Pharmacological Targeting of STING-Dependent IL-6 Production in Cancer Cells

Activation of the STING pathway upon genotoxic treatment of cancer cells has been shown to lead to anti-tumoral effects, mediated through the acute production of interferon (IFN)-β. Conversely, the pathway also correlates with the expression of NF-κB-driven pro-tumorigenic genes, but these associations are only poorly defined in the context of genotoxic treatment, and are thought to correlate with a chronic engagement of the pathway. We demonstrate here that half of the STING-expressing cancer cells from the NCI60 panel rapidly increased expression of pro-tumorigenic IL-6 upon genotoxic DNA damage, often independent of type-I IFN responses. While preferentially dependent on canonical STING, we demonstrate that genotoxic DNA damage induced by camptothecin (CPT) also drove IL-6 production through non-canonical STING signaling in selected cancer cells. Consequently, pharmacological inhibition of canonical STING failed to broadly inhibit IL-6 production induced by CPT, although this could be achieved through downstream ERK1/2 inhibition. Finally, prolonged inhibition of canonical STING signaling was associated with increased colony formation of MG-63 cells, highlighting the duality of STING signaling in also restraining the growth of selected cancer cells. Collectively, our findings demonstrate that genotoxic-induced DNA damage frequently leads to the rapid production of pro-tumorigenic IL-6 in cancer cells, independent of an IFN signature, through canonical and non-canonical STING activation; this underlines the complexity of STING engagement in human cancer cells, with frequent acute pro-tumorigenic activities induced by DNA damage. We propose that inhibition of ERK1/2 may help curb such pro-tumorigenic responses to DNA-damage, while preserving the anti-proliferative effects of the STING-interferon axis.

Differences of Enzymatic Activity During Composting and Vermicomposting of Sewage Sludge Mixed With Straw Pellets

The study aims were focused on profiling eight hydrolytic enzymes by fluorescence method using a multifunctional modular reader and studying the proportion of basic microorganism groups during composting and vermicomposting of sewage sludge mixed with straw pellets in several proportions (0, 25, 50, 75, and 100%). The greatest decrease in enzymatic activity occurred in the first half of composting and vermicomposting. After 4 months of these processes, the least enzymatic activity was observed in the sludge with 50% and also 25% straw addition, indicating that straw is an important means for the rapid production of mature compost from sewage sludge. Enzymatic activity was usually less in the presence of earthworms than in the control treatment because some processes took place in the digestive tract of the earthworm. For the same reason, we observed reduced enzyme activity during fresh feedstock vermicomposting than precomposted material. The final vermicompost from fresh feedstocks exhibited less microbial biomass, and few fungi and G− bacteria compared to precomposted feedstock. The enzymatic activity during composting and vermicomposting of sewage sludge and their mixtures stabilized at the following values: β-D-glucosidase—50 μmol MUFG/h/g dw, acid phosphatase—200 μmol MUFP/h/g dw, arylsulphatase—10 μmol MUFS/h/g dw, lipase—1,000 μmol MUFY/h/g dw, chitinase—50 μmol MUFN/h/g dw, cellobiohydrolase—20 μmol MUFC/h/g dw, alanine aminopeptidase—50 μmol AMCA/h/g dw, and leucine aminopeptidase—50 μmol AMCL/h/g dw. At these and lesser values, these final products can be considered mature and stable.

Green and Highly-Efficient Microwave Synthesis Route for Sulfur/Carbon Composite for Li-S Battery

Multiporous carbons (MPCs) are prepared using ZnO as a hard template and biomass pyrolysis oil as the carbon source. It is shown that the surface area, pore volume, and mesopore/micropore ratio of the as-prepared MPCs can be easily controlled by adjusting the ZnO/oil ratio. Sulfur/MPC (S/MPC) composite is prepared by blending sulfur powder with the as-prepared MPCs followed by microwave heating at three different powers (100 W/200 W/300 W) for 60 s. The unique micro/mesostructure characteristics of the resulting porous carbons not only endow the S/MPC composite with sufficient available space for sulfur storage, but also provide favorable and efficient channels for Li-ions/electrons transportation. When applied as the electrode material in a lithium-ion battery (LIB), the S/MPC composite shows a reversible capacity (about 500 mAhg−1) and a high columbic efficiency (>95%) after 70 cycles. Overall, the method proposed in this study provides a simple and green approach for the rapid production of MPCs and S/MPC composite for high-performance LIBs.

Humoral responses following SARS–CoV–2 breakthrough infection in COVID–19 vaccinated individuals

Numerous studies have shown that a prior SARS–CoV–2 infection can greatly enhance the antibody response to COVID–19 vaccination, with this so called ′hybrid immunity′ leading to greater neutralization breadth against SARS–CoV–2 variants of concern. However, little is known about how breakthrough infection (BTI) in COVID–19 vaccinated individuals will impact the magnitude and breadth of the neutralizing antibody response. Here, we compared neutralizing antibody responses between unvaccinated and COVID–19 vaccinated individuals (including both AZD1222 and BNT162b2 vaccinees) who have been infected with the delta (B.1.617.2) variant. Rapid production of Spike-reactive IgG was observed in the vaccinated group providing evidence of effective vaccine priming. Overall, potent cross-neutralizing activity against current SARS–CoV–2 variants of concern was observed in the BTI group compared to the infection group. This study provides important insights into population immunity where transmission levels remain high.

Ultrapotent and Broad Neutralization of SARS-CoV-2 Variants by Modular, Tetravalent, Bi-paratopic Antibodies

Neutralizing antibodies (nAbs) that target the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein (S-protein) are promising therapeutics for COVID-19. However, natural bivalent nAbs suffer from limited potency and are vulnerable to SARS-CoV-2 variants with mutated RBDs. We report a novel format that enables modular assembly of bi-paratopic, tetravalent nAbs with antigen-binding sites from two distinct nAbs. The diabody-Fc-Fab format consists of a central Fc with a bivalent diabody fused to its N-terminus and two Fabs fused to its C-terminus. The diabody and Fab modules do not interfere with each other, and thus, any diabody can be combined with any Fab in a facile manner. We engineered a diabody-Fc-Fab that contained the paratopes of two distinct nAbs derived from a phage-displayed library of synthetic Abs. The tetravalent nAb was purified in high yields with methods used to produce conventional IgGs, and it exhibited favorable biophysical characteristics comparable to those of approved therapeutic antibodies. The tetravalent nAb bound to the S-protein trimer at least 100-fold more tightly than the bivalent IgGs (apparent KD <1 pM). Most importantly, the tetravalent nAb exhibited extremely high potencies in neutralization assays across a panel of pseudoviruses representing seven natural SARS-CoV-2 variants (IC50 <5 ng/mL), including several that resisted IgGs and are known to evade approved IgG drugs. Taken together, our results showed that the tetravalent diabody-Fc-Fab is a robust, modular platform for rapid production of drug-grade nAbs with potencies and breadth of coverage that far exceed those of conventional bivalent IgGs.

Research on the key design and application of the device of opening cover and ring cutting and digging flesh of Mandari

Aiming at the problems of low efficiency and poor sanitation of manual cover opening and meat digging in the production of common orange tea, a multi station full-automatic meat digging machine for common orange tea is designed to realize the automatic cover opening and pulp separation device of green orange, so as to meet the production of common orange tea in Xinhui District of Jiangmen. The mechanism of peel stress and pulp separation damage of green citrus in the process of mechanical meat digging was studied. The state of peel contact deformation area of green Citrus under the extrusion of meat digging knife was analyzed by simulation, and the structural and operating parameters of de jacking and opening mechanism and meat digging mechanism were optimized. When the extrusion force was less than 35.6n, the peel damage rate could be minimized. Compared with manual operation, it reduces labor cost and production cost, saves processing time, and realizes automatic and rapid production.

Rapid production of SaCas9 in plant-based cell-free lysate for activity testing

Cas9 nucleases have become the most versatile tool for genome editing projects in a broad range of organisms. The recombinant production of Cas9 nuclease is desirable for in vitro activity assays or the preparation of ribonucleoproteins (RNPs) for DNA-free genome editing approaches. For the rapid production of Cas9, we explored the use of a recently established cell-free lysate from tobacco (Nicotiana tabacum L.) BY-2 cells. Using this system, the 130-kDa Cas9 nuclease from Staphylococcus aureus (SaCas9) was produced and subsequently purified via affinity chromatography. The purified apoenzyme was supplemented with ten different sgRNAs, and the nuclease activity was confirmed by the linearization of plasmid DNA containing cloned DNA target sequences.