ABSTRACT
We investigate the impact of solar activity changes on mortality from cardiovascular causes of death in the period 1994-2011 in the Czech Republic. This period coincides with the time of solar cycle no. 23 and the surrounding minima when there was an unusually low level of solar activity. We use long-period daily time series of numbers of deaths by cause, solar activity indices (the relative sunspot number, and the intensity of solar radio flux), geomagnetic indices (Kp-the planetary index that indicates the fluctuation rate of horizontal components of the geomagnetic field, the Auroral Electrojet, and the disturbance storm time), and physical parameters describing the ionospheric effects (the critical frequency of the ionospheric F2 layer and the content of free electrons in the ionosphere). The results of the analysis confirm the hypothesis that there is no direct correlation between the geomagnetic solar index, Kp, and the number of deaths from acute myocardial infarction (code I21) or brain stroke (code I64) during the maxima of the solar cycle. On the other hand, the ionospheric parameters explain a greater part of the variability in the number of deaths for acute myocardial infarction or brain stroke than the model with solar parameters. The analysis shows that, because the values are geographically specific, the ionospheric parameters may describe the variability in the number of deaths from cardiovascular causes better than the solar indices. The cardiovascular diseases thus respond to the changes in the solar activity and to abnormal solar events indirectly through a concentration of electrical charges in the earth's environment.
Subject(s)
Cardiovascular Diseases/mortality , Ecosystem , Solar Activity , Cardiovascular Diseases/diagnosis , Cause of Death , Cluster Analysis , Cosmic Radiation , Czech Republic/epidemiology , Electromagnetic Fields , Humans , Linear Models , Myocardial Infarction/diagnosis , Myocardial Infarction/mortality , Prognosis , Risk Factors , Solar Energy , Stochastic Processes , Stroke/diagnosis , Stroke/mortality , Time FactorsABSTRACT
Production of small RNAs by ribonuclease III Dicer is a key step in microRNA and RNA interference pathways, which employ Dicer-produced small RNAs as sequence-specific silencing guides. Further studies and manipulations of microRNA and RNA interference pathways would benefit from identification of small-molecule modulators. Here, we report a study of a fluorescence-based in vitro Dicer cleavage assay, which was adapted for high-throughput screening. The kinetic assay can be performed under single-turnover conditions (35 nM substrate and 70 nM Dicer) in a small volume (5 µL), which makes it suitable for high-throughput screening in a 1536-well format. As a proof of principle, a small library of bioactive compounds was analyzed, demonstrating potential of the assay.
Subject(s)
High-Throughput Screening Assays , Ribonuclease III/genetics , Ribonuclease III/metabolism , Spectrometry, Fluorescence/methods , Animals , Cell Line , Drug Discovery , Humans , Reproducibility of Results , Ribonuclease III/antagonists & inhibitors , Small Molecule Libraries , Substrate SpecificityABSTRACT
Processing bodies (P-bodies) are dynamic cytoplasmic structures involved in mRNA degradation, but the mechanism that governs their formation is poorly understood. In this paper, we address a role of Like-Sm (LSm) proteins in formation of P-bodies and provide evidence that depletion of nuclear LSm8 increases the number of P-bodies, while LSm8 overexpression leads to P-body loss. We show that LSm8 knockdown causes relocalization of LSm4 and LSm6 proteins to the cytoplasm and suggest that LSm8 controls nuclear accumulation of all LSm2-7 proteins. We propose a model in which redistribution of LSm2-7 to the cytoplasm creates new binding sites for other P-body components and nucleates new, microscopically visible structures. The model is supported by prolonged residence of two P-body proteins, DDX6 and Ago2, in P-bodies after LSm8 depletion, which indicates stronger interactions between these proteins and P-bodies. Finally, an increased number of P-bodies has negligible effects on microRNA-mediated translation repression and nonsense mediated decay, further supporting the view that the function of proteins localized in P-bodies is independent of visible P-bodies.
Subject(s)
Cell Nucleus/metabolism , Cytoplasmic Granules/metabolism , N-Terminal Acetyltransferase C/physiology , RNA Processing, Post-Transcriptional , Ribonucleoprotein, U4-U6 Small Nuclear/physiology , Autoantigens/metabolism , DEAD-box RNA Helicases/metabolism , Humans , Microscopy, Fluorescence , N-Terminal Acetyltransferase C/metabolism , Protein Transport , Proto-Oncogene Proteins/metabolism , RNA-Binding Proteins/metabolism , Recombinant Fusion Proteins/metabolism , Ribonucleoprotein, U4-U6 Small Nuclear/metabolism , Ribonucleoproteins, Small Nuclear/metabolismABSTRACT
More than a decade has passed since the discovery of RNA interference (RNAi), an eukaryotic sequence-specific degradation of mRNA induced by complementary double-stranded RNA (dsRNA). RNAi became a common tool for controlled down-regulation of gene expression in cultured cells, as well as in various model organisms. This review summarizes RNAi-based tools for silencing genes in living mammals, which include: (i) transgenic RNAi strategies, where RNAi is triggered by a transgene transmitted through the germline and (ii) approaches, where an RNAi trigger is delivered into an adult animal.
