Molecular Dynamics Analysis Reveals Structural Insights into Mechanism of Nicotine N-Demethylation Catalyzed by Tobacco Cytochrome P450 Mono-Oxygenase

CYP82E4, a cytochrome P450 monooxygenase, has nicotine N-demethylase (NND) activity, which mediates the bioconversion of nicotine into nornicotine in senescing tobacco leaves. Nornicotine is a precursor of the carcinogen, tobacco-specific nitrosamine. CYP82E3 is an ortholog of CYP82E4 with 95% sequence identity, but it lacks NND activity. A recent site-directed mutagenesis study revealed that a single amino acid substitution, i.e., cysteine to tryptophan at the 330 position in the middle of protein, restores the NND activity of CYP82E3 entirely. However, the same amino acid change caused the loss of the NND activity of CYP82E4. To determine the mechanism of the functional turnover of the two molecules, four 3D structures, i.e., the two molecules and their corresponding cys–trp mutants were modeled. The resulting structures exhibited that the mutation site is far from the active site, which suggests that no direct interaction occurs between the two sites. Simulation studies in different biological scenarios revealed that the mutation introduces a conformation drift with the largest change at the F-G loop. The dynamics trajectories analysis using principal component analysis and covariance analysis suggests that the single amino acid change causes the opening and closing of the transfer channels of the substrates, products, and water by altering the motion of the F-G and B-C loops. The motion of helix I is also correlated with the motion of both the F-G loop and the B-C loop and; the single amino acid mutation resulted in the curvature of helix I. These results suggest that the single amino acid mutation outside the active site region may have indirectly mediated the flexibility of the F-G and B-C loops through helix I, causing a functional turnover of the P450 monooxygenase

Pulmonary delivery of mucus-traversing PF127-modified silk fibroin nanoparticles loading with quercetin for lung cancer therapy

The mucosal barrier remains a major barrier in the pulmonary drug delivery system, as mucociliary clearance in the airway accelerates the removal of inhaled nanoparticles (NPs). Herein, we designed and developed the inhalable Pluronic F127-modified silk fibroin NPs loading with quercetin (marked as QR-SF (PF127) NPs), aiming to solve the airway mucus barrier and improve the cancer therapeutic effect of QR. The PF127 coating on the SF NPs could attenuate the interaction between NPs and mucin proteins, thus facilitating the diffusion of SF(PF127) NPs in the mucus layer. The QR-SF (PF127) NPs had particle sizes of approximately 200 nm with negatively charged surfaces and showed constant drug release properties. Fluorescence recovery after photobleaching (FRAP) assay and transepithelial transport test showed that QR-SF (PF127) NPs exhibited superior mucus-penetrating ability in artificial mucus and monolayer Calu-3 cell model. Notably, a large amount of QR-SF (PF127) NPs distributed uniformly in the mice airway section, indicating the good retention of NPs in the respiratory tract. The mice melanoma lung metastasis model was established, and the therapeutic effect of QR-SF (PF127) NPs was significantly improved in vivo. PF127-modified SF NPs may be a promising strategy to attenuate the interaction with mucin proteins and enhance mucus penetration efficiency in the pulmonary drug delivery system

Increased CO2 concentrations increasing water use efficiency and improvement PSII function of mulberry seedling leaves under drought stress

In this paper, the effects of different CO2 concentrations (400 and 800 μmol·mol−1) on photosynthetic gas exchange and chlorophyll fluorescence of mulberry (Morus alba L.) seedling leaves under drought stress were studied using an artificial climate chamber. The results showed that under non-drought conditions, the stomatal concentrations (Gs) and transpiratration rate (Tr) of mulberry seedling leaves decreased slightly with increased CO2 concentrations, but the net photosynthetic rate (Pn) increased significantly. Under mild drought stress (5 days of drought), higher CO2 concentrations significantly affected the photosynthetic gas exchange of mulberry seedling leaves, but had little effect on chlorophyll fluorescence. Under severe drought stress (10 das of drought), increased CO2 concentrations not only alleviated drought stress by increasing the WUE of mulberry seedling leaves, but also significantly increased its PSII photochemical activity, which promoted electron transfer on the PSII acceptor side. In conclusion, increased CO2 concentrations could raise the WUE of mulberry seedling leaves under normal water conditions and drought stress, and this effect was more significant under drought stress. Under severe drought stress, increased CO2 concentrations improved the drought resistance of mulberry seedlings by improving their PSII function

Popular culture in the making of anti-imperialist and nationalist sentiments in Sichuan

Existing Western scholarship on the rights recovery movement in Sichuan mainly focuses on the role played by elites. This article argues that popular culture, in the form of folk stories, songs, and children’s primers, also contributed to that movement by shaping and expressing popular anti-imperialist attitudes. Its analysis of primers available in late Qing Sichuan and popular stories about the activities of foreigners prevalent in the early 1900s serves to reveal a rich local cultural milieu of time-nurtured anti-imperialist sentiment among common people, which broadly influenced local political action. The protests over the Jiangbei mining concession encompassed both elite and ordinary people, although each group understood the issue differently