The future of integrated structural biology.

Instruct-ERIC, "the European Research Infrastructure Consortium for Structural biology research," is a pan-European distributed research infrastructure making high-end technologies and methods in structural biology available to users. Here, we describe the current state-of-the-art of integrated structural biology and discuss potential future scientific developments as an impulse for the scientific community, many of which are located in Europe and are associated with Instruct. We reflect on where to focus scientific and technological initiatives within the distributed Instruct research infrastructure.

Vitamin E-stabilized polyethylene shows similar survival rates at minimum 7-year follow-up compared to conventional polyethylene in primary total knee arthroplasty.

Purpose: The aim of this cross-sectional study was to compare survival, clinical and radiographic results of total knee arthroplasty (TKA) with vitamin E-stabilized polyethylene (VEPE) or conventional polyethylene (CPE) at a minimum of 7-year follow-up.

Methods: Patients who underwent primary TKA between 2011 and 2015, receiving the same cemented rotating platform knee design with VEPE or CPE tibial inserts, were identified. Patients were contacted for clinical and radiographic follow-up.

Towards cost-effective side-chain isotope labelling of proteins expressed in human cells.

Side chain isotope labelling is a powerful tool to study protein structure and interactions by NMR spectroscopy. H,C labelling of side-chain methyl groups in a deuterated background allows studying large molecules, while side-chain aromatic groups are highly sensitive to the interaction with ligands, drugs, and other proteins. In E.

A systematic study on the effect of protonation and deuteration on electron spin / in a cellular context.

In recent years, DEER experiments in pulsed EPR have garnered interest for their precise distance distribution insights in cellular and buffer setups. These measurements linked to electron spin / values of the labelled sample are impacted by the cellular environment being fully protonated or deuterated, as demonstrated in the present study.

H, C and N assignment of self-complemented MrkA protein antigen from Klebsiella pneumoniae.

Klebsiella pneumoniae (Kp) poses an escalating threat to public health, particularly given its association with nosocomial infections and its emergence as a leading cause of neonatal sepsis, particularly in low- and middle-income countries (LMICs). Host cell adherence and biofilm formation of Kp is mediated by type 1 and type 3 fimbriae whose major fimbrial subunits are encoded by the fimA and mrkA genes, respectively. In this study, we focus on MrkA subunit, which is a 20 KDa protein whose 3D molecular structure remains elusive.

Structural aspects of iron‑sulfur protein biogenesis: An NMR view.

Over the last decade, structural aspects involving iron‑sulfur (Fe/S) protein biogenesis have played an increasingly important role in understanding the high mechanistic complexity of mitochondrial and cytosolic machineries maturing Fe/S proteins. In this respect, solution NMR has had a significant impact because of its ability to monitor transient protein-protein interactions, which are abundant in the networks of pathways leading to Fe/S cluster biosynthesis and transfer, as well as thanks to the developments of paramagnetic NMR in both terms of new methodologies and accurate data interpretation. Here, we review the use of solution NMR in characterizing the structural aspects of human Fe/S proteins and their interactions in the framework of Fe/S protein biogenesis.

Kuntscher nail removal: Revision of a failed hip resurfacing above an incarcerated Kuntscher nail. A case report of a smart nail extraction technique through a custom-made instrumentation.

Removal of a retained and osseointegrated intramedullary femoral nail can represent a considerable problem, especially in the case of contemporary total hip arthroplasty or, even worse, in the case of revision hip arthroplasty. Usually, complex and invasive surgical techniques are required to remove incarcerated Kuntscher nails. We described a case of an incarcerated Kuntscher nail, inserted 39 years before, in a 75-year-old woman waiting for a revision total hip arthroplasty of a failed metal-on-metal hip resurfacing.

Short- to Mid-Term Clinical and Radiological Results of Selective Laser Melting Highly Porous Titanium Cup in Primary Total Hip Arthroplasty.

(1) : The aim of this study was to evaluate short- to mid-term clinical and radiological results in patients undergoing primary total hip arthroplasty (THA) with the use of a Selective Laser Melting 3D-printed highly porous titanium acetabular cup (Jump System Traser, Permedica Orthopaedics). (2) : We conducted a retrospective study and collected prospective data on 125 consecutive patients who underwent primary THA with the use of highly porous titanium cup. Each patient was evaluated preoperatively and postoperatively with a clinical and radiological assessment.

Controlling the incorporation of fluorinated amino acids in human cells and its structural impact.

Fluorinated aromatic amino acids (FAAs) are promising tools when studying protein structure and dynamics by NMR spectroscopy. The incorporation FAAs in mammalian expression systems has been introduced only recently. Here, we investigate the effects of FAAs incorporation in proteins expressed in human cells, focusing on the probability of incorporation and its consequences on the F NMR spectra.

Biochemical and cellular characterization of the CISD3 protein: Molecular bases of cluster release and destabilizing effects of nitric oxide.

The NEET proteins, an important family of iron-sulfur (Fe-S) proteins, have generated a strong interest due to their involvement in diverse diseases such as cancer, diabetes, and neurodegenerative disorders. Among the human NEET proteins, CISD3 has been the least studied, and its functional role is still largely unknown. We have investigated the biochemical features of CISD3 at the atomic and in cellulo levels upon challenge with different stress conditions i.

Hip arthrodesis conversion to dual mobility total hip arthroplasty in a young adult using a custom-made highly-porous titanium monobloc cup: A case report.

Introduction: Conversion of hip arthrodesis to total hip arthroplasty (THA) is an effective but challenging procedure to restore hip functionality and improve quality of life. Herein, a case of arthrodesis takedown to a dual mobility THA with a custom-made 3D-printed cup.

Presentation Of Case: Due to higher risk of instability and loosening for the altered and atrophic anatomy, a custom-made highly-porous titanium dual mobility monobloc cup has been used for a cementless conversion THA in 31-year-old patient with hip arthrodesis.

Targeting the Main Protease (M, nsp5) by Growth of Fragment Scaffolds Exploiting Structure-Based Methodologies.

The main protease M, nsp5, of SARS-CoV-2 (SCoV2) is one of its most attractive drug targets. Here, we report primary screening data using nuclear magnetic resonance spectroscopy (NMR) of four different libraries and detailed follow-up synthesis on the promising uracil-containing fragment Z604 derived from these libraries. Z604 shows time-dependent binding.

Ligand-Based Competition Binding by Real-Time F NMR in Human Cells.

The development of more effective drugs requires knowledge of their bioavailability and binding efficacy directly in the native cellular environment. In-cell nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for investigating ligand-target interactions directly in living cells. However, the target molecule may be NMR-invisible due to interactions with cellular components, while observing the ligand by H NMR is impractical due to the cellular background.

Enlarging the scenario of site directed F labeling for NMR spectroscopy of biomolecules.

The possibility of using selectively incorporated F nuclei for NMR spectroscopic studies has retrieved increasing interest in recent years. The high gyromagnetic ratio of F and its absence in native biomolecular systems make this nucleus an interesting alternative to standard H NMR spectroscopy. Here we show how we can attach a label, carrying a F atom, to protein tyrosines, through the use of a specific three component Mannich-type reaction.

Tony W. Keller (1937-2023).

Tony Keller, a pioneer in the field of Nuclear Magnetic Resonance (NMR) spectroscopy, passed away on October 27, 2023, at the age of 86 in Spiez, Switzerland. His work and vision were essential to the development and commercialization of NMR spectrometers for many areas of scientific research.

Evolution and New Generation of Dual Mobility Cups.

Although total hip arthroplasty (THA) is considered a successful procedure, hip dislocation remains the main cause of early failure. Dual mobility cups (DMCs) have been shown to significantly reduce the dislocation rate in both primary and revision THAs. During the past several decades, DMCs have evolved in design and fixation interface.

Understanding the Molecular Basis of the Multiple Mitochondrial Dysfunctions Syndrome 2: The Disease-Causing His96Arg Mutation of BOLA3.

Multiple mitochondrial dysfunctions syndrome type 2 with hyperglycinemia (MMDS2) is a severe disorder of mitochondrial energy metabolism, associated with biallelic mutations in the gene encoding for BOLA3, a protein with a not yet completely understood role in iron-sulfur (Fe-S) cluster biogenesis, but essential for the maturation of mitochondrial [4Fe-4S] proteins. To better understand the role of BOLA3 in MMDS2, we have investigated the impact of the p.His96Arg (c.

Is Modular Dual Mobility Superior to Standard Bearings for Reducing Dislocation Risk after Primary Total Hip Arthroplasty? A Retrospective Comparative Multicenter Study.

Background: Dual mobility (DM) has been proven to reduce dislocation risk after total hip arthroplasty (THA). In the last decade modular DM (modDM) constructs have been introduced to allow the use of DM articulation with standard cementless acetabular shells. However, clinical evidence of modDM effectiveness is still low in primary THA and concerns about implant-related complications are increasing.

Structural Plasticity of NFU1 Upon Interaction with Binding Partners: Insights into the Mitochondrial [4Fe-4S] Cluster Pathway.

In humans, the biosynthesis and trafficking of mitochondrial [4Fe-4S] clusters is a highly coordinated process that requires a complex protein machinery. In a mitochondrial pathway among various proposed to biosynthesize nascent [4Fe-4S] clusters, two [2Fe-2S] clusters are converted into a [4Fe-4S] cluster on a ISCA1-ISCA2 complex. Along this pathway, this cluster is then mobilized from this complex to mitochondrial apo recipient proteins with the assistance of accessory proteins.

Unraveling the mechanism of [4Fe-4S] cluster assembly on the N-terminal cluster binding site of NUBP1.

[4Fe-4S] cluster assembly in human cytosol requires both a [2Fe-2S] cluster chaperone being able to donate two [2Fe-2S] clusters and an electron donor providing two electrons to reductively couple the two [2Fe-2S] clusters into a [4Fe-4S] cluster. The mechanism through which the cytosolic [4Fe-4S] cluster assembly works is still not defined. Here, we show that a hetero-tetrameric complex formed by two molecules of cluster-reduced [2Fe-2S] -anamorsin and one molecule of dimeric cluster-oxidized [2Fe-2S] -GLRX3 orchestrates the assembly of a [4Fe-4S] cluster on the N-terminal cluster binding site of the cytosolic protein NUBP1.

Direct Expression of Fluorinated Proteins in Human Cells for F In-Cell NMR Spectroscopy.

In-cell NMR spectroscopy is a powerful approach to study protein structure and function in the native cellular environment. It provides precious insights into the folding, maturation, interactions, and ligand binding of important pharmacological targets directly in human cells. However, its widespread application is hampered by the fact that soluble globular proteins often interact with large cellular components, causing severe line broadening in conventional heteronuclear NMR experiments.