CD62L as target receptor for specific gene delivery into less differentiated human T lymphocytes

Chimeric antigen receptor (CAR)-expressing T cells are a complex and heterogeneous gene therapy product with variable phenotype compositions. A higher proportion of less differentiated CAR T cells is usually associated with improved antitumoral function and persistence. We describe in this study a novel receptor-targeted lentiviral vector (LV) named 62L-LV that preferentially transduces less differentiated T cells marked by the L-selectin receptor CD62L, with transduction rates of up to 70% of CD4+ and 50% of CD8+ primary T cells. Remarkably, higher amounts of less differentiated T cells are transduced and preserved upon long-term cultivation using 62L-LV compared to VSV-LV. Interestingly, shed CD62L neither altered the binding of 62L-LV particles to T cells nor impacted their transduction. The incubation of 2 days of activated T lymphocytes with 62L-LV or VSV-LV for only 24 hours was sufficient to generate CAR T cells that controlled tumor growth in a leukemia tumor mouse model. The data proved that potent CAR T cells can be generated by short-term ex vivo exposure of primary cells to LVs. As a first vector type that preferentially transduces less differentiated T lymphocytes, 62L-LV has the potential to circumvent cumbersome selections of T cell subtypes and offers substantial shortening of the CAR T cell manufacturing process

Validation of a Horizontal Standard for the Determination of Linear Alkylbenzene Sulphonates (LAS) by Liquid Chromatography with Fluorescence Detection (LC-FLD) and Liquid Chromatography with Mass Selective Detection (LC-MS) in a European Intercomparison Exercise

Project HORIZONTAL is interdisciplinary aiming at a harmonisation and horizontal standardisation of test procedures, in particular for sludge, soils and biowastes. In the context of this standardization project, a series of draft technical specification were designed upon an extensive desk study, fine-tuned after expert consultations and finally validated in international intercomparisons exercise. This report summarises the work performed within the validation study of the draft standard for the determination of linear alkylbenzene sulphonates (LAS) in soils, sludge and treated bio-waste using liquid chromatography with fluorescence detection (LC-FLD) and liquid chromatography with mass selective detection (LC-MS). It further explains the underlying statistical concept for the calculation of reproducibility and repeatability from intercomparisons data. In addition all single values, results of the statistical evaluation as well as background information on the validation materials used are described and explained.JRC.H.5-Rural, water and ecosystem resource

Integrators for highly oscillatory Hamiltonian systems: an homogenization approach

International audienceWe introduce a systematic way to construct symplectic schemes for the numerical integration of a large class of highly oscillatory Hamiltonian systems. The bottom line of our construction is to consider the Hamilton-Jacobi form of the Newton equations of motion, and to perform a two-scale expansion of the solution, for small times and high frequencies. The approximation obtained for the solution is then used as a generating function, from which the numerical scheme is derived. Several options for the derivation are presented, some of them also giving rise to non symplectic variants. The various integrators obtained are tested and compared to several existing algorithms. The numerical results demonstrate their efficiency

Language Mechanisms for Controlling and Mitigating Timing Channels

We propose a new language-based approach to mitigating timing channels. In this language, well-typed programs provably leak only a bounded amount of information over time through external timing channels. By incorporating mechanisms for predictive mitigation of timing channels, this approach also permits a more expressive programming model. Timing channels arising from interaction with underlying hardware features such as instruction caches are controlled. Assumptions about the underlying hardware are explicitly formalized, supporting the design of hardware that efficiently controls timing channels. One such hardware design is modeled and used to show that timing channels can be controlled in some simple programs of real-world significance.This work has been supported by a grant from the Office of Naval Research (ONR N000140910652), by two grants from the NSF: 0424422 (the TRUST center), and 0964409, and by MURI grant FA9550-12-1-0400, administered by the US Air Force. This research is also sponsored by the Air Force Research Laboratory

Effects of EGb 761® Ginkgo biloba extract on mitochondrial function and oxidative stress

As major sources of reactive oxygen species (ROS), mitochondrial structures are exposed to high concentrations of ROS and may therefore be particularly susceptible to oxidative damage. Mitochondrial damage could play a pivotal role in the cell death decision. A decrease in mitochondrial energy charge and redox state, loss of transmembrane potential (depolarization), mitochondrial respiratory chain impairment, and release of substances such as calcium and cytochrome c all contribute to apoptosis. These mitochondrial abnormalities may constitute a part of the spectrum of chronic oxidative stress in Alzheimer's disease. Accumulation of amyloid beta (Abeta) in form of senile plaques is also thought to play a central role in the pathogenesis of Alzheimer's disease mediated by oxidative stress. In addition, increasing evidence shows that Abeta generates free radicals in vitro, which mediate the toxicity of this peptide. In our study, PC12 cells were used to examine the protective features of EGb 761(definition see editorial) on mitochondria stressed with hydrogen peroxide and antimycin, an inhibitor of complex III. In addition, we investigated the efficacy of EGb 761 in Abeta-induced MTT reduction in PC12 cells. Moreover, we examined the effects of EGb 761 on ROS levels and ROS-induced apoptosis in lymphocytes from aged mice after in vivo administration. Here, we will report that EGb 761 was able to protect mitochondria from the attack of hydrogen peroxide, antimycin and Abeta. Furthermore, EGb 761 reduced ROS levels and ROS-induced apoptosis in lymphocytes from aged mice treated orally with EGb 761 for 2 weeks. Our data further emphasize neuroprotective properties of EGb 761, such as protection against Abeta-toxicity, and antiapoptotic properties, which are probably due to its preventive effects on mitochondria

Lattice worldline representation of correlators in a background field

We use a discrete worldline representation in order to study the continuum limit of the one-loop expectation value of dimension two and four local operators in a background field. We illustrate this technique in the case of a scalar field coupled to a non-Abelian background gauge field. The first two coefficients of the expansion in powers of the lattice spacing can be expressed as sums over random walks on a d-dimensional cubic lattice. Using combinatorial identities for the distribution of the areas of closed random walks on a lattice, these coefficients can be turned into simple integrals. Our results are valid for an anisotropic lattice, with arbitrary lattice spacings in each direction.Comment: 54 pages, 14 figure