Characterization of Aspergillus oryzae glycoside hydrolase family 43 β-xylosidase expressed in Escherichia coli

This is the first report of glycoside hydrolase family 43 β-xylosidase from Aspergillus oryzae. To characterize this enzyme, the recombinant enzyme was expressed in Escherichia coli. Unlike known β-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline pH.     

Detection of norovirus RNA in bivalve molluscs by using bacteria-culture-employed method (A3T method)

Norovirus (NV) RNA has rarely been detected in foods despite the use of highly sensitive methods such as RT-PCR and real-time RT-PCR. In the modified method (A3T method) reported previously, a bacterial culture process was introduced into the standard protocol for NV detection to remove some inhibitor(s) present in food ingredients. To confirm the efficiency of the A3T method and to examine NV contamination in bivalve molluscs, we tried to detect NV RNA in bivalve molluscs on the market and in oyster samples associated with foodborne outbreaks by using the standard method and the A3T method. NV RNAs were detected in 20 samples (18.0%) of 111 bivalve molluscs, including oysters, on the market by use of the A3T method, while only one sample (0.9%) was positive according to the standard method. NV RNA was also detected in 10 of 35 oyster samples related to foodborne outbreaks by the A3T method. Those results show that the A3T method is suitable for the detection of NV in bivalve molluscs in general laboratories.     

Protective Role of Glutathione in the Hippocampus after Brain Ischemia

Stroke is a major cause of death worldwide, leading to serious disability. Post-ischemic injury, especially in the cerebral ischemia-prone hippocampus, is a serious problem, as it contributes to vascular dementia. Many studies have shown that in the hippocampus, ischemia/reperfusion induces neuronal death through oxidative stress and neuronal zinc (Zn²⁺) dyshomeostasis. Glutathione (GSH) plays an important role in protecting neurons against oxidative stress as a major intracellular antioxidant. In addition, the thiol group of GSH can function as a principal Zn²⁺ chelator for the maintenance of Zn²⁺ homeostasis in neurons. These lines of evidence suggest that neuronal GSH levels could be a key factor in post-stroke neuronal survival. In neurons, excitatory amino acid carrier 1 (EAAC1) is involved in the influx of cysteine, and intracellular cysteine is the rate-limiting substrate for the synthesis of GSH. Recently, several studies have indicated that cysteine uptake through EAAC1 suppresses ischemia-induced neuronal death via the promotion of hippocampal GSH synthesis in ischemic animal models. In this article, we aimed to review and describe the role of GSH in hippocampal neuroprotection after ischemia/reperfusion, focusing on EAAC1.     

Evolving autonomous specialization in congested path formation task of robotic swarms

Redundancy in the number of robots is a fundamental feature of robotic swarms to confer robustness, flexibility, and scalability. However, robots tend to interfere with each other in a case, where multiple robots gather in a spatially limited environment. The aim of this paper is to understand how a robotic swarm develops an effective strategy to manage congestion. The controllers of the robots are obtained by an evolutionary robotics approach. The strategy of managing congestion is observed in the process of generating a collective path of robots visiting two landmarks alternately. The robotic swarm exhibits autonomous specialization that the robots traveling inside the path activate the LEDs, while the robots in the outer side deactivate them. We found that the congestion is regulated in an emergent way of autonomous specialization by the result of an artificial evolution.     

New superconducting lead cuprate with 1201 structure, (Pb0.5B0.5) (SrLa)CuOz

New Pb-based superconducting cuprate with the 1201 structure has been synthesized in the (Pb_1 − yB_y)(Sr_2 − xLa_x)CuO_z system. From X-ray powder diffraction study, the almost-single 1201 phase sample is found to be obtained at x = 1.0 and y = 0.5 for the nominal composition of (Pb_1 − yB_y)(Sr_2 − xLa_x)CuO_z. The crystal structure has a tetragonal symmetry with the lattice parameters of a = 0.3779 nm and c = 0.8654 nm. The sample shows an onset of superconductivity at about 32 K as measured by the temperature dependence of the DC magnetic susceptibility.     

Nanostructure of pure iron anodically oxidized in borate buffer solution and annealed by infrared radiation

The behavior of oxide film on pure iron passivated in a borate buffer solution and subsequently radiated by infrared light (IR) was investigated in comparing to that by just IR annealing without passivation, and was evaluated by film structure, etc. The effect of thermal annealing over 250 degrees C was observed with gamma-Fe2O3 grain growth and sharp increase in surface roughness, film thickness and oxygen content. An ellipsometric parameter of tan psi was sensitively reflected by annealing effect, and tan psi curve had a shoulder at 150 degrees C for 5 min and a peak of tan psi was shifted from 350 nm to 450 nm in wavelength. This shift was also caused by the formation of gamma-Fe2O3, because the peak was also observed in tan psi of the bulk Fe2O3 family. Passivation effects at 800 mV prior to IR annealing on thickness and oxygen content changed at 150 degrees C, and decreased tan psi at 350 nm and excessive film growth over 250 degrees C, and increased oxygen content under 100 degrees C and surface roughness at 50-250 degrees C. The terrace width with atomic scale flatness was slightly increase by passivation prior to IR annealing at 50-250 degrees C, and the maximum terrace width reached larger than 10 nm by passivation and IR annealing at 100 degrees C for 30 min.     

Synthesis of New Superconducting Pb-Based 1222 Cuprates Containing Sulfur in the (Pb0.75S0.25)Sr2(Eu2.0−x Ce x )Cu2O z System

We have synthesized new Pb-based 1222 cuprates containing sulfur in the (Pb_0.75S_0.25)Sr₂(Eu_2.0?x Ce _x )Cu₂O _z system. X-ray diffraction study shows that almost the single 1222 phase samples can be obtained for a considerable wide area of 0.3≤x≤1.2 in the composition of (Pb_0.75S_0.25)Sr₂(Eu_2.0?x Ce _x )Cu₂O _z . Next, the sample with x=0.4 after annealing under high-pressure O₂ is discovered to show an onset of the resistivity drop at about 31 K and to show an onset of diamagnetic signal at about 29 K. These phenomena clearly indicate that the sample is a new superconductor.     

Humido-sensitive conducting polymer films and applications to linear actuators

Free-standing films made of poly(3,4-ethylenedioxythiophene) doped with poly(4-styrenesulfonate) (PEDOT/PSS) with various PSS contents were newly prepared by casting water dispersion of the PEDOT/PSS colloidal particles in the presence of an extra PSS. Electrical conductivity, morphology, water vapor sorption, and electro-active polymer actuating behavior of the resulting films were investigated by means of four-point method, atomic force microscope (AFM), sorption isotherm, and electromechanical analyses. The maximum contraction of the film by application of an electric field increased with increasing both PSS content and relative humidity (RH), where the value attained 7% at 70% RH for the film with 93% of PSS. Since the isothermal sorption curve of the film was less dependent on the PSS content, the significant increase of the film contraction was explained by two mechanisms: (i) the extra PSS prevented from hydrogen bonding between adjacent PEDOT/PSS particles that suppressed dimensional changes of the film; and (ii) the higher the RH, the larger the degree of water vapor sorption, which led to the large film contraction by desorption of water vapor via Joule heating. On the basis of this phenomenon linear actuators utilizing PEDOT/PSS films were successfully developed and applied to leverage actuator and Braille cell.     

Ex Vivo Expanded and Activated Natural Killer Cells Prolong the Overall Survival of Mice with Glioblastoma-like Cell-Derived Tumors

Glioblastoma (GBM) is the leading malignant intracranial tumor and is associated with a poor prognosis. Highly purified, activated natural killer (NK) cells, designated as genuine induced NK cells (GiNKs), represent a promising immunotherapy for GBM. We evaluated the anti-tumor effect of GiNKs in association with the programmed death 1(PD-1)/PD-ligand 1 (PD-L1) immune checkpoint pathway. We determined the level of PD-1 expression, a receptor known to down-regulate the immune response against malignancy, on GiNKs. PD-L1 expression on glioma cell lines (GBM-like cell line U87MG, and GBM cell line T98G) was also determined. To evaluate the anti-tumor activity of GiNKs in vivo, we used a xenograft model of subcutaneously implanted U87MG cells in immunocompromised NOG mice. The GiNKs expressed very low levels of PD-1. Although PD-L1 was expressed on U87MG and T98G cells, the expression levels were highly variable. Our xenograft model revealed that the retro-orbital administration of GiNKs and interleukin-2 (IL-2) prolonged the survival of NOG mice bearing subcutaneous U87MG-derived tumors. PD-1 blocking antibodies did not have an additive effect with GiNKs for prolonging survival. GiNKs may represent a promising cell-based immunotherapy for patients with GBM and are minimally affected by the PD-1/PD-L1 immune evasion axis in GBM.     

Possible Involvement of the Upregulation of ΔNp63 Expression Mediated by HER2-Activated Aryl Hydrocarbon Receptor in Mammosphere Maintenance

Cancer stem cells (CSCs) contribute to the drug resistance, recurrence, and metastasis of breast cancers. Recently, we demonstrated that HER2 overexpression increases mammosphere formation via the activation of aryl hydrocarbon receptor (AHR). In this study, the objective was to identify the mechanism underlying mammosphere maintenance mediated by HER2 signaling-activated AHR. We compared the chromatin structure of AHR-knockout (AHRKO) HER2-overexpressing MCF-7 (HER2-5) cells with that of wild-type HER2-5 cells; subsequently, we identified TP63, a stemness factor, as a potential target gene of AHR. ΔNp63 mRNA and protein levels were higher in HER2-5 cells than in HER2-5/AHRKO cells. Activation of HER2/HER3 signaling by heregulin treatment increased ΔNp63 mRNA levels, and its induction was decreased by AHR knockdown in HER2-5 cells. The results of the chromatin immunoprecipitation assay revealed an interaction between AHR and the intronic region of TP63, which encodes ΔNp63. A luciferase reporter gene assay with the intronic region of TP63 showed that AHR expression increased reporter activity. Collectively, our findings suggest that HER2-activated AHR upregulates ΔNp63 expression and that this signaling cascade is involved in CSC maintenance in HER2-expressing breast cancers.