The composition of phenolic compounds in Chinese olive (Canarium album L.) cultivars and their contribution to the anti-inflammatory properties of the cultivars.

Objective: This study aimed to explore the phenolic compounds (PCs) present in three Chinese olive (Canarium album L.) cultivars and the contribution of these PCs to the anti-inflammatory activities of the cultivars. Methods: Ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap/mass spectrometry (UPLC-Q-Exactive/MS) was used to identify and quantify the PCs present in three Chinese olive cultivars, "Na zhong," "Tan xiang," and "Xiang zhong". 2,2-diphenyl-1-picrylhydrazyl (DPPH); 2,2'-azinobis (3-ethylbenzothiazoline 6-sulfonate) (ABTS); and oxygen radical absorption capacity (ORAC) assays were used to assess the antioxidant activities of the PCs. Furthermore, we analyzed the anti-inflammatory action of these PCs using lipopolysaccharide (LPS)-induced RAW264.7 cells. Results: A total of 44 PCs were identified in the three cultivars. Of these, 17 PCs were previously unidentified in Chinese olive. Among the cultivars, the free phenolics (FPs) of "Tan xiang" showed the strongest antioxidant activity. All cultivars have shown significant inhibition of TNF-α and IL-6 production. Clustering correlation analysis showed galloyl-bis-HHDP-glucose and paeonol have significant anti-inflammatory ability in FPs. Quininic, galloylquinic acid, 4-hydroxycinnamic acid and gallic acid hexoside have shown significant inhibition of IL-6 production in BPs. Furthermore, gallic acid, catechin, syringic acid, and nobiletin exhibit negative correlation in FPs and positive correlation in BPs of cytokine production, while corilagin and methyl ellagic acid pentoside exhibited opposite correlation. Conclusion: In summary, this study contributed to the literature on PCs in Chinese olives and the potential health benefits of FPs and BPs.

The Dynamic Poly(A) Tail Acts as a Signal Hub in mRNA Metabolism.

In eukaryotes, mRNA metabolism requires a sophisticated signaling system. Recent studies have suggested that polyadenylate tail may play a vital role in such a system. The poly(A) tail used to be regarded as a common modification at the 3' end of mRNA, but it is now known to be more than just that. It appears to act as a platform or hub that can be understood in two ways. On the one hand, polyadenylation and deadenylation machinery constantly regulates its dynamic activity; on the other hand, it exhibits the ability to recruit RNA-binding proteins and then interact with diverse factors to send various signals to regulate mRNA metabolism. In this paper, we outline the main complexes that regulate the dynamic activities of poly(A) tails, explain how these complexes participate polyadenylation/deadenylation process and summarize the diverse signals this hub emit. We are trying to make a point that the poly(A) tail can metaphorically act as a "flagman" who is supervised by polyadenylation and deadenylation and sends out signals to regulate the orderly functioning of mRNA metabolism.

Impact of the Cellular Environment on Adenosine Triphosphate Conformations.

The cytoplasm is an environment crowded by macromolecules and filled with metabolites and ions. Recent experimental and computational studies have addressed how this environment affects protein stability, folding kinetics, and protein-protein and protein-nucleic acid interactions, though its impact on metabolites remains largely unknown. Here we show how a simulated cytoplasm affects the conformation of adenosine triphosphate (ATP), a key energy source and regulatory metabolite present at high concentrations in cells. Analysis of our all-atom model of a small volume of the Escherichia coli cytoplasm when contrasted with ATP modeled in vitro or resolved with protein structures deposited in the Protein Data Bank reveals that ATP molecules bound to proteins in cell form specific pitched conformations that are not observed at significant concentrations in the other environments. We hypothesize that these interactions evolved to fulfill functional roles when ATP interacts with protein surfaces.

Overexpressing CrePAPS Polyadenylate Activity Enhances Protein Translation and Accumulation in Chlamydomonas reinhardtii.

The alga Chlamydomonas reinhardtii is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of C. reinhardtii strains producing genetically engineered recombinant therapeutic protein have been reported. However, owing to extremely low protein expression efficiency, none have been applied for industrial purposes. Improving protein expression efficiency at the molecular level is, therefore, a priority. The 3'-end poly(A) tail of mRNAs is strongly correlated with mRNA transcription and protein translation efficiency. In this study, we identified a canonical C. reinhardtii poly(A) polymerase (CrePAPS), verified its polyadenylate activity, generated a series of overexpressing transformants, and performed proteomic analysis. Proteomic results demonstrated that overexpressing CrePAPS promoted ribosomal assembly and enhanced protein accumulation. The accelerated translation was further verified by increased crude and dissolved protein content detected by Kjeldahl and bicinchoninic acid (BCA) assay approaches. The findings provide a novel direction in which to exploit photosynthetic green algae as a recombinant protein expression platform.

Chemical synthesis of left arm of Chlamydomonas reinhardtii mitochondrial genome and in vivo functional analysis.

Chlamydomonas reinhardtii is a photosynthetic eukaryote showing great industrial potential. The synthesis and in vivo function of the artificial C. reinhardtii genome not only promotes the development of synthetic biology technology but also supports industries that utilize this algae. Mitochondrial genome (MtG) is the smallest and simplest genome of C. reinhardtii that suits synthetic exploration. In this article, we designed and assembled a synthetic mitochondria left arm (syn-LA) genome sharing >92% similarity to the original mitochondria genome (OMtG) left arm, transferred it into the respiratory defect strain cc-2654, screened syn-LA containing transformants from recovered dark-growth defects using PCR amplification, verified internal function of syn-LA via western blot, detected heteroplasmic ratio of syn-LA, tried promoting syn-LA into homoplasmic status with paromomycin stress, and discussed the main limitations and potential solutions for this area of research. This research supports the functionalization of a synthetic mitochondrial genome in living cells. Although further research is needed, this article nevertheless provides valuable guidance for the synthesis of eukaryotic organelle genomes and opens possible directions for future research.