Discovery of potentially biased agonists of mu-opioid receptor (MOR) through molecular docking, pharmacophore modeling, and MD simulation.

Opioids are well known for their potent analgesic efficacy and severe side effects. Studies have shown that analgesic effects are mediated by the downstream G-protein-dependent pathway of the µ-opioid receptor (MOR), and another ß-arrestin-dependent pathway mediates side effects such as respiratory depression, constipation and tolerance etc. TRV130 is a biased ligand for G-protein-dependent pathway, which has high analgesia and has fewer side effects than morphine. In this study, the structure similarity search was performed on the IBSSC database using Oliceridine (TRV130) and PZM21 as templates. The 3D structure-based pharmacophore model was built and combined molecular docking prediction mode was selected to filter out small molecules, Finally, based on affinity prediction, four candidate molecules were obtained. Molecular dynamics simulations explored the detailed interaction mechanism of proteins with small molecules under dynamics. These results suggest that these candidate molecules are potential MOR agonists.

Overexpression of soybean GmPLDγ enhances seed oil content and modulates fatty acid composition in transgenic Arabidopsis.

Phospholipase D (PLD) hydrolyzes the phosphodiester bond of glycerophospholipids to yield phosphatidic acid (PA) and a free headgroup. PLDs are important for plant growth, development, and responses to external stresses. However, their roles in triacylglycerol (TAG) synthesis are still unclear. Here, we report that a soybean (Glycine max) PLDγ (GmPLDγ) is involved in glycerolipid turnover and seed oil production. GmPLDγ was targeted to mitochondria and exhibited PLD activity that was activated by oleate and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Overexpression of GmPLDγ (abbreviated GmPLDγ-OE) in Arabidopsis thaliana resulted in enhanced seed weight, elevated levels of TAGs with 18-, 20-, and 22-carbon fatty acids (FAs), and altered oil-body morphology. Furthermore, the levels of membrane lipids in vegetative tissues decreased significantly, whereas no overt changes were found in mature seeds except for a decrease in the digalactosyldiacylglycerol (DGDG) level in the GmPLDγ-OE lines. Additionally, the expression of genes involved in glycerolipid metabolism was significantly upregulated in developing siliques in GmPLDγ-OE lines. Together, our data indicate a regulatory role for GmPLDγ in TAG synthesis and fatty-acid remodeling, highlighting the importance of mitochondria-directed glycerophospholipid homeostasis in seed oil accumulation.

Genome-wide analysis and functional characterization of Acyl-CoA:diacylglycerol acyltransferase from soybean identify GmDGAT1A and 1B roles in oil synthesis in Arabidopsis seeds.

Acyl-CoA:diacylglycerol acyltransferase (DGAT) is a key enzyme in the Kennedy pathway of triacylglycerol (TAG) synthesis. It catalyzes the acyl-CoA-dependent acylation of sn-1, 2-diacylglycerol to form TAG. DGATs in soybean (Glycine max) have been reported, but their functions are largely unclear. Here we cloned three members of DGAT1 and four members of DGAT2 family from soybean, named GmDGAT1A to GmDGAT1C, and GmDGAT2A to GmDGAT2D, respectively. GmDGAT1A and GmDGAT1C were expressed at a high level in immature seeds, GmDGAT2B in mature seeds, and GmDGAT2C in older leaves. The seven genes were transformed into the H1246 quadruple mutant yeast strain, in which GmDGAT1A, GmDGAT1B, GmDGAT1C, GmDGAT2A, and GmDGAT2B had the ability to produce TAG. Six genes were transformed into Arabidopsis respectively, and constitutive expression of GmDGAT1A and GmDGAT1B resulted in an increase in oil content at the cost of reduced protein content in seeds. Overexpression of GmDGAT1A produced heavier weight of individual seed, but did not affect the weight of total seeds from a plant. Our results reveal the functions of soybean DGATs in seed oil synthesis using transgenic Arabidopsis. The implications for the biotechnological modification of the oil contents in soybeans by altering DGAT expression are discussed.

The clinical effect of rehabilitation following arthroscopic rotator cuff repair: A meta-analysis of early versus delayed passive motion.

BACKGROUND: The argument on the recommended rehabilitation protocol following arthroscopic rotator cuff repair remains to be resolved. So this meta-analysis was presented to evaluate the differences of clinical effects between the 2 distinct rehabilitation protocols after arthroscopic rotator cuff repair. METHODS: The PubMed, Cochrane Library, Web of Science, and EMBASE were systematically searched. Only randomized controlled trials (RCTs) published up to July 25, 2017, comparing early passive motion (EPM) versus delayed passive motion (DPM) rehabilitation protocols following arthroscopic rotator cuff repair were identified. The primary outcomes included range of motion and healing rate, while the secondary outcomes were Constant score, American Shoulder and Elbow Society (ASES) score, and Simple Shoulder Test (SST) score. The exclusion criteria contained biochemical trials, reviews, case reports, retrospective studies, without mention about passive motion exercise, no assessment of outcomes mentioned above, and no comparison of EPM and DPM rehabilitation protocols. RESULTS: Eight RCTs with 671 patients were enrolled in this study. The EPM resulted in improved shoulder forward flexion at short term, mid-term, and long-term follow-ups. The EPM group was superior to the DPM group in terms of external rotation (ER) at short-term and mid-term follow-ups. However, the DPM performed better long-term ASES score. These 2 protocols were equivalent in terms of ER at long term, ASES score at mid-term, SST score, Constant score, and healing rate. After excluding 2 RCTs that examined only small- and medium-sized tears, the pooled results of healing rate decreased from 82.4% to 76.6% in the EPM and 86.9% to 85.9% in the DPM. CONCLUSION: The meta-analysis suggests that the EPM protocol results in superior ROM recovery after arthroscopic rotator cuff repair but may adversely affect the shoulder function, which should be supported by further research. The healing rate at long-term follow-up is not clearly affected by the type of rehabilitation, but the EPM protocol might result in lower rates of tendon healing in the shoulder with large-sized tendon tears.

Homology modeling and screening of new 14α-demethylase inhibitor (DMI) fungicides based on optimized expression of CYP51 from Ustilago maydis in Escherichia coli.

Ustilago maydis infection is a serious disease affecting corn crops worldwide. Sterol 14α-demethylase (CYP51) is one of the key enzymes of sterol biosynthesis and an effective target of antifungal drugs. To further study the interaction between CYP51 and drugs and exploit more specific 14α-demethylase inhibitor (DMI) fungicides for U. maydis, in this study homology modeling of CYP51 from U. maydis (UmCYP51) templated as the eukaryotic orthologues (the human CYP51) and screening of new DMI fungicides based on optimized expression were carried out for the first time. In addition, XF-113 and ZST-4 were screened by analyzing the spectral characteristics between the purified UmCYP51-35 and fungicides. These results provide a theoretical basis and new ideas for efficient design and development of new antifungal drugs.