The Identification and Gene Mapping of Spotted Leaf Mutant spl43 in Rice.

Our study investigates the genetic mechanisms underlying the spotted leaf phenotype in rice, focusing on the spl43 mutant. This mutant is characterized by persistent reddish-brown leaf spots from the seedling stage to maturity, leading to extensive leaf necrosis. Using map-based cloning, we localized the responsible locus to a 330 Kb region on chromosome 2. We identified LOC_Os02g56000, named OsRPT5A, as the causative gene. A point mutation in OsRPT5A, substituting valine for glutamic acid, was identified as the critical factor for the phenotype. Functional complementation and the generation of CRISPR/Cas9-mediated knockout lines in the IR64 background confirmed the central role of OsRPT5A in controlling this trait. The qPCR results from different parts of the rice plant revealed that OsRPT5A is constitutively expressed across various tissues, with its subcellular localization unaffected by the mutation. Notably, we observed an abnormal accumulation of reactive oxygen species (ROS) in spl43 mutants by examining the physiological indexes of leaves, suggesting a disruption in the ROS system. Complementation studies indicated OsRPT5A's involvement in ROS homeostasis and catalase activity regulation. Moreover, the spl43 mutant exhibited enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo), highlighting OsRPT5A's role in rice pathogen resistance mechanisms. Overall, our results suggest that OsRPT5A plays a critical role in regulating ROS homeostasis and enhancing pathogen resistance in rice.

Antagonistic activity of selenium-enriched Bifidobacterium breve against Clostridioides difficile.

Probiotics have the potential to be used in the prevention of Clostridioides difficile infection (CDI). In this study, selenium (Se)-enriched Bifidobacterium breve YH68-Se was obtained under optimal culture conditions with single-factor and response surface optimization. The overall environmental resistance of YH68-Se was superior to that of the parental strain YH68, mainly reflected in the substantial improvement of antioxidant activity and gastrointestinal tolerance. YH68-Se dramatically inhibited C. difficile growth, spore, biofilm, toxin production, and virulence gene expression, rapidly disrupted C. difficile cell membrane permeability and integrity, and altered the membrane proton motive force (PMF), induced a large outflow of intracellular substances and eventually caused bacterial death. The main factor inducing this process originated from the lactic acid (LD) in YH68-Se. In addition, the LD production of YH68 increased with increasing selenite concentration and was accompanied by enhanced activities of thioredoxin reductase (TrxR), glutathione peroxidase (GSH-Px), and increased concentration of autoinducer-2 (AI-2), which may be the crucial factors contributing to the outstanding probiotic properties of YH68-Se and their potent antagonism of C. difficile. KEY POINTS: • Compared with the parental strain B. breve YH68, the environmental resistance of YH68-Se was improved. • YH68-Se was able to produce more lactic acid, which suppressed the important physiological activities of C. difficile and rapidly disrupted their cell membrane structures. • Sodium selenite in the suitable concentration range gradually increases the yield of lactic acid and phenylacetic acid, increased the concentration of autoinducer-2, and enhanced the activities of antioxidant enzymes TrxR and GSH-Px in YH68.

CO2-Low Interfacial Tension Viscoelastic Fluid Synergistic Flooding in Tight Reservoirs.

Tight oil reservoirs have poor physical properties, insufficient formation energy, and low natural productivity. CO2 flooding is an important technical mean that enhances the oil recovery of dense reservoirs and achieves effective CO2 sequestration, but strong heterogeneity of the tight oil reservoir usually results in gas channeling and poor enhanced oil recovery effect. The existing methods to prevent gas channeling are mainly to use the small-molecule amine system and the polymer gel system to plug fracture and high permeability channels. The small-molecular amine system has low flash points and pollutes the environment and the polymer gel has poor injectivity and great damage to the formation, which limit their large-scale application. Therefore, a new viewpoint of CO2-low interfacial tension viscoelastic fluid synergistic flooding for enhanced oil recovery in a tight oil reservoir was made. The performance of low interfacial tension viscoelastic fluid (GOBT) was studied. The injectivity and oil displacement effect of CO2-GOBT synergistic flooding were evaluated, and the mechanism of CO2-GOBT synergistic flooding was discussed. The experimental results showed that 0.4% GOBT is a low interfacial tension viscoelastic fluid, which has strong adaptability to the salinity water of tight oil reservoirs (6788-80,000 mg/L), good viscosity stability at different pHs, excellent capacity to emulsify crude oil, and the ability to improve reservoir water wettability. CO2 alternating 0.4% GOBT flooding has good injection ability in cores (K a = 0.249 mD), and injecting 0.4% GOBT can effectively increase the injection pressure of subsequent CO2 flooding. CO2 alternating 0.4% GOBT flooding can effectively improve water flooding recovery in tight sandstone reservoirs, which is better than CO2 flooding and 0.4% GOBT flooding in both homogeneous and heterogeneous conditions. The mechanisms of CO2 alternating 0.4% GOBT flooding to enhance the oil recovery include that GOBT and CO2 foam block high permeability layers, shunt and sweep low permeability layers, and GOBT emulsify and wash oil. CO2 partially dissolving in GOBT synergistically enhances the core water wettability, which improves GOBT injectability, emulsification, and stripping ability to residual oil.

Study on Well Spacing Optimization in a Tight Sandstone Gas Reservoir Based on Dynamic Analysis.

Compared to the shale gas and coalbed methane in China, tight gas has been recently considered as a priority in the exploration and exploitation of unconventional gas resources. In the development of a tight gas field, how to enhance the gas recovery is a prevalent topic. Unlike the conventional gas reservoir, the ultimate gas recovery is not only determined by the geological characteristics but is also affected by other factors such as well drainage area and well spacing design. For tight sandstone reservoirs, the gas recovery can be improved by increasing the drainage area. Moreover, the well drainage area is closely associated with well spacing. Therefore, effective drainage area estimation and well spacing optimization are essential aspects for tight gas exploitation. In this paper, a new optimization workflow is established, which combined dynamic analysis and numerical simulation techniques. First, through interference well test results and production data dynamic analysis, the total gas production can be expressed and predicted. Then the well density can be optimized by the economic evaluation method. Meanwhile, a numerical model is built up to determine the optimal well spacing. This new optimization workflow can provide guidance to the operators of tight gas fields where the interference well test results are available and several years of production data are collected. Furthermore, in the case of the Sulige gas field, the single well drainage area is estimated and the optimal well pattern is obtained by the established approach. The results indicate that the well pattern of 500 m × 600 m is most reasonable for the pilot gas field.

IGFBP2 upregulates ZEB1 expression and promotes hepatocellular carcinoma progression through NF-κB signaling pathway.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most lethal cancers owing to the high metastasis rate. The molecular mechanism underlying HCC progression remains unclear. AIMS: We aimed to explore the function and mechanism of action of insulin-like growth factor binding protein 2 (IGFBP2) in HCC. METHODS: Expression of IGFBP2 was evaluated with western blotting and reverse transcription polymerase chain reaction (RT-PCR). Loss- and gain-function assays were conducted to evaluate the effects of IGFBP2 on HCC cell proliferation, migration, and invasion. Signaling pathways were screened with a dual-fluorescein reporting system, and levels of epithelial and mesenchymal markers were measured after altering IGFBP2 expression. Cell fractionation analysis was conducted to evaluate the nuclear translocation of p65. RESULTS: IGFBP2 expression was upregulated in HCC tissues, predicted worse prognosis, and was associated with strong metastatic potentials. IGFBP2 depletion significantly inhibited HCC cell proliferation, migration, and invasion, whereas IGFBP2 overexpression showed reverse phenotypes. The underlying mechanism involved IGFBP2-mediated nuclear localization of p65, which activated nuclear factor kappa B (NF-κB) and zinc finger E-Box binding homeobox 1 (ZEB1) transcription via binding to the gene promoter. CONCLUSION: This study for the first time identifies IGFBP2 as a novel therapeutic target in HCC that activates the NF-κB-ZEB1 signaling axis and promotes HCC tumorigenesis.