Role of Dicer Enzyme in the Regulation of Store Operated Calcium Entry (SOCE) in CD4+ T Cells

Background/Aims: Activation of T cell receptors (TCRs) in CD4+ T cells leads to a cascade of signalling reactions including increase of intracellular calcium (Ca2+) levels with subsequent Ca2+ dependent stimulation of gene expression, proliferation, cell motility and cytokine release. The increase of cytosolic Ca2+ results from intracellular Ca2+ release with subsequent activation of store-operated Ca2+ entry (SOCE). Previous studies suggested miRNAs are required for the development and functions of CD4+ T cells. An enzyme called Dicer is required during the process of manufacturing mature miRNAs from the precursor miRNAs. In this study, we explored whether loss of Dicer in CD4+ T cells affects SOCE and thus Ca2+ dependent regulation of cellular functions. Methods: We tested the expression of Orai1 by q-RT-PCR and flow cytometry. Further, we measured SOCE by an inverted phase-contrast microscope with the Incident-light fluorescence illumination system using Fura-2. Intracellular Ca2+ was also measured by flow cytometry using Ca2+ sensitive dye Fluo-4. Results: We found that in Dicer deficient (DicerΔ/Δ) mice Orai1 was downregulated at mRNA and protein level in CD4+ T cells. Further, SOCE was significantly smaller in DicerΔ/Δ CD4+ T cells than in CD4+ T cells isolated from wild-type (Dicerfl/fl) mice. Conclusion: Our data suggest that miRNAs are required for adequate Ca2+ entry into CD4+ T cells and thus triggering of Ca2+ sensitive immune functions

Metabolism and distribution of p,p'-DDT during flight of the white-crowned sparrow, Zonotrichia leucophrys

This study evaluated the interactions of flight, fasting, and 1,1,1-trichloro-bis(4-chlorophenyl)ethane (p,p′-DDT) loading on residue metabolism and distribution in recently exposed white-crowned sparrows (Zonotrichia leucophrys). Female sparrows were dosed with 5 mg p,p′-DDT per kg body weight over 3 d. Following 1 d of recovery, sparrows were flown in a wind tunnel for up to 140 min, in 15-min blocks. Food was withheld from the start of the flight period until birds were euthanized. DDT, 1,1-dichloro-2,2-bis(4 chlorophenyl)ethane (DDD), and 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (DDE) were present in all tissues examined. 1-Chloro-2,2-bis(4-chlorophenyl)ethene (DDµ), 1,1-bis(4-chlorophenyl)ethane (p,p′-DDη), and 2,2-bis(4-chlorophenyl)ethanol (p,p′-DDOH) were not found. Fasting did not significantly affect the rate of residue increase over time in any of the tissues examined. When sparrows flew and fasted simultaneously, fasting seldom contributed to an increase in tissue residues. However, the length of time flown was significantly correlated with increasing toxicant concentrations in the brain, kidney, and liver, effectively demonstrating the potential for brief flights to enhance mobilization of DDT and its metabolites. Dose, flight, and fasting also increased residues in brain tissue. These contaminant redistributions may have important ramifications on the stresses experienced by migratory birds

Effects of the Fibrolytic Enzyme Cattle-ASE?äó on Growth of Prepuberal Romosinuano Crossbred Heifers

Last updated: 6/12/200

Stocker Performance of Romosinuano Crossbred Steers Grazing Bermudagrass and Receiving a Supplement

Last updated: 6/12/200

Nesprin-2-dependent ERK1/2 compartmentalisation regulates the DNA damage response in vascular smooth muscle cell ageing

Prelamin A accumulation and persistent DNA damage response (DDR) are hallmarks of vascular smooth muscle cell (VSMC) ageing and dysfunction. Although prelamin A is proposed to interfere with DNA repair, our understanding of the crosstalk between prelamin A and the repair process remains limited. The extracellular signal-regulated kinases 1 and 2 (ERK1/2) have emerged as key players in the DDR and are known to enhance ataxia telangiectasia-mutated protein (ATM)activity at DNA lesions, and in this study, we identified a novel relationship between prelamin A accumulation and ERK1/2 nuclear compartmentalisation during VSMC ageing. We show both prelamin A accumulation and increased DNA damage occur concomitantly, before VSMC replicative senescence, and induce the localisation of ERK1/2 to promyelocytic leukaemia protein nuclear bodies (PML NBs) at the sites of DNA damage via nesprin-2 and lamin A interactions. Importantly, VSMCs treated with DNA damaging agents also displayed prelamin A accumulation and ERK compartmentalisation at PML NBs, suggesting that prelamin A and nesprin-2 are novel components of the DDR. In support of this, disruption of ERK compartmentalisation at PML NBs, by either depletion of nesprin-2 or lamins A/C, resulted in the loss of ATM from DNA lesions. However, ATM signalling and DNA repair remained intact after lamins A/C depletion, whereas nesprin-2 disruptionablated downstream Chk2 activation and induced genomic instability. We conclude that lamins A/C and PML act as scaffolds to organise DNA-repair foci and compartmentalise nesprin-2/ERK signalling. However, nesprin-2/ERK signalling fidelity, but not their compartmentalisation at PML NBs, is essential for efficient DDR in VSMCs

MicroRNAs control the apoptotic threshold in primed Pluripotent stem cells through regulation of BIM

Mammalian primed pluripotent stem cells have been shown to be highly susceptible to cell death stimuli due to their low apoptotic threshold, but how this threshold is regulated remains largely unknown. Here we identify microRNA (miRNA)-mediated regulation as a key mechanism controlling apoptosis in the post-implantation epiblast. Moreover, we found that three miRNA families, miR-20, miR-92, and miR-302, control the mitochondrial apoptotic machinery by fine-tuning the levels of expression of the proapoptotic protein BIM. These families therefore represent an essential buffer needed to maintain cell survival in stem cells that are primed for not only differentiation but also cell death

The EPICS Software Framework Moves from Controls to Physics

The Experimental Physics and Industrial Control System (EPICS), is an open-source software framework for high-performance distributed control, and is at the heart of many of the world’s large accelerators and telescopes. Recently, EPICS has undergone a major revision, with the aim of better computing supporting for the next generation of machines and analytical tools. Many new data types, such as matrices, tables, images, and statistical descriptions, plus users’ own data types, now supplement the simple scalar and waveform types of the former EPICS. New computational architectures for scientific computing have been added for high-performance data processing services and pipelining. Python and Java bindings have enabled powerful new user interfaces. The result has been that controls are now being integrated with modelling and simulation, machine learning, enterprise databases, and experiment DAQs. We introduce this new EPICS (version 7) from the perspective of accelerator physics and review early adoption cases in accelerators around the world

Effects of Thermal Process Parameters on Mechanical Interlayer Strength for Additively Manufactured Ultem 9085

The effects of the envelope temperature on the microstructure and mechanical strength of Ultem 9085 fused deposition modeling (FDM) components were studied. A customized build chamber was developed for a commercial 3D printer in order to control the envelope temperature during printing. Test specimens were printed in the vertical direction because their mechanical strength exhibited the greatest dependence on inter-layer adhesion and neck development. A delay was introduced between two layers in each specimen in order to create a weak region where the neck was not expected to fully develop. However, none of the specimens failed in this region. Mechanical testing revealed that neck growth was highly dependent on the envelope temperature, and the strength was shown to vary significantly (20%) based on the envelope temperature. The variability of the mechanical strength also decreased as the envelope temperature increased. Thermal imaging revealed that the cooling rate of the specimens was consistent regardless of the envelope temperature. Fracture analysis confirmed that higher envelope temperatures improved the amount of neck growth and inter-layer adhesion in the specimens. This study showed that increasing the envelope temperature created parts with higher strengths and improved consistencies

Far Ultraviolet Absolute Flux of alpha Virginis

We present the far ultraviolet spectrum of alpha Virginis taken with EURD spectrograph on-board MINISAT-01. The spectral range covered is from ~900 to 1080 A with 5 A spectral resolution. We have fitted Kurucz models to IUE spectra of alpha Vir and compared the extension of the model to our wavelengths with EURD data. This comparison shows that EURD fluxes are consistent with the prediction of the model within 20-30%, depending on the reddening assumed. EURD fluxes are consistent with Voyager observations but are ~60% higher than most previous rocket observations of alpha Vir.Comment: 13 pages, 4 figures. Submitted to The Astrophysical Journa

Current and Nascent SETI Instruments in the Radio and Optical

Here we describe our ongoing efforts to develop high-performance and sensitive instrumentation for use in the search for extra-terrestrial intelligence (SETI). These efforts include our recently deployed Search for Extraterrestrial Emissions from Nearby Developed Intelligent Populations Spectrometer (SERENDIP V.v) and two instruments currently under development; the Heterogeneous Radio SETI Spectrometer (HRSS) for SETI observations in the radio spectrum and the Optical SETI Fast Photometer (OSFP) for SETI observations in the optical band. We will discuss the basic SERENDIP V.v instrument design and initial analysis methodology, along with instrument architectures and observation strategies for OSFP and HRSS. In addition, we will demonstrate how these instruments may be built using low-cost, modular components and programmed and operated by students using common languages, e.g. ANSI C