eCRUIS captures RNA-protein interaction in vitro and in vivo.

As an information bridge between DNA and protein, RNA regulates cellular processes and gene expression in various ways. From its synthesis to degradation, RNA is associated with a range of RNA-binding proteins. Therefore, it is necessary to develop innovative methods to study the interaction between RNA and proteins. Previously, we developed an RNA-centric method, called CRISPR-based RNA-United Interacting System (CRUIS), to capture RNA-protein interaction in cells. On this basis, here we develop an enhanced CRUIS (eCRUIS) by combining the power of dCas13d and the engineered promiscuous ligase TurboID. The current version allows us to rapidly label RNA-binding proteins on the target RNA within 30 minutes, potentially for in vivo use. By introducing bait-assay with exogenous RNA, we confirm that eCRUIS can effectively label RNA-binding proteins on bait RNA in a short time. eCRUIS provides a broader range of in vitro and in vivo applications for studying RNA-protein interactions.

Cytoophidia coupling adipose architecture and metabolism.

Tissue architecture determines its unique physiology and function. How these properties are intertwined has remained unclear. Here we show that the metabolic enzyme CTP synthase (CTPS) form filamentous structures termed cytoophidia along the adipocyte cortex in Drosophila adipose tissue. Loss of cytoophidia, whether due to reduced CTPS expression or a point mutation that specifically abrogates its polymerization ability, causes impaired adipocyte adhesion and defective adipose tissue architecture. Moreover, CTPS influences integrin distribution and dot-like deposition of type IV collagen (Col IV). Col IV-integrin signaling reciprocally regulates the assembly of cytoophidia in adipocytes. Our results demonstrate that a positive feedback signaling loop containing both cytoophidia and integrin adhesion complex couple tissue architecture and metabolism in Drosophila adipose tissue.

Drosophila intestinal homeostasis requires CTP synthase.

CTP synthase (CTPS) senses all four nucleotides and forms filamentous structures termed cytoophidia in all three domains of life. How CTPS and cytoophidia function in a developmental context, however, remains underexplored. We report that CTPS forms cytoophidia in a subset of cells in the Drosophila midgut. We found that cytoophidia exist in intestinal stem cells (ISC) and enteroblasts in similar proportions. Both refeeding after starvation and feeding with dextran sulfate sodium (DSS) induce ISC proliferation and elongate cytoophidia. Knockdown of CTPS inhibits ISC proliferation. Remarkably, disruption of CTPS cytoophidia inhibits DSS-induced ISC proliferation. Taken together, these data suggest that both the expression level and the filament-form property of CTPS are crucial for intestinal homeostasis in Drosophila.

Drought stress introduces growth, physiological traits and ecological stoichiometry changes in two contrasting Cunninghamia lanceolata cultivars planted in continuous-plantation soils.

BACKGROUND: The decrease in Cunninghamia lanceolata (Lamb.) production on continuously planted soil is an essential problem. In this study, two-year-old seedlings of two cultivars (a normal cultivar, NC, and a super cultivar, SC) were grown in two types of soil (not planted (NP) soil; continuously planted (CP) soil) with three watering regimes, and the interactive effects on plant growth and physiological traits were investigated in a greenhouse experiment. The water contents of the soil in the control (CK) (normal water content), medium water content (MWC) and low water content (LWC) treatments reached 75-80 %, 45-50 % and 20-25 % of the field water capacity, respectively. RESULTS: The results indicated that the CP soil had a negative effect on growth and physiological traits and that the LWC treatment caused even more severe and comprehensive negative effects. In both cultivars, the CP soil significantly decreased the height increment (HI), basal diameter increment (DI), dry matter accumulation (DMA), net photosynthetic rate (Pn), total chlorophyll content (TChl), carotenoid content (Caro) and photosynthetic nitrogen use efficiency (PNUE). Compared to the NP soil, the CP soil also decreased the proline and soluble protein contents, nitrogen use efficiency (NUE) and phosphorus use efficiency (PUE) and increased the nitrogen:phosphorus ratio in roots, stems and leaves. The LWC treatment decreased growth and photosynthesis, changed ecological stoichiometry, induced oxidative stress, promoted water use efficiency and damaged chloroplast ultrastructure. Significant increases in ascorbate peroxidase (APX), peroxidase (POD), soluble protein and proline contents were found in the LWC treatment. Compared with the NC, the SC was more tolerant to the CP soil and water stress, as indicated by the higher levels of DMA, Pn, and WUE. After exposure to the CP soil and watering regimes, the decreases in biomass accumulation and gas exchange were more pronounced. CONCLUSIONS: The combination of drought and CP soil may have detrimental effects on C. lanceolata growth, and low water content enhances the impacts of CP soil stress on C. lanceolata seedlings. The superiority of the SC over the NC is significant in Chinese fir plantation soil. Therefore, continuously planted soil can be utilized to cultivate improved varieties of C. lanceolata and maintain water capacity. This can improve their growth and physiological performance to a certain extent.

STAMP: Spatio-Temporal Association Mapping of Proteins.

We describe a tool, Spatio-Temporal Association Mapping of Proteins (STAMP), for identifying protein interactomes via proximity labeling. For a proof-of-principle study, we use cytidine 5'-triphosphate synthase (CTPS) as an example. CTPS, a metabolic enzyme, forms filamentous structures termed cytoophidia in various tissues. We apply STAMP to a variety of developmental stages and tissues in Drosophila including adult ovaries. Using a cell-specific GAL4 driver, we verify that TurboID can biotinylate the bait protein CTPS, making possible the identification of protein-protein interactions (PPIs) in individual cells. Using the wild-type and mutant CTPS as bait proteins, STAMP results in two distinct sets of proximate proteomes. Our results suggest that STAMP is a feasible tool to catch in vivo PPIs in situ at a defined spatiotemporal resolution.

Fat body-specific reduction of CTPS alleviates HFD-induced obesity.

Obesity induced by high-fat diet (HFD) is a multi-factorial disease including genetic, physiological, behavioral, and environmental components. Drosophila has emerged as an effective metabolic disease model. Cytidine 5'-triphosphate synthase (CTPS) is an important enzyme for the de novo synthesis of CTP, governing the cellular level of CTP and the rate of phospholipid synthesis. CTPS is known to form filamentous structures called cytoophidia, which are found in bacteria, archaea, and eukaryotes. Our study demonstrates that CTPS is crucial in regulating body weight and starvation resistance in Drosophila by functioning in the fat body. HFD-induced obesity leads to increased transcription of CTPS and elongates cytoophidia in larval adipocytes. Depleting CTPS in the fat body prevented HFD-induced obesity, including body weight gain, adipocyte expansion, and lipid accumulation, by inhibiting the PI3K-Akt-SREBP axis. Furthermore, a dominant-negative form of CTPS also prevented adipocyte expansion and downregulated lipogenic genes. These findings not only establish a functional link between CTPS and lipid homeostasis but also highlight the potential role of CTPS manipulation in the treatment of HFD-induced obesity.

The high rate of obesity has created a global health burden by leading to increased rates of chronic diseases like diabetes and cardiovascular disease. Tackling this issue is complicated as it is influenced by many factors, including genetics, behaviour and environment. To better understand the biochemical changes that underly metabolic issues in a simpler setting, scientists can study fruit flies in the laboratory. These insects share many genes with humans and have similar responses to a high-fat diet. Previous research identified an enzyme, called CTP synthase (CTPS), which is produced in large amounts by the liver and fat tissue in mammals, and the equivalent in fruit flies, known as the fat body. Multiple CTPS molecules can combine to form long strands of protein called cytoophidia, which have been seen in organisms ranging from humans to bacteria. Recent results showed that the fruit fly equivalent of CTPS drives fat cells to stick together, which is necessary to maintain and form fat tissue. However, it is not clear if altering the levels of CTPS can affect the response to a high-fat diet. To address this, Liu, Zhang, Wang et al. studied fruit flies on a high-fat diet, showing that this increased the production of CTPS. When the flies were treated to deplete levels of CTPS in the fat body, they had less body weight gain, smaller fat cells and lower amounts of fats in the body. Genetically modified flies with a version of CTPS that was unable to form cytoophidia also showed fewer signs of obesity, indicating how the enzyme might influence the response to dietary fats. These findings further implicate CTPS in the cause of obesity and help to understand its role. However, it remains to be seen if this also applies to humans. If this is the case, drugs that block the activity of CTPS could help to reduce the impact of a high-fat diet on public health.

Cinnamaldehyde regulates mitochondrial quality against hydrogen peroxide induced apoptosis in mouse lung mesenchymal stem cells via the PINK1/Parkin signaling pathway.

Background: Idiopathic pulmonary fibrosis (IPF) is a fatal respiratory disease without effective treatments. Mitochondrial dysfunction weakens the ability of mesenchymal stem cells (MSCs) to repair the distal lung epithelium, which is a probable pathogenesis of IPF. In previous research, we found that cinnamaldehyde (CA) can maintain the mitochondrial morphology of MSCs. Methods: This present study evaluated the effect and mechanism of CA on murine lung MSCs using the hydrogen peroxide model. Antioxidant effects and mitochondrial function were determined using flow cytometry. The mRNA levels of mitochondrial dynamics and the expressions of autophagy-related proteins were also detected. Results: CA can increase the levels of SOD, MMP and ATP, decrease the rate of ROS and apoptosis, and restore the mitochondrial structure. CA can also improve the mRNA expression of MFN1, MFN2, FIS1, DRP1, OPA1, and PGC-1α, increase the expression of LC3 II and p62 and promote the PINK1/Parkin signaling pathway. Our results demonstrated that CA can control mitochondrial quality and avoid apoptosis, which may be associated with the regulation of the PINK1/Parkin signaling pathway.

Efficacy of Yiqi Wenyang Huoxue method in treating adult bronchial asthma: A protocol for systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Bronchial asthma (BA) is a chronic inflammatory disease of the airway, which has the characteristics of recurrent attacks and difficult to cure. Glucocorticoid and bronchodilator are the primary treatment drugs for asthma. Although the treatment has made some progress, the control status is still not ideal. According to clinical reports, the Yi-qi Wen-yang Huo-xue method (YQWYHXM) of Traditional Chinese Medicine (TCM) is safe and effective in the treatment of BA, but there is not enough evidence to prove it. Based on it, we conducted this systematic evaluation. METHODS: Eight databases and Clinical trial registries (Embase, Cochrane, PubMed, CNKI, CBM, Wanfang, VIP, China Clinical Trial Registry and Clinical Trails) were searched from the establishment of those until January 22, 2021 with the following terms for retrieval: BS, TCM, Chinese medicinal herb, Chinese herbal medicine and randomized controlled trial. Data analysis was performed by 2 researchers using RevMan 5.3 and SATA 16.0 separately from the Cochrane Collaboration. RESULTS: This study will be able to provide definitive evidence to clarify all the suspicions we seek, confirming the effectiveness of YQWYHXM in the treatment of adults with BA. CONCLUSION: This study will prove that YQWYHXM is a safe and effective TCM adjuvant therapy for BA. REGISTRATION: Efficacy of Yiqi Wenyang Huoxue method in Treating Adult Bronchial Asthma. ASTHMA: A protocol for Systematic Review and Meta-Analysis of Randomized Controlled Trials. PROSPERO 2021 CRD42021256791. Available from: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021256791.

Highly effective proximate labeling in Drosophila.

The protein-protein interaction (PPI) is a basic strategy for life to operate. The analysis of PPIs in multicellular organisms is very important but extremely challenging because PPIs are particularly dynamic and variable among different development stages, tissues, cells, and even organelles. Therefore, understanding PPI needs a good resolution of time and space. More importantly, understanding in vivo PPI needs to be realized in situ. Proximity-based biotinylation combined with mass spectrometry (MS) has emerged as a powerful approach to study PPI networks and protein subcellular compartmentation. TurboID, the newly engineered promiscuous ligase, has been reported to label proximate proteins effectively in various species. In Drosophila, we systematically apply TurboID-mediated biotinylation in a wide range of developmental stages and tissues, and demonstrate the feasibility of TurboID-mediated labeling system in desired cell types. For a proof-of-principle, we use the TurboID-mediated biotinylation coupled with MS to distinguish CTP synthase with or without the ability to form filamentous cytoophidia, retrieving two distinct sets of proximate proteomes. Therefore, this makes it possible to map PPIs in vivo and in situ at a defined spatiotemporal resolution, and demonstrates a referable resource for cytoophidium proteome in Drosophila.

The atlas of cytoophidia in Drosophila larvae.

In 2010, cytidine 5'-triphosphate synthase (CTPS) was reported to form the filamentous or serpentine structure in Drosophila, which we termed the cytoophidium. In the last decade, CTPS filaments/cytoophidia have been found in bacteria, budding yeast, human cells, mice, fission yeast, plants, and archaea, indicating that this mechanism is highly conserved in evolution. In addition to CTPS, other metabolic enzymes have been identified to have the characteristics of forming cytoophidia or similar advanced structures, demonstrating that this is a basic strategy of cells. Nevertheless, our understanding of the physiological function of the cytoophidium remains incomplete and elusive. Here, we took the larva of Drosophila melanogaster as a model to systematically describe the localization and distribution of cytoophidia in different tissues during larval development. We found that the distribution pattern of CTPS cytoophidia is dynamic and heterogenic in larval tissues. Our study provides a road map for further understanding of the function and regulatory mechanism of cytoophidia.

Cellulose nanocrystals reinforced foamed nitrile rubber nanocomposites.

Research on foamed nitrile rubber (NBR)/cellulose nanocrystals (CNs) nanocomposites is rarely found in the literatures. In this paper, CNs suspension and NBR latex was mixed to prepared the foamed NBR/CNs nanocomposites. We found that the CNs mainly located in the cell walls, effectively reinforcing the foamed NBR. The strong interaction between the CNs and NBR matrix restricted the mobility of NBR chains surrounding the CNs, hence increasing the crosslink density of the NBR matrix. CNs exhibited excellent reinforcement on the foamed NBR: a remarkable increase nearly 76% in the tensile strength of the foamed nanocomposites was achieved with a load of only 15 phr CNs. Enhanced mechanical properties make the foamed NBR/CNs nanocomposites a promising damping material for industrial applications with a potential to reduce the petroleum consumption.