Prediction of Drug Synergism between Peptides and Antineoplastic Drugs Paclitaxel, 5-Fluorouracil,and Doxorubicin Using In Silico Approaches

Chemotherapy is the main treatment for most early-stage cancers; nevertheless, its efficacy is usually limited by drug resistance, toxicity, and tumor heterogeneity. Cell-penetrating peptides (CPPs) are small peptide sequences that can be used to increase the delivery rate of chemotherapeutic drugs to the tumor site, therefore contributing to overcoming these problems and enhancing the efficacy of chemotherapy. The drug combination is another promising strategy to overcome the aforementioned problems since the combined drugs can synergize through interconnected biological processes and target different pathways simultaneously. Here, we hypothesized that different peptides (P1–P4) could be used to enhance the delivery of chemotherapeutic agents into three different cancer cells (HT-29, MCF-7, and PC-3). In silico studies were performed to simulate the pharmacokinetic (PK) parameters of each peptide and antineoplastic agent to help predict synergistic interactions in vitro. These simulations predicted peptides P2–P4 to have higher bioavailability and lower T_max, as well as the chemotherapeutic agent 5-fluorouracil (5-FU) to have enhanced permeability properties over other antineoplastic agents, with P3 having prominent accumulation in the colon. In vitro studies were then performed to evaluate the combination of each peptide with the chemotherapeutic agents as well as to assess the nature of drug interactions through the quantification of the Combination Index (CI). Our findings in MCF-7 and PC-3 cancer cells demonstrated that the combination of these peptides with paclitaxel (PTX) and doxorubicin (DOXO), respectively, is not advantageous over a single treatment with the chemotherapeutic agent. In the case of HT-29 colorectal cancer cells, the combination of P2–P4 with 5-FU resulted in synergistic cytotoxic effects, as predicted by the in silico simulations. Taken together, these findings demonstrate that these CPP6-conjugates can be used as adjuvant agents to increase the delivery of 5-FU into HT-29 colorectal cancer cells. Moreover, these results support the use of in silico approaches for the prediction of the interaction between drugs in combination therapy for cancer.     

Chitosan biobased and intelligent films: Monitoring pH variations

Many factors can influence the shelf-life product, such as pH, water activity, nutrient levels available oxygen, etc. Variations in food pH could occur due to microorganism growth and chemical reactions that may impact flavour, consistency and shelf-life. The objective of this paper was to obtain and characterize the fast pH-colourimetric indicator device, applying a simple manufacturing technique, using food grade and biodegradable materials. The intelligent film was based on natural compounds as chitosan and anthocyanin (pH-colourimetric indicator). Chitosan intelligent films (C-ATH, 2.0 g/100 g) were obtained incorporating anthocyanin (1.0 g/100 g) in matrix films. Initially, a dark violet colour was observed in chitosan films dried. When immersed in different pH buffers, the colour range of the films varied from pink (in acid pH) to bluish-green (in neutral pH) and to violet (in basic pH). The water solubility and water vapour transmission rate of C-ATH were decreased to 60% and 48%, respectively, as compared to control film (CF, without indicator). The mechanical properties such as tensile strength and rigidity were maintained, and the elongation at break was reduced to 47% compared to CF. The advantages of this system were the simple manufacturing process, biodegradability and usage of natural and safe compounds.     

Nonstarter lactobacilli isolated from soft and semihard Argentinean cheeses: genetic characterization and resistance to biological barriers

Nonstarter lactic acid bacteria isolated from Argentinean cheeses were identified and characterized by focusing on their resistance to biological barriers, along with other physiological features of potential interest, in the search for future probiotic organisms. Lactobacilli were enumerated and isolated from semihard and soft cheeses made with multistrain Streptococcus thermophilus starters. Lactobacilli counts in 1-week-old cheeses were between 10(5) and 10(7) CFU/g and then reached 10(7) CFU/ g in all 1-month samples, while streptococci were always above 10(9) CFU/g. A total number of 22 lactobacilli isolates were retained, identified, and characterized by in vitro tests. Species identity was determined by carbohydrate metabolism and species-specific PCR assays. Genetic diversity was explored by random amplified polymorphic DNA (RAPD) PCR analysis. The Lactobacillus strains were assigned to the species L. casei, L. plantarum, L. rhamnosus, L. curvatus, L. fermentum, and L. perolens. All the strains studied tolerated 25 ppm of lysozyme, and most of them showed resistance to 0.3% bile. After incubation in gastric solution (pH 2.0), counts decreased by several log units, ranging from 3.2 to 7.0. The strains were able to grow in the presence of bile salts, but only three isolates were capable of deconjugation. The nonstarter lactobacilli that were assayed fermented the prebiotic substrates (especially lactulose and inulin). Some strains showed high cell hydrophobicity and beta-galactosidase activity, as well as inhibitory activity against pathogenic bacteria. It was concluded that most of the lactobacilli isolated in this study demonstrated resistance to biological barriers and physiological characteristics compatible with probiotic properties, which make them suitable for further research in in vivo studies aimed at identifying new probiotic organisms.     

Enzymatic de-glycosylation of rutin improves its antioxidant and antiproliferative activities

Bioavailability and biological properties of flavonoid glycosides can be improved after the enzymatic hydrolysis of specific glycosyl groups. In this study, we evaluate the antioxidant and antiproliferative potential of rutin after enzymatic hydrolysis performed by α-l-rhamnosidases (hesperidinase from Penicillium sp. and naringinase from Penicillium decumbens) previously heated at 70 °C for 30 min to inactivate the undesirable β-d-glucosidase activity. The highest in vitro antioxidant activity determined by DPPH radical scavenging was achieved with rutin hydrolyzed by hesperidinase. Rutin was predominantly bioconverted into quercetin-3-glucoside. There was no statistical difference between xanthine oxidase inhibition by rutin before and after hydrolysis. However, in vitro inhibitory activity against ten human tumor cell lines showed that hydrolyzed rutin exerted a more potent antiproliferative effect than quercetin and rutin on various cancer cell lines, specially glioma, and ovarian and breast adenocarcinomas. These results indicate that quercetin-3-glucoside could be a promising functional derivative obtained by rutin hydrolysis.     

Synthesis of poly(urethane-urea) varnish bearing azobenzene chromophoric pendants

A new diol with azoaromatic pendant was prepared by N-phenyl-4-amido-3,4-dichloromaleimide with 2-mercaptoethanol in the presence of NaOH, and used to obtain photosensible poly(urethane-urea) varnish. A poly(urethane-urea) varnish bearing azobenzene chromophores, based on a poly(ethylene adipate)diol (average molecular weight—2000), 4,4′-dibenzyldiisocyanate, diethylene glycol, trimethylolpropane, and afore-mentioned diol, were prepared and characterized. The polymers were characterized by FTIR spectroscopy, thermal analysis (DMA, DSC, and TGA), and the photochromic behavior by UV irradiation of thin films was discussed.     

Microstructure and flow behaviour during backward extrusion of semi-solid 7075 aluminium alloy

The microstructure and flow behaviour during thixo backward extrusion of 7075 aluminium alloy were investigated. Reheating the steel die and the aluminium billet placed into the die at the same time using an induction furnace provides rapidly a very homogeneous microstructure suitable for thixoforming. During thixoextrusion, despite the high solid fraction, the solid globules are weakly connected and slide over each other without any plastic deformation. The flow remains quasi homogeneous resulting in homogeneous induced microstructure of the component.     

Structural design of carbon/epoxy bio‐inspired wind turbine blade using fluid/structure simulation

The purpose of this paper is to present the structural design procedure of a low‐speed, horizontal axis, bio‐inspired wind turbine blade made of carbon/epoxy. The methodology initiates with the mechanical characterization of the carbon fiber composite material. An aerodynamic simulation using Computational Fluid Dynamics (CFD) method is performed in order to obtain the pressure distribution profile of the blade. This result is coupled with a Finite Element Analysis (FEA) to carry out an iterative design process through a Fluid‐Structure Interaction (FSI) simulation. Different stacking sequences of laminates are evaluated to find a configuration which allows balance between aerodynamic and dynamic inertial loads, ensuring an almost undeformed geometry during wind turbine's operation. The final structural design of the blade consists in six regions with different laminates. These are balanced and symmetric with distinct thickness characteristics and stacking sequences, which vary in three different orientations: 0^°, ± 45^° and 90^°, achieving a minimum deflection at the tip close to 3.11 cm, and a total weight of 3.6 kg of a 1.8 m radius blade, even with the restrictions imposed by the non‐conventional geometry. Copyright © 2016 John Wiley & Sons, Ltd.     

Molds and Resists Studies for Nanoimprint Lithography of Electrodes in Low-Voltage Polymer Thin-Film Transistors

A low-cost patterning of electrodes was investigated looking forward to replacing conventional photolithography for the processing of low-operating voltage polymeric thin-film transistors. Hard silicon, etched by sulfur hexafluoride and oxygen gas mixture, and flexible polydimethylsiloxane imprinting molds were studied through atomic force microscopy (AFM) and field emission gun scanning electron microscopy. The higher the concentration of oxygen in reactive ion etching, the lower the etch rate, sidewall angle, and surface roughness. A concentration around 30 % at 100 mTorr, 65 W and 70 sccm was demonstrated as adequate for submicrometric channels, presenting a reduced etch rate of 176 nm/min. Imprinting with positive photoresist AZ1518 was compared to negative SU-8 2002 by optical microscopy and AFM. Conformal results were obtained only with the last resist by hot embossing at 120 °C and 1 kgf/cm² for 2 min, followed by a 10 min post-baking at 100 °C. The patterning procedure was applied to define gold source and drain electrodes on oxide-covered substrates to produce bottom-gate bottom-contact transistors. Poly(3-hexylthiophene) (P3HT) devices were processed on high-κ titanium oxynitride (TiO _x N _y ) deposited by radiofrequency magnetron sputtering over indium tin oxide-covered glass to achieve low-voltage operation. Hole mobility on micrometric imprinted channels may approach amorphous silicon (～0.01 cm²/V s) and, since these devices operated at less than 5 V, they are not only suitable for electronic applications but also as sensors in aqueous media.     

Incremental forms of Schapery’s model: convergence and inversion to simulate strain controlled ramps

Schapery’s nonlinear viscoelastic model is written in incremental form, and three different approximations of nonlinearity functions in the time increment are systematically analysed with respect to the convergence rate. It is shown that secant slope is the best approximation of the time shift factor, leading to significantly higher convergence rate. This incremental form of the viscoelastic model, Zapas’ model for viscoplasticity, supplemented with terms accounting for damage effect is used to predict inelastic behaviour of material in stress controlled tests. Then the incremental formulation is inverted to simulate stress development in ramps where strain is the input parameter. A comparison with tests shows good ability of the model in inverted form to predict stress–strain response as long as the applied strain is increasing. However, in strain controlled ramps with unloading, the inverted model shows unrealistic hysteresis loops. This is believed to be a proof of the theoretically known incompatibility of the stress and strain controlled formulations for nonlinear materials. It also shows limitations of material models identified in stress controlled tests for use in strain controlled tests.