China, Internet Freedom, and U.S. Policy

This report discusses Congressional interest in how Internet use in the People's Republic of China (PRC) is tied to human rights concerns in several ways: as a U.S. policy tool for promoting rights in China; though use of the Internet political dissidents and political repression; the role of U.S. Internet companies in spreading freedom and complying with PRC censorship; and the development of U.S. Internet freedom policies globally

Induction of Autophagy by Cystatin C: A Mechanism That Protects Murine Primary Cortical Neurons and Neuronal Cell Lines

Cystatin C (CysC) expression in the brain is elevated in human patients with epilepsy, in animal models of neurodegenerative conditions, and in response to injury, but whether up-regulated CysC expression is a manifestation of neurodegeneration or a cellular repair response is not understood. This study demonstrates that human CysC is neuroprotective in cultures exposed to cytotoxic challenges, including nutritional-deprivation, colchicine, staurosporine, and oxidative stress. While CysC is a cysteine protease inhibitor, cathepsin B inhibition was not required for the neuroprotective action of CysC. Cells responded to CysC by inducing fully functional autophagy via the mTOR pathway, leading to enhanced proteolytic clearance of autophagy substrates by lysosomes. Neuroprotective effects of CysC were prevented by inhibiting autophagy with beclin 1 siRNA or 3-methyladenine. Our findings show that CysC plays a protective role under conditions of neuronal challenge by inducing autophagy via mTOR inhibition and are consistent with CysC being neuroprotective in neurodegenerative diseases. Thus, modulation of CysC expression has therapeutic implications for stroke, Alzheimer's disease, and other neurodegenerative disorders

Value-Driven Analysis of New Paradigms in Space Architectures: An Ilities-Based Approach

Current commercial, civil, and military space architecture designs perform exquisitely and reliably. However, today’s architecture paradigms are also characterized by expensive launches, large and expensive high-performance spacecraft, long development cycles, and wide variations in ground architectures. While current assets provide high-quality services, and future assets are slated to improve performance within the same design frameworks, proposed future architectures may not be capitalizing on technology improvements, system innovations, or policy alternatives explored during the last two decades. This paper identifies five “trends” along which space architectures may develop, aimed at granting systems several “ilities,” such as resiliency, robustness, flexibility, scalability, and affordability. The trends examined include: commercialization of space, significant reductions in launch costs and the development of hybrid or reusable launch systems, development of on-orbit infrastructure and servicing, aggregation or disaggregation of orbital assets, and the automation and standardization of ground architectures. Further refinement of these key technological and system trends could result in major paradigm shifts in the development and fielding of space operations as well as lead to space architecture designs in the future that are radically different from those today. Within the framework of systems engineering ilities and risk management, this paper reviews current literature surrounding these new change trends and justifies their potential to cause significant paradigm shifts. By examining the work and research conducted so far through an ilities-based approach, systems engineers can more fully appreciate the value being offered by these trends

Sleeping Beauty Transposition of Chimeric Antigen Receptors Targeting Receptor Tyrosine Kinase-Like Orphan Receptor-1 (ROR1) into Diverse Memory T-Cell Populations.

T cells modified with chimeric antigen receptors (CARs) targeting CD19 demonstrated clinical activity against some B-cell malignancies. However, this is often accompanied by a loss of normal CD19+ B cells and humoral immunity. Receptor tyrosine kinase-like orphan receptor-1 (ROR1) is expressed on sub-populations of B-cell malignancies and solid tumors, but not by healthy B cells or normal post-partum tissues. Thus, adoptive transfer of T cells specific for ROR1 has potential to eliminate tumor cells and spare healthy tissues. To test this hypothesis, we developed CARs targeting ROR1 in order to generate T cells specific for malignant cells. Two Sleeping Beauty transposons were constructed with 2nd generation ROR1-specific CARs signaling through CD3ζ and either CD28 (designated ROR1RCD28) or CD137 (designated ROR1RCD137) and were introduced into T cells. We selected for T cells expressing CAR through co-culture with γ-irradiated activating and propagating cells (AaPC), which co-expressed ROR1 and co-stimulatory molecules. Numeric expansion over one month of co-culture on AaPC in presence of soluble interleukin (IL)-2 and IL-21 occurred and resulted in a diverse memory phenotype of CAR+ T cells as measured by non-enzymatic digital array (NanoString) and multi-panel flow cytometry. Such T cells produced interferon-γ and had specific cytotoxic activity against ROR1+ tumors. Moreover, such cells could eliminate ROR1+ tumor xenografts, especially T cells expressing ROR1RCD137. Clinical trials will investigate the ability of ROR1-specific CAR+ T cells to specifically eliminate tumor cells while maintaining normal B-cell repertoire

CysC induces an increase in the number of autophagic vesicles (AVs) in neuronal cells.

<p>Electron micrographs of primary rat cortical neurons (<b>A</b>) incubated for 12 hours in either B27-supplemented or deprived medium in the absence or presence of 0.75 µM CysC. The total number of AVs <i>per</i> cell was counted for at least 20 cells/condition, and the average number of vesicles <i>per</i> cell is shown (<b>B</b>). Data are the mean and SEM. Differences between CysC treated and untreated samples were calculated by Student's t-test.</p

CysC induces autophagy via the mTOR signaling pathway in either serum-containing or serum-free medium.

<p><b>A.</b> N2a cells were incubated for 12 hrs in serum-containing or serum-free medium in the presence or absence of CysC. Cell lysate proteins were separated by gel electrophoresis and blotted with antibodies to P70S6 kinase, p-P70S6 kinase (Thr389), or β-tubulin. Representative images of Western blot analysis are presented. <b>B.</b> The intensity of the bands was measured, and p-P70S6 protein levels were calculated relative to total P70S6 values showing a decrease in the level of P70S6 kinase phosphorylation. Data are the mean and SEM of 6 experiments. For serum containing groups the F and P values determined by one way ANOVA were 9.07 and 0.006 and for serum deprived groups were 10.18 and 0.005.</p