Viral Small Interfering RNAs Target Host Genes to Mediate Disease Symptoms in Plants

The Cucumber mosaic virus (CMV) Y-satellite RNA (Y-Sat) has a small non-protein-coding RNA genome that induces yellowing symptoms in infected Nicotiana tabacum (tobacco). How this RNA pathogen induces such symptoms has been a longstanding question. We show that the yellowing symptoms are a result of small interfering RNA (siRNA)-directed RNA silencing of the chlorophyll biosynthetic gene, CHLI. The CHLI mRNA contains a 22-nucleotide (nt) complementary sequence to the Y-Sat genome, and in Y-Sat-infected plants, CHLI expression is dramatically down-regulated. Small RNA sequencing and 5′ RACE analyses confirmed that this 22-nt sequence was targeted for mRNA cleavage by Y-Sat-derived siRNAs. Transformation of tobacco with a RNA interference (RNAi) vector targeting CHLI induced Y-Sat-like symptoms. In addition, the symptoms of Y-Sat infection can be completely prevented by transforming tobacco with a silencing-resistant variant of the CHLI gene. These results suggest that siRNA-directed silencing of CHLI is solely responsible for the Y-Sat-induced symptoms. Furthermore, we demonstrate that two Nicotiana species, which do not develop yellowing symptoms upon Y-Sat infection, contain a single nucleotide polymorphism within the siRNA-targeted CHLI sequence. This suggests that the previously observed species specificity of Y-Sat-induced symptoms is due to natural sequence variation in the CHLI gene, preventing CHLI silencing in species with a mismatch to the Y-Sat siRNA. Taken together, these findings provide the first demonstration of small RNA-mediated viral disease symptom production and offer an explanation of the species specificity of the viral disease

Design of Group IIA Secreted/Synovial Phospholipase A2 Inhibitors: An Oxadiazolone Derivative Suppresses Chondrocyte Prostaglandin E2 Secretion

Group IIA secreted/synovial phospholipase A2 (GIIAPLA2) is an enzyme involved in the synthesis of eicosanoids such as prostaglandin E2 (PGE2), the main eicosanoid contributing to pain and inflammation in rheumatic diseases. We designed, by molecular modeling, 7 novel analogs of 3-{4-[5(indol-1-yl)pentoxy]benzyl}-4H-1,2,4-oxadiazol-5-one, denoted C1, an inhibitor of the GIIAPLA2 enzyme. We report the results of molecular dynamics studies of the complexes between these derivatives and GIIAPLA2, along with their chemical synthesis and results from PLA2 inhibition tests. Modeling predicted some derivatives to display greater GIIAPLA2 affinities than did C1, and such predictions were confirmed by in vitro PLA2 enzymatic tests. Compound C8, endowed with the most favorable energy balance, was shown experimentally to be the strongest GIIAPLA2 inhibitor. Moreover, it displayed an anti-inflammatory activity on rabbit articular chondrocytes, as shown by its capacity to inhibit IL-1β-stimulated PGE2 secretion in these cells. Interestingly, it did not modify the COX-1 to COX-2 ratio. C8 is therefore a potential candidate for anti-inflammatory therapy in joints

First results on the search for chameleons with the KWISP detector at CAST

We report on a first measurement with a sensitive opto-mechanical force sensor designed for the direct detection of coupling of real chameleons to matter. These dark energy candidates could be produced in the Sun and stream unimpeded to Earth. The KWISP detector installed on the CAST axion search experiment at CERN looks for tiny displacements of a thin membrane caused by the mechanical effect of solar chameleons. The displacements are detected by a Michelson interferometer with a homodyne readout scheme. The sensor benefits from the focusing action of the ABRIXAS X-ray telescope installed at CAST, which increases the chameleon flux on the membrane. A mechanical chopper placed between the telescope output and the detector modulates the incoming chameleon stream. We present the results of the solar chameleon measurements taken at CAST in July 2017, setting an upper bound on the force acting on the membrane of 80pN at 95% confidence level. The detector is sensitive for direct coupling to matter 104 = ßm = 108, where the coupling to photons is locally bound to ß¿ = 1011

Search for Dark Matter Axions with CAST-CAPP

The CAST-CAPP axion haloscope, operating at CERN inside the CAST dipole magnet, has searched for axions in the 19.74 $\mu$eV to 22.47 $\mu$eV mass range. The detection concept follows the Sikivie haloscope principle, where Dark Matter axions convert into photons within a resonator immersed in a magnetic field. The CAST-CAPP resonator is an array of four individual rectangular cavities inserted in a strong dipole magnet, phase-matched to maximize the detection sensitivity. Here we report on the data acquired for 4124 h from 2019 to 2021. Each cavity is equipped with a fast frequency tuning mechanism of 10 MHz/min between 4.774 GHz and 5.434 GHz. In the present work, we exclude axion-photon couplings for virialized galactic axions down to $g_{a{\gamma}{\gamma}} = 8 \times {10^{-14}}$ $GeV^{-1}$ at the 90% confidence level. The here implemented phase-matching technique also allows for future large-scale upgrades.Comment: 24 pages, 5 figures, Published version available with Open Access at https://www.nature.com/articles/s41467-022-33913-

First results of the CAST-RADES haloscope search for axions at 34.67 µeV

We present results of the Relic Axion Dark-Matter Exploratory Setup (RADES), a detector which is part of the CERN Axion Solar Telescope (CAST), searching for axion dark matter in the 34.67 µeV mass range. A radio frequency cavity consisting of 5 sub-cavities coupled by inductive irises took physics data inside the CAST dipole magnet for the first time using this filter-like haloscope geometry. An exclusion limit with a 95% credibility level on the axion-photon coupling constant of ga¿ ¿ 4 × 10-13 GeV-1 over a mass range of 34.6738 µeV < ma< 34.6771 µeV is set. This constitutes a significant improvement over the current strongest limit set by CAST at this mass and is at the same time one of the most sensitive direct searches for an axion dark matter candidate above the mass of 25 µeV. The results also demonstrate the feasibility of exploring a wider mass range around the value probed by CAST-RADES in this work using similar coherent resonant cavities. © 2021, The Author(s)

Expression of Linear and Novel Circular Forms of an INK4/ARF-Associated Non-Coding RNA Correlates with Atherosclerosis Risk

Human genome-wide association studies have linked single nucleotide polymorphisms (SNPs) on chromosome 9p21.3 near the INK4/ARF (CDKN2a/b) locus with susceptibility to atherosclerotic vascular disease (ASVD). Although this locus encodes three well-characterized tumor suppressors, p16INK4a, p15INK4b, and ARF, the SNPs most strongly associated with ASVD are ∼120 kb from the nearest coding gene within a long non-coding RNA (ncRNA) known as ANRIL (CDKN2BAS). While individuals homozygous for the atherosclerotic risk allele show decreased expression of ANRIL and the coding INK4/ARF transcripts, the mechanism by which such distant genetic variants influence INK4/ARF expression is unknown. Here, using rapid amplification of cDNA ends (RACE) and analysis of next-generation RNA sequencing datasets, we determined the structure and abundance of multiple ANRIL species. Each of these species was present at very low copy numbers in primary and cultured cells; however, only the expression of ANRIL isoforms containing exons proximal to the INK4/ARF locus correlated with the ASVD risk alleles. Surprisingly, RACE also identified transcripts containing non-colinear ANRIL exonic sequences, whose expression also correlated with genotype and INK4/ARF expression. These non-polyadenylated RNAs resisted RNAse R digestion and could be PCR amplified using outward-facing primers, suggesting they represent circular RNA structures that could arise from by-products of mRNA splicing. Next-generation DNA sequencing and splice prediction algorithms identified polymorphisms within the ASVD risk interval that may regulate ANRIL splicing and circular ANRIL (cANRIL) production. These results identify novel circular RNA products emanating from the ANRIL locus and suggest causal variants at 9p21.3 regulate INK4/ARF expression and ASVD risk by modulating ANRIL expression and/or structure

First results of the CAST-RADES haloscope search for axions at 34.67 μeV

We present results of the Relic Axion Dark-Matter Exploratory Setup (RADES), a detector which is part of the CERN Axion Solar Telescope (CAST), searching for axion dark matter in the 34.67μeV mass range. A radio frequency cavity consisting of 5 sub-cavities coupled by inductive irises took physics data inside the CAST dipole magnet for the first time using this filter-like haloscope geometry. An exclusion limit with a 95% credibility level on the axion-photon coupling constant of gaγ & 4 × 10−13 GeV−1 over a mass range of 34.6738μeV < ma < 34.6771μeV is set. This constitutes a significant improvement over the current strongest limit set by CAST at this mass and is at the same time one of the most sensitive direct searches for an axion dark matter candidate above the mass of 25μeV. The results also demonstrate the feasibility of exploring a wider mass range around the value probed by CAST-RADES in this work using similar coherent resonant cavitiesWe wish to thank our colleagues at CERN, in particular Marc Thiebert from the coating lab, as well as the whole team of the CERN Central Cryogenic Laboratory for their support and advice in speci c aspects of the project. We thank Arefe Abghari for her contributions as the project's summer student during 2018. This work has been funded by the Spanish Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) under project FPA-2016-76978-C3-2-P and PID2019-108122GB-C33, and was supported by the CERN Doctoral Studentship programme. The research leading to these results has received funding from the European Research Council and BD, JG and SAC acknowledge support through the European Research Council under grant ERC-2018-StG-802836 (AxScale project). BD also acknowledges fruitful discussions at MIAPP supported by DFG under EXC-2094 { 390783311. IGI acknowledges also support from the European Research Council (ERC) under grant ERC-2017-AdG-788781 (IAXO+ project). JR has been supported by the Ramon y Cajal Fellowship 2012-10597, the grant PGC2018-095328-B-I00(FEDER/Agencia estatal de investigaci on) and FSE-GA2017-2019-E12/7R (Gobierno de Aragón/FEDER) (MINECO/FEDER), the EU through the ITN \Elusives" H2020-MSCA-ITN-2015/674896 and the Deutsche Forschungsgemeinschaft under grant SFB-1258 as a Mercator Fellow. CPG was supported by PROMETEO II/2014/050 of Generalitat Valenciana, FPA2014-57816-P of MINECO and by the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreements 690575 and 674896. AM is supported by the European Research Council under Grant No. 742104. Part of this work was performed under the auspices of the US Department of Energy by Lawrence Livermore National Laboratory under Contract No. DE-AC52-07NA27344