Batch Pyrolysis and Co-Pyrolysis of Beet Pulp and Wheat Straw.

Granulated beet pulp and wheat straw, first separately and then mixed in a weight ratio of 50/50%, underwent a pyrolysis process in a laboratory batch generator with process temperatures of 400 and 500 °C. The feedstock's chemical composition and the pyrolysis products' chemical composition (biochar and pyrolysis gas) were analysed. A synergistic effect was observed in the co-pyrolysis of the combined feedstock, which occurred as an increase the content of the arising gas in relation to the total weight of the products. and as a reduction of bio-oil content. The maximum gas proportion was 21.8% at 500 °C and the minimum between 12.6% and 18.4% for the pyrolysis of individual substrates at 400 °C. The proportions of the gases, including CO, CO2, CH4, H2, and O2, present in the resulting synthesis gases were also analysed. The usage of a higher pyrolysis final temperature strongly affected the increase of the CH4 and H2 concentration and the decrease of CO2 and CO concentration in the pyrolysis gas. The highest percentage of hydrogen in the synthesis gas, around 33%vol, occurred at 500 °C during co-pyrolysis.

Drosophila miR2 primarily targets the m7GpppN cap structure for translational repression.

Understanding the molecular mechanism(s) of how miRNAs repress mRNA translation is a fundamental challenge in RNA biology. Here we use a validated cell-free system from Drosophila embryos to investigate how miR2 inhibits translation initiation. By screening a library of chemical m7GpppN cap structure analogs, we identified defined modifications of the triphosphate backbone that augment miRNA-mediated inhibition of translation initiation but are "neutral" toward general cap-dependent translation. Interestingly, these caps also augment inhibition by 4E-BP. Kinetic dissection of translational repression and miR2-induced deadenylation shows that both processes proceed largely independently, with establishment of the repressed state involving a slow step. Our data demonstrate a primary role for the m7GpppN cap structure in miRNA-mediated translational inhibition, implicate structural determinants outside the core eIF4E-binding region in this process, and suggest that miRNAs may target cap-dependent translation through a mechanism related to the 4E-BP class of translational regulators.

Protein disaggregation by the AAA+ chaperone ClpB involves partial threading of looped polypeptide segments.

The ring-forming AAA+ chaperone ClpB cooperates with the DnaK chaperone system to reactivate aggregated proteins. With the assistance of DnaK, ClpB extracts unfolded polypeptides from aggregates via substrate threading through its central channel. Here we analyze the processing of mixed aggregates consisting of protein fusions of misfolded and native domains. ClpB-DnaK reactivated all aggregated fusion proteins with similar efficiency, without unfolding native domains, demonstrating that partial threading of the misfolded moiety is sufficient to solubilize aggregates. Reactivation by ClpB-DnaK occurred even when two stably folded domains flanked the aggregated moiety, indicating threading of internal substrate segments. In contrast with the related AAA+ chaperone ClpC, ClpB lacks a robust unfolding activity, enabling it to sense the conformational state of substrates. ClpB rings are highly unstable, which may facilitate dissociation from trapped substrates during threading.

A genetic module regulates the melanization response of Anopheles to Plasmodium.

Two modes of refractoriness to Plasmodium, ookinete lysis and melanization, are known in the malaria vector, Anopheles gambiae. Melanization, a potent insect immune response, is manifested in a genetically selected refractory strain and in susceptible mosquitoes that are depleted of specific C-type lectins (CTLs). Here we use a systematic in vivo RNA interference-mediated reverse genetic screen and other recent results to define a melanization-regulating genetic module or network. It encompasses at least 14 genes, including those that encode five Easter-like clip domain serine proteases and four Masquerade-like serine protease homologues of the mosquito CLIPB and CLIPA subfamilies respectively. We show that several but not all CLIPB genes promote Plasmodium melanization, exhibiting partial functional overlap and synergy. We also report that several CLIPA genes have contrasting roles: CLIPA8 is essential for parasite melanization, while three other CLIPAs are novel synergistic inhibitors of this response. Importantly, the roles of certain CLIPAs and CLIPBs are strain specific, indicating that this network may differ between strains. Finally, we provide evidence that in susceptible mosquitoes melanization induced by knockdown of either CTL4 or CLIPA2/CLIPA5 directly kills ookinetes, in contrast to refractory mosquitoes where it merely disposes of dead parasites.