Exploring the stereochemistry of CXCR4-peptide recognition and inhibiting HIV-1 entry with D-peptides derived from chemokines

Chemokine receptor CXCR4 plays an important role in the immune system and the cellular entry of human immunodeficiency virus type 1 (HIV-1). To probe the stereospecificity of the CXCR4-ligand interface, d-amino acid peptides derived from natural chemokines, viral macrophage inflammatory protein II (vMIP-II) and stromal cell-derived factor-1alpha (SDF-1alpha), were synthesized and found to compete with (125)I-SDF-1alpha and monoclonal antibody 12G5 binding to CXCR4 with potency and selectivity comparable with or higher than their l-peptide counterparts. This was surprising because of the profoundly different side chain topologies between d- and l-enantiomers, which circular dichroism spectroscopy showed adopt mirror image conformations. Further direct binding experiments using d-peptide labeled with fluorescein (designated as FAM-DV1) demonstrated that d- and l-peptides shared similar or at least overlapping binding site(s) on the CXCR4 receptor. Structure-activity analyses of related peptide analogs of mixed chiralities or containing alanine replacements revealed specific residues at the N-terminal half of the peptides as key binding determinants. Acting as CXCR4 antagonists and with much higher biological stability than l-counterparts, the d-peptides showed significant activity in inhibiting the replication of CXCR4-dependent HIV-1 strains. These results show the remarkable stereochemical flexibility of the CXCR4-peptide interface. Further studies to understand the mechanism of this unusual feature of the CXCR4 binding surface might aid the development of novel CXCR4-binding molecules like the d-peptides that have high affinity and stability.     

Allelopathic control of cyanobacterial blooms by periphyton biofilms

Periphyton biofilms are natural mixtures comprised of photoautotrophic and heterotrophic complex microorganisms. In this work, the inhibition effects of periphyton biofilms on cyanobacterial blooms were studied in pilot and field trials. Results show that the cyanobacterial species responsible for the blooms had an upper nutrient concentration threshold, below which it could not effectively compete with other organisms in the periphyton. The disappearance of the cyanobacterial blooms was due to the allelopathy between the cyanobacteria and periphyton biofilm. In particular, it was found that the periphyton biofilm could produce water-soluble allelochemicals such as indole and 3-oxo-α-ionone to significantly inhibit the growth of the cyanobacteria. These allelochemicals are able to damage the thylakoid membranes of the cyanobacteria, interrupt the electron transport in photosystem II, decrease effective quantum yields, and eventually lead to the failure of photosynthesis. A comprehensive discussion on the ecological consequences of these findings is also presented. This work demonstrates the potential of periphyton biofilm to be used as an environmentally friendly ecological engineering solution for (i) the control of cyanobacterial blooms and (ii) a transitional means for the construction of beneficial conditions for ecosystem restoration. In addition, this work provides significant insights into the competitive relationships between algae and biofilms.     

Distinct Kinetic and Spatial Patterns of Protein Kinase C (PKC)- and Epidermal Growth Factor Receptor (EGFR)-dependent Activation of Extracellular Signal-regulated Kinases 1 and 2 by Human Nicotinic Acid Receptor GPR109A

Nicotinic acid (niacin) has been widely used as a lipid-lowering drug for several decades, and recently, orphan G protein-coupled receptor GPR109A has been identified as a receptor for niacin. Mechanistic investigations have shown that, upon niacin activation, GPR109A couples to a G_i protein and inhibits adenylate cyclase activity, leading to inhibition of liberation of free fatty acid. However, the underlying molecular mechanisms for GPR109A signaling remain largely unknown. Using CHO-K1 cells stably expressing GPR109A and A431 cells, which are a human epidermoid cell line with high levels of endogenous expression of functional GPR109A receptors, we found that activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) by niacin was rapid, peaking at 5 min, and was significantly blocked by pertussis toxin. Furthermore, time course experiments with different kinase inhibitors demonstrated that GPR109A induced ERK1/2 activation via the matrix metalloproteinase/epidermal growth factor receptor transactivation pathway at both early and later time points (2–5 min); this pathway was distinct from the PKC pathway-mediated ERK1/2 phosphorylation that occurs at early time points (≤2 min) in response to niacin. Overexpression of Gβγ subunit scavengers βARK1-CT and the Gα subunit of transducin led to a significant reduction of ERK1/2 phosphorylation, suggesting a critical role for βγ subunits in GPR109A-activated ERK1/2 phosphorylation. Using arrestin-2/3-specific siRNA and an internalization-deficient GPR109A mutant, we found that arrestin-2 and arrestin-3 were not involved in GPR109A-mediated ERK1/2 activation. In conclusion, our findings demonstrate that upon binding to niacin GPR109A receptors initially activate G_i, leading to dissociation of the Gβγ subunit from activated G_i, and subsequently induce ERK1/2 activation via two distinct pathways, one PKC-dependent pathway occurring at a peak time of ≤2 min and the other matrix metalloproteinase-dependent growth factor receptor transactivation occurring at both early and later time points (2–5 min).     

The spatial pattern and dispersion of Lygodium microphyllum in the Everglades wetland ecosystem

Since 1958, L. microphyllum (Old World Climbing Fern), which originated from the Old World Tropics, has become a nuisance exotic and has rapidly spread and is being established system-wide in extremely remote and undisturbed areas such as the Florida Everglades. Of particular concern is that L. microphyllum is disrupting, at an alarming rate, the flora and fauna of the native ecosystem at the same time that a major 8.4 billion dollar Everglades restoration program is trying to enhance these same attributes. This research utilized IKONOS satellite data to map L. microphyllum within the 58,000-ha Loxahatchee National Wildlife Refuge wetland in south Florida. Results show that approximately 11.6% of the tree/shrub vegetation within the impoundment has been infected by L. microphyllum. These data were then utilized to explore the spatial spread patterns of L. microphyllum within the Refuge. Results suggest that L. microphyllum is more likely to establish on the southeast side of a tree/shrub island and then spread to the northwest, which corresponds to the prevailing wind direction in south Florida. Spatial pattern analysis of L. microphyllum spread indicated that it is correlated with the density and spatial distribution of tree/shrub island vegetation. It appears that the dispersion of L. microphyllum is density dependent, which can be expressed as a logistic function and has a catastrophic threshold of 160 m of mean distance between tree/shrub islands in the Everglades. It is predicted that 38% (or 1910 ha) of tree/shrub islands in the Refuge will be invaded by L. microphyllum by 2012. Tree islands in the Everglades wetland could be considered similar to oceanic islands throughout the world that are notoriously vulnerable to biological invasions.     

用构建的新型BmNPV载体在家蚕高效表达人β-干扰素

家蚕核多角体病毒(Bombyx mori Nuclear Polyhedrosis Virus.BmNPV)和家蚕细胞已成功地用来大量生产具有生物活性的重组蛋白。但是BmNPV的通用载体的类型较少。因此,本实验构建了BmNPV新型载体pBm92,该载体将多角体蛋白基因的起始密码ATG改变为ATT,然后在多角体蛋白基因的 12位外连接有5个外源基因的克隆位点。将HuIFN-β基因克隆在多角体蛋白基因的 12位后,构建了pBmIFN 12;同时构建HuIFN-β克隆在-3位后的转移栽体pBmIFN-3。将两种转移载体DNA分别与BmNPV基因组DNA共转染Bm-N细胞。利用重组病毒不产生多角体蛋白的特征,筛选重组病毒。用HuIFN-β基因探针与重组病毒DNA进行杂交鉴定。重组病毒BmIFN 12感染Bm-N细胞,其上清IFN活性最高时可达2.0×10~6IU/ml,将BmIFN 12注射5龄家蚕虫体,表达水平为50×10~7IU/ml,是HuIFN-β基因克隆在多角体蛋白基因的-3位后获得的重组病毒的表达量的2～4倍。家蚕体生产的rHulFN-β为糖基化蛋白具有天然HuIFN-β的抗原性。     

Coupled Attitude-Orbit Dynamics and Control for an Electric Sail in a Heliocentric Transfer Mission

The paper discusses the coupled attitude-orbit dynamics and control of an electric-sail-based spacecraft in a heliocentric transfer mission. The mathematical model characterizing the propulsive thrust is first described as a function of the orbital radius and the sail angle. Since the solar wind dynamic pressure acceleration is induced by the sail attitude, the orbital and attitude dynamics of electric sails are coupled, and are discussed together. Based on the coupled equations, the flight control is investigated, wherein the orbital control is studied in an optimal framework via a hybrid optimization method and the attitude controller is designed based on feedback linearization control. To verify the effectiveness of the proposed control strategy, a transfer problem from Earth to Mars is considered. The numerical results show that the proposed strategy can control the coupled system very well, and a small control torque can control both the attitude and orbit. The study in this paper will contribute to the theory study and application of electric sail.     

巨枝叶绿体基因组密码子偏好性分析

该文针对巨桉叶绿体基因组序列,选取其中长于300 nt且以AUG为起始密码子的43个非重复基因作为研究对象,采用CodonW1.4.2软件分析巨桉叶绿体基因组的密码子使用偏好性。结果表明:第3位密码子的平均GC含量为27.97%; ENC的变化范围为39.49～61.00,平均为47.04; RSCU1的密码子有31个,其中29个以A/U结尾;中性分析显示,GC12与GC3无显著相关;回归分析未达到显著性水平; ENCplot分析发现,大部分基因落在曲线上或附近;对应分析表明第1轴的贡献率为17.68%,第2轴的贡献率为11.49%,第3轴、第4轴的贡献率分别为8.00%和5.76%,前4轴累计贡献率达42.93%,第1轴与GC、ENC、CAI达到极显著相关。上述分析结果表明,巨桉叶绿体基因组的密码子偏好较弱,密码子第3位偏好以A或U结尾,选择和突变在巨桉叶绿体基因组密码子偏好中起相对均衡的作用,最终确定UUG、CUU、GUU、UCC、UCA、ACA、UAU、UAA、CAU、AAU、AGA和GGA 12个高频高表达密码子为最优密码子。这为转化叶绿体基因密码子优化,提高表达效率和改良巨桉目标性状奠定了坚实基础。     

基于遥感生态指数的新疆玛纳斯湖湿地生态变化评价

以2000、2006和2016年3期的Landsat遥感影像为基础数据源,结合前人研究成果和实地考察,借助RSEI指数,对玛纳斯湖湿地生态环境进行监测和评价。结果表明:利用主成分分析技术集成植被指数、湿度分量、地表温度和土壤指数建立的RSEI指数具有一定的适用性,可较好的对玛纳斯湖湿地生态环境质量状况及其时空变化进行监测和评价。2000、2006和2016年RSEI指数均值分别为0.227、0.183、0.234,对RSEI指数进行分级处理后,发现流域生态环境质量"较差"等级居于主导地位,"优"等级有所增加,湿地生态环境质量向好的方向发展。气候变化和人类活动共同作用于玛纳斯湖湿地产生的生态环境效应,日益增强的人类活动是湿地退化的主要原因,多年来粗放型的农业发展使玛纳斯河流域人口、经济与生态环境之间的关系严重失调。从优化水资源配置角度提出对主要源流流域水土资源大规模开发的同时,应重视尾闾湖泊湿地的生态价值与可持续发展。