Mechanism of folate deficiency-induced apoptosis in mouse embryonic stem cells: Cell cycle arrest/apoptosis in G1/G0 mediated by microRNA-302a and tumor suppressor gene Lats2

Deficiencies in maternal diet, such as inadequate intake of folate, can inhibit normal development and lead to developmental defects. MicroRNAs (miRNAs) may play a role in mediating the effects of folate deficiency in the growing mammalian embryo, although conclusive evidences to support that possibility are not yet available. The goal of the present study was to investigate whether and how folate deprivation alters the properties of mouse embryonic stem cells (mESCs) in culture. For this purpose, mESCs were cultured in folate-deficient or complete culture medium. The results show that folate-deficient mESCs have a significantly higher rate of apoptosis, accumulate in G0/G1 and fail to proliferate. Expression profiling revealed several miRs and many mRNAs are differently expressed in folate-deficient cells. RT-PCR data confirmed differential expressions of 12 miRNAs in folate-deficient cells. Furthermore, bioinformatics analyses and in vitro studies suggested that miR-302a plays a critical role in mediating the effects of folate on cell proliferation and cell cycle-specific apoptosis by targeting Lats2 gene. Together, these results suggest that the effects of folate deficiency on mammalian development may be mediated by miRNAs that regulate proliferation and/or cell cycle progression in ESCs.     

Kinetic, structural and molecular docking studies on the inhibition of tyrosinase induced by arabinose

Tyrosinase plays a central role in biological pigment formation, and hence knowledge of tyrosinase catalytic mechanisms and regulation may have medical, cosmetic, and agricultural applications. We found in this study that arabinose significantly inhibited tyrosinase, and this was accompanied by conformational changes in enzyme structure. Kinetic analysis showed that arabinose-mediated inactivation followed first-order kinetics, and single and multiple classes of rate constants were measured. Arabinose displayed a mixed-type inhibitory mechanism with K(i)=0.22±0.07 mM. Measurements of intrinsic and ANS-binding fluorescence showed that arabinose induced tyrosinase to unfold and expose inner hydrophobic regions. We simulated the docking between tyrosinase and arabinose (binding energies were -26.28 kcal/mol for Dock6.3 and -2.02 kcal/mol for AutoDock4.2) and results suggested that arabinose interacts mostly with His61, Asn260, and Met280. The present strategy of predicting tyrosinase inhibition by simulation of docking by hydroxyl groups may prove useful in screening for potential tyrosinase inhibitors, as shown here for arabinose.     

Boosting Polysulfide Redox Kinetics by Graphene‐Supported Ni Nanoparticles with Carbon Coating

Lithium–sulfur batteries have attracted extensive attention because of their high energy density. However, their application is still impeded by the inherent sluggish kinetics and solubility of intermediate products (i.e., polysulfides) of the sulfur cathode. Herein, graphene‐supported Ni nanoparticles with a carbon coating are fabricated by directly carbonizing a metal–organic framework/graphene oxide composite, which is then dispersed on a commercial glass fiber membrane to form a separator with electrocatalytic activity. In situ analysis and electrochemical investigation demonstrate that this modified separator can effectively suppress the shuttle effect and regulate the catalytic conversion of intercepted polysulfides, which is also confirmed by density functional theory calculations. It is found that Ni–C sites can chemically interact with polysulfides and stabilize the radical S₃^?? through Ni? S bonds to enable fast dynamic equilibrium with S₆^2?, while Ni nanoparticles reduce the oxidation barrier of Li₂S and accelerate ion/electron transport. As a result, the corresponding lithium–sulfur battery shows a high cycle stability (88% capacity retention over 100 cycles) even with a high sulfur mass loading of 8 mg cm^?2 and lean electrolyte (6.25 µ L mg^?1). Surprisingly, benefitting from the improved kinetics, the battery can work well at ?50 °C, which is rarely achieved by conventional Li–S batteries.     

Chemical synthesis,microbial transformation and biological evaluation of tetrahydroprotoberberines as dopamine D1/D2 receptor ligands

Dopamine D1/D2 receptors are important targets for drug discovery in the treatment of central nervous system diseases. To discover new and potential D1/D2 ligands, 17 derivatives of tetrahydroprotoberberine (THPB) with various substituents were prepared by chemical synthesis or microbial transformation using Streptomyces griseus ATCC 13273. Their functional activities on D1 and D2 receptors were determined by cAMP assay and calcium flux assay. Seven compounds showed high activity on D1/D2 receptor with low IC₅₀ values less than 1?µM. Especially, top compound 5 showed strong antagonistic activity on both D1 and D2 receptor with an IC₅₀ of 0.391 and 0.0757?µM, respectively. Five compounds displayed selective antagonistic activity on D1 and D2 receptor. The SAR studies revealed that (1) the hydroxyl group at C-9 position plays an important role in keeping a good activity and small or fewer substituents on ring D of THPBs may also stimulate their effects, (2) the absence of substituents at C-9 position tends to be more selective for D2 receptor, and (3) hydroxyl substitution at C-2 position and the substitution at C-9 position may facilitate the conversion of D1 receptor from antagonist to agonist. Molecular docking simulations found that Asp 103/Asp 114, Ser 107/Cys 118, and Trp 285/ Trp 386 of D1/ D2 receptors are the key residues, which have strong interactions with the active D1/D2 compounds and may influence their functional profiles.     

A PGD Pregnancy Achieved by Embryo Copy Number Variation Sequencing with Confirmation by Non-Invasive Prenatal Diagnosis

Aneuploidy rates in embryos produced by assisted reproduc- tive technologies commonly exceed 50%, particularly in couples where the woman is of advanced maternal age or has experienced repeated implantation failure （Harper et al., 2012）. Pre-implantation genetic diagnosis （PGD） conducted at many fertility clinics worldwide is now a frontline treatment for infertile couples with a poor prognosis for pregnancy as well as couples where one partner is a carrier of a balanced translocation （Munne, 2012）.     

pH值对FOXO1、GDF-8基因反义RNA寡核苷酸吸收的影响

研究不同pH值下,口服靶向FOXO1与GDF-8基因反义RNA寡核苷酸药物对药效的影响.化学合成靶向FOXO1与GDF-8基因的有效反义RNA寡核苷酸片段,调节药物PH值,通过灌胃方式给药,给药20 d后处死小鼠,进行体重,腿部肌肉增长情况分析.提取腿部肌肉组织总RNA,用real time PCR检测FOXO1与GDF-8基因的表达.结果表明,服用RNA oligos的试验组小鼠腿部肌肉重量均比对照组肌肉重量增长快.其中pH为7.0的RNA oligos的效果比pH为5.0的效果好, pH9.0的RNA oligos的作用效果最弱.与小鼠体重变化结果一致,real time PCR检测实验组FOXO1与GDF-8基因转录水平较对照组均有明显下降,其中服用pH为7.0的RNA oligos的下降最多.因此,口服靶向FOXO1与GDF-8基因的反义RNA寡核苷酸药物的最适pH值应为中性偏酸.     

Kinetics Absorption Characteristics of Ferrous Glycinate in SD Rats and Its Impact on the Relevant Transport Protein

Ferrous glycinate (Fe-Gly) maintains high bioavailability in animals, but its exact absorption mechanism is still unknown. Here, we studied on the absorption kinetics of ferrous glycinate and its impact on the relevant transport protein in Sprague-Dawley (SD) rats. A total of 72 SD rats (male, BW 100?±?6.25 g) were randomly allotted to three treatments. These treatments were perfused with 1 mL of normal saline, ferrous sulfate (FeSO₄), and ferrous glycinate (71.35 mg/L as iron) separately. Four rats were selected from each treatment for collection of blood from the tails at certain times (15, 30, 45, 60, 75, 90, 120, 240, and 360 min) after gavage. Moreover, other six rats selected from each treatment were slaughtered for sampling after gavage at 2, 4, and 6 h to evaluate the expression of intestinal transport protein. Pharmacokinetic parameters of iron were determined by one-compartmental analysis. Compared with FeSO₄, the peak plasma concentration of iron (C _max) is higher in the rats given gavage with Fe-Gly (P?<?0.05). Four hours after gavage with Fe-Gly, the expression of divalent metal transporter 1 (DMT1) in the duodenum is significantly decreased (P?<?0.05), but the expression of ferroportin 1 (Fpn1) is significantly increased (P?<?0.05). This study indicates that Fe-Gly as iron sources can be absorbed more and utilized faster than FeSO₄, and they had different effects on the expression of intestinal transport protein.