Visualizing Intramolecular Dynamics of Membrane Proteins

Membrane proteins play important roles in biological functions, with accompanying allosteric structure changes. Understanding intramolecular dynamics helps elucidate catalytic mechanisms and develop new drugs. In contrast to the various technologies for structural analysis, methods for analyzing intramolecular dynamics are limited. Single-molecule measurements using optical microscopy have been widely used for kinetic analysis. Recently, improvements in detectors and image analysis technology have made it possible to use single-molecule determination methods using X-rays and electron beams, such as diffracted X-ray tracking (DXT), X-ray free electron laser (XFEL) imaging, and cryo-electron microscopy (cryo-EM). High-speed atomic force microscopy (HS-AFM) is a scanning probe microscope that can capture the structural dynamics of biomolecules in real time at the single-molecule level. Time-resolved techniques also facilitate an understanding of real-time intramolecular processes during chemical reactions. In this review, recent advances in membrane protein dynamics visualization techniques were presented.     

Hematopoietic–Mesenchymal Signals Regulate the Properties of Mesenchymal Stem Cells

It is well known that the properties of hematopoietic stem/progenitor cells (HSCs), such as their self-renewal ability and multipotency, are maintained through interactions with mesenchymal stem/stromal cells (MSCs). MSCs are rare cells that are present in the bone marrow and are useful for clinical applications due to their functional ability. To obtain the necessary number of cells, MSCs must be cultured to expand, but this causes a remarkable decrease in stem cell properties, such as multipotency and proliferation ability. In this study, we show that the c-Mpl signal, which is related to the maintenance of hematopoietic stem cells, has an important effect on the proliferation and differentiation ability of MSCs. Utilizing a co-culture system comprising MSCs and HSCs, it is suggested that signaling from hematopoietic cells to MSCs supports cell proliferation. Interestingly, the enhanced proliferation ability of the HSCs was decreased in c-Mpl knock-out HSCs (c-Mpl-KO). In addition, the MSCs co-cultured with c-Mpl-KO HSCs had reduced MSC marker expression (PDGFRa and Sca-1) compared to the MSCs co-cultured with c-Mpl-wild-type HSCs. These results suggest that a hematopoietic–mesenchymal signal exists, and that the state of the HSCs is important for the stability of MSC properties.     

Determination of three dimensional in situ stress from core discing based on analysis of principal tensile stress

To determine all of the components of in situ stress from core discing, both the directions and magnitudes of the principal in situ stresses must be determined for a disc of a given thickness. In this study, we analyzed the direction and magnitude of tensile stress below an HQ core stub for 11 core lengths using stress conditions under which core discing is likely to occur. First, based on an analysis of the direction of tensile stress below the core stub, we propose a method for determining directions of in situ stress from the height distribution at the periphery of the end surface of a disc. This method can be used with a disc of any thickness. Next, based on an analysis of the magnitude of tensile stress in the central part of a core, we propose a linear criterion for core discing, which can be applied to a core of any length. This criterion was in good agreement with an empirical formula obtained previously in laboratory experiments. By combining information on the direction of in situ stress and the linear criterion for core discing, we propose a method for determining all of the components of in situ stress from core discing under the assumption that vertical stress is given by the overburden stress. Finally, these methods were applied to discs obtained from a field where hydraulic fracturing was performed to measure horizontal stresses. The results showed that the azimuths of the principal stresses estimated from core discing were consistent with those of the principal horizontal stresses determined by hydraulic fracturing and that while the magnitudes of the principal horizontal stresses estimated from core discing showed a large scatter, they were similar to those determined by hydraulic fracturing.     

Simulation study of eutrophication in Hiroshima Bay: Simulation of particulate and dissolved organic matter using cyclic transformation of carbon

A phytoplankton model, which includes a carbon cycle combined with a constant current obtained by the computation of tidal currents, was applied to the coastal region (Hiroshima Bay, Japan). The transformation rate constants for the carbon cycle were experimentally evaluated.The seasonal variations of particulate and dissolved organic matter by the simulation model were reasonably consistent with the field data. Carbon concentration increased by primary production in the upper layer was approx. 10 times that augmented by loading of carbon from land at the shore in summer. Sinking plays an important role in the elimination of particulate organic matter. On the other hand, horizontal advection and turbulent diffusion are important factors for the diminution of dissolved organic matter.     

Homogenized elastic-viscoplastic behavior of plate-fin structures at high temperatures: Numerical analysis and macroscopic constitutive modeling

In this study, homogenized elastic-viscoplastic behavior of an ultra-fine plate-fin structure fabricated for compact heat exchangers is investigated. First, the homogenized behavior is numerically analyzed using a fully implicit mathematical homogenization scheme of periodic elastic-inelastic solids. A power-law creep relation is assumed to represent the viscoplasticity of base metals at high temperatures. The plate-fin structure is thus shown to exhibit significant anisotropy as well as noticeable compressibility in both the elastic and viscoplastic ranges of the homogenized behavior. Second, a non-linear rate-dependent macroscopic constitutive model is developed using the quadratic yield function proposed for anisotropic compressible plasticity. The resulting constitutive model is shown to be successful for simulating the anisotropy, compressibility, and rate dependency in the homogenized behavior in multi-axial stress states.     

Oxidation products of cyanidin 3-O-β-d-glucoside with a free radical initiator

Recently, we have reported that anthocyanins show strong antioxidative activity, but no attention has been paid to anthocyanins from the viewpoint of the reaction mechanism of alkylperoxyl radicals; therefore, we investigated the reaction products of antioxidative anthocyanins (cyanidin 3-O-β-d-glucoside). Cyanidin 3-O-β-d-glucoside was reacted with 2,2′-azobis(2,4-dimet hylvaleronitrile) to generate the alkylperoxyl radicals, and the reaction products were isolated by high-performance liquid chromatography. The products were identified as 4,6-dihydroxy-2-O-β-d-glucosyl-3-oxo-2,3-dihydrobenzofuran and protocatechuic acid. Based on reaction products, the antioxidative mechanism of cyanidin 3-O-β-d-glucoside may be different from that of α-tocopherol; cyanidin 3-O-β-d-glucoside would produce another radical scavenger, as it would break down the structure and scavenge the radicals.     

Quantitative structural analysis of twin boundary in alpha-Zn7Sb2O12 using HAADF STEM method

The structure and composition of the 1/4{110} twin boundary in alpha-Zn7Sb2O12 have been determined by using quantitative high-angle annular dark field scanning transmission electron microscopy (HAADF STEM) analysis. The noise in the experimental HAADF STEM images is reduced by using the maximum entropy method and average processing, and the parameters used in dynamical simulations are experimentally determined. From the analysis, it has been found that octahedral sites in the twin boundary slightly shift parallel to the [110] direction, and a reduction of the Sb concentration at the octahedral sites on the plane adjacent to the twin boundary was detected. The reduction was measured from three regions in the same twin boundary, and the Sb concentrations were 4 +/- 3, 8 +/- 3 and 19 +/-2 at% from 33 at%.     

Contact and friction between catheter and blood vessel

Therapeutic vascular catheterization techniques are sometimes hampered by the frictional forces between the blood vessel and the catheter, when contact points of the vessel are changing and deforming during the movement of the catheter. The goal of the present study was to characterize frictional interactions between the blood vessel wall and the catheter using experimental and numerical analysis. First, the frictional force was measured with an experimental apparatus that uses a ball and flattened porcine aorta to simulate frictional forces between the catheter and the vessel. Second, catheter motion was characterized by two-dimensional numerical calculations based on the experimental results. Experimental analysis demonstrated that slip occurred and that friction coefficient between the vessel and the catheter and the deformation of the specimen were small when the contact between the ball and the aorta occurred at a small angle. The compliance of the specimen in the normal direction obtained by the experiment was by far larger than that calculated according to the Hertzian contact theory. Numerical analysis shows that this difference of the parameter of the vessel, which must be determined accurately in surgical simulator, could affect the trajectory of the catheter.     

Experimental and simulation study of undesirable short-period deformation in piezoelectric deformable x-ray mirrors

To construct adaptive x-ray focusing optics whose optical parameters can be varied while performing wavefront correction, ultraprecise piezoelectric deformable mirrors have been developed. We computationally and experimentally investigated undesirable short-period deformation caused by piezoelectric actuators adhered to the substrate during mirror deformation. Based on the results of finite element method analysis, shape measurements, and the observation of x-ray reflection images, a guideline is developed for designing deformable mirrors that do not have short-period deformation errors.