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PIWI-interacting RNAs (piRNAs) are important for ensuring the integrity of the germline. 3'-terminal 2'-O-methylation is essential for piRNA maturation and to protect them from degradation. HENMT1 (HEN Methyltransferase 1) carries out the 2'-O-methylation, which is of key importance for piRNA stability and functionality. However, neither the structure nor the catalytic mechanism of mammalian HENMT1 have been studied. We have constructed a catalytic-competent HENMT1 complex using computational approaches, in which Mg2+ is primarily coordinated by four evolutionary conserved residues, and is further auxiliary coordinated by the 3'-O and 2'-O on the 3'-terminal nucleotide of the piRNA. Our study suggests that metal has limited effects on substrate and cofactor binding but is essential for catalysis. The reaction consists of deprotonation of the 2'-OH to 2'-O and a methyl transfer from SAM to the 2'-O. The methyl transfer is spontaneous and fast. Our in-depth analysis suggests that the 2'-OH may be deprotonated before entering the active site or it may be partially deprotonated at the active site by His800 and Asp859, which are in a special alignment that facilitates the proton transfer out of the active site. Furthermore, we have developed a detailed potential reaction scenario indicating that HENMT1 is Mg2+ utilizing but is not a Mg2+ dependent enzyme.

We investigate the effect of the optimized aging processing on magnetism and mechanical property of the sintered Dy-doped Nd–Fe–B permanent magnet. The experimental results show that the magnetism, especially intrinsic coercivity, of the optimized aged Dy-doped Nd–Fe–B magnet is more excellent than that of the sintered one, but the former's strength and hardness are lower than that of the latter. It was observed that the optimized aged Dy-doped Nd–Fe–B magnet have more uniform grain size, thinner (Nd, Dy)-rich boundary phase. By means of the EBSD technology, the number of larger angle grain boundaries in the optimized aged Dy-doped Nd–Fe–B magnet is more than that of the sintered one. The reasons for the increased intrinsic coercivity and decreased mechanical properties of the optimized aged Dy-doped Nd–Fe–B magnet are also discussed. [ABSTRACT FROM AUTHOR]

Methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFD2) is a new drug target that is expressed in cancer cells but not in normal adult cells, which provides an Achilles heel to selectively kill cancer cells. Despite the availability of crystal structures of MTHFD2 in the inhibitor- and cofactor-bound forms, key information is missing due to technical limitations, including (a) the location of absolutely required Mg2+ ion, and (b) the substrate-bound form of MTHFD2. Using computational modeling and simulations, we propose that two magnesium ions are present at the active site whereby (i) Arg233, Asp225, and two water molecules coordinate [Formula: see text], while [Formula: see text] together with Arg233 stabilize the inorganic phosphate (Pi); (ii) Asp168 and three water molecules coordinate [Formula: see text], and [Formula: see text] further stabilizes Pi by forming a hydrogen bond with two oxygens of Pi; (iii) Arg201 directly coordinates the Pi; and (iv) through three water-mediated interactions, Asp168 contributes to the positioning and stabilization of [Formula: see text], [Formula: see text] and Pi. Our computational study at the empirical valence bond level allowed us also to elucidate the detailed reaction mechanisms. We found that the dehydrogenase activity features a proton-coupled electron transfer with charge redistribution connected to the reorganization of the surrounding water molecules which further facilitates the subsequent cyclohydrolase activity. The cyclohydrolase activity then drives the hydration of the imidazoline ring and the ring opening in a concerted way. Furthermore, we have uncovered that two key residues, Ser197/Arg233, are important factors in determining the cofactor (NADP+/NAD+) preference of the dehydrogenase activity. Our work sheds new light on the structural and kinetic framework of MTHFD2, which will be helpful to design small molecule inhibitors that can be used for cancer treatment.

The heart is one of the organs most vulnerable to sepsis. This review describes the general characteristics of sepsis-induced cardiomyopathy and the main pathogenesis of myocardial dysfunction in sepsis. Levosimendan is a novel drug for treatment of sepsis-induced myocardial dysfunction. This review also elaborates on the pathogenesis of levosimendan, including the mechanisms of its anti-inflammatory effects, improvement of myocardial ischaemia, increased synthesis of nitric oxide, vascular endothelial cell protection, increased myocardial contractility, improved diastolic function, and inhibition of hypoxia-inducible factor-1α expression. Many clinical studies have proven that levosimendan effectively prevents myocardial dysfunction in sepsis. In addition to the widespread use of levosimendan in patients with heart failure, the role of levosimendan in the treatment of patients with sepsis-induced cardiomyopathy will be increasingly studied and applied in the future.

Aims: β-Catenin accumulation promotes proliferation. However, the correlation between proliferation of colorectal epithelium and β-catenin in type 2 diabetes mellitus (DM) patients remains unclear. Methods: Colorectal epithelium samples from distal ends of colorectal adenocarcinomas without histological aberrances were divided into two groups: DM patients with type 2 DM for more than 1 year (n = 27) and non-DM patients without hyperglycemia (n = 20). Samples from patients without colorectal epithelial disease or hyperglycemia served as a control group (n = 6). Proliferative index was calculated as the percentage of proliferating cell nuclear antigen positive cells. Wnt/β-catenin signaling was assessed immunohistochemically and phosphorylation of β-catenin was assessed by immunofluorescence. Results: Compared with the non-DM or control group, the proliferative index and expression of lactate dehydrogenase A and Wnt/β-catenin signaling were significantly higher in the DM group(all p < 0.01). The proliferative index correlated positively with β-catenin expression (Spearman correlation coefficient = 0.55; p < 0.01). Reduced phosphorylation at serine 33/37 and increased phosphorylation at serine 675 of β-catenin were detected in the DM group (all p < 0.01). Conclusions: Enhanced proliferation, accompanied by increased aerobic glycolysis, was detected in colorectal epithelium of patients with diabetes. β-Catenin accumulation with altered phosphorylation correlated with the proliferative changes. [ABSTRACT FROM AUTHOR]

Shaker-like K + outward rectifying channel (SKOR) is involved in mediating long-distance K + transport from roots to shoots. In this study, a Shaker-like outward K + channel gene CmSKOR (GenBank accession number MF447462) was isolated from melon (Cucumis melo L.). Phylogenetic analysis showed that CmSKOR belongs to the SKOR-subfamily in the Shaker-like K + channel family. Electrophysiological experiments indicated that CmSKOR was a K + -permeable channel with low affinity. Expressed in Xenopus oocytes, CmSKOR displayed classical Shaker-like outwardly rectifying K + currents. Confocal imaging of a CmSKOR - yellow fluorescent fusion protein (YFP) in transgenic Nicotiana tabacum leaves indicated that CmSKOR was located in the plasma membrane. Transcript analysis showed CmSKOR predominantly expressed in melon roots and with lower abundance in stem and leaves. In addition, both external K + and NaCl treatment could up-regulate the expression of CmSKOR in melon and enhance the K + content in shoot. Constitutive overexpressed CmSKOR in Arabidopsis thaliana, the transgenic plants showed changes in root length in MS plates, displayed higher maximum photochemical efficiency of PSII (Fv/Fm), higher fresh and dry weight, and accumulation of K + in shoot, together with the changes of transcript amount of CmSKOR with NaCl treatments in mixture substrate. In conclusion, it was proposed that CmSKOR may play the role on distributing K + to the shoot in melon and its constitutive expression in Arabidopsis improved saline tolerance by maintaining K + homeostasis in the plant., (Copyright © 2018 Elsevier B.V. All rights reserved.)