Biochemical and structural characterization of β-lactamases tem-180 and tem-201

With the constant development of new antibiotics, selective pressure is a force to reckon when investigating antibiotic resistance. Although advantageous for medical treatments, it leads to increasing resistance. It is essential to use more potent and toxic antibiotics. Enzymes capable of hydrolyzing antibiotics are among the most common ways of resistance and TEM variants have been detected in several resistant isolates. Due to the rapid evolution of these variants, complex phenotypes have emerged and the need to understand their biological activity becomes crucial. To investigate the biochemical properties of TEM-180 and TEM-201 several computational methodologies have been used, allowing the comprehension of their structure and catalytic activity, which translates into their biological phenotype. In this work we intent to characterize the interface between these proteins and the several antibiotics used as ligands. We performed explicit solvent molecular dynamics (MD) simulations of these complexes and studied a variety of structural and energetic features. The interfacial residues show a distinct behavior when in complex with different antibiotics. Nevertheless, it was possible to identify some common Hot Spots among several complexes – Lys73, Tyr105 and Glu166. The structural changes that occur during the Molecular Dynamic (MD) simulation lead to the conclusion that these variants have an inherent capacity of adapting to the various antibiotics. This capability might be the reason why they can hydrolyze antibiotics that have not been described until now to be degraded by TEM variants. The results obtained with computational and experimental methodologies for the complex with Imipenem have shown that in order to this type of enzymes be able to acylate the antibiotics, they need to be capable to protect the ligand from water molecules.

Structural, physical, and chemical modifications induced by microwave heating on native agar-like galactans

Native agars from Gracilaria vermiculophylla produced in sustainable aquaculture systems (IMTA) were extracted under conventional (TWE) and microwave (MAE) heating. The optimal extracts from both processes were compared in terms of their properties. The agars’ structure was further investigated through Fourier transform infrared and NMR spectroscopy. Both samples showed a regular structure with an identical backbone, β-D-galactose (G) and 3,6-anhydro-α-L-galactose (LA) units; a considerable degree of methylation was found at C6 of the G units and, to a lesser extent, at C2 of the LA residues. The methylation degree in the G units was lower for MAEopt agar; the sulfate content was also reduced. MAE led to higher agar recoveries with drastic extraction time and solvent volume reductions. Two times lower values of [η] and Mv obtained for the MAEopt sample indicate substantial depolymerization of the polysaccharide backbone; this was reflected in its gelling properties; yet it was clearly appropriate for commercial application in soft-texture food products.

Structural modification of TiO2 nanorod films with an influence on the photovoltaic efficiency of a dye-sensitized solar cell (DSSC).

TiO2 nanorod films have been deposited on ITO substrates by dc reactive magnetron sputtering technique. The structures of these nanorod films were modified by the variation of the oxygen pressure during the sputtering process. Although all these TiO2 nanorod films deposited at different oxygen pressures show an anatase structure, the orientation of the nanorod films varies with the oxygen pressure. Only a very weak (101) diffraction peak can be observed for the TiO2 nanorod film prepared at low oxygen pressure. However, as the oxygen pressure is increased, the (220) diffraction peak appears and the intensity of this diffraction peak is increased with the oxygen pressure. The results of the SEM show that these TiO2 nanorods are perpendicular to the ITO substrate. At low oxygen pressure, these sputtered TiO2 nanorods stick together and have a dense structure. As the oxygen pressure is increased, these sputtered TiO2 nanorods get separated gradually and have a porous structure. The optical transmittance of these TiO2 nanorod films has been measured and then fitted by OJL model. The porosities of the TiO2 nanorod films have been calculated. The TiO2 nanorod film prepared at high oxygen pressure shows a high porosity. The dye-sensitized solar cells (DSSCs) have been assembled using these TiO2 nanorod films prepared at different oxygen pressures as photoelectrode. The optimum performance was achieved for the DSSC using the TiO2 nanorod film with the highest (220) diffraction peak and the highest porosity.

Study of optical and structural properties of Cu2ZnSnS4 thin films

Cu2ZnSnS4 is a promising semiconductor to be used as absorber in thin film solar cells. In this work, we investigated optical and structural properties of Cu2ZnSnS4 thin films grown by sulphurization of metallic precursors deposited on soda lime glass substrates. The crystalline phases were studied by X-ray diffraction measurements showing the presence of only the Cu2ZnSnS4 phase. The studied films were copper poor and zinc rich as shown by inductively coupled plasma mass spectroscopy. Scanning electron microscopy revealed a good crystallinity and compactness. An absorption coefficient varying between 3 and 4×104cm−1 was measured in the energy range between 1.75 and 3.5 eV. The band gap energy was estimated in 1.51 eV. Photoluminescence spectroscopy showed an asymmetric broad band emission. The dependence of this emission on the excitation power and temperature was investigated and compared to the predictions of the donor-acceptor-type transitions and radiative recombinations in the model of potential fluctuations. Experimental evidence was found to ascribe the observed emission to radiative transitions involving tail states created by potential fluctuations.

Tensile behaviour of a structural adhesive at high temperatures by the extended finite element method

Component joining is typically performed by welding, fastening, or adhesive-bonding. For bonded aerospace applications, adhesives must withstand high-temperatures (200°C or above, depending on the application), which implies their mechanical characterization under identical conditions. The extended finite element method (XFEM) is an enhancement of the finite element method (FEM) that can be used for the strength prediction of bonded structures. This work proposes and validates damage laws for a thin layer of an epoxy adhesive at room temperature (RT), 100, 150, and 200°C using the XFEM. The fracture toughness (G Ic ) and maximum load ( ); in pure tensile loading were defined by testing double-cantilever beam (DCB) and bulk tensile specimens, respectively, which permitted building the damage laws for each temperature. The bulk test results revealed that decreased gradually with the temperature. On the other hand, the value of G Ic of the adhesive, extracted from the DCB data, was shown to be relatively insensitive to temperature up to the glass transition temperature (T g ), while above T g (at 200°C) a great reduction took place. The output of the DCB numerical simulations for the various temperatures showed a good agreement with the experimental results, which validated the obtained data for strength prediction of bonded joints in tension. By the obtained results, the XFEM proved to be an alternative for the accurate strength prediction of bonded structures.

Ligand-Induced structural changes in TEM‑1 probed by molecular dynamics and relative binding free energy calculations

The TEM family of enzymes has had a crucial impact on the pharmaceutical industry due to their important role in antibiotic resistance. Even with the latest technologies in structural biology and genomics, no 3D structure of a TEM- 1/antibiotic complex is known previous to acylation. Therefore, the comprehension of their capability in acylate antibiotics is based on the protein macromolecular structure uncomplexed. In this work, molecular docking, molecular dynamic simulations, and relative free energy calculations were applied in order to get a comprehensive and thorough analysis of TEM-1/ampicillin and TEM-1/amoxicillin complexes. We described the complexes and analyzed the effect of ligand binding on the overall structure. We clearly demonstrate that the key residues involved in the stability of the ligand (hot-spots) vary with the nature of the ligand. Structural effects such as (i) the distances between interfacial residues (Ser70−Oγ and Lys73−Nζ, Lys73−Nζ and Ser130−Oγ, and Ser70−Oγ−Ser130−Oγ), (ii) side chain rotamer variation (Tyr105 and Glu240), and (iii) the presence of conserved waters can be also influenced by ligand binding. This study supports the hypothesis that TEM-1 suffers structural modifications upon ligand binding.

Optimization study of hybrid spot-welded/bonded single-lap joints

Joining of components with structural adhesives is currently one of the most widespread techniques for advanced structures (e.g., aerospace or aeronautical). Adhesive bonding does not involve drilling operations and it distributes the load over a larger area than mechanical joints. However, peak stresses tend to develop near the overlap edges because of differential straining of the adherends and load asymmetry. As a result, premature failures can be expected, especially for brittle adhesives. Moreover, bonded joints are very sensitive to the surface treatment of the material, service temperature, humidity and ageing. To surpass these limitations, the combination of adhesive bonding with spot-welding is a choice to be considered, adding a few advantages like superior static strength and stiffness, higher peeling and fatigue strength and easier fabrication, as fixtures during the adhesive curing are not needed. The experimental and numerical study presented here evaluates hybrid spot-welded/bonded single-lap joints in comparison with the purely spot-welded and bonded equivalents. A parametric study on the overlap length (LO) allowed achieving different strength advantages, up to 58% compared to spot-welded joints and 24% over bonded joints. The Finite Element Method (FEM) and Cohesive Zone Models (CZM) for damage growth were also tested in Abaqus® to evaluate this technique for strength prediction, showing accurate estimations for all kinds of joints.

A wireless sensor network platform for structural health monitoring: enabling accurate and synchronized measurements through COTS+custom-based design

Structural health monitoring has long been identified as a prominent application of Wireless Sensor Networks (WSNs), as traditional wired-based solutions present some inherent limitations such as installation/maintenance cost, scalability and visual impact. Nevertheless, there is a lack of ready-to-use and off-the-shelf WSN technologies that are able to fulfill some most demanding requirements of these applications, which can span from critical physical infrastructures (e.g. bridges, tunnels, mines, energy grid) to historical buildings or even industrial machinery and vehicles. Low-power and low-cost yet extremely sensitive and accurate accelerometer and signal acquisition hardware and stringent time synchronization of all sensors data are just examples of the requirements imposed by most of these applications. This paper presents a prototype system for health monitoring of civil engineering structures that has been jointly conceived by a team of civil, and electrical and computer engineers. It merges the benefits of standard and off-the-shelf (COTS) hardware and communication technologies with a minimum set of custom-designed signal acquisition hardware that is mandatory to fulfill all application requirements.

Adherend thickness effect on the tensile fracture toughness of a structural adhesive using an optical data acquisition method

Adhesive bonding is nowadays a serious candidate to replace methods such as fastening or riveting, because of attractive mechanical properties. As a result, adhesives are being increasingly used in industries such as the automotive, aerospace and construction. Thus, it is highly important to predict the strength of bonded joints to assess the feasibility of joining during the fabrication process of components (e.g. due to complex geometries) or for repairing purposes. This work studies the tensile behaviour of adhesive joints between aluminium adherends considering different values of adherend thickness (h) and the double-cantilever beam (DCB) test. The experimental work consists of the definition of the tensile fracture toughness (GIC) for the different joint configurations. A conventional fracture characterization method was used, together with a J-integral approach, that take into account the plasticity effects occurring in the adhesive layer. An optical measurement method is used for the evaluation of crack tip opening and adherends rotation at the crack tip during the test, supported by a Matlab® sub-routine for the automated extraction of these quantities. As output of this work, a comparative evaluation between bonded systems with different values of adherend thickness is carried out and complete fracture data is provided in tension for the subsequent strength prediction of joints with identical conditions.

Mechanical and thermal characterization of a structural polyurethane adhesive modified with thermally expandable particles

Thermally expandable particles (TEPs) are used in a wide variety of applications by industry mainly for weight reduction and appearance improvement for thermoplastics, inks, and coatings. In adhesive bonding, TEPs have been used for recycling purposes. However, TEPs might be used to modify structural adhesives for other new purposes, such as: to increase the joint strength by creating an adhesive functionally modified along the overlap of the joint by gradual heating and/or to heal the adhesive in case of damage. In this study, the behaviour of a structural polyurethane adhesive modified with TEPs was investigated as a preliminary study for further investigations on the potential of TEPs in adhesive joints. Tensile bulk tests were performed to get the tensile properties of the unmodified and TEPs-modified adhesive, while Double Cantilever Beam (DCB) test was performed in order to evaluate the resistance to mode I crack propagation of unmodified and TEPs-modified adhesive. In addition, in order to investigate the behaviour of the particles while encapsulated in adhesives, a thermal analysis was done. Scanning electron microscopy (SEM) was used to examine the fracture surface morphology of the specimens. The fracture toughness of the TEPs-modified adhesive was found to increase by addition of TEPs, while the adhesive tensile strength at yield decreased. The temperature where the particles show the maximum expansion varied with TEPs concentration, decreasing with increasing the TEPs content.

Acetyl-L-Carnitine Prevents Methamphetamine-Induced Structural Damage on Endothelial Cells via ILK-Related MMP-9 Activity

Methamphetamine (METH) is a potent psychostimulant highly used worldwide. Recent studies evidenced the involvement of METH in the breakdown of the blood-brain-barrier (BBB) integrity leading to compromised function. The involvement of the matrix metalloproteinases (MMPs) in the degradation of the neurovascular matrix components and tight junctions (TJs) is one of the most recent findings in METH-induced toxicity. As BBB dysfunction is a pathological feature of many neurological conditions, unveiling new protective agents in this field is of major relevance. AcetylL-carnitine (ALC) has been described to protect the BBB function in different paradigms, but the mechanisms underling its action remain mostly unknown. Here, the immortalized bEnd.3 cell line was used to evaluate the neuroprotective features of ALC in METH-induced damage. Cells were exposed to ranging concentrations of METH, and the protective effect of ALC 1 mM was assessed 24 h after treatment. F-actin rearrangement, TJ expression and distribution, and MMPs activity were evaluated. Integrin-linked kinase (ILK) knockdown cells were used to assess role of ALC in ILK mediated METHtriggered MMPs’ activity. Our results show that METH led to disruption of the actin filaments concomitant with claudin-5 translocation to the cytoplasm. These events were mediated by MMP-9 activation in association with ILK overexpression. Pretreatment with ALC prevented METH-induced activation of MMP-9, preserving claudin-5 location and the structural arrangement of the actin filaments. The present results support the potential of ALC in preserving BBB integrity, highlighting ILK as a new target for the ALC therapeutic use.

Influence of structural features on Portuguese toddler child care quality

Whereas child care quality has been extensively studied in the U.S., there is much less information about the quality of child care in other countries.With one of the highest maternal employment rates in Europe, it is important to examine child care in Portugal. Thirty toddler classrooms in child care centers were observed. The purpose of this studywas to determine whether structural features account for overall toddler child care quality. Results showed younger and better-paid teachers provided better toddler child care quality. Space available per childwas not a statistically significant predictor of toddler child care quality.Overall quality results suggest some issues to be addressed by early education policy makers and indicate the need to promote quality in Portuguese toddler child care programs.

Hard real-time multiprocessor scheduling resilient to core failures

Presented at 21st IEEE International Conference on Embedded and Real-Time Computing Systems and Applications (RTCSA 2015). 19 to 21, Aug, 2015, pp 122-131. Hong Kong, China.

The effect of adhesive thickness on the mechanical behavior of a structural polyurethane adhesive

One parameter that influences the adhesively bonded joints performance is the adhesive layer thickness. Hence, its effect has to be investigated experimentally and should be taken into consideration in the design of adhesive joints. Most of the results from literature are for typical structural epoxy adhesives which are generally formulated to perform in thin sections. However, polyurethane adhesives are designed to perform in thicker sections and might have a different behavior as a function of adhesive thickness. In this study, the effect of adhesive thickness on the mechanical behavior of a structural polyurethane adhesive was investigated. The mode I fracture toughness of the adhesive was measured using double-cantilever beam (DCB) tests with various thicknesses of the adhesive layer ranging from 0.2 to 2 mm. In addition, single lap joints (SLJs) were fabricated and tested to assess the influence of adhesive thickness on the lap-shear strength of the adhesive. An increasing fracture toughness with increasing adhesive thickness was found. The lap-shear strength decreases as the adhesive layer gets thicker, but in contrast to joints with brittle adhesives the decrease trend was less pronounced.

Structural adhesives modified with thermally expandable particles

In this study, the behaviour of two structural adhesives modified with thermally expandable particles (TEPs) was investigated as a preliminary study for further investigations on the potential of TEPs in adhesive joints. Tensile bulk tests were performed to get the tensile properties of the adhesives and TEPs-modified adhesives. In order to determine the expansion temperature of the particles while encapsulated in these particular adhesive systems, the variation of the volume of adhesive samples modified with different TEPs concentration as a function of temperature was measured. Further, the possibility of any chemical interactions between TEPs and adhesives matrix in the TEPs-modified specimens was verified by a Fourier transform infrared spectroscopy analysis. Finally, the fracture surfaces of the unmodified and TEPs-modified specimens, as well as the dispersion and the morphology of the particles, were examined by a scanning electron microscopy analysis. It was found that the stiffness of the TEPs-modified adhesives is not affected by incorporation of TEPs in the adhesives matrix, while the tensile yield strength decreased by increasing the wt% TEPs content. In applications of such particular materials (TEPs-modified adhesives), the temperature should be controlled to stay between 90°C and 120°C in order to obtain the highest expansion ratio. At a lower temperature, not all the particles will expand, and above, the TEPs will deteriorate and as a result the TEPs-modified adhesives will deteriorate.