Evasion of intracellular host defence by hepatitis C virus

Viral infection of mammalian cells rapidly triggers intracellular signalling events leading to interferon alpha/beta production and a cellular antiviral state. This 'host response' is our first line of immune defence against infection as it imposes several barriers to viral replication and spread. Hepatitis C virus (HCV) evades the host response through a complex combination of processes that include signalling interference, effector modulation and continual viral genetic variation. These evasion strategies support persistent infection and the spread of HCV. Defining the molecular mechanisms by which HCV regulates the host response is of crucial importance and may reveal targets for novel therapeutic strategies.     

LNA-mediated microRNA silencing in non-human primates

microRNAs (miRNAs) are small regulatory RNAs that are important in development and disease and therefore represent a potential new class of targets for therapeutic intervention. Despite recent progress in silencing of miRNAs in rodents, the development of effective and safe approaches for sequence-specific antagonism of miRNAs in vivo remains a significant scientific and therapeutic challenge. Moreover, there are no reports of miRNA antagonism in primates. Here we show that the simple systemic delivery of a unconjugated, PBS-formulated locked-nucleic-acid-modified oligonucleotide (LNA-antimiR) effectively antagonizes the liver-expressed miR-122 in non-human primates. Acute administration by intravenous injections of 3 or 10 mg kg(-1) LNA-antimiR to African green monkeys resulted in uptake of the LNA-antimiR in the cytoplasm of primate hepatocytes and formation of stable heteroduplexes between the LNA-antimiR and miR-122. This was accompanied by depletion of mature miR-122 and dose-dependent lowering of plasma cholesterol. Efficient silencing of miR-122 was achieved in primates by three doses of 10 mg kg(-1) LNA-antimiR, leading to a long-lasting and reversible decrease in total plasma cholesterol without any evidence for LNA-associated toxicities or histopathological changes in the study animals. Our findings demonstrate the utility of systemically administered LNA-antimiRs in exploring miRNA function in rodents and primates, and support the potential of these compounds as a new class of therapeutics for disease-associated miRNAs.     

2'-O methylation of the viral mRNA cap evades host restriction by IFIT family members

Cellular messenger RNA (mRNA) of higher eukaryotes and many viral RNAs are methylated at the N-7 and 2'-O positions of the 5' guanosine cap by specific nuclear and cytoplasmic methyltransferases (MTases), respectively. Whereas N-7 methylation is essential for RNA translation and stability, the function of 2'-O methylation has remained uncertain since its discovery 35 years ago. Here we show that a West Nile virus (WNV) mutant (E218A) that lacks 2'-O MTase activity was attenuated in wild-type primary cells and mice but was pathogenic in the absence of type I interferon (IFN) signalling. 2'-O methylation of viral RNA did not affect IFN induction in WNV-infected fibroblasts but instead modulated the antiviral effects of IFN-induced proteins with tetratricopeptide repeats (IFIT), which are interferon-stimulated genes (ISGs) implicated in regulation of protein translation. Poxvirus and coronavirus mutants that lacked 2'-O MTase activity similarly showed enhanced sensitivity to the antiviral actions of IFN and, specifically, IFIT proteins. Our results demonstrate that the 2'-O methylation of the 5' cap of viral RNA functions to subvert innate host antiviral responses through escape of IFIT-mediated suppression, and suggest an evolutionary explanation for 2'-O methylation of cellular mRNA: to distinguish self from non-self RNA. Differential methylation of cytoplasmic RNA probably serves as an example for pattern recognition and restriction of propagation of foreign viral RNA in host cells.     

Structural basis of RNA recognition and activation by innate immune receptor RIG-I

Retinoic-acid-inducible gene-I (RIG-I; also known as DDX58) is a cytoplasmic pathogen recognition receptor that recognizes pathogen-associated molecular pattern (PAMP) motifs to differentiate between viral and cellular RNAs. RIG-I is activated by blunt-ended double-stranded (ds)RNA with or without a 5'-triphosphate (ppp), by single-stranded RNA marked by a 5'-ppp and by polyuridine sequences. Upon binding to such PAMP motifs, RIG-I initiates a signalling cascade that induces innate immune defences and inflammatory cytokines to establish an antiviral state. The RIG-I pathway is highly regulated and aberrant signalling leads to apoptosis, altered cell differentiation, inflammation, autoimmune diseases and cancer. The helicase and repressor domains (RD) of RIG-I recognize dsRNA and 5'-ppp RNA to activate the two amino-terminal caspase recruitment domains (CARDs) for signalling. Here, to understand the synergy between the helicase and the RD for RNA binding, and the contribution of ATP hydrolysis to RIG-I activation, we determined the structure of human RIG-I helicase-RD in complex with dsRNA and an ATP analogue. The helicase-RD organizes into a ring around dsRNA, capping one end, while contacting both strands using previously uncharacterized motifs to recognize dsRNA. Small-angle X-ray scattering, limited proteolysis and differential scanning fluorimetry indicate that RIG-I is in an extended and flexible conformation that compacts upon binding RNA. These results provide a detailed view of the role of helicase in dsRNA recognition, the synergy between the RD and the helicase for RNA binding and the organization of full-length RIG-I bound to dsRNA, and provide evidence of a conformational change upon RNA binding. The RIG-I helicase-RD structure is consistent with dsRNA translocation without unwinding and cooperative binding to RNA. The structure yields unprecedented insight into innate immunity and has a broader impact on other areas of biology, including RNA interference and DNA repair, which utilize homologous helicase domains within DICER and FANCM.     

Relations between Karl Popper and Michael Polanyi

Karl Popper and Michael Polanyi grew up in central Europe and, having escaped from Nazism, went on to pursue academic careers in Britain where they wrote prolifically on science and politics. Popper and Polanyi corresponded with each other, and met for discussions in the late 1940s and early 50s, but they seldom referred to each other in their publications. This article examines their correspondence so as to produce a picture of their intellectual relations. The most important of the letters was one that Popper wrote in 1952, which we reproduce in its entirety, indicating his dissatisfaction with ideas that Polanyi had expressed in a paper of that year, ‘The Stability of Beliefs’. In this paper, Polanyi used the example of the framework of Zande witchcraft to shed analogical light on science and other systems of belief, arguing that ‘frameworks of belief’ equip their adherents with intellectual powers whose use reinforces commitment to the framework, inoculating adherents against criticism. Polanyi’s 1952 paper and his 1951 and 1952 Gifford Lectures (to which that paper is intimately tied) are the first articulation of Polanyi’s sharp rejection of the modern critical philosophical tradition that by implication included Popper’s philosophical ideas. The 1952 paper is also part of Polanyi’s constructive philosophical effort to set forth a fiduciary philosophy emphasizing commitment. Popper regarded Polanyi’s position as implying cognitive relativism and irrationalism, and from the time of Polanyi’s 1952 paper their personal relationship became strained. Discord between them became publicly manifest when Polanyi subtitled his book Personal Knowledge (1958), Towards a post-critical philosophy, and Popper lambasted the idea of a ‘post-critical’ philosophy in his Preface in The Logic of Scientific Discovery (1959).     

Unclassed matrix shading and optimal ordering in hierarchical cluster analysis

A method is presented for the graphic display of proximity matrices as a complement to the common data analysis techniques of hierarchical clustering. The procedure involves the use of computer generated shaded matrices based on unclassed choropleth mapping in conjunction with a strategy for matrix reorganization. The latter incorporates a combination of techniques for seriation and the ordering of binary trees.Partial support for this research was provided by NIJ Grant #82-IJ-CX-0019 and NSF Grant #SES82-06067. The authors wish to acknowledge the assistance of Professors L.J. Hubert, R.G. Golledge, and W.R. Tobler.     

A large discontinuity in the mid-twentieth century in observed global-mean surface temperature

Data sets used to monitor the Earth's climate indicate that the surface of the Earth warmed from approximately 1910 to 1940, cooled slightly from approximately 1940 to 1970, and then warmed markedly from approximately 1970 onward. The weak cooling apparent in the middle part of the century has been interpreted in the context of a variety of physical factors, such as atmosphere-ocean interactions and anthropogenic emissions of sulphate aerosols. Here we call attention to a previously overlooked discontinuity in the record at 1945, which is a prominent feature of the cooling trend in the mid-twentieth century. The discontinuity is evident in published versions of the global-mean temperature time series, but stands out more clearly after the data are filtered for the effects of internal climate variability. We argue that the abrupt temperature drop of approximately 0.3 degrees C in 1945 is the apparent result of uncorrected instrumental biases in the sea surface temperature record. Corrections for the discontinuity are expected to alter the character of mid-twentieth century temperature variability but not estimates of the century-long trend in global-mean temperatures.     

Supernova SN 2011fe from an exploding carbon-oxygen white dwarf star

Type Ia supernovae have been used empirically as 'standard candles' to demonstrate the acceleration of the expansion of the Universe even though fundamental details, such as the nature of their progenitor systems and how the stars explode, remain a mystery. There is consensus that a white dwarf star explodes after accreting matter in a binary system, but the secondary body could be anything from a main-sequence star to a red giant, or even another white dwarf. This uncertainty stems from the fact that no recent type Ia supernova has been discovered close enough to Earth to detect the stars before explosion. Here we report early observations of supernova SN 2011fe in the galaxy M101 at a distance from Earth of 6.4 megaparsecs. We find that the exploding star was probably a carbon-oxygen white dwarf, and from the lack of an early shock we conclude that the companion was probably a main-sequence star. Early spectroscopy shows high-velocity oxygen that slows rapidly, on a timescale of hours, and extensive mixing of newly synthesized intermediate-mass elements in the outermost layers of the supernova. A companion paper uses pre-explosion images to rule out luminous red giants and most helium stars as companions to the progenitor.