Attaching Different Kinds of Proteinaceous Nanospheres to a Variety of Fabrics Using Ultrasound Radiation

The application of a rapid, non-destructive, cost-effective technique such as ultrasonic emulsification for the coating of different textiles was explored. The technical benefits for this research were the generation of multifunctional materials and their combinations through environmentally friendly processing technologies. We have shown for the first time that ultrasonic waves can be used to coat proteinaceous micro- and nanospheres (PM) of BSA (Bovine Serum Albumin) protein and casein on the surface of cotton and polyester (PE) fabrics. The creation and the anchoring of the microbubbles to the fabrics were performed by a one-step reaction, and the process is usually stopped after 3 min. The PM of bovine serum albumin (BSA) bonded to cotton and polyester fabrics has shown stability for ∼9 months. The PMs were shown to be attached more strongly to the polyester than to the cotton, and sustained stronger washing conditions on PE. The diameter of the BSA and the casein spheres on cotton was in the range of 0.8–1.0 μm, while on the PE it varied between 60 and 120 nm.     

The Autophagy-Related Organelle Autophagoproteasome Is Suppressed within Ischemic Penumbra

The peri-infarct region, which surrounds the irreversible ischemic stroke area is named ischemic penumbra. This term emphasizes the borderline conditions for neurons placed within such a critical region. Area penumbra separates the ischemic core, where frank cell loss occurs, from the surrounding healthy brain tissue. Within such a brain region, nervous matter, and mostly neurons are impaired concerning metabolic conditions. The classic biochemical marker, which reliably marks area penumbra is the over-expression of the heat shock protein 70 (HSP70). However, other proteins related to cell clearing pathways are modified within area penumbra. Among these, autophagy proteins like LC3 increase in a way, which recapitulates Hsp70. In contrast, components, such as P20S, markedly decrease. Despite apparent discrepancies, the present study indicates remarkable overlapping between LC3 and P20S redistribution within area penumbra. In fact, the amount of both proteins is markedly reduced within vacuoles. Specifically, a massive loss of LC3 + P20S immuno-positive vacuoles (autophagoproteasomes) is reported here. This represents the most relevant sub-cellular alteration here described in cell clearing pathways within area penumbra. The functional significance of these findings remains to be determined and it will take a novel experimental stream to decipher the fine-tuning of such a phenomenon.     

Drug-Induced Lysosomal Impairment Is Associated with the Release of Extracellular Vesicles Carrying Autophagy Markers

Amiodarone is a cationic amphiphilic drug used as an antiarrhythmic agent. It induces phospholipidosis, i.e., the accumulation of phospholipids within organelles of the endosomal–lysosomal system. Extracellular vesicles (EVs) are membrane-enclosed structures released by any type of cell and retrieved in every fluid of the body. EVs have been initially identified as a system to dispose cell waste, but they are also considered to be an additional manner to transmit intercellular signals. To understand the role of EVs in drug-induced phospholipidosis, we investigated EVs release in amiodarone-treated HEK-293 cells engineered to produce fluorescently labelled EVs. We observed that amiodarone induces the release of a higher number of EVs, mostly of a large/medium size. EVs released upon amiodarone treatment do not display significant morphological changes or altered size distribution, but they show a dose-dependent increase in autophagy associated markers, indicating a higher release of EVs with an autophagosome-like phenotype. Large/medium EVs also show a higher content of phospholipids. Drugs inducing lysosomal impairment such as chloroquine and bafilomycin A1 similarly prompt a higher release of EVs enriched in autophagy markers. This result suggests a mechanism associated with amiodarone-induced lysosomal impairment more than a connection with the accumulation of specific undigested substrates. Moreover, the implementation of the lysosomal function by overexpressing TFEB, a master gene regulator of lysosomal biogenesis, prevents the amiodarone-induced release of EVs, suggesting that this could be a feasible target to attenuate drug-induced abnormalities.     

Quality Changes and Tocopherols and γ-Orizanol Concentrations in Rice Bran Oil During the Refining Process

The quality changes and the concentrations of tocopherols and γ-oryzanol, during successive steps of rice bran oil refining (RBO), were studied. For this purpose, samples of crude, degummed, neutralized, bleached, dewaxed and deodorized RBO were taken from an industrial plant and analyzed. The moisture, pH, acidity, peroxide value and unsaponifiable matter, were determined. The fatty acid composition was evaluated by GC, and the concentrations of tocopherols and γ-oryzanol were determined using HPLC with fluorescence and UV–Vis detection, respectively. To identify γ-oryzanol components, fractions of the HPLC eluant were collected and analyzed using mass spectrometry. Oil refining reduced the peroxide value and acidity to 1 and 3% of the values obtained in crude RBO, respectively. The fatty acid composition were not significantly altered during refining. The concentrations of the tocopherols in RBO followed the order α > (β + γ) > δ. The total concentration of tocopherols was 26 mg/100 g, and remained practically unaltered during refining. Up to nine components were distinguished in γ-oryzanol. After collecting the elution fractions, up to six components were identified by electrospray mass spectrometry. Refining reduced the total concentration of γ-oryzanol to 2% of its initial value.     

Blends of polyamide 6 and epichlorohydrin elastomers. I. Graft copolymerization in the melt blending

With the purpose of improving the mechanical properties of the polyamides, the possibility of combining polyamides with elastomers has been used. The low compatibility of the resulting blends leads to deficient mechanical properties, and therefore, it is necessary to add the compatibilizer to the mixture or to produce the compatibilizer during the melting mixture. Usually, at least one of the components must contain a reactive functional groups. In the present work, blends of polyamide 6 (PA 6) and epichlorohydrin elastomers, polyepichlorohydrin (PEPI), and the equimolar copolymer poly(epichlorohydrin‐co‐ethylene oxide), ECO, with different compositions were prepared by mechanical mixture using a Banbury‐type mixer. The blends were characterized by rheological measurements, the Molau test, elemental analysis, Infrared Spectroscopy by Diffuse Reflectance, Transmission Electron Microscopy, and X‐ray Diffractometry. The blends of PA 6 with PEPI and ECO are heterogeneous, showing a morphology of elastomer particles dispersed in the polyamide matrix. The results of rheological measurements and the Molau test indicate a graft copolymerization in the interface between the polyamide and the elastomer, PA 6‐g‐elastomer, whose concentration decreases with the elastomer content. It was found that the grafting of PEPI and PA 6 changed the diffraction pattern of PA 6. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 72: 1827–1833, 1999     

Steroid bioconversion: Towards green processes

There is an increasing trend towards reducing the use of organic solvents in industry due to environmental constraints and the adoption of green chemistry guidelines. To overcome the low volumetric productivity of aqueous bioconversion systems involving sparingly water soluble hydrophobic compounds, processes are being developed and designed to incorporate green solvent such as supercritical fluids, ionic liquids and natural oils, and liquid polymers, among others as an alternative to organic solvents. Moreover, processes are developed and redesigned to use/reuse chemicals and reagents derived from waste or renewable feed stocks in order to diminish E-factors.In this work, the use of green solvents as key components in the bioconversion media for a multi-step microbial bioconversion was assessed in a suspended whole cell system, combined with the use of by-products as raw materials, ultimately used as carbon source for cell growth and as sterol substrate for bioconversion. The model system is the selective cleavage of the side-chain of β-sitosterol performed by free resting cells of Mycobacterium sp. NRRL B-3805, a well-established industrial multi-enzymatic process involving the use of nine catabolic enzymes in a 14-step metabolic pathway.Bioconversion yields in silicone media were higher than the ones obtained in polyethylene glycol (PEG), polypropylene glycol (PPG) and ionic liquids, as well as in dioctyl phthalate (DOP), an organic solvent that has previously been shown to allow high conversion yields. Total conversion of 12 mM substrate in silicone media was consistently obtained at the end of 120-h bioconversion runs. Similar bioconversion profiles were attained during a 50-fold scale-up, maintaining constant the power consumption per unit of volume.     

Effectual dispersion of carbon nanofibers in polyetherimide composites and their mechanical and tribological properties

The use of proliferation of nanotechnology in commercial applications is driving requirements for minimal chemical processing and simple processes in industry. Carbon nanofiber (CNF) products possess very high purity levels without the need of purification processing before use and are in growing demand for this quality. Polyetherimide (PEI) has excellent mechanical and thermal performance, but its high viscosity makes its nanocomposites processing very challenging. In this study, a facile melt‐mixing method was used to fabricate PEI nanocomposites with as received and physically treated CNFs. The dispersion of CNFs was characterized by scanning electron microscopy, transmitted optical microscopy, and electrometer with large‐area electrodes. The results showed that the facile and powerful melt‐mixing method is effective in homogeneously dispersing CNFs in the PEI matrix. The flexural and tribological characteristics were investigated and the formation of spatial networks of CNFs and weak interfacial bonding were considered as competitive factors to enhanced flexural properties. The composites with 1.0 wt% CNFs showed flexural strength and toughness increased by more than 50 and 550%, respectively, but showed very high wear rate comparable with that of pure PEI. The length of the CNFs also exerted great influences on both mechanical and tribological behaviors. POLYM. ENG. SCI., 50:1914–1922, 2010. © 2010 Society of Plastics Engineers     

Friction stir welding between extrusions and laminates

Current market demands drive companies to innovate their production techniques to improve products and simultaneously hold down costs. In the search for solutions aimed at an optimization of production processes, semi-finished products in aluminium alloys very definitely play a significant role. Moreover, the assembly of extruded parts with laminates may result in considerable difficulties, especially in relation to distortions or generated residual stress and process productivity. A solution to these issues may be found in the application of friction stir welding (FSW), characterized by low thermal input and high productivity. The asymmetry of the FSW process determines a different thermal input to the edges of the pieces to be joined. This aspect can be exploited when it is necessary to join pieces characterized by a very different thermal lag, as occurs in the case of joining extruded products and sheeting. This study aims to identify optimal FW parameters, placing particular attention on heat transferred to materials so as to provide an adequate thermal input that will allow for compensation of the different thermal capacity of the pieces in question.     

Countercurrent supercritical fluid extraction of grape-spirit

A procedure for the recovery of aromatic extracts from raw grape-spirit (alcohol strength 73 vol.%) by means of a countercurrent supercritical fluid extraction (CC-SFE) on a pilot plant scale is presented. The effect of the solvent-to-feed ratio at 7, 8, 10, and 20 kg_CO2/kg_Feed on the extraction of flavor compounds, operating at constant pressure of 100 bar and extraction temperature of 50 °C have been studied. Increasing the solvent-to-feed ratio from 7 to 20 kg_CO2/kg_Feed, the total volatile compounds yield decreases from 64 to 5% and that of ethanol from 85 to 49%. The highest concentration of volatile compounds has been obtained at 7 and 8 kg_CO2/kg_Feed solvent-to-feed ratio. Mixtures containing grape-spirit (alcohol strength 40 vol.%) added with 30% (v/v) of different extracts have been tested and judged by a panel. Sensory evaluation indicated as preferred the mixtures added with the extracts obtained at lowest solvent-to-feed ratio.     

Sofosbuvir Selects for Drug-Resistant Amino Acid Variants in the Zika Virus RNA-Dependent RNA-Polymerase Complex In Vitro

The nucleotide analog sofosbuvir, licensed for the treatment of hepatitis C, recently revealed activity against the Zika virus (ZIKV) in vitro and in animal models. However, the ZIKV genetic barrier to sofosbuvir has not yet been characterized. In this study, in vitro selection experiments were performed in infected human hepatoma cell lines. Increasing drug pressure significantly delayed viral breakthrough (p = 0.029). A double mutant in the NS5 gene (V360L/V607I) emerged in 3 independent experiments at 40–80 µM sofosbuvir resulting in a 3.9 ± 0.9-fold half- maximal inhibitory concentration (IC₅₀) shift with respect to the wild type (WT) virus. A triple mutant (C269Y/V360L/V607I), detected in one experiment at 80 µM, conferred a 6.8-fold IC₅₀ shift with respect to the WT. Molecular dynamics simulations confirmed that the double mutant V360L/V607I impacts the binding mode of sofosbuvir, supporting its role in sofosbuvir resistance. Due to the distance from the catalytic site and to the lack of reliable structural data, the contribution of C269Y was not investigated in silico. By a combination of sequence analysis, phenotypic susceptibility testing, and molecular modeling, we characterized a double ZIKV NS5 mutant with decreased sofosbuvir susceptibility. These data add important information to the profile of sofosbuvir as a possible lead for anti-ZIKV drug development.