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Subgenomic flavivirus RNA binds the mosquito DEAD/H-box helicase ME31B and determines Zika virus transmission by Aedes aegypti
Göertz, Giel P. ; Bree, Joyce W.M. van; Hiralal, Anwar ; Fernhout, Bas M. ; Steffens, Carmen ; Boeren, Sjef ; Visser, Tessa M. ; Vogels, Chantal B.F. ; Abbo, Sandra R. ; Fros, Jelke J. ; Koenraadt, Constantianus J.M. ; Oers, Monique M. van; Pijlman, Gorben P. - \ 2019
Proceedings of the National Academy of Sciences of the United States of America 116 (2019)38. - ISSN 0027-8424 - p. 19136 - 19144.
Aedes aegypti - Purification - RNA-affinity - Subgenomic flavivirus RNA - Transmission - Zika virus
Zika virus (ZIKV) is an arthropod-borne flavivirus predominantly transmitted by Aedes aegypti mosquitoes and poses a global human health threat. All flaviviruses, including those that exclusively replicate in mosquitoes, produce a highly abundant, noncoding subgenomic flavivirus RNA (sfRNA) in infected cells, which implies an important function of sfRNA during mosquito infection. Currently, the role of sfRNA in flavivirus transmission by mosquitoes is not well understood. Here, we demonstrate that an sfRNA-deficient ZIKV (ZIKVΔSF1) replicates similar to wild-type ZIKV in mosquito cell culture but is severely attenuated in transmission by Ae. aegypti after an infectious blood meal, with 5% saliva-positive mosquitoes for ZIKVΔSF1 vs. 31% for ZIKV. Furthermore, viral titers in the mosquito saliva were lower for ZIKVΔSF1 as compared to ZIKV. Comparison of mosquito infection via infectious blood meals and intrathoracic injections showed that sfRNA is important for ZIKV to overcome the mosquito midgut barrier and to promote virus accumulation in the saliva. Next-generation sequencing of infected mosquitoes showed that viral small-interfering RNAs were elevated upon ZIKVΔSF1 as compared to ZIKV infection. RNA-affinity purification followed by mass spectrometry analysis uncovered that sfRNA specifically interacts with a specific set of Ae. aegypti proteins that are normally associated with RNA turnover and protein translation. The DEAD/H-box helicase ME31B showed the highest affinity for sfRNA and displayed antiviral activity against ZIKV in Ae. aegypti cells. Based on these results, we present a mechanistic model in which sfRNA sequesters ME31B to promote flavivirus replication and virion production to facilitate transmission by mosquitoes.
Membrane separation and characterisation of lignin and its derived products obtained by a mild ethanol organosolv treatment of rice straw
Moniz, Patrícia ; Serralheiro, Cláudia ; Matos, Cristina T. ; Boeriu, Carmen G. ; Frissen, Augustinus E. ; Duarte, Luís C. ; Roseiro, Luísa B. ; Pereira, Helena ; Carvalheiro, Florbela - \ 2018
Process Biochemistry 65 (2018). - ISSN 1359-5113 - p. 136 - 145.
Added value compounds - Combined pretreatment - Lignin - Phenolic compounds - Purification
An organosolv process using ethanol-water was optimized in order to recover high quality lignin from rice-straw previously pre-treated by autohydrolysis at 210 °C. The results showed a selective and appreciable removal of lignin under very mild conditions and the highest delignification yield occurred at 30 °C. The lignin extracts were characterised using capillary zone electrophoresis (CZE), size exclusion chromatography (SEC), Fourier transform infrared spectroscopy (FT-IR) and 31P-NMR, and two-dimensional heteronuclear single quantum correlation NMR spectroscopy (2D-HSQC NMR), which enabled the identification of low molecular weight lignins with a syringyl/guaiacyl ratio of about 0.74 containing phenolic compounds with potential bioactive properties. In order to separate the target compounds, membrane technology has been used and an enriched extract containing value-added phenolic compounds such as tricin, vanillin, ferulic acid and p-coumaric acid was obtained. High membrane efficiency (around 80%) was obtained for target compounds.
Red mud as secondary source for critical raw materials - purification of rare earth elements by liquid/liquid extraction
Ujaczki, Éva ; Zimmermann, Yannick ; Gasser, Christoph ; Molnár, Mónika ; Feigl, Viktória ; Lenz, Markus - \ 2017
Journal of Chemical Technology and Biotechnology 92 (2017)10. - ISSN 0268-2575 - p. 2683 - 2690.
Critical raw materials - Purification - Rare earth - Red mud - Systematic approach
BACKGROUND: Critical raw materials (CRM) are crucial to Europe's economy and essential to maintaining and improving our quality of life due to their usage for production of many devices. Red mud is generated from alumina production where bauxite is digested in hot sodium hydroxide solution during the Bayer process. Red mud can contain considerable amounts of CRM such as rare earth elements (REEs). In the present study, purification of CRM from perturbing, co-extracted elements such as Fe and Al from red mud hydrochloric acid leachates was evaluated. RESULTS: A first purification was achieved by removing Fe (>87%) from the acidic leachate using precipitation with NaOH. REEs as well as Al were hardly removed by precipitation (21%, and 33%, resp.). A second purification was achieved using liquid/liquid extraction (LLE) with di-(2-ethylhexyl)phosphoric acid (D2EHPA). Here, four explanatory variables (i.e. LLE organic/aqueous ratio, D2EHPA concentration in kerosene, stripping acid organic/aqueous ratio, HCl concentration) were studied. Finally, the optimal extraction conditions maximizing the economic potential (total metal extracted×economic value of the respective metal) of CRM were determined using a design of experiment approach. CONCLUSION: The experimentally determined economic potential extracted corresponded well to the prediction (88%; to the predictions, maximum recovery of 17.18±0.59 US $ t-1). Ultimately, more than 40% of the overall REEs (>62% of the leachable REEs) in red mud were purified using LLE, whereas Al was successfully rejected from the concentrate (∼5% of the overall Al present).
Purification and immuno-gold labeling of lily mottle virus from lily leaves
Zhang, Yubao ; Wang, Yajun ; Xie, Zhongkui ; Wang, Ruoyu ; Guo, Zhihong ; Werf, Wopke van der; Wang, Le - \ 2016
Journal of Virological Methods 232 (2016). - ISSN 0166-0934 - p. 33 - 38.
Immuno-gold labeling assay - Lily mottle virus - Purification - Transmission electron microscope
Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.