PHENOTYPIC ANALYSIS OF MICE DEFICIENT FOR Ly6C1/Ly6C2

Ly6C comprises two homologous components of Ly6C1 and Ly6C2, and the expression of either of the Ly6C molecules defines unique functional subsets of monocytes. Ly6C is also expressed by other immune cell types, including Aire-expressing medullary thymic epithelial cells. Because the role of Ly6C expression in determining the functional subsets remains unclear, we generated mice deficient for both Ly6C1 and Ly6C2 with CRISPR-Cas9–mediated deletion. Mice deficient for Ly6C1/Ly6C2 showed no major alterations in the subsets and function of monocyte and other immune cells, including the cells involved in the dextran sulfate sodium salt–induced colitis model. By generating the mice deficient for Ly6C1 alone, we have also investigated the expression pattern of Ly6C1 and Ly6C2 in immune cells. Except for medullary thymic epithelial cells and CD4 single-positive T cells, immune cells predominantly expressed Ly6C2. Thus, despite the importance as a marker with a unique differential expression pattern, the Ly6C molecules have no major impact on determining the functional subsets and maintaining immune homeostasis

Tumorigenesis of Epstein–Barr Virus-Positive Epithelial Cell Lines Derived from Gastric Tissues in the SCID Mouse

AbstractTo study the tumorigenesis of Epstein–Barr virus (EBV)-positive epithelial cell lines GT38 and GT39 derived from human gastric tissues, we inoculated these cells under the skin of severe combined immunodeficient (SCID) mice. The development of tumors was observed in each of the mice about 2 months after the inoculation. The tumors were diagnosed with undifferentiated carcinoma by hematoxylin/eosin staining. EBV-encoded small RNA1 was detected in the paraffin-embedded tumor sections. The tumor cells had human chromosome. The circular, but not linear, EBV DNA was detected in the tumors. The molecular sizes of EBV DNA termini were the same as that of the inoculated GT38 or GT39 cells. The expressions of EBV nuclear antigen 2 and latent membrane protein 1 reduced in the tumors. Transcripts of BamHI C and W promoters in latency III were detected in the tumors and the cultured cells in vitro. The tumor cells were passaged from one SCID mouse to other SCID mice and to cultures in vitro. This is the first evidence that the EBV-positive epithelial cell lines produced tumors in the SCID mouse

Effect of irradiation hardening on brittle fracture performance of box-shaped blanket fabricated by F-82H

Fusion reactor blankets designed for DEMO are based on a water cooled ceramic breeder. The unstable fracture of the box-shaped blanket would occur by the superimposition of three factors as follows: (1) it is the reduction of fracture toughness for structural materials, such as F-82H, caused by the significant irradiation hardening, (2) it is the formation of cracks which act as the initiation point of the fracture, and (3) it is the excessive tensile load caused by the inner pressure to In-Box loss of coolant accident (LOCA). In the present study, application of Weibull stress criterion to fracture assessment is to investigate the effect of irradiation hardening on the brittle fracture performance of box-shaped blanket fabricated by F-82H. Both structural analysis of the box-shaped blanket simulating In-Box LOCA and deformation behavior analysis of irradiated toughness specimens were conducted by elastic-plastic finite element analysis (FEA). Weibull stress values calculated from FEA of irradiated toughness specimens were higher than those from FEA of irradiated structural components. It, therefore, is considered that the irradiation hardening of structural materials would not promote the brittle fracture at the corner of rectangular coolant tubes of the blankets caused by the load of In-Box LOCA

Comparative study on the effect of human BST-2/Tetherin on HIV-1 release in cells of various species

In this study, we first demonstrate that endogenous hBST-2 is predominantly expressed on the plasma membrane of a human T cell line, MT-4 cells, and that Vpu-deficient HIV-1 was less efficiently released than wild-type HIV-1 from MT-4 cells. In addition, surface hBST-2 was rapidly down-regulated in wild-type but not Vpu-deficient HIV-1-infected cells. This is a direct insight showing that provirus-encoded Vpu has the potential to down-regulate endogenous hBST-2 from the surface of HIV-1-infected T cells. Corresponding to previous reports, the aforementioned findings suggested that hBST-2 has the potential to suppress the release of Vpu-deficient HIV-1. However, the molecular mechanism(s) for tethering HIV-1 particles by hBST-2 remains unclear, and we speculated about the requirement for cellular co-factor(s) to trigger or assist its tethering ability. To explore this possibility, we utilize several cell lines derived from various species including human, AGM, dog, cat, rabbit, pig, mink, potoroo, and quail. We found that ectopic hBST-2 was efficiently expressed on the surface of all analyzed cells, and its expression suppressed the release of viral particles in a dose-dependent manner. These findings suggest that hBST-2 can tether HIV-1 particles without the need of additional co-factor(s) that may be expressed exclusively in primates, and thus, hBST-2 can also exert its function in many cells derived from a broad range of species. Interestingly, the suppressive effect of hBST-2 on HIV-1 release in Vero cells was much less pronounced than in the other examined cells despite the augmented surface expression of ectopic hBST-2 on Vero cells. Taken together, our findings suggest the existence of certain cell types in which hBST-2 cannot efficiently exert its inhibitory effect on virus release. The cell type-specific effect of hBST-2 may be critical to elucidate the mechanism of BST-2-dependent suppression of virus release

Tip dependence of three-dimensional scanning force microscopy images of calcite-water interfaces investigated by simulation and experiments

In this study, we have investigated the influence of the tip on the three-dimensional scanning force microscopy (3D-SFM) images of calcite-water interfaces by experiments and simulations. We calculated 3D force images by simulations with the solvent tip approximation (STA), Ca, CO(3)and OH tip models. For all the 3D images, thezprofiles at the surface Ca and CO(3)sites alternately show oscillatory peaks corresponding to the hydration layers. However, the peak heights and spacings become larger when the mechanical stability of the tip becomes higher. For analyzing thexyslices of the 3D force images, we developed the extended STA (E-STA) model which allowed us to reveal the strong correlation between the hydration structure just under the tip and the atomic-scale force contrasts. Based on these understandings on the image features showing the strong tip dependence, we developed a method for objectively estimating the similarity between 3D force images. With this method, we compared the simulated images with the three experimentally obtained ones. Among them, two images showed a relatively high similarity with the image obtained by the simulation with the Ca or the CO(3)tip model. Based on these agreements, we characterized the hydration structure and mechanical stability of the experimentally used tips. The understanding and methodology presented here should help us to derive accurate information on the tip and the interfacial structure from experimentally obtained 3D-SFM images.Peer reviewe

Improvements in fundamental performance of liquid-environment atomic force microscopy with true atomic resolution

Recently, there have been significant advancements in liquid-environment atomic force microscopy (AFM) with true atomic resolution. The technical advancements are followed by a rapid expansion of its application area. Examples include subnanometer-scale imaging of biological systems and three-dimensional measurements of water distributions (i.e., hydration structures) and fluctuating surface structures. However, to continue this progress, we should improve the fundamental performance of liquid-environment dynamic-mode AFM. The present AFM technique does not allow real-time imaging of atomic-scale dynamic phenomena at a solid-liquid interface. This has hindered atomic-level understanding of crystal growth and dissolution, catalytic reactions and metal corrosion processes. Improvement in force sensitivity is required not only for such a high-speed imaging but also for various surface property measurements using a high-resolution AFM technique. In this review, we summarize recent works on the improvements in the force sensitivity and operation speed of atomic-resolution dynamic-mode AFM for liquid-environment applications. © 2015 The Japan Society of Applied Physics.Embargo Period 12 month