Loop-Mediated Isothermal Amplification for Influenza A (H5N1) Virus

We describe a 1-step reverse-transcription loop-mediated isothermal amplification assay for detection of highly pathogenic avian influenza A (H5N1) viruses. The assay was tested by using a panel of highly pathogenic H5N1 subtypes isolated over the past 10 years and clinical specimens. The assay produced negative results for all non-H5N1 subtypes

Benzene at 1GHz. Magnetic field-induced fine structure

The deuterium NMR spectrum of benzene-d6 in a high field spectrometer (1 GHz protons) exhibits a magnetic field-induced deuterium quadrupolar splitting ??. The magnitude of ?? observed for the central resonance is smaller than that observed for the 13C satellite doublets ???. This difference, ?(??) = ??? ? ??, is due to unresolved fine structure contributions to the respective resonances. We determine the origins of and simulate this difference, and report pulse sequences that exploit the connectivity of the peaks in the 13C and 2H spectra to determine the relative signs of the indirect coupling, JCD, and ??. The positive sign found for ?? is consonant with the magnetic field biasing of an isolated benzene molecule—the magnetic energy of the aromatic ring is lowest for configurations where the C6 axis is normal to the field. In the neat liquid the magnitude of ?? is decreased by the pair correlations in this prototypical molecular liquid

Hexagonal dielectric resonators and microcrystal lasers

We study long-lived resonances (lowest-loss modes) in hexagonally shaped dielectric resonators in order to gain insight into the physics of a class of microcrystal lasers. Numerical results on resonance positions and lifetimes, near-field intensity patterns, far-field emission patterns, and effects of rounding of corners are presented. Most features are explained by a semiclassical approximation based on pseudointegrable ray dynamics and boundary waves. The semiclassical model is also relevant for other microlasers of polygonal geometry.Comment: 12 pages, 17 figures (3 with reduced quality

Repeatability of ectopic beats from 48-hr ambulatory electrocardiography: The Atherosclerosis Risk in Communities (ARIC) Study

Background: The purpose of this study was to characterize the repeatability of ectopic beats, defined by premature atrial contractions (PACs) and premature ventricular contractions (PVCs), on ambulatory electrocardiogram (aECG) monitoring and evaluate the effect of length of aECG monitoring on the repeatability estimates. Methods: This analysis includes 95 randomly selected participants from the Atherosclerosis Risk in Communities Study (ARIC; 2011–2013). The participants wore a Holter monitor for two, 48-hr periods separated by a mean of 38 days following an identical, standardized protocol. We divided each 48-hr recording into 3-, 6-, 12-, and 24-hr recording periods and calculated intraclass correlation coefficients (ICCs) for PACs and PVCs and also as a percentage of the corresponding total of recorded beats per hour among these periods. Results: All participants had ≥1 PAC during the 48-hr recordings, and only two participants had no PVCs. ICCs were >0.83 for all indices and recording lengths ≥12 hrs. ICCs were intermediate for 6-hr recordings (range 0.80–0.83) and lower for 3-hr recordings (range 0.74–0.80). The ratio of the between- to within-participant variation increased with recording length. Conclusion: Repeatability of PACs and PVCs was excellent for recording lengths of 6–24 hr and fair for 3 hr. Repeatability varies over shorter duration recordings within the 48-hr recording period, and thus the present results have implications for detection algorithms for ectopic beats and can facilitate epidemiologic and clinical applications in which knowledge of measurement variability and misclassification are needed

Mini viral RNAs act as innate immune agonists during influenza virus infection

We thank the High-Throughput Genomics Group at the Wellcome Trust Centre for Human Genetics (funded by Wellcome Trust grant 090532/Z/09/Z) for the generation of adapter-ligated mvRNA sequencing data. This work was supported by the Wellcome Trust grant 098721/Z/12/Z, the joint Wellcome Trust and Royal Society grant 206579/Z/17/Z and a Netherlands Organization for Scientific Research (NWO) grant 825.11.029 to A.J.W.t.V.; EPA Cephalosporin Junior Research Fellowship to D.L.V.B.; support by the Intramural Research Program of NIAID, NIH, to E.d.W.; Research Grants Council of the Hong Kong Special Administrative Region, China, project no. T11-705/14N and a Croucher Senior Research Fellowship to L.L.M.P.; and Medical Research Council (MRC) programme grants MR/K000241/1 and MR/R009945/1 to E.F. and studentship to J.C.L.The molecular processes that determine the outcome of influenza virus infection in humans are multifactorial and involve a complex interplay between host, viral and bacterial factors1. However, it is generally accepted that a strong innate immune dysregulation known as ‘cytokine storm’ contributes to the pathology of infections with the 1918 H1N1 pandemic or the highly pathogenic avian influenza viruses of the H5N1 subtype2,3,4. The RNA sensor retinoic acid-inducible gene I (RIG-I) plays an important role in sensing viral infection and initiating a signalling cascade that leads to interferon expression5. Here, we show that short aberrant RNAs (mini viral RNAs (mvRNAs)), produced by the viral RNA polymerase during the replication of the viral RNA genome, bind to and activate RIG-I and lead to the expression of interferon-β. We find that erroneous polymerase activity, dysregulation of viral RNA replication or the presence of avian-specific amino acids underlie mvRNA generation and cytokine expression in mammalian cells. By deep sequencing RNA samples from the lungs of ferrets infected with influenza viruses, we show that mvRNAs are generated during infection in vivo. We propose that mvRNAs act as the main agonists of RIG-I during influenza virus infection.PostprintPeer reviewe