Atomic Force Microscopy and Voltammetric Investigation of Quadruplex Formation between a Triazole-Acridine Conjugate and Guanine-Containing Repeat DNA Sequences

The interactions of the Tetrahymena telomeric repeat sequence d(TG4T) and the polyguanylic acid (poly(G)) sequence with the quadruplex-targeting triazole-linked acridine ligand GL15 were investigated using atomic force microscopy (AFM) at a highly oriented pyrolytic graphite and voltammetry at a glassy carbon electrode. GL15 interacted with both sequences, in a time dependent manner, and G-quadruplex formation was detected. AFM showed the adsorption of quadruplexes as small d(TG4T) and poly(G) spherical aggregates and large quadruplex-based poly(G) assemblies, and voltammetry showed the decrease and disappearance of GL15 and guanine oxidation peak currents and appearance of the G-quadruplex oxidation peak. The GL15 molecule strongly stabilized and accelerated G-quadruplex formation in both Na+ and K+ ion-containing solution, although only K+ promoted the formation of perfectly aligned tetra-molecular G-quadruplexes. The small-molecule complex with the d(TG4T) quadruplex is discrete and approximately globular, whereas the G-quadruplex complex with poly(G) is formed at a number of points along the length of the polynucleotide, analogous to beads on a string.Financial support from Fundação para a Ciência e Tecnologia (FCT), Grant SFRH/BPD/92726/2013 (A.-M. Chiorcea-Paquim), Project Grant (A.D.R. Pontinha), projects PTDC/SAU-BMA/118531/2010, PTDC/QEQ-MED/0586/2012, PEst-C/EME /UI0285/2013 and CENTRO-07-0224-FEDER-002001 (MT4MOBI) (co-financed by the European Community Fund FEDER), FEDER funds through the program COMPETE - Programa Operacional Factores de Competitividade is gratefully acknowledged. Work in the S.N. laboratory was supported by Programme Grant No. C129/A4489, from Cancer Research UK, and by the FP6 framework grant “Molecular Cancer Medicine” from the EU. S.S. was a Maplethorpe Fellow of The University of London.Peer reviewe

Mecanismos redox de fármacos e deteção eletroquímica in situ de danos causados ao DNA

Tese de doutoramento em Bioquímica, na especialidade de Tecnologia Bioquímic

Silica Aerogel-Rubber Composite: A Sustainable Alternative for Buildings’ Thermal Insulation

Silica aerogel composites with recycled tire rubber have been synthesized and evaluated for their potential use for thermal protection in buildings. The present work describes for the first time the preparation of silica-based aerogel composites containing recycled rubber tires reinforced with polyvinyl butyral (PVB) by hot pressing. The developed composite was extensively characterized regarding its physical, morphological, thermal and mechanical features, and the results showed their properties were relevant, leading to composites with different properties/performances. The obtained bulk density values were satisfactory, down to 474 kg·m−3, and very good thermal properties were achieved, namely, thermal conductivity as low as 55 mW·m−1·K−1 for composites with silica aerogel, recycled tire rubber and PVB. The most promising composites were those based on low bulk density and thermal conductivity values, and they were thermally stable, indicating their suitability for thermal insulation applications

Thermomechanical Performance Assessment of Sustainable Buildings' Insulating Materials under Accelerated Ageing Conditions

The reliable characterization of insulation materials in relevant environmental conditions is crucial, since it strongly influences the performance (e.g., thermal) of building elements. In fact, their properties may vary with the moisture content, temperature, ageing degradation, etc. Therefore, in this work, the thermomechanical behaviour of different materials was compared when subjected to accelerated ageing. Insulation materials that use recycled rubber in their composition were studied, along with others for comparison: heat-pressed rubber, rubber_cork composites, aerogel_rubber composite (developed by the authors), silica aerogel, and extruded polystyrene. The ageing cycles comprised dry-heat, humid-heat, and cold conditions as the stages, during cycles of 3 and 6 weeks. The materials' properties after ageing were compared with the initial values. Aerogel-based materials showed superinsulation behaviour and good flexibility due to their very high porosity and reinforcement with fibres. Extruded polystyrene also had a low thermal conductivity but exhibited permanent deformation under compression. In general, the ageing conditions led to a very slight increase in the thermal conductivity, which vanished after drying of the samples in an oven, and to a decrease in Young's moduli

Aerogel Composites Produced from Silica and Recycled Rubber Sols for Thermal Insulation

Hydrophobic rubber-silica aerogel panels (21.5 × 21.5 × 1.6 cm3) were fabricated from silica and rubber sols and reinforced with several fiber types (recycled tire textile fibers, polyester blanket, silica felt, glass wool). A recycled rubber sol was prepared using peracetic acid and incorporated for the first time in TEOS-based sol-gel chemistry. The composites exhibited good thermal stability up to 400 °C and very low thermal conductivity, in the superinsulation range when using polyester fibers (16.4 ± 1.0 mW·m−1·K−1), and of 20–30 mW·m−1·K−1 for the remaining fibers. They could also endure cyclic compression loads with near full recovery, thus showing very promising properties for insulation of buildings

In situ evaluation of anticancer drug methotrexate-DNA interaction using a DNA-electrochemical biosensor and AFM characterization

An in situ evaluation of the dsDNA-methotrexate (MTX) interaction was performed by voltammetry using a DNA-electrochemical biosensor and characterized by atomic force microscopy (AFM) at a highly oriented pyrolytic graphite (HOPG) surface. Electrochemical experiments in incubated solutions showed that the interaction of MTX with dsDNA leads to modifications to the dsDNA structure in a time-dependent manner. The AFM images show reorganization of the DNA self-assembled network on the surface of the HOPG electrode upon binding methotrexate and the formation of a more densely packed and slightly thicker MTX-dsDNA lattice with a large number of aggregates embedded into the network film. The intercalation of MTX between complementary base pairs of dsDNA lead to the increase of purine oxidation peaks due to the unwinding of the dsDNA. The dsDNA-electrochemical biosensor and the purinic homo-polynucleotide single stranded sequences of guanosine and adenosine, poly[G] and poly[A]-electrochemical biosensors, were used to investigate and understand the interaction between MTX and dsDNA.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Silica Aerogel-Polycaprolactone Scaffolds for Bone Tissue Engineering

Silica aerogel is a material composed of SiO2 that has exceptional physical properties when utilized for tissue engineering applications. Poly-ε-caprolactone (PCL) is a biodegradable polyester that has been widely used for biomedical applications, namely as sutures, drug carriers, and implantable scaffolds. Herein, a hybrid composite of silica aerogel, prepared with two different silica precursors, tetraethoxysilane (TEOS) or methyltrimethoxysilane (MTMS), and PCL was synthesized to fulfil bone regeneration requirements. The developed porous hybrid biocomposite scaffolds were extensively characterized, regarding their physical, morphological, and mechanical features. The results showed that their properties were relevant, leading to composites with different properties. The water absorption capacity and mass loss were evaluated as well as the influence of the different hybrid scaffolds on osteoblasts’ viability and morphology. Both hybrid scaffolds showed a hydrophobic character (with water contact angles higher than 90°), low swelling (maximum of 14%), and low mass loss (1–7%). hOB cells exposed to the different silica aerogel-PCL scaffolds remained highly viable, even for long periods of incubation (7 days). Considering the obtained results, the produced hybrid scaffolds may be good candidates for future application in bone tissue engineering