Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice

We have previously developed an efficient and safe transfection method for the kidney in mice: renal suction-mediated transfection. In this study, we verified the detailed characteristics of transgene expression and plasmid DNA (pDNA) in mice to develop therapeutic strategies and application to gene function analysis in the kidney. After naked pDNA was administered intravenously, the right kidney was immediately suctioned by a tissue suction device. We examined the spatial distribution of transgene expression and pDNA in the suctioned kidney using tissue clearing by CUBIC, ClearT2, and Scale SQ reagents. Spatial distribution analysis showed that pDNA was transfected into extravascular cells and sufficiently delivered to the deep renal cortex. In addition, we revealed that transgene expression occurred mainly in peritubular fibroblasts of the suctioned kidney by tissue clearing and immunohistochemistry. Next, we confirmed the periods of pDNA uptake and activation of transcription factors nuclear factor-κB and activator protein 1 by luciferase assays. Moreover, the use of a pCpG-free plasmid enabled sustained transgene expression in the suctioned kidney. In conclusion, analyses of the spatial distribution and immunostaining of the section suggest that pDNA and transgene expression occurs mainly in peritubular fibroblasts of the suctioned kidney. In addition, we clarified some factors for efficient and/or sustained transgene expression in the suctioned kidney

Internal Promotion and the Effect of Board Monitoring: A Comparison of Japan and the United States

This paper analyses two pronounced features of Japanese corporate governance--large corporate boards almost entirely composed of insiders and the tendency to appoint CEOs through internal promotions. It is often argued that Japanese boards are less effective in monitoring CEOs than U.S. boards which tends to be composed of a small number of directors, majority of which are outsiders. I show that Japanese corporate governance exhibits less inefficiencies than U.S. corporate governance. I further discuss the recent changes in Japanese corporate governance and provide theoretical explanation that they do not necessarily enhance board monitoring

Electoral Redistricting, Incumbency Advantage, and Endogenous Candidate Selection

In this paper, we examine the effects of redrawing district boundaries (redistricting) on political parties� candidate selection strategies. This study is distinct from previous studies in two regards. First, we focus on a unique natural experimental setting in Australia, where a non-partisan committee determines redistricting exogenously to the causal mechanism constituting the process of candidate selection. Second, we develop a formal model of endogenous candidate selection, which assumes strategic interactions by a party that won a seat in the previous election and a party that was defeated, and examine how an exogenous shock (i.e., redistricting) affects their decisions. Our empirical results based on the Australian Lower House election data for the period of 1990-2010 are consistent with an incumbency disadvantage hypothesis deduced from our model; namely, the magnitude of redistricting significantly increases the probability that the winning party will replace their incumbent legislator with a fresh candidate

Decomposition of Valence and Paradox of Primary Election

When does a political party decide to hold a primary election to choose a candidate for the next election, and when does an incumbent decide to announce his/her political retirement? Furthermore, if they make decisions interactively, how do their decisions affect the quality � �valence� � of the candidate who ends up running for the party? In this paper, we examine these questions by modeling a simple strategic game between a party and an incumbent in the context of Australia where political parties take important roles in selecting candidates and making policies. Specifically, by decomposing the valence into three factors, we show that primary elections may facilitate (potentially) high-quality incumbents to retire unexpectedly. We call it a �paradox of primary elec

Derivation of Mesenchymal Stromal Cells from Pluripotent Stem Cells through a Neural Crest Lineage using Small Molecule Compounds with Defined Media

<div><p>Neural crest cells (NCCs) are an embryonic migratory cell population with the ability to differentiate into a wide variety of cell types that contribute to the craniofacial skeleton, cornea, peripheral nervous system, and skin pigmentation. This ability suggests the promising role of NCCs as a source for cell-based therapy. Although several methods have been used to induce human NCCs (hNCCs) from human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), further modifications are required to improve the robustness, efficacy, and simplicity of these methods. Chemically defined medium (CDM) was used as the basal medium in the induction and maintenance steps. By optimizing the culture conditions, the combination of the GSK3β inhibitor and TGFβ inhibitor with a minimum growth factor (insulin) very efficiently induced hNCCs (70–80%) from hPSCs. The induced hNCCs expressed cranial NCC-related genes and stably proliferated in CDM supplemented with EGF and FGF2 up to at least 10 passages without changes being observed in the major gene expression profiles. Differentiation properties were confirmed for peripheral neurons, glia, melanocytes, and corneal endothelial cells. In addition, cells with differentiation characteristics similar to multipotent mesenchymal stromal cells (MSCs) were induced from hNCCs using CDM specific for human MSCs. Our simple and robust induction protocol using small molecule compounds with defined media enabled the generation of hNCCs as an intermediate material producing terminally differentiated cells for cell-based innovative medicine.</p></div