Investigation of Brain Function-Related Myokine Secretion by Using Contractile 3D-Engineered Muscle

Brain function-related myokines, such as lactate, irisin, and cathepsin B (CTSB), are upstream factors that control brain-derived neurotrophic factor (BDNF) expression and are secreted from skeletal muscle by exercise. However, whether irisin and CTSB are secreted by muscle contraction remains controversial. Three-dimensional (3D)-engineered muscle (3D-EM) may help determine whether skeletal muscle contraction leads to the secretion of irisin and CTSB, which has never been identified with the addition of drugs in conventional 2D muscle cell cultures. We aimed to investigate the effects of electrical pulse stimulation (EPS)-evoked muscle contraction on irisin and CTSB secretion in 3D-EM. The 3D-EM, which consisted of C2C12 myoblasts and type-1 collagen gel, was allowed to differentiate for 2 weeks and divided into the control and EPS groups. EPS was applied at 13 V, 66 Hz, and 2 msec for 3 h (on: 5 s/off: 5 s). Irisin and CTSB secretion into the culture medium was measured by Western blotting. Irisin secretion was significantly increased following EPS (p < 0.05). However, there was no significant difference in CTSB secretion between the two groups. The present study suggests that irisin may be a contractile muscle-derived myokine, but CTSB is not secreted by EPS-evoked muscle contractile stimulation in 3D-EM.     

LIF–IGF Axis Contributes to the Proliferation of Neural Progenitor Cells in Developing Rat Cerebrum

In rodent models, leukemia inhibitory factor (LIF) is involved in cerebral development via the placenta, and maternal immune activation is linked to psychiatric disorders in the child. However, whether LIF acts directly on neural progenitor cells (NPCs) remains unclear. This study performed DNA microarray analysis and quantitative RT-PCR on the fetal cerebrum after maternal intraperitoneal or fetal intracerebral ventricular injection of LIF at day 14.5 (E14.5) and determined that the expression of insulin-like growth factors (IGF)-1 and -2 was induced by LIF. Physiological IGF-1 and IGF-2 levels in fetal cerebrospinal fluid (CSF) increased from E15.5 to E17.5, following the physiological surge of LIF levels in CSF at E15.5. Immunostaining showed that IGF-1 was expressed in the cerebrum at E15.5 to E19.5 and IGF-2 at E15.5 to E17.5 and that IGF-1 receptor and insulin receptor were co-expressed in NPCs. Further, LIF treatment enhanced cultured NPC proliferation, which was reduced by picropodophyllin, an IGF-1 receptor inhibitor, even under LIF supplementation. Our findings suggest that IGF expression and release from the NPCs of the fetal cerebrum in fetal CSF is induced by LIF, thus supporting the involvement of the LIF–IGF axis in cerebral cortical development in an autocrine/paracrine manner.     

Clinical Implications and Molecular Characterization of Drebrin-Positive,Tumor-Infiltrating Exhausted T Cells in Lung Cancer

T cells express an actin-binding protein, drebrin, which is recruited to the contact site between the T cells and antigen-presenting cells during the formation of immunological synapses. However, little is known about the clinical implications of drebrin-expressing, tumor-infiltrating lymphocytes (TILs). To address this issue, we evaluated 34 surgical specimens of pathological stage I–IIIA squamous cell lung cancer. The immune context of primary tumors was investigated using fluorescent multiplex immunohistochemistry. The high-speed scanning of whole-slide images was performed, and the tissue localization of TILs in the tumor cell nest and surrounding stroma was automatically profiled and quantified. Drebrin-expressing T cells were characterized using drebrin⁺ T cells induced in vitro and publicly available single-cell RNA sequence (scRNA-seq) database. Survival analysis using the propensity scores revealed that a high infiltration of drebrin⁺ TILs within the tumor cell nest was independently associated with short relapse-free survival and overall survival. Drebrin⁺ T cells induced in vitro co-expressed multiple exhaustion-associated molecules. The scRNA-seq analyses confirmed that the exhausted tumor-infiltrating CD8⁺ T cells specifically expressed drebrin. Our study suggests that drebrin-expressing T cells present an exhausted phenotype and that tumor-infiltrating drebrin⁺ T cells affect clinical outcomes in patients with resectable squamous cell lung cancer.     

Anti-Inflammatory Action of Dexmedetomidine on Human Microglial Cells

Neuroinflammation, where inflammatory cytokines are produced in excess, contributes to the pathogenesis of delirium. Microglial cells play a central role in neuroinflammation by producing and releasing inflammatory cytokines in response to infection, tissue damage and neurodegeneration. Dexmedetomidine (DEX) is a sedative, which reduces the incidence of delirium. Thus, we hypothesized that DEX may alleviate delirium by exhibiting anti-inflammatory action on microglia. In the present study, we investigated the anti-inflammatory action of DEX on human microglial HMC3 cells. The results indicated that DEX partially suppressed the IL-6 and IL-8 production by lipopolysaccharide (LPS)-stimulated HMC3 cells as well as the phosphorylation of p38 MAPK and IκB and the translocation of NF-κB. Furthermore, DEX substantially suppressed IL-6 and IL-8 production by unstimulated HMC3 cells as wells as the phosphorylation of p38 MAPK and IκB and the translocation of NF-κB. These observations suggest that DEX exhibits anti-inflammatory action on not only LPS-stimulated but also unstimulated microglial cells via the suppression of inflammatory signaling and cytokine production.     

Estimation of surface properties of grafted layers formed on low- and high-density polyethylene plates by photografting of methacrylic acid and acrylic acid at different monomer concentrations and temperatures

An investigation was carried out on estimation of hydrophilicity, wettability and water-absorptivity, and surface analysis by X-ray photoelectron spectroscopy of the low- and high-density polyethylene (LDPE and HDPE) plates photografted with methacrylic acid (MAA) and acrylic acid (AA) at different monomer concentrations or temperatures. Wettability of the MAA-grafted LDPE and HDPE plates increased with grafted amounts, and became constant when the substrate surfaces were fully covered with the grafted polymer chains. On the other hand, for the AA-grafted LDPE and HDPE plates, wettability had the maximum value, and then gradually decreased against the grafted amount probably due to aggregation of grafted PAA chains, although the surfaces were covered with grafted PAA chains at lower grafted amounts compared with grafted PMAA chains. Water-absorptivity sharply increased at lower grafted amounts due to formation of shorter grafted polymer chains for photografting at lower monomer concentrations or due to restriction of the location of grafting to the outer surface region for photografting at lower temperatures. Therefore, for photograftings of AA or onto the HDPE plates, the substrate surfaces were covered with grafted polymer chains and the grafted layers formed possessed higher water-absorptivity at lower grafted amounts. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

The effects of 5-OP-RU stereochemistry on its stability and MAIT-MR1 axis

Mucosal-associated invariant T (MAIT) cells are an abundant subset of innate-like T lymphocytes. MAIT cells are activated by microbial riboflavin-derived antigens, such as 5-(2-oxopropylideneamino)-6-d -ribitylaminouracil (5-OP-RU), when presented by the major histocompatibility complex (MHC) class I-related protein (MR1). We have synthesized all stereoisomers of 5-OP-RU to investigate the effects of its stereochemistry on the MR1-dependent MAIT cell activation and MR1 upregulation. The analysis of MAIT cell activation by these 5-OP-RU isomers revealed that the stereocenters at the 2’- and 3’-OH groups in the ribityl tail are crucial for the recognition of MAIT-TCR, whereas that of 4’-OH group does not significantly affect the regulation of MAIT cell activity. Furthermore, kinetic analysis of complex formation between the ligands and MR1 suggested that 5-OP-RU forms a covalent bond to MR1 in cells within 1 hour. These findings provide guidelines for designing ligands that regulate MAIT cell functions.     

Yield improvement in progressive freeze-concentration by partial melting of ice

Progressive freeze-concentration (PFC) was applied for the concentration of sucrose solutions with various concentrations (0.3-40%), a model pear (La France) flavor solution, tomato juice (5.3 Brix) and apple juice (12-15 Brix). When the solute concentration was low with low osmotic pressure as in the case of the model pear flavor solution, the apparent partition coefficient (K_app) between the ice and the concentrated liquid phase was low to give a high yield. For samples with high solute concentrations, however, K_app increased with an increase in solute concentration to reduce the yield in PFC operation. In this case, the partial melting of ice was effective. In this method, the ice crystal formed after PFC was melted gradually with time to collect the melted fractions with intervals. Then the initial fractions were found to contain the higher amount of solute. Therefore, the yield could be improved to a necessary level (>90%) by recovering these fractions with the higher solute concentration. The recovered melted ice fraction may be mixed with the feed in the succeeding batch of PFC operation to avoid dilution of the concentrate. The partial ice melting will extend the practical applicability of PFC substantially.     

Application of chemical interaction between (Fe,Cr) oxides and Mo oxide at high temperature for self-healing intelligence on nuclear fuel cladding in LWRs

A novel scheme for a bilayer coating with self-healing ability is proposed in this study. The candidate materials for the coatings and the potential self-healing reaction are assessed in high-temperature aqueous environments and high-temperature air. The pure Cr₂O₃ layer and the composite of Cr₂O₃ and MoO₃ are the candidate materials for the outer layer and inner layer, respectively, due to their compatibility under normal condition and fabricability. Fe₂O₃–MoO₃ reactions exhibit a potential ability to heal the cracks because of a high reaction rate under normal condition. The self-healing process proceeds via the following mechanism under normal condition: Fe₂O₃ (a corrosion product in the coolant) diffuses into the cracks on the coating and reacts with MoO₃ (inner layer) to produce the insoluble Fe₂(MoO₄)₃, which deposits and repairs the cracks. In the loss-of-coolant accident (LOCA) situation, Cr₂O₃–MoO₃ reaction is expected to strengthen the adhesion of the coating.     

Auto-oxidation products of astaxanthin

The auto-oxidation products of astaxanthin were investigated. Astaxanthin was allowed to react with atmospheric oxygen at 55°C in the dark for 35 days. A series of oxidative cleavage products, 7-apoastaxanthinal (1), 9-apoastaxanthinone (2), 11-apoastaxanthinal (3), 13-apoastaxanthinone (4), 15-apoastaxanthinal (5), 14'-apoastaxanthinal (6), 12'-apoastaxanthinal (7), 10'-apoastaxanthinal (8), and 8'-apoastaxanthinal (9), were identified. Among them, 3 and 6 were isolated and characterized for the first time. Cleavage of the double bond in astaxanthin was discussed on the basis of the calculation of the stable molecular energy.     

Transparent conductive oxide‐less three‐dimensional cylindrical dye‐sensitized solar cell fabricated with flexible metal mesh electrode

A cylindrical transparent conductive oxide‐less dye‐sensitized solar cell (DSSC) consisting of glass tube/stainless steel mesh–TiO₂–dye/gel electrolytes/Pt‐Ti rod having capability of self‐light trapping is reported. Replacing the glass tube with heat‐shrinkable tube to reduce electrolyte gap and optical loss due to light transmission and reflection led to the enhancement in the power conversion efficiency from 2.61% to 3.91%. Profiling of the current distribution measured by laser beam‐induced current exhibited nearly the same current in the axial and radial directions, suggesting that light reflection on a cylindrical DSSC does not affect the efficiency seriously. Optimized best DSSC in this novel device architecture gave a short‐circuit current density of 11.94 mA/cm², an open‐circuit voltage of 0.71 V and a fill factor of 0.66 leading to the power conversion efficiency of 5.58% at AM 1.5 under simulated solar irradiation. Copyright © 2011 John Wiley & Sons, Ltd.