Identification of a Ribonucleoprotein Intermediate of Tomato Mosaic Virus RNA Replication Complex Formation

The replication of eukaryotic positive-strand RNA virus genomes occurs in the membrane-bound RNA replication complexes. Previously, we found that the extract of evacuolated tobacco BY-2 protoplasts (BYL) is capable of supporting the translation and subsequent replication of the genomic RNAs of plant positive-strand RNA viruses, including Tomato mosaic virus (ToMV). Here, to dissect the process that precedes the formation of ToMV RNA replication complexes, we prepared membrane-depleted BYL (mdBYL), in which the membranes were removed by centrifugation. In mdBYL, ToMV RNA was translated to produce the 130-kDa and 180-kDa replication proteins, but the synthesis of any ToMV-related RNAs did not occur. When BYL membranes were added back to the ToMV RNA-translated mdBYL after the termination of translation with puromycin, ToMV RNA was replicated. Using a replication-competent ToMV derivative that encodes the FLAG-tagged 180-kDa replication protein, it was shown by affinity purification that a complex that contained the 130-kDa and 180-kDa proteins and ToMV genomic RNA was formed after translation in mdBYL. When the complex was mixed with BYL membranes, ToMV RNA was replicated, which suggests that this ribonucleoprotein complex is an intermediate of ToMV RNA replication complex formation. We have named this ribonucleoprotein complex the “pre-membrane-targeting complex.” Our data suggest that the formation of the pre-membrane-targeting complex is coupled with the translation of ToMV RNA, while posttranslationally added exogenous 180-kDa protein and replication templates can contribute to replication and can be replicated, respectively. Based on these results, we discuss the mechanisms of ToMV RNA replication complex formation

Probing Optimal Reaction Energy for Synthesis of Element 119 from ⁵¹V+²⁴⁸Cm Reaction with Quasielastic Barrier Distribution Measurement

International audienceThe quasielastic barrier distribution of 51V+248Cm was extracted by measuring the excitation function of quasielastic backscattering using a gas-filled recoil ion separator, GARIS-III. The obtained barrier distribution is well explained by the coupled-channels calculation, indicating a significant effect of the rotational excitation of deformed 248Cm. From the measured average Coulomb barrier height and deformation parameters of 248Cm, the side-collision energy leading to a compact configuration of colliding nuclei was obtained. The relation between the side collision energy and the excitation function of the evaporation-residue cross sections in the 48Ca+248Cm system was evaluated as a reference for the 51V+248Cm case. The optimal reaction energy to synthesize a new element 119 at the 51V+248Cm fusion reaction (3n and 4n channels) was estimated with an aid of these experimental data

Additional file 1 of Chest CT findings in severe acute respiratory distress syndrome requiring V-V ECMO: J-CARVE registry

Additional file 1: SMethods. Figure S1. Representative images of each of the characteristic pulmonary opacities on chest computed tomography scans. Figure S2. Distribution of registered patients by years. Figure S3. Cumulative proportion of the duration (h) between chest computed tomography examinations and initiation of veno-venous extracorporeal membrane oxygenation support. Figure S4. Characteristics of the chest computed tomography findings according to the mechanical ventilation–extracorporeal membrane oxygenation support duration and the underlying etiology of the acute respiratory distress syndrome. Figure S5. Survival curve of the chest computed tomography findings related to changes outside of the pulmonary opacity (excluding subcutaneous emphysema). Figure S6. Survival curve of participants with and without traction bronchiectasis separately according to the underlying etiology of acute respiratory distress syndrome. Table S1. Concordance rates between two evaluators. Table S2. Basic information of the participating hospitals. Table S3. Characteristics of chest computed tomography findings. Table S4. Results of multivariate Cox regression analysis of the relationship between V-V ECMO support initiation and 90-day in-hospital mortality. Table S5. Results of multivariate logistic regression analysis for successful ECMO liberation