Cellular Deformations Induced by Conical Silicon Nanowire Arrays Facilitate Gene Delivery

Engineered cell–nanostructured interfaces generated by vertically aligned silicon nanowire (SiNW) arrays have become a promising platform for orchestrating cell behavior, function, and fate. However, the underlying mechanism in SiNW-mediated intracellular access and delivery is still poorly understood. This study demonstrates the development of a gene delivery platform based on conical SiNW arrays for mechanical cell transfection, assisted by centrifugal force, for both adherent and nonadherent cells in vitro. Cells form focal adhesions on SiNWs within 6 h, and maintain high viability and motility. Such a functional and dynamic cell–SiNW interface features conformational changes in the plasma membrane and in some cases the nucleus, promoting both direct penetration and endocytosis; this synergistically facilitates SiNW-mediated delivery of nucleic acids into immortalized cell lines, and into difficult-to-transfect primary immune T cells without pre-activation. Moreover, transfected cells retrieved from SiNWs retain the capacity to proliferate—crucial to future biomedical applications. The results indicate that SiNW-mediated intracellular delivery holds great promise for developing increasingly sophisticated investigative and therapeutic tools. © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinhei

Setting Financial Accounting Standards for the Twenty-first Century

This paper was prepared for the purpose of facilitating discussion at the Symposium on Financial Reporting and Standard Setting sponsored by the American Institute of Certified Public Accountants. The authors of the paper believe that the changing environment necessitates a thorough reexamination of financial accounting standard-setting processes in the United States. The following observations underlie the propositions offered in the paper: * The increasing demand for and availability of alternative information is decreasing the demand for traditional financial accounting information. * The conventional historical-cost-based accounting model is becoming increasingly inadequate for decision-support purposes

Efficient non-viral CAR-T cell generation via silicon-nanotube-mediated transfection

Cell-based immunotherapy such as chimeric antigen receptor (CAR)-T therapy holds great promise in treating cancer and other diseases; but the current viral-based method represents a significant cost and safety hurdle. Here, we show for the first time successful CAR transfection into primary T cells via vertically aligned silicon nanotube (SiNT) arrays. SiNT-mediated transfection achieves comparable or even higher delivery efficiency (20–37%) and expression efficiency (18–24%) to that achieved by electroporation. Scanning electron microscopy imaging after focused ion beam milling demonstrated the tight T cell–SiNT interface. The induced membrane invaginations and the proximity between individual SiNTs and the nucleus might enhance endocytic pathways, and enable direct delivery of CAR construct into the nucleus, thus resulting in higher CAR expression efficiency. SiNT-interfacing also results in faster proliferation of T cells compared to cells transfected by electroporation; nonactivated T (N_SiNT) cells undergo higher numbers of cell division than pre-activated ones (A_SiNT). By co-culturing with target lymphoma Raji cells, we prove that SiNT-transfected CAR-T cells can suppress Raji cell growth, indicated by significant increase in effector:target (E:T) ratio (by up to 30.7-fold). While SiNTs induce an overall upregulation of cytokine production in T cells, N_SiNT T cells exhibited high increase in secretion of IFNc and IL-6, and relatively high in TNFa, which could contribute to their enhanced killing ability (∼96% cytotoxicity), demonstrated by their stronger inhibition on target Raji cells through luciferase assay. The results demonstrate the capacity of SiNT-mediated transfection of generating effective anti-lymphoma CAR-T cells. Considering the growing potential of cell-based therapies, we expect that a non-viral nanoinjection platform such as ours will facilitate the full realization of their therapeutic promise.Yaping Chen, Melanie Mach, Ali-Reza Shokouhi, Hao Zhe Yoh, David C. Bishop, Takahide Murayama, Koukou Suu, Yasuhiro Morikawa, Simon C. Barry, Kenneth Micklethwaite, Roey Elnathan, Nicolas H. Voelcke

What have we learned? Themes from the literature on best-practice benchmarking

The aim of this paper is to review the important themes in the literature on benchmarking, and to assess the contribution to knowledge provided thus far. Four themes from the literature are highlighted: studies of the nature of benchmarking practice; criticisms of benchmarking; evaluating the effectiveness of benchmarking; and the notion of best practice. The authors argue that, while the literature does include a few critical analyses of the practice and impacts of benchmarking, it is dominated by relatively descriptive, and even prescriptive, accounts of the ‘realities’ of benchmarking. The longer-term qualitative and quantitative) effects and perspectives of diverse stakeholders on benchmarking are largely neglected. This selective focus has implications for the contribution of benchmarking research to practice, for example, in terms of the role of internal organizational capabilities in analysing the performance of processes and managing communications and change. The authors advocate areas of future research to improve theoretical understanding of benchmarking and thoroughly evaluate its impact within the context of performance management