Effects of neural refractoriness on spatio-temporal variability in spike initiations with Electrical stimulation.

In this paper, the effects of neural refractoriness on action potential (spike) initiations with electrical stimulation are investigated using computer modeling and simulation techniques. The computational model was composed of a myelinated nerve fiber with 50 nodes of Ranvier, each consisting of stochastic sodium and potassium channels, making it possible to represent the fluctuations of spike initiation. A series of two-pulse stimuli was presented by a stimulating electrode above the central (26th) node of Ranvier. The amplitude of the first (masker) pulse stimulus was set such that the masker pulse stimulus evoked spikes on each trial, while that of the second (probe) pulse stimulus was set such that the probe pulse stimulus evoked spikes on a half of trials, threshold values. Then the transmembrane potentials in response to the probe pulse stimulus were recorded at each node (i.e., 1-50 nodes) in order to determine the spike initiation node and time. From the observation of the spike initiation node and time, a spatio-temporal histogram as well as a spatial variability and a temporal variability of spike initiations was generated which allowed us to interpret fluctuations in spike initiation node and time. It was shown that the distribution of spike initiations tended to become greater spatially and longer temporally as the masker-probe intervals (MPIs) of the two-pulse stimuli shortened. It was also shown that the number of activated sodium channels as functions of space and time tended to become smaller due to inactivation of sodium channels and varied spatially and temporally as MPIs shortened. These findings may imply that the stochastic sodium channels during a relative refractory period may contribute to enhancing the fluctuations in spike initiations, and give us an insight into encoding information with electric stimuli to improve the performance of the prosthetic devices, especially cochlear implants.     

Thermal buckling of cross-ply laminated composite and sandwich plates according to a global higher-order deformation theory

A two-dimensional global higher-order deformation theory is presented for thermal buckling of cross-ply laminated composite and sandwich plates. By using the method of power series expansion of continuous displacement components, a set of fundamental governing equations which can take into account the effects of both transverse shear and normal stresses is derived through the principle of virtual work. Several sets of truncated Mth-order approximate theories are applied to solve the eigenvalue problems of a simply supported multilayered plate. Modal transverse shear and normal stresses can be calculated by integrating the three-dimensional equations of equilibrium in the thickness direction, and satisfying the continuity conditions at the interface between layers and stress boundary conditions at the external surfaces. Numerical results are compared with those of the published three-dimensional layerwise theory in which both in-plane and normal displacements are assumed to be C⁰ continuous in the continuity conditions at the interface between layers. Effects of the difference of displacement continuity conditions between the three-dimensional layerwise theory and the global higher-order theory are clarified in thermal buckling problems of multilayered composite plates.     

Shape from Shading with Interreflections Under a Proximal Light Source: Distortion-Free Copying of an Unfolded Book

We address the problem of recovering the 3D shape of an unfolded book surface from the shading information in a scanner image. This shape-from-shading problem in a real world environment is made difficult by a proximal, moving light source, interreflections, specular reflections, and a nonuniform albedo distribution. Taking all these factors into account, we formulate the problem as an iterative, non-linear optimization problem. Piecewise polynomial models of the 3D shape and albedo distribution are introduced to efficiently and stably compute the shape in practice. Finally, we propose a method to restore the distorted scanner image based on the reconstructed 3D shape. The image restoration experiments for real book surfaces demonstrate that much of the geometric and photometric distortions are removed by our method.     

GH-16 Type β-1,3-Glucanase from Lysobacter sp. MK9-1 Enhances Antifungal Activity of GH-19 Type Chitinase,and Its Glucan-binding Domain Binds to Fungal Cell-wall

The GH-16 type β-1,3-glucanase (BgluC16MK) gene of Lysobacter sp. MK9-1 was cloned to study its antifungal activities. BgluC16MK displays amino acid sequence similarity with GluC from L. enzymogenes strain N4-7. BgluC16MK includes a signal sequence, a catalytic domain and carbohydrate-binding module family 6-type β-glucan binding domain (B-GBD). The expression of the BgluC16MK gene in Escherichia coli without the signal sequence resulted in antifungal activity at a dose of 0.6-0.8 nmol/disk. However, BgluC16MK displayed antifungal activity at a dose of 0.025 nmol/disk in combination with Chi19MK. Substrate-specific assay revealed that purified BgluC16MK hydrolyzed insoluble curdlan more readily than the soluble substrate. Furthermore, to explore the binding selectivity of B-GBD of BgluC16MK, we constructed a fusion protein (B-GBD-GFP) using the B-GBD and green fluorescent protein. The activity of the fusion protein against various substrates indicates that B-GBD was selective for glucans with β-1,3-linkages. An additional study demonstrated the binding ability of B-GBD-GFP to the cell-wall of living fungi, such as T. reesei and Aspergillus oryzae. These findings suggest that BgluC16MK can be utilized to generate antifungal enzyme preparations and that the fusion protein B-GBD-GFP can be used to identify the fungal cell surface structure using β-glucans.     

Macrophage Activation Markers Predict Liver-Related Complications in Primary Biliary Cholangitis

Primary biliary cholangitis (PBC) has a wide variation in clinical presentation and course. There is no significant correlation between these symptoms and the disease stage, although patients with more advanced stages generally have more symptoms. It is important to develop biomarkers in order to identify patients with an increased risk of complications and end-stage liver disease. This study investigated surrogate markers for risk estimation of PBC-related complications, including a study population of 77 patients with PBC who underwent liver biopsy and were measured for serum levels of macrophage activation markers, soluble CD163 (sCD163), soluble mannose receptor (sMR), and zonulin. Patients with PBC were divided into symptomatic (Group S, n = 20) and asymptomatic (Group A, n = 57) groups. The correlations of histological stages based on both Scheuer and Nakanuma classifications with the three serum markers were investigated. The Nakanuma classification involves grading for liver fibrosis and bile duct loss. The three biomarkers were assessed for their diagnostic ability to identify patients with PBC having high risk of developing complications. The predictive factors of these complications were examined as well. Group S had significantly higher serum sMR (p = 0.011) and sCD163 (p = 0.048) levels versus Group A. A composite index of sMR and sCD163 measurements had significantly better prediction performance than sCD163 alone (p = 0.012), although not when compared to sMR alone (p = 0.129). Serum sMR was an independent factor for developing complications on both univariate (Odds ratio (OR) = 30.20, 95% confidence interval (95% CI): 3.410–267.0, p = 0.00220), and multivariate (OR = 33.70, 95% CI: 3.6600–311.0, p = 0.0019) analyses. Patients with PBC having sMR of ≥56.6 had a higher incidence of clinical complications versus those with a sMR of <56.6. Serum sMR predicts the development of complications in patients with PBC. sMR plus sCD163 showed better predictive power than either marker alone, although the addition of sCD163 did not improve the predictive power of sMR. Future prospective studies are required in order to validate the findings of the present study.     

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.     

Development of an Enzyme-Linked Immunosorbent Assay for Specific Detection of Mulberroside A in Mulberry (Morus alba L.) Using Anti-mulberroside A Polyclonal Antibody

An enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal antibody (PAb) against mulberroside A (MuA), a major active component found in the root bark of mulberry (Morus alba L.). MuA was conjugated with the carrier protein bovine serum albumin for immunization to rabbits. The results showed that the antibodies were specific only for MuA and have very low specificity for its aglycone, oxyresveratrol. The ELISA assay was suitable for quantitating MuA in the range of 0.17–15.62 μg/mL with a relative standard deviation (RSD) of less than 5 % for both intra- and inter-assay precision levels. The recovery rates of MuA in the samples were in the range of 97.6–101.4 % with a RSD of less than 5 %. The developed ELISA exhibited a good correlation value (R ²?=?0.994) with the standard high-performance liquid chromatography method in the crude extracts of plant samples. These results suggest that the developed ELISA method using PAb against MuA can be applied to determine MuA content with high specificity, rapidity, and simplicity in mulberry samples. The developed ELISA method described could prove to be useful as an analytical tool for quality control of mulberry and their products.     

Determination of residual avoparcin in chicken muscle by HPLC with ultraviolet and amperometric detection

An analytical method was developed for determination of residual avoparcin in chicken muscle by measuring alpha- and beta-avoparcin, major components of the pharmaceutical preparation avoparcin, using HPLC with UV and amperometric detectors. The analytical HPLC was run on a Cosmosil 5C18-AR column (4.6 mm x 25 cm) with a gradient formed from A: 2.5% acetic acid, 0.01 mol/L sodium heptane sulfonic acid-acetonitrile (88.5:11.5) (pH 4.0) and B: 2.5% acetic acid-acetonitrile (10:90), using UV and amperometric detection (AMD) with glassy-carbon electrode (+900 mV). Avoparcin was extracted from chicken muscle by homogenization with methanol-0.2 mol/L sulfuric acid (6:4) followed by centrifugation after pH adjustment to 4 with 1 mol/L sodium hydroxide. The supernatant was evaporated to dryness, and the residue was dissolved in water. The aqueous layer was adjusted to pH 4 by adding 1 mol/L sodium hydroxide. Then it was purified on a Sep-Pak tC18 plus ENV cartridge. The cartridge was washed with water, and retained substances were eluted with 50% methanol. The eluate was evaporated to dryness under reduced pressure. The residue was dissolved in water and determined by HPLC. Recoveries of avoparcin spiked in chicken muscle were 73.1-88.1% at levels of 2-10 micrograms/g. The detection limits were 0.5 microgram/g (UV) and 0.2 microgram/g (AMD).     

Sar1 Affects the Localization of Perilipin 2 to Lipid Droplets

Lipid droplets (LDs) are intracellular organelles that are ubiquitous in many types of cells. The LD core consists of triacylglycerols (TGs) surrounded by a phospholipid monolayer and surface proteins such as perilipin 2 (PLIN2). Although TGs accumulate in the phospholipid bilayer of the endoplasmic reticulum (ER) and subsequently nascent LDs buds from ER, the mechanism by which LD proteins are transported to LD particles is not fully understood. Sar1 is a GTPase known as a regulator of coat protein complex Ⅱ (COPⅡ) vesicle budding, and its role in LD formation was investigated in this study. HuH7 human hepatoma cells were infected with adenoviral particles containing genes coding GFP fused with wild-type Sar1 (Sar1 WT) or a GTPase mutant form (Sar1 H79G). When HuH7 cells were treated with oleic acid, Sar1 WT formed a ring-like structure around the LDs. The transient expression of Sar1 did not significantly alter the levels of TG and PLIN2 in the cells. However, the localization of PLIN2 to the LDs decreased in the cells expressing Sar1 H79G. Furthermore, the effects of Sar1 on PLIN2 localization to the LDs were verified by the suppression of endogenous Sar1 using the short hairpin RNA technique. In conclusion, it was found that Sar1 has some roles in the intracellular distribution of PLIN2 to LDs in liver cells.     

Numerical simulation of deflecting flow in a symmetric enlarged channel

Numerical results of three-dimensional separated flow and heat transfer in a rectangular channel with a suddenexpansion are presented in this paper.Numerical simulations of Navier-Stokes and energy equations are carriedout using the finite difference method.The results of three-dimensional calculations are compared with thetwo-dimensional ones,and effects of the aspect ratio of channel upon the flow are shown.The transition fromsymmetric to asymmetric flow appears at lower Reynolds number as increasing the aspect ratio.The details oflocal heat transfer characteristics in two different separated flow regions on two downstream walls are clarified.Two-dimensionality of the flow and heat transfer almost disappears for the aspect ratio considered.