection of the gene of interest inevitably depends on the solubility of its translated product. Here we report the construction of the pInSALect vector, which provides strict reading frame selection without concomitant selection for protein solubility or folding. This plasmid incorporates the cis-splicing VMA intein sequence from Saccharomyces cerevisiae to facilitate the post-translational self-excision of the protein of interest, thereby eliminating potential aggregation problems. Results from two libraries of chimeric glycinamide ribonucleotide formyltransferases confirm the superior performance of pInSALect over existing reading frame selection systems.     

Catalyst fragmentation during propylene polymerization: Part I. The effects of grain size and structure

Fragmentation of support/catalyst particles during propylene polymerization in the gas phase is analyzed via a mathematical model including energy and mass transfer with chemical reaction processes. The rupture phenomenon is considered specifically by the model, and evaluated as it proceeds in time, Two different regions are recognized in the polymerizing particle at fragmentation time: an inner core resembling the original solid support/catalyst structure, and an external set of layers where most of the polymerization occurs. Model predictions concerning the effects of fragmentation on polymerization are discussed. The influence of different degrees of fragmentation on thermal runaways and monomer availability at active sites located inside the support/catalyst/polymer complex is shown. Monomer concentration profiles inside the growing particles are explained in terms of the combined fragmentation-polymerization interaction. Results show a strong influence of catalyst structure on critical phenomena during early polymerization stages, and suggest the possibility of controlling critical parameters via the definition of fragment structure at catalyst preparation time.