Protein crystallization still remains mostly an empirical science, as the production of crystals with the required quality for X-ray analysis is dependent on the intensive screening of the best protein crystallization and crystal’s derivatization conditions. Herein, this demanding step was addressed by the development of a high-throughput and low-budget microfluidic platform consisting of an ion exchange membrane (117 Nafion® membrane) sandwiched between a channel layer (stripping phase compartment) and a wells layer (feed phase compartment) forming 75 independent micro-contactors. This microfluidic device allows for a simultaneous and independent screening of multiple protein crystallization and crystal derivatization conditions, using Hen Egg White Lysozyme (HEWL) as the model protein and Hg2+ as the derivatizing agent. This microdevice offers well-regulated crystallization and subsequent crystal derivatization processes based on the controlled transport of water and ions provided by the 117 Nafion® membrane. Diffusion coefficients of water and the derivatizing agent (Hg2+) were evaluated, showing the positive influence of the protein drop volume on the number of crystals and crystal size. This microfluidic system allowed for crystals with good structural stability and high X-ray diffraction quality and, thus, it is regarded as an efficient tool that may contribute to the enhancement of the proteins’ crystals structural resolution.

Advances in concrete technology during the last decades have resulted in the development of materials with enhanced mechanical properties, such as High Strength Concrete (HSC), Fibre Reinforced Concrete (FRC) and Ultra-High Performance Fibre Reinforced Concrete (UHPFRC). The application of these materials in flat slabs, which are a popular structural solution in Reinforced Concrete (RC) buildings worldwide, has the potential of significantly reducing raw material consumption by enabling the design of slenderer and therefore lighter structures. However, flat slabs are susceptible to punching shear failure, which is a complex phenomenon that remains challenging, even though significant efforts have been made to experimentally study it. For advanced concrete materials (HSC, FRC and UHPFRC), the challenge is further accentuated by the continuous and rapid development of these materials. With the purpose of identifying and highlighting gaps in the published literature, a review of tests with HSC, FRC and UHPFRC slab-column connections in non-seismic and seismic loading applications is presented in this paper. It is shown that future research directions in this field include, among others, testing thicker slabs, HSC slabs with higher concrete compressive strength, HSC combined with FRC and several more cases related to seismic loading conditions.

Being able to efficiently mitigate the effects of blast loads on structures, sacrificial cladding solutions are increasingly used to protect structural elements from the effects of accidental explosions and/or terrorist attacks. The present study analyses the loss of effectiveness of a deterministically designed sacrificial cladding when variability in the material properties and uncertainties in the mechanical model are considered. The results of an experimental campaign are used to validate the numerical models that allow the deterministic design of a sacrificial cladding which successfully improves the blast resistant capabilities of a given structural element. Nonetheless, it is shown that, taking into account the probabilistic variability of key parameters is of vital importance when designing sacrificial cladding solutions, since, when not properly designed for the structural element it intends to protect, adding a sacrificial cladding might negatively impact its blast resistant capabilities. Additionally, it is concluded that the deterministic approach might be against safety. In the reported case study, when comparing the admissible charge weight yielding from the deterministic and probabilistic approaches, one verifies that the former allows a higher charge weight.

Abstract Fiber reinforced polymer (FRP) composites can be efficient for flexural strengthening of flat slabs if debonding of the FRP is postponed. However, with the increase of the flexural capacity, the flat slab becomes more susceptible to punching shear failure. In this context, four flexural or simultaneous flexural and punching shear retrofitting systems are investigated in this study to strengthen a flexure-deficient flat slab. Externally Bonded Reinforcement on Grooves (EBROG) and externally bonded reinforcement (EBR) methods are used for flexural strengthening in two cases: slabs without punching shear reinforcement and with post-installed shear bolts as shear reinforcement. According to the results, flexural strengthening of the slab using the EBR and EBROG techniques increased its load capacity by 12% and 21%, respectively. Simultaneous flexural and shear strengthening of the slab using the EBROG technique was the most effective, leading to a 57% enhancement of the load capacity. For specimens whose failure was governed by punching, comparing the results with code predictions showed that Eurocode and ACI (and the respective guide documents fib bulletin 90 and ACI 440.2R) overestimated the capacity of these specimens. In cases where failure was governed by flexure, a simple application of the yield line theory predicted reasonably well the load capacity of the specimens.

Microstructured transparent conductive oxides (TCOs) have shown great potential as photonic electrodes in photovoltaic (PV) applications, providing both optical and electrical improvements in the solar cells’ performance due to: (1) strong light trapping effects that enhance broadband light absorption in PV material and (2) the reduced sheet resistance of the front illuminated contact. This work developed a method for the fabrication and optimization of wavelength-sized indium zinc oxide (IZO) microstructures, which were soft-patterned on flexible indium tin oxide (ITO)-coated poly(ethylene terephthalate) (PET) substrates via a simple, low-cost, versatile, and highly scalable colloidal lithography process. Using this method, the ITO-coated PET substrates patterned with IZO micro-meshes provided improved transparent electrodes endowed with strong light interaction effects—namely, a pronounced light scattering performance (diffuse transmittance up to  50%). In addition, the photonic-structured IZO mesh allowed a higher volume of TCO material in the electrode while maintaining the desired transparency, which led to a sheet resistance reduction (by  30%), thereby providing further electrical benefits due to the improvement of the contact conductance. The results reported herein pave the way for a new class of photonic transparent electrodes endowed with mechanical flexibility that offer strong potential not only as advanced front contacts for thin-film bendable solar cells but also for a much broader range of optoelectronic applications.

This paper deals with the determination of upper and lower bounds for the three-dimensional undrained stability of shallow tunnels. The tunnel is circular and a distance between its face and its lining is considered. The soil shear strength is modeled using the Tresca criterion. Results of the upper and lower bounds of the stability number are presented, for several geometrical and resistance configurations and their comparison with previous results is made, showing the clear improvement obtained. Finally, equations approaching the stability number are proposed.

Journal of Trace Elements in Medicine and Biology, 68 (2021) 126837. doi:10.1016/j.jtemb.2021.126837

Ion beam analysis of fluorine has applications in research on teeth and bones, materials science, geochemistry and archaeometry. A novel PIGE (particle induced gamma-ray emission) standard free methodology for fluorine content determination for in-depth heterogeneous samples based on the excitation function of the 19F(p,p’$\gamma$)19F nuclear reaction is presented. New precise cross section measurements of this reaction in the proton energy range 2.1 to 4.1 MeV have been performed. In addition, the ERYA-Profiling code, a computer program specially developed for PIGE analysis of in-depth heterogeneous samples, employed this new excitation function in a case study where different fluorine simulated depth profiles probed the capability of insight into fluorine distributions in a given sample, showing the potential of PIGE analysis.

We present relativistic ab initio calculations of fundamental parameters for atomic selenium, based on the Multiconfiguration Dirac-Fock method. In detail, fluorescence yields and subshell linewidths, both of K shell, as well as K$\beta$ to K$\alpha$ intensity ratio are provided, showing overall agreement with previous theoretical calculations and experimental values. Relative intensities were evaluated assuming the same ionization cross-section for the K-shell hole states, leading to a statistical distribution of these initial states. A method for estimating theoretical linewidths of X-ray lines, where the lines are composed by a multiplet of fine-structure levels that are spread in energy, is proposed. This method provides results that are closer to K$\alpha$1,2 experimental width values than the usual method, although slightly higher discrepancies occur for the K$\beta$1,3 lines. This indicates some inaccuracies in the calculation of Auger rates that have a higher contribution for partial linewidths of the subshells involved in the K$\beta$1,3 profile. Apart from this, the calculated value of K$\beta$ to K$\alpha$ intensity ratio, which is less sensitive to Auger rates issues, is in excellent agreement with recommended values.

Nuclear Inst. and Methods in Physics Research, A, 1014 (2021) 165701. doi:10.1016/j.nima.2021.165701

The SOUL, or heme-binding protein HBP/SOUL, family represents a group of evolutionary conserved putative heme-binding proteins that contains a number of members in animal, plant andbacterial species. The structures of the murine form of HEBP1, or p22HBP, and the human form of HEBP2, or SOUL, have been determined in 2006 and 2011 respectively. In this work we discuss the structures of HEBP1 and HEBP2 in light of new X-ray data for heme bound murine HEBP1. The interaction between tetrapyrroles and HEBP1, initially proven to be hydrophobic in nature, was thought to also involve electrostatic interactions between heme propionate groups and positively charged amino acid side chains. However, the new X-ray structure, and results from murine HEBP1 variants and human HEBP1, confirm the hydrophobic nature of the heme-HEBP1 interaction, resulting in Kd values in the low nanomolar range, and rules out any electrostatic stabilization. Results from NMR relaxation time measurements for human HEBP1 describe a rigid globular protein with no change in motional regime upon heme binding. X-ray structures deposited in the PDB for human HEBP2 are very similar to each other and to the new heme-bound murine HEBP1 X-ray structure (backbone rmsd ca. 1 {\AA}). Results from a HSQC spectrum centred on the histidine side chain N$δ$-proton region for HEBP2 confirm that HEBP2 does not bind heme via H42 as no chemical shift differences were observed upon heme addition for backbone NH and N$δ$ protons. A survey of the functions attributed to HEBP1 and HEBP2 over the last 20 years span a wide range of cellular pathways. Interestingly, many of them are specific to higher eukaryotes, particularly mammals and a potential link between heme release under oxidative stress and human HEBP1 is also examined using recent data. However, at the present moment, trying to relate function to the involvement of heme or tetrapyrrole binding, specifically, makes little sense with our current biological knowledge and can only be applied to HEBP1, as HEBP2 does not interact with heme. We suggest that it may not be justified to call this very small family of proteins, heme-binding proteins. The family may be more correctly called “the SOUL family of proteins related to cellular fate” as, even though only HEBP1 binds heme tightly, both proteins may be involved in cell survival and/or proliferation.