A rapid and sensitive assay for quantification of siRNA efficiency and specificity

RNA Interference has rapidly emerged as an efficient procedure for knocking down gene expression in model systems. However, cross-reactivity, whereby multiple genes may be simultaneously targeted by a single short interfering RNA (siRNA), can potentially jeopardize correct interpretation of gene function. As such, it is essential to test the specificity of a siRNA prior to a full phenotypic analysis. To this end, we have adapted a reporter-based assay harnessing the sensitivity of luciferase activity to provide a quantitative readout of relative RNAi efficacy and specificity. We have tested different siRNAs directed against Thymosin β4 (Tβ4); determined their effectiveness at silencing Tβ4 and have both excluded off-target silencing of the Tβ4 homologue Thymosin β10 (Tβ10) and demonstrated partial knockdown of Tβ10 despite significant (12/23; 52%) sequence mismatch. This assay system is applicable to any RNAi study where there is a risk of targeting homologous genes and to the monitoring of off-target effects at the genome level following microarray expression profiling

The orchestrated cellular and molecular responses of the kidney to endotoxin define a precise sepsis timeline

Sepsis is a dynamic state that progresses at variable rates and has life-threatening consequences. Staging patients along the sepsis timeline requires a thorough knowledge of the evolution of cellular and molecular events at the tissue level. Here, we investigated the kidney, an organ central to the pathophysiology of sepsis. Single-cell RNA-sequencing in a murine endotoxemia model revealed the involvement of various cell populations to be temporally organized and highly orchestrated. Endothelial and stromal cells were the first responders. At later time points, epithelial cells upregulated immune-related pathways while concomitantly downregulating physiological functions such as solute homeostasis. Sixteen hours after endotoxin, there was global cell-cell communication failure and organ shutdown. Despite this apparent organ paralysis, upstream regulatory analysis showed significant activity in pathways involved in healing and recovery. This rigorous spatial and temporal definition of murine endotoxemia will uncover precise biomarkers and targets that can help stage and treat human sepsis

Development and Verification of a Non-Linear Look-Up Table Model for RF Silicon BJTs

This work presents, for the first time, a large-signal look-up table based model for the forward-active region of RF Si BJTs. The state functions are obtained by integrating the device’s bias-dependent intrinsic elements, extracted from de-embedded multi-bias S-parameter measurements. The model accuracy is evaluated using vectorial large-signal measurements

United States Bureau of Mines Information Circular 8012

Report issued by the U.S. Bureau of Mines discussing mining methods of the Rosiclare Fluorspar mine and mill. As stated in the introduction, "the publication covers exploration, development, mining and milling practices, mine-drainage problems, and other activities and is presented as part of the Bureau of Mines series of publications on mining and milling practices in various parts of the United States" (p. 1). This report includes maps, tables, illustrations, and photographs

Compact diode-pumped passively Q-switched tunable Er-Yb double-clad fiber laser

Highly-efficient repetitive passive Q-switching of a cladding-pumped Er-Yb fiber laser has been demonstrated by employing an external cavity configuration containing a Co2+:ZnS crystal as saturable absorber. Energies up to 60µJ in pulses of duration as short as 3.5ns (FWHM), corresponding to a peak power >10kW, have been generated, and the maximum slope efficiency with respect to absorbed pump power was 13%. Using a bulk diffraction grating in Littrow configuration to provide wavelength selective feedback, the passively Q-switched fiber laser was tuned over 31nm from 1532nm to 1563nm. The prospects for further improvement in performance will be discussed