<Superscript>57</Superscript>Fe emission Mössbauer spectroscopy following dilute implantation of <Superscript>57</Superscript>Mn into In <Subscript>2</Subscript>O<Subscript>3</Subscript>

Emission Mössbauer spectroscopy has been utilised to characterize dilute ⁵⁷Fe impurities in In ₂O₃ following implantation of ⁵⁷Mn (T _1/2 = 1.5 min.) at the ISOLDE facility at CERN. From stoichiometry considerations, one would expect Fe to adopt the valence state 3 + , substituting In ³⁺, however the spectra are dominated by spectral lines due to paramagnetic Fe²⁺. Using first principle calculations in the framework of density functional theory (DFT), the density of states of dilute Fe and the hyperfine parameters have been determined. The hybridization between the 3d-band of Fe and the 2p band of oxygen induces a spin-polarized hole on the O site close to the Fe site, which is found to be the cause of the Fe²⁺ state in In ₂O₃. Comparison of experimental data to calculated hyperfine parameters suggests that Fe predominantly enters the 8b site rather than the 24d site of the cation site in the Bixbyite structure of In ₂O₃. A gradual transition from an amorphous to a crystalline state is observed with increasing implantation/annealing temperature.     

Engineering Protein Venoms as Self-Assembling CXCR4-Targeted Cytotoxic Nanoparticles

Protein venoms are effective cytotoxic molecules that when conveniently targeted to tumoral markers can be exploited as promising anticancer drugs. Here, it is explored whether the structurally unrelated melittin, gomesin, and CLIP71 could be functionally active when engineered, in form of GFP fusions, as self-assembling multimeric nanoparticles. Incorporated in modular constructs including a C-terminal polyhistidine tag and an N-terminal peptidic ligand of the cytokine receptor CXCR4 (overexpressed in more than 20 human neoplasias), these venoms are well produced in recombinant bacteria as proteolytically stable regular nanoparticles ranging between 12 and 35 nm. Being highly fluorescent, these materials selectively penetrate, label, and kill CXCR4⁺ tumor cells in a CXCR4-dependent fashion. The obtained data support the concept of recombinant venoms as promising drugs, through the precise formulation as tumor-targeted nanomaterials for selective theragnostic applications in CXCR4⁺ cancers.     

Lagrangian Decomposition for large-scale two-stage stochastic mixed 0-1 problems

In this paper we study solution methods for solving the dual problem corresponding to the Lagrangian Decomposition of two-stage stochastic mixed 0-1 models. We represent the two-stage stochastic mixed 0-1 problem by a splitting variable representation of the deterministic equivalent model, where 0-1 and continuous variables appear at any stage. Lagrangian Decomposition (LD) is proposed for satisfying both the integrality constraints for the 0-1 variables and the non-anticipativity constraints. We compare the performance of four iterative algorithms based on dual Lagrangian Decomposition schemes: the Subgradient Method, the Volume Algorithm, the Progressive Hedging Algorithm, and the Dynamic Constrained Cutting Plane scheme. We test the tightness of the LD bounds in a testbed of medium- and large-scale stochastic instances.     

Efficient bioactive oligonucleotide-protein conjugation for cell-targeted cancer therapy

Oligonucleotide-protein conjugates have important applications in biomedicine. Simple and efficient methods are described for the preparation of these conjugates. Specifically, we describe a new method in which a bifunctional linker is attached to thiol-oligonucleotide to generate a reactive intermediate that is used to link to the protein. Having similar conjugation efficacy compared with the classical method in which the bifunctional linker is attached first to the protein, this new approach produces significantly more active conjugates with higher batch to batch reproducibility. In a second approach, direct conjugation is proposed using oligonucleotides carrying carboxyl groups. These methodologies have been applied to prepare nanoconjugates of an engineered nanoparticle protein carrying a T22 peptide with affinity for the CXCR4 chemokine receptor and oligomers of the antiproliferative nucleotide 2’-deoxy-5-fluorouridine in a very efficient way. The protocols have potential uses for the functionalization of proteins, amino-containing polymers or amino-lipids in order to produce complex therapeutic nucleic acid delivery systems.     

A Native Ternary Complex Trapped in a Crystal Reveals the Catalytic Mechanism of a Retaining Glycosyltransferase

Glycosyltransferases (GTs) comprise a prominent family of enzymes that play critical roles in a variety of cellular processes, including cell signaling, cell development, and host–pathogen interactions. Glycosyl transfer can proceed with either inversion or retention of the anomeric configuration with respect to the reaction substrates and products. The elucidation of the catalytic mechanism of retaining GTs remains a major challenge. A native ternary complex of a GT in a productive mode for catalysis is reported, that of the retaining glucosyl‐3‐phosphoglycerate synthase GpgS from M. tuberculosis in the presence of the sugar donor UDP‐Glc, the acceptor substrate phosphoglycerate, and the divalent cation cofactor. Through a combination of structural, chemical, enzymatic, molecular dynamics, and quantum‐mechanics/molecular‐mechanics (QM/MM) calculations, the catalytic mechanism was unraveled, thereby providing a strong experimental support for a front–side substrate‐assisted S_Ni‐type reaction.     

57Fe Emission Mössbauer Study on Gd3Ga5O12 implanted with dilute57Mn

⁵⁷Fe emission Mössbauer spectroscopy has been applied to study the lattice location and properties of Fe in gadolinium gallium garnet Gd₃Ga₅O₁₂ (GGG) single crystals in the temperature interval 300 – 563 K within the extremely dilute (<10^?4 at.%) regime following the implantation of⁵⁷Mn (T¹_/2= 1.5 min.) at ISOLDE/CERN. These results are compared with earlier Mössbauer spectroscopy study of Fe-doped gadolinium gallium garnet Gd₃Ga₅O₁₂(GGG), with implantation fluences between 8×10¹⁵ and 6×10¹⁶ atoms cm^?2. Three Fe components are observed in the emission Mössbauer spectra: (i) high spin Fe²⁺ located at damage sites due to the implantation process, (ii) high spin Fe³⁺ at substitutional tetrahedral Ga sites, and (iii) interstitial Fe, probably due to the recoil imparted on the daughter^57?Fe nucleus in the β^? decay of⁵⁷Mn. In contrast to high fluence⁵⁷Fe implantation studies the Fe³⁺ ions are found to prefer the tetrahedral Ga site over the octahedral Ga site. No annealing stages are evident in the temperature range investigated. Despite the very low concentration, high-spin Fe³⁺ shows fast spin relaxation, presumably due to an indirect interaction between nearby gadolinium atoms.     

Sub-lattice polarization states in anti-ferroelectrics and their relaxation process

We report studies of quasi-remanent polarization states in Pb_0.99Nb_0.02[(Zr_0.57Sn_0.43)_0.94Ti_0.06]_0.98O₃ (PNZST) anti-ferroelectric ceramics and investigation of their relaxation effects using unique in-situ electrically activated time-resolved Synchrotron X-ray powder diffraction (SXPD) and ¹¹⁹Sn Mössbauer Spectroscopy (MS). The SXPD patterns are consistent with a phase transition from quasi-tetragonal perovskite in 0 V relaxed anti-ferroelectric state to rhombohedral distortion in ferroelectric state under saturating applied voltages of ±2 kV. The observed quasi-remanent polarization relaxation processes are due to the fact that tetragonal to rhombohedral distortion does not occur at the applied voltage required to access the quasi-remanent polarization states, and the tetragonal symmetry restored after the removal of the applied electric field is preserved. Since these quasi-remanent polarization states were seen as possibly suitable for memory applications, the implications of this study are that anti-ferroelectrics are more feasible for multi-state dynamic random access memories (DRAM), while their application to non-volatile memories requires development of more sophisticated “read-out” protocols, possibly involving dc electrical biasing.     

Nanostructure Empowers Active Tumor Targeting in Ligand‐Based Molecular Delivery

Cell‐selective targeting is expected to enhance effectiveness and minimize side effects of cytotoxic agents. Functionalization of drugs or drug nanoconjugates with specific cell ligands allows receptor‐mediated selective cell delivery. However, it is unclear whether the incorporation of an efficient ligand into a drug vehicle is sufficient to ensure proper biodistribution upon systemic administration, and also at which extent biophysical properties of the vehicle may contribute to the accumulation in target tissues during active targeting. To approach this issue, structural robustness of self‐assembling, protein‐only nanoparticles targeted to the tumoral marker CXCR4 is compromised by reducing the number of histidine residues (from six to five) in a histidine‐based architectonic tag. Thus, the structure of the resulting nanoparticles, but not of building blocks, is weakened. Upon intravenous injection in animal models of human CXCR4⁺ colorectal cancer, the administered material loses the ability to accumulate in tumor tissue, where it is only transiently found. It instead deposits in kidney and liver. Therefore, precise cell‐targeted delivery requires not only the incorporation of a proper ligand that promotes receptor‐mediated internalization, but also, unexpectedly, its maintenance of a stable multimeric nanostructure that ensures high ligand exposure and long residence time in tumor tissue.