A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR

Intrinsically disordered proteins (IDPs) that lack stable conformations and are highly flexible have attracted the attention of biologists. Therefore, the development of a systematic method to identify polypeptide regions that are unstructured in solution is important. We have designed an “indirect/reflected” detection system for evaluating the physicochemical properties of IDPs using nuclear magnetic resonance (NMR). This approach employs a “chimeric membrane protein”-based method using the thermostable membrane protein PH0471. This protein contains two domains, a transmembrane helical region and a C-terminal OB (oligonucleotide/oligosaccharide binding)-fold domain (named NfeDC domain), connected by a flexible linker. NMR signals of the OB-fold domain of detergent-solubilized PH0471 are observed because of the flexibility of the linker region. In this study, the linker region was substituted with target IDPs. Fifty-three candidates were selected using the prediction tool POODLE and 35 expression vectors were constructed. Subsequently, we obtained ¹⁵N-labeled chimeric PH0471 proteins with 25 IDPs as linkers. The NMR spectra allowed us to classify IDPs into three categories: flexible, moderately flexible, and inflexible. The inflexible IDPs contain membrane-associating or aggregation-prone sequences. This is the first attempt to use an indirect/reflected NMR method to evaluate IDPs and can verify the predictions derived from our computational tools.     

Correction to: Designing cost-efficient inspection schemes for stochastic streamflow environment using an effective Hamiltonian approach

Optimization and Engineering -     

Manufacturing of advanced smart tooling for metal forming

Forming is widely used due to its high efficiency in material utilization and its high production rate in general. Most forming processes control the geometry of final products through a set of tooling. The increasing demands on lightweight products have challenged the performances and functionalities of tooling. This paper provides a systematic review of recent advancements related to tooling performance and functionalities, including tool materials, tooling fabrication processes (e.g., machining, heat treatment, coating, surface texturing, and additive manufacturing), sensing, and data analytics. Finally, recommendations on future research directions for metal forming tooling are provided.     

NMR-Guided Repositioning of Non-Steroidal Anti-Inflammatory Drugs into Tight Junction Modulators

Bioavailability is a major bottleneck in the clinical application of medium molecular weight therapeutics, including protein and peptide drugs. Paracellular transport of these molecules is hampered by intercellular tight junction (TJ) complexes. Therefore, safe chemical regulators for TJ loosening are desired. Here, we showed a potential application of select non-steroidal anti-inflammatory drugs (NSAIDs) as TJ modulators. Based on our previous observation that diclofenac and flufenamic acid directly bound various PDZ domains with a broad specificity, we applied solution nuclear magnetic resonance techniques to examine the interaction of other NSAIDs and the first PDZ domain (PDZ1) of zonula occludens (ZO)-1, ZO-1(PDZ1). Inhibition of ZO-1(PDZ1) is expected to provide loosening of the epithelial barrier function because the domain plays a crucial role in maintaining TJ integrity. Accordingly, diclofenac and indomethacin were found to decrease the subcellular localization of claudin (CLD)-2 but not occludin and ZO-1 at the apicolateral intercellular compartment of Madin–Darby canine kidney (MDCK) II cells. These NSAIDs exhibited 125–155% improved paracellular efflux of fluorescein isothiocyanate insulin for the Caco-2 cell monolayer. We propose that these NSAIDs can be repurposed as drug absorption enhancers for peptide drugs.     

Investigation on twisting distortion of thin plate stiffened structure under welding

The welding distortion of thin plate was selected as research subject, which causes loss of dimensional control, structural integrity and increases fabrication costs for straightening. To study the characteristic of the welding distortion on thin plate structures, experiments were conducted. The test model was a thin plate stiffened structure and a large twisting distortion was observed. The welding distortion of the same structure was analyzed as a large deformation problem using a thermal elastic plastic FEM and an elastic FEM based on the concept of inherent deformation. The computed results by both methods showed the twisting distortion which is a typical buckling type deformation and the magnitude of this distortion agreed well with the experimental measurement.     

添加钛、铌、锰、硫及镁对亚共晶灰铸铁初生奥氏体形核及生长的影响

共晶铸铁在共晶凝固时,熔体中碳的状态,即“C的微小集合体”已经被深入研究.然而时于与初生奥氏体密切相关的铁原子的状态,即“铁的微小集合体”却缺乏研究.本文作者提出,在亚共晶灰铸铁熔体中,初晶γ的形核受“Fe的富集部”的影响,而“Fe的富集部”的形成及消失与“C的微小集合体”有关.研究表明,初生奥氏体的形核过冷度、形核数和晶核生长形貌随工艺条件如过热度、保温时间和冷却速率而变化.本文讨论了在熔体的过热度、保温时间和冷却速率固定的条件下,钛、铌、锰、硫和镁这五种添加元素对亚共晶灰铸铁初晶γ的凝固过程的影响.     

Rapidly solidified thick stainless cast iron alloy deposit with niobium carbide particles produced by plasma spraying

In plasma spraying process, spray material is heated, melted and accelerated by a high temperature flame. Low-pressure plasma spraying can produce rapidly solidified thick materials because alloy droplets accumulate successively on the substrate, and solidify at a cooling rate in the range of 10⁵–10⁸ ks^?1. Depending on the cooling conditions of the substrate and on the alloy composition, deposits can be produced as metastable phases or extremely fine crystalline phases. Plasma spraying is an attractive method for the production of deposits with in-situ formed fine particles. In recent years, much attention has been paid to stainless cast iron with vanadium carbide, which is attractive for use in metal molds and pump parts, due to its high wear resistance and high corrosion resistance. In the present work, stainless cast iron alloy powder was low-pressure plasma sprayed to produce stainless cast iron base alloy deposits with finely dispersed niobium carbide particles. The as-sprayed deposit and deposits obtained by heat treatment of the as-sprayed deposit had niobium carbide particles. The carbide particles in the as-sprayed deposit produced on a water-cooled substrate were finer than that in the as-sprayed deposit produced on a non-cooled substrate. With increasing heat treatment temperature up to 1273 K, the carbide particles coarsened. The as-sprayed deposit produced on a non-cooled substrate had higher hardness than the heat-treated deposits.     

Surface structure and visible light photocatalytic activity of titanium–calcium hydroxyapatite modified with Cr(III)

The synthetic titanium–calcium hydroxyapatite (Ti–CaHap) particles were treated with different concentrations of aqueous Cr(NO₃)₃·9H₂O solution and the materials obtained were characterized by a variety of conventional techniques. The crystal structure and particle morphology of Ti–CaHap were essentially not altered by treating with Cr(III) solution. With increasing the Cr(III) concentration, the amount of Cr(III) in the products was increased and that of Ca(II) was decreased. XPS results revealed that the surface state of Cr of Ti–CaHap was trivalent. These facts allow us to infer that the Cr(III) was doped by substitution of surface Ca(II) of Ti–CaHap. Besides, IR results proved that increasing the Cr(III) concentration developed the surface Cr–OH band while the surface Ti–OH and P–OH bands of Ti–CaHap vanished. This imply that the formation of surface P–O-Cr(OH)₂ and Ti–O–Cr(OH)₂ groups, resulting the Cr(OH)₃-like layer on the surface of Ti–CaHap particles. The Cr(III)-doped Ti–CaHap possessed the absorption peaks at 446 and 623 nm in vis range in addition to the UV absorption of charge transfer transition of O^2? → Ti⁴⁺. The vis absorption peaks developed on raising the Cr(III) concentration. The photocatalytic decomposition of acetaldehyde into CO₂ over Cr(III)-doped Ti–CaHap was detected under vis irradiation and the activity was lowered by the formation of Cr(OH)₃-like layer on the particle surface.