The Cellulosome is an intricate macromolecular protein complex that centralizes the cellulolytic efforts of many anaerobic microorganisms through the promotion of enzyme synergy and protein stability. The assembly of numerous carbohydrate processing enzymes into a macromolecular multiprotein structure results from the interaction of enzyme-borne dockerin modules with repeated cohesin modules present in noncatalytic scaffold proteins, termed scaffoldins. Cohesin–dockerin (Coh-Doc) modules are typically classified into different types, depending on structural conformation and cellulosome role. Thus, type I Coh-Doc complexes are usually responsible for enzyme integration into the cellulosome, while type II Coh-Doc complexes tether the cellulosome to the bacterial wall. In contrast to other known cellulosomes, cohesin types from Bacteroides cellulosolvens, a cellulosome-producing bacterium capable of utilizing cellulose and cellobiose as carbon sources, are reversed for all scaffoldins, i.e., the type II cohesins are located on the enzyme-integrating primary scaffoldin, whereas the type I cohesins are located on the anchoring scaffoldins. It has been previously shown that type I B. cellulosolvens interactions possess a dual-binding mode that adds flexibility to scaffoldin assembly. Herein, we report the structural mechanism of enzyme recruitment into B. cellulosolvens cellulosome and the identification of the molecular determinants of its type II cohesin–dockerin interactions. The results indicate that, unlike other type II complexes, these possess a dual-binding mode of interaction, akin to type I complexes. Therefore, the plasticity of dual-binding mode interactions seems to play a pivotal role in the assembly of B. cellulosolvens cellulosome, which is consistent with its unmatched complexity and size.

Mechanical anchorage devices have the potential to efficiently improve the bond behavior of Carbon Fiber-Reinforced Polymers (CFRP) Externally Bonded (EB) onto steel with an adhesive. Existing relevant studies, however, have been very limited. Against this background, experimental and numerical studies on the double overlapped CFRP-to-steel bonded joints with an end mechanical anchorage were carried out in the present study. Two types of CFRP laminates (i.e., SIKA CFRP and the other type made in Harbin Institute of Technology – HIT) were used in the bond tests, with or without end mechanical anchorage devices. The test results showed that the end mechanical anchorage had marginal effect on the bond-slip relationship between EB CFRP and steel as well as debonding load, but could increase the ultimate load. It was also found that CFRP laminate from HIT had a better bond performance than that from SIKA, in terms of both debonding load and ultimate load. Using ABAQUS, Finite Element (FE) model was established on such bonded joints and the accuracy was verified with test results. The verified FE model was adopted in a parametric study to further clarify the effect of key parameters on the bond behavior of such bonded joints. © 2021 Elsevier Ltd

In this work, we present a novel application of the full-field energy-dispersive X-ray fluorescence (EDXRF) imaging system based on a MicroPattern Gaseous Detector (2D-THCOBRA) in the cultural heritage field. The detector has an intrinsic imaging capability with spatial resolution of 400μmFWHM, and is energy sensitive, presenting an energy resolution of approximately 1keVFWHM at 5.9keV. The full-field XRF scanner based on the 2D-THCOBRA detector allows mapping the distribution of elements in large area samples with high detection efficiency (75 % at 5.9keV), being a very promising choice for elemental mapping analysis of large area cultural heritage samples. In this work, we have demonstrated the imaging capabilities of the full-field XRF scanner and used it to assess the restoration of a Portuguese faience piece.

Nowadays, the use of the Digital Image Correlation (DIC) technique has spread and it is being used in several engineering areas to measure displacements. The available data obtained from the DIC measurement to evaluate the bond performance between a Carbon fibre Reinforced Polymer (CFRP) externally bonded to a timber substrate is scarce. From the existing data obtained with other materials, this contactless technique revealed to be quite useful but its accuracy with other well-established techniques, such as the use of electric strain gauges is not well understood yet. Therefore, the current work aims to study the accuracy of 2D DIC measurements with the measurements obtained from the use of strain gauges within a low-cost perspective. To that end, several CFRP-to-timber bonded joints were tested under the single-lap shear test and different bonding techniques were considered as well. Some flaws intrinsically derived from the DIC measurements that complicate the bond assessment, such as the fluctuations in the generated displacements field, are identified, and to bypass this problem a new methodology is proposed. This new methodology is based on two different closed-form solutions that, after defining the local and global bond behaviours of different CFRP-to-timber bonded joints, allowed to eliminate the fluctuations found from the DIC measurements, facilitating the estimation and the comprehension of the full debonding process of the CFRP-to-timber joints, which was achieved with a good proximity to the homologous debonding process derived from the strain gauge measurements. © 2020

Carbon fibre reinforced polymer (CFRP) composite systems are widely used to strengthen reinforced concrete (RC) structures through bonding strips/sheets on the concrete surface – externally bonded reinforcement (EBR) technique, or through strips/rods bonded inside slits in the concrete cover – near-surface mounted (NSM) technique. Although both techniques provide high strength increases, it is usually not possible to use the CFRPs’ full strength due to premature debonding, especially with EBR. This limitation can be overcome when using CREatE (continuous reinforcement embedded at ends) technique (developed by the last three authors), which consists of (i) bonding the central part of the CFRP strip (as in EBR), or the CFRP rod (as in NSM), and (ii) anchoring both ends of the strip/rod inside the concrete section, after a transition curve, enhancing its anchorage capacity. However, all these techniques are susceptible to fire, due to the polymeric nature of CFRP materials and epoxy adhesives. This paper presents the results of an experimental study regarding the fire behaviour of RC slab strips strengthened with EBR, NSM and CREatE techniques, in which the influence of applying different fire protection systems was investigated. The specimens were strengthened with those systems and simultaneously subjected to a service load and the ISO 834 fire curve. The following main results were obtained: (i) without fire protection, the CREatE technique presented the highest fire resistance due to the better anchorage of the CFRP; (ii) when protected, the NSM technique presented higher fire resistance compared to EBR and CREatE techniques; and (iii) “critical” temperatures were proposed for each technique, 1.0Tg, 2.5Tg, and 3.0Tg for EBR, NSM and CREatE techniques, respectively. © Fédération Internationale du Béton (fib) – International Federation for Structural Concrete.

After more than one hundred and thirty thousand protein structures determined by X-ray crystallography{,} the challenge of protein crystallization for 3D structure determination remains. In the quest for additives for efficient protein crystallization{,} inorganic materials emerge as an alternative. Magnetic particles (MPs) are versatile inorganic materials{,} easy to use{,} modify and manipulate in a wide range of biological assays. The potential of using functionalised MPs as crystallization chaperones for protein crystallization was shown in this work. MPs with distinct coatings were rationally designed to promote protein crystallization by affinity-triggered heterogeneous nucleation. Hen egg white lysozyme (HEWL) and trypsin{,} were crystallized in the presence of MPs either bare or coated with a polysaccharide (chitin) or a protein (casein){,} respectively. The addition of MPs was characterized in terms of bound protein to the MPs{,} crystal morphology{,} time-lapse of crystal emergence{,} crystallization yield fold change and crystal diffraction quality for structure determination. The MPs additives have shown to bind to the respective target protein{,} and to promote nucleation and crystal growth without compromising crystal morphology. On the other hand{,} MPs addition led to faster detectable crystal emergence and up to 13 times higher crystallization yield{,} addressing some the challenges in protein crystallization{,} the main bottleneck of macromolecular crystallography. Structure determination of the protein crystallized in the presence of MPs revealed that the structural characteristics of the protein remained unchanged{,} as shown by the superposition with PDB annotated proteins. Moreover{,} and unlike most reported cases{,} it was possible to exclude the inhibitor benzamidine during trypsin crystallisation{,} which is a remarkable result opening new prospects in enzyme engineering and drug design. Our results show that MPs coated with affinity ligands to target proteins can be used as controlled and tailor-made crystallization inducers.

The test of independence of two groups of variables is addressed in the case where the sample size N is considered randomly distributed. This assumption may lead to a more realist testing procedure since in many situations the sample size is not known in advance. Three sample schemes are considered where N may have a Poisson, Binomial or Hypergeometric distribution. For the case of two groups with p1 and p2 variables, it is shown that when either p1 or p2 (or both) are even the exact distribution corresponds to a finite or an infinite mixture of Exponentiated Generalized Integer Gamma distributions. In these cases a computational module is made available for the cumulative distribution function of the test statistic. When both p1 and p2 are odd, the exact distribution of the test statistic may be represented as a finite or an infinite mixture of products of independent Beta random variables whose density and cumulative distribution functions do not have a manageable closed form. Therefore, a computational approach for the evaluation of the cumulative distribution function is provided based on a numerical inversion formula originally developed for Laplace transforms. When the exact distribution is represented through infinite mixtures, an upper bound for the error of truncation of the cumulative distribution function is provided. Numerical studies are developed in order to analyze the precision of the results and the accuracy of the upper bounds proposed. A simulation study is provided in order to assess the power of the test when the sample size N is considered randomly distributed. The results are compared with the ones obtained for the fixed sample size case.