Discovery and evaluation of novel FAAH inhibitors in neuropathic pain model

Conceptual design and modification of urea moiety in chemotype PF-3845/04457845, the bench marking irreversible inhibitor of fatty acid amide hydrolase (FAAH), led to discovery of a novel nicotinamide-based lead 12a having reversible mechanism of action. Focused SAR around the pyridine heterocycle (Ar) in 12a (Tables 1 and 2) resulted into four shortlisted compounds, (?)-12a, (?)-12i, (?)-12l–m. The required (?)-enantiomers were obtained via diastereomeric resolution of a novel chiral dissymmetric intermediate 15. Based on comparative profile of FAAH potency, metabolic stability in liver microsome, liability of inhibiting major hCYP450 isoforms, rat PK, and brain penetration ability, two SAR optimized compounds, (?)-12l and (?)-12m, were selected for efficacy study in rat model of chemotherapy-induced peripheral neuropathy (CIPN). Both the compounds exhibited dose related antihyperalgesic effects, when treated with 3–30?mg/kg po for 7?days. The effects at 30?mg/kg are comparable to that of PF-04457845 (10?mg/kg) and Tramadol (40?mg/kg).     

Optimization of Pre-transplantation Conditions to Enhance the Efficacy of Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) are considered a potential tool for cell based regenerative therapy due to their immunomodulatory property, differentiation potentials, trophic activity as well as large donor pool. Poor engraftment and short term survival of transplanted MSCs are recognized as major limitations which were linked to early cellular ageing, loss of chemokine markers during ex vivo expansion, and hyper-immunogenicity to xeno-contaminated MSCs. These problems can be minimized by ex vivo expansion of MSCs in hypoxic culture condition using well defined or xeno-free media i.e., media supplemented with growth factors, human serum or platelet lysate. In addition to ex vivo expansion in hypoxic culture condition using well defined media, this review article describes the potentials of transient adaptation of expanded MSCs in autologous serum supplemented medium prior to transplantation for long term regenerative benefits. Such transient adaptation in autologous serum supplemented medium may help to increase chemokine receptor expression and tissue specific differentiation of ex vivo expanded MSCs, thus would provide long term regenerative benefits.     

Effects of Magnesium Supplementation on Testosterone Levels of Athletes and Sedentary Subjects at Rest and after Exhaustion

This study was performed to assess how 4 weeks of magnesium supplementation and exercise affect the free and total plasma testosterone levels of sportsmen practicing tae kwon do and sedentary controls at rest and after exhaustion. The testosterone levels were determined at four different periods: resting before supplementation, exhaustion before supplementation, resting after supplementation, and exhaustion after supplementation in three study groups, which are as follows: Group 1—sedentary controls supplemented with 10 mg magnesium per kilogram body weight. Group 2—tae kwon do athletes practicing 90–120 min/day supplemented with 10 mg magnesium per kilogram body weight. Group 3—tae kwon do athletes practicing 90–120 min/day receiving no magnesium supplements. The free plasma testosterone levels increased at exhaustion before and after supplementation compared to resting levels. Exercise also increased testosterone levels relative to sedentary subjects. Similar increases were observed for total testosterone. Our results show that supplementation with magnesium increases free and total testosterone values in sedentary and in athletes. The increases are higher in those who exercise than in sedentary individuals.     

乌鲁木齐山地草地生态系统服务价值变化评估——基于遥感与GIS

草地生态系统服务功能是指草地生态系统及其生态过程所形成和维持的人类赖以生存的自然环境及其有效性,中国干旱半干旱地区的草地退化严重制约着生态系统服务功能的。以乌鲁木齐市山地草地为研究靶区,利用遥感图像(Landsat TM/OLI)划分出7种不同草地类型并计算其生态系统服务价值(Ecosystem Service Values, ESV)。使用空间统计方法、网格分析、空间自相关等方法进一步分析研究区山地草地退化程度和ESV变化。结果表明:21年中,乌鲁木齐市草地生态系统服务价值呈现波动性下降趋势,年均减少0.27×10~7元。生态损失主要是由山地荒漠草地(7.44%)、山地荒漠草原草地(10.71%)、山地草甸草原草地(18.32%)和高寒草甸草地(40.69%)退化造成。单项生态系统服务价值中,草地对调节服务(土壤形成)和供给服务的贡献率较高。草地ESVs Moran平均值为0.869(1994—2015年),并且草地ESV与相邻网格(2 km×2 km)具有更高的空间相关性。草地ESV具有明显的空间分布特征,高ESV区基本上分布在南山山区,低ESV区主要分布在平原草地区,博格达山草地亚区呈现随机分布格局。研究区气候变化对草地的影响十分显著,平均气温、平均地表温度、湿度、蒸散量、降水量和风速对草地ESV变化均有一定的影响,过度放牧、不合理垦荒、药用植物过度开发、城市化等人为活动仍然是驱动草地退化的主要因子,因此亟需采取措施保育乌鲁木齐市山地草原生态系统。     

Polarization-Dependent Confocal Imaging of Individual Ag Nanorods and Nanoparticles

Polarization-dependent inelastic optical scattering (IOS) of individual Ag nanorods and nanoparticles are studied by confocal imaging. Stronger IOS is observed at two ends of the nanorod with laser polarizing parallel to the rod long axis while the IOS images of Ag nanoparticles elongate along laser polarization direction. The correlation between the far-field IOS image and near-field spatial distributions of the nanostructures′ electric field can be obtained. The IOS imaging is demonstrated to be an effective technique to study the optical properties of metal nanostructures, which also provides information for their applications in surface-enhanced Raman scattering.     

Testosterone Levels in Athletes at Rest and Exhaustion: Effects of Calcium Supplementation

The effects of 4 weeks of calcium supplementation on free- and total testosterone levels were established in active and sedentary adult males at rest and exhaustion. Thirty healthy male athletes were equally divided into three study groups, as follows: Group 1—non-exercising subjects receiving 35 mg calcium/kg body weight; Group 2—subjects receiving 35 mg calcium/kg body weight undergoing training routines for 90 min/day, 5 days a week and Group 3—subjects undergoing training routines for 90 min/day, 5 days a week. The testosterone levels were determined before and after supplementation, at rest and following a hard training routine. The plasma free- and total testosterone levels increased at exhaustion before and after supplementation relative to resting values (p?<?0.05). This was also true when active subjects were compared to inactive subjects (p?<?0.05). Our results show that training results in increased testosterone levels in athletes and that the increase is greater if accompanied by calcium supplementation, which may be useful for increasing overall athletic performance.     

Live fluorescence and transmission-through-dye microscopic study of actinomycin D-induced apoptosis and apoptotic volume decrease

The effect of actinomycin D on HeLa cells was studied by live fluorescence and transmission-through-dye microscopy—a recently developed technique that permits volume measurements in live cells. In particular, it is well suited for the observation and quantification of the apoptotic volume decrease (AVD), which is widely viewed as an essential feature of apoptosis. The main results from our study are as follows. (1) Apoptosis caused in HeLa cells by actinomycin D proceeds in two morphologically distinct stages: the early stage is characterized by extensive blebbing, and the late stage by a more compact shape. The loss of mitochondrial membrane potential occurs at about the same time as blebbing, and chromatin condensation follows 30–90 min later. Caspase-3 and 7 become activated during the late stage. (2) Because blebbing occurs before activation of caspase-3, it has to be initiated by a different mechanism. Although blebbing is one of the earliest observable changes, it can be selectively inhibited without affecting other apoptotic reactions. (3) The majority of cells experience a temporary volume increase after the appearance of blebs. Eventually, AVD takes over and the cells shrink by approximately 40 % of their initial volume; the volume loss becomes noticeable at the end of the blebbing phase and continues through the late stage. Sometimes, at the end of long incubations, shrinkage gives way to swelling, possibly indicating secondary necrosis. (4) Both early and late apoptosis are accompanied by intracellular accumulation of Na⁺, while low-sodium medium prevents apoptosis. Except for a partial protective effect of quinine, all of the tested blockers of Na⁺, K⁺ and Cl^? channels failed to prevent apoptosis or AVD.     

Determination of the Role of DDX3 a Factor Involved in Mammalian RNAi Pathway Using an shRNA-Expression Library

RNA interference (RNAi) is an endogenous RNA-destruction phenomenon induced by certain double-stranded RNAs (dsRNAs). In RNAi, dsRNAs are processed into small interfering RNAs (siRNAs) which in turn trigger the cleavage of the target mRNA. Here, using a short hairpin RNA-expression library, we identified a DEAD-box helicase 3, DDX3, as an essential factor involved in RNAi pathway and revealed that DDX3 is colocalized with Ago2, an essential factor in RNAi pathway that cleaves target mRNA. Results of experiments with a dominant negative mutant of DDX3 further confirmed that this factor affects the RNAi activity. Together, DDX3 functions to assure mammalian RNAi pathway. Together, our results indicate that DDX3 is a new key molecule to understand the molecular mechanism underlying RNAi pathway in mammals.     

Mastl is required for timely activation of APC/C in meiosis I and Cdk1 reactivation in meiosis II

In mitosis, the Greatwall kinase (called microtubule-associated serine/threonine kinase like [Mastl] in mammals) is essential for prometaphase entry or progression by suppressing protein phosphatase 2A (PP2A) activity. PP2A suppression in turn leads to high levels of Cdk1 substrate phosphorylation. We have used a mouse model with an oocyte-specific deletion of Mastl to show that Mastl-null oocytes resume meiosis I and reach metaphase I normally but that the onset and completion of anaphase I are delayed. Moreover, after the completion of meiosis I, Mastl-null oocytes failed to enter meiosis II (MII) because they reassembled a nuclear structure containing decondensed chromatin. Our results show that Mastl is required for the timely activation of anaphase-promoting complex/cyclosome to allow meiosis I exit and for the rapid rise of Cdk1 activity that is needed for the entry into MII in mouse oocytes.