Structure-function studies of an engineered scaffold protein derived from stefin A. I: Development of the SQM variant

Non-antibody scaffold proteins are used for a range of applications, especially the assessment of protein–protein interactions within human cells. The search for a versatile, robust and biologically neutral scaffold previously led us to design STM (stefin A triple mutant), a scaffold derived from the intracellular protease inhibitor stefin A. Here, we describe five new STM-based scaffold proteins that contain modifications designed to further improve the versatility of our scaffold. In a step-by-step approach, we introduced restriction sites in the STM open reading frame that generated new peptide insertion sites in loop 1, loop 2 and the N-terminus of the scaffold protein. A second restriction site in ‘loop 2’ allows substitution of the native loop 2 sequence with alternative oligopeptides. None of the amino acid changes interfered significantly with the folding of the STM variants as assessed by circular dichroism spectroscopy. Of the five scaffold variants tested, one (stefin A quadruple mutant, SQM) was chosen as a versatile, stable scaffold. The insertion of epitope tags at varying positions showed that inserts into loop 1, attempted here for the first time, were generally well tolerated. However, N-terminal insertions of epitope tags in SQM had a detrimental effect on protein expression

Dorsal Visual Pathway Changes in Patients with Comitant Extropia

BACKGROUND: Strabismus is a disorder in which the eyes are misaligned. Persistent strabismus can lead to stereopsis impairment. The effect of strabismus on human brain is not unclear. The present study is to investigate whether the brain white structures of comitant exotropia patients are impaired using combined T1-weighted imaging and diffusion tensor imaging (DTI). PRINCIPAL FINDINGS: Thirteen patients with comitant strabismus and twelve controls underwent magnetic resonance imaging (MRI) with acquisition of T1-weighted and diffusion tensor images. T1-weighted images were used to analyze the change in volume of white matter using optimized voxel-based morphology (VBM) and diffusion tensor images were used to detect the change in white matter fibers using voxel-based analysis of DTI in comitant extropia patients. VBM analysis showed that in adult strabismus, white matter volumes were smaller in the right middle occipital gyrus, right occipital lobe/cuneus, right supramarginal gyrus, right cingulate gyrus, right frontal lobe/sub-gyral, right inferior temporal gyrus, left parahippocampa gyrus, left cingulate gyrus, left occipital lobe/cuneus, left middle frontal gyrus, left inferior parietal lobule, and left postcentral gyrus, while no brain region with greater white matter volume was found. Voxel-based analysis of DTI showed lower fractional anisotropy (FA) values in the right middle occipital gyrus and right supramarginal gyrus in strabismus patients, while brain region with increased FA value was found in the right inferior frontal gyrus. CONCLUSION: By combining VBM and voxel-based analysis of DTI results, the study suggests that the dorsal visual pathway was abnormal or impaired in patients with comitant exotropia

Effects of MoO3 + C on Crystallization and Radiative Heat Transfer of CaO–SiO2–B2O3-Based Glassy Fluoride-Free Mold Fluxes

The fluorine in traditional mold fluxes could be harmful in steel-plant environments. Accordingly, fluoride-free mold fluxes have received great attention in recent years. In this work, a method to adjust the crystallization and radiative heat transfer of fluoride-free mold fluxes is proposed. MoO3 and C mixtures (mass ratio: 4:1) were added into CaO–SiO2–B2O3-based mold fluxes and produced MoB, Mo2CB and Mo2C foreign particles. The influences of foreign particles on the radiation of glassy CaO–SiO2–B2O3-based mold fluxes were investigated by measuring the transmissivity of a glassy disk for light in the wavenumber range of 300 to 2500 nm. It was found that transmissivity in all wavenumber ranges were reduced and extinction coefficients were enhanced by the scattering of foreign particles. The effect of foreign particles on crystallization (devitrification) of bulk glassy CaO–SiO2–B2O3-based mold fluxes was also investigated. The crystallization mechanism of glassy mold fluxes disks is mainly surface crystallization. The introduction of foreign particles induced heterogeneous nucleation and the crystallization mechanism of the bulk sample with MoO3% = 2% changed into bulk crystallization

Crystallization Kinetics of CaO-SiO2(CaO/SiO2=1)-TiO2-10 mass%B2O3 Glassy Slag by Differential Thermal Analysis

Crystallization characteristics of the CaO-SiO2-TiO2-10%B2O3 glassy slag at w(CaO)/w(SiO2)=1 have been studied by Differential Thermal Analysis (DTA) and Matusita-Sakka method. Crystallization products have been distinguished by employing X-ray diffraction (XRD) and Scanning Electron Microscopy equipped with Energy Dispersive Spectroscopy (SEM-EDS). As the TiO2 content is within 10-18%, the crystal phase precipitated is mainly CaSiO3, and the effective activation energies for crystal growth increase with the increase of TiO2 content. Crystallization mechanism for CaSiO3 shifted from surface crystallization to one-dimensional growth with increase of TiO2. As the TiO2 content in slag further increases to 22% and 26%, CaTiSiO5 becomes the predominant crystal phase precipitated, and the effective activation energies for crystal growth decrease with the increase of TiO2 content. Crystallization mechanism for CaTiSiO5 is mainly surface crystallization. Therefore, with the increase of TiO2 content, the crystallization ability of the CaO-SiO2-TiO2-10%B2O3 glass system decreases initially and then increases

Effects of MoO₃ + C on Crystallization and Radiative Heat Transfer of CaO–SiO₂–B₂O₃-Based Glassy Fluoride-Free Mold Fluxes

The fluorine in traditional mold fluxes could be harmful in steel-plant environments. Accordingly, fluoride-free mold fluxes have received great attention in recent years. In this work, a method to adjust the crystallization and radiative heat transfer of fluoride-free mold fluxes is proposed. MoO3 and C mixtures (mass ratio: 4:1) were added into CaO−SiO2−B2O3-based mold fluxes and produced MoB, Mo2CB and Mo2C foreign particles. The influences of foreign particles on the radiation of glassy CaO−SiO2−B2O3-based mold fluxes were investigated by measuring the transmissivity of a glassy disk for light in the wavenumber range of 300 to 2500 nm. It was found that transmissivity in all wavenumber ranges were reduced and extinction coefficients were enhanced by the scattering of foreign particles. The effect of foreign particles on crystallization (devitrification) of bulk glassy CaO−SiO2−B2O3-based mold fluxes was also investigated. The crystallization mechanism of glassy mold fluxes disks is mainly surface crystallization. The introduction of foreign particles induced heterogeneous nucleation and the crystallization mechanism of the bulk sample with MoO3% = 2% changed into bulk crystallization

Identifying Oxide and CaS Non-Metallic Inclusions in Steel with Raman Spectroscopy

Identification and characterization play an essential role in the study of non-metallic inclusions (NMIs) in monitoring the steel casting process and product quality. This study uses Raman spectroscopy, a versatile, relatively fast, and non-destructive technique for characterizing oxide and sulfide-based inclusion phases in steel samples. The relative Raman peak intensities and positions were used to qualitatively estimate the inclusion phases of Al2O3, CA, C12A7, C3A, MgO·Al2O3, and CaS. For a quantitative estimation, FESEM-EDS (field emission scanning electron microscope with energy dispersive X-ray spectroscopy) was used to evaluate the inclusion phases in the samples. This study demonstrates that Raman spectroscopy can be used to rapidly estimate the content of Al2O3, magnesium aluminate spinel (MgO·Al2O3), and the calcium aluminate phases of CA, C3A, and C12A7 in steel samples to complement established characterization techniques for NMIs analysis such as FESEM-EDS