Structural Basis for Certain Naturally Occurring Bioflavonoids to Function as Reducing Co-Substrates of Cyclooxygenase I and II

Background

Recent studies showed that some of the dietary bioflavonoids can strongly stimulate the catalytic activity of cyclooxygenase (COX) I and II in vitro and in vivo, presumably by facilitating enzyme re-activation. In this study, we sought to understand the structural basis of COX activation by these dietary compounds.

Methodology/Principal Findings

A combination of molecular modeling studies, biochemical analysis and site-directed mutagenesis assay was used as research tools. Three-dimensional quantitative structure-activity relationship analysis (QSAR/CoMFA) predicted that the ability of bioflavonoids to activate COX I and II depends heavily on their B-ring structure, a moiety known to be associated with strong antioxidant ability. Using the homology modeling and docking approaches, we identified the peroxidase active site of COX I and II as the binding site for bioflavonoids. Upon binding to this site, bioflavonoid can directly interact with hematin of the COX enzyme and facilitate the electron transfer from bioflavonoid to hematin. The docking results were verified by biochemical analysis, which reveals that when the cyclooxygenase activity of COXs is inhibited by covalent modification, myricetin can still stimulate the conversion of PGG₂ to PGE₂, a reaction selectively catalyzed by the peroxidase activity. Using the site-directed mutagenesis analysis, we confirmed that Q189 at the peroxidase site of COX II is essential for bioflavonoids to bind and re-activate its catalytic activity.

Conclusions/Significance

These findings provide the structural basis for bioflavonoids to function as high-affinity reducing co-substrates of COXs through binding to the peroxidase active site, facilitating electron transfer and enzyme re-activation.     

Identification and characterization of a leucine-rich repeat kinase 2 (LRRK2) consensus phosphorylation motif

Mutations in LRRK2 (leucine-rich repeat kinase 2) have been identified as major genetic determinants of Parkinson's disease (PD). The most prevalent mutation, G2019S, increases LRRK2's kinase activity, therefore understanding the sites and substrates that LRRK2 phosphorylates is critical to understanding its role in disease aetiology. Since the physiological substrates of this kinase are unknown, we set out to reveal potential targets of LRRK2 G2019S by identifying its favored phosphorylation motif. A non-biased screen of an oriented peptide library elucidated F/Y-x-T-x-R/K as the core dependent substrate sequence. Bioinformatic analysis of the consensus phosphorylation motif identified several novel candidate substrates that potentially function in neuronal pathophysiology. Peptides corresponding to the most PD relevant proteins were efficiently phosphorylated by LRRK2 in vitro. Interestingly, the phosphomotif was also identified within LRRK2 itself. Autophosphorylation was detected by mass spectrometry and biochemical means at the only F-x-T-x-R site (Thr 1410) within LRRK2. The relevance of this site was assessed by measuring effects of mutations on autophosphorylation, kinase activity, GTP binding, GTP hydrolysis, and LRRK2 multimerization. These studies indicate that modification of Thr1410 subtly regulates GTP hydrolysis by LRRK2, but with minimal effects on other parameters measured. Together the identification of LRRK2's phosphorylation consensus motif, and the functional consequences of its phosphorylation, provide insights into downstream LRRK2-signaling pathways.     

CD8+ DC,but Not CD8?DC,Isolated from BCG-Infected Mice Reduces Pathological Reactions Induced by Mycobacterial Challenge Infection

Background

Tuberculosis is a mycobacterial infection causing worldwide public health problems but the available vaccine is far from ideal. Type-1 T cell immunity has been shown to be critical for host defence against tuberculosis infection, but the role of dendritic cell (DC) subsets in pathogenesis of mycobacterial infection remains unclear.

Methodology/Principal Findings

We examined the effectiveness of dendritic cell (DC) subsets in BCG-infected mice in generating immune responses beneficial for pathogen clearance and reduction of pathological reactions in the tissues following challenge infection. Our data showed that only the adoptive transfer of the subset of CD8α⁺ DC isolated from infected mice (iCD8⁺ DC) generated significant protection, demonstrated by less mycobacterial growth and pathological changes in the lung and liver tissues in iCD8⁺ DC recipients than sham-treated control mice. The adoptive transfer of the CD8α⁻DC from the infected mice (iCD8⁻ DC) not only failed to reduce bacterial growth, but enhanced inflammation characterized by diffuse heavy cellular infiltration. Notably, iCD8⁻ DC produced significantly higher levels of IL-10 than iCD8⁺ DC and promoted more Th2 cytokine responses in in vitro DC-T cell co-culture and in vivo adoptive transfer experiments.

Conclusions/Significance

The data indicate that in vivo BCG-primed CD8⁺ DC is the dominant DC subset in inducing protective immunity especially for reducing pathological reactions in infected tissues. The finding has implications for the rational improvement of the prophylactic and therapeutic approaches for controlling tuberculosis infection and related diseases.     

Altered sperm tsRNAs in aged male contribute to anxiety‐like behavior in offspring

Parental age at first pregnancy is increasing worldwide. The offspring of aged father has been associated with higher risk of several neuropsychiatric disorders, such as schizophrenia and autism, but the underlying mechanism remains elusive. Here we report that advanced paternal age in mice alters the profile of transfer RNA‐derived small RNAs (tsRNAs). Injection of sperm tsRNAs from aged male mice into zygotes induced anxiety‐like behaviors in F1 males. RNA sequencing of the cerebral cortex and hippocampus of those F1 male mice altered the gene expression of dopaminergic synapse and neurotrophin. tsRNAs from aged male mice injection also altered the neuropsychiatry‐related gene expression in two‐cell and blastocyst stage embryos. More importantly, the sperm tsRNA profile changes significantly during aging in human. The up‐regulated sperm tsRNA target genes were involved in neurogenesis and nervous system development. These results suggest that aging‐related changes of sperm tsRNA may contribute to the intergenerational transmission of behavioral traits.     

NaCl对渗透胁迫下三角叶滨藜光合作用和水分状况的调节

以溶液培养的三角叶滨藜(Atriplex triangularis)为材料, 测定分析了在PEG诱导的渗透胁迫条件下, 适量的NaCl对其光合作用和水分吸收的影响, 以探讨环境溶液中NaCl对植物适应干旱的影响。结果表明, PEG诱导的渗透胁迫导致三角叶滨藜植株吸水困难、叶绿素含量降低、光合系统受损、生长受抑制、生物量减少; 而在PEG渗透胁迫的处理液中添加10–40 mmol·L^–1NaCl可以明显降低植株水势和叶片渗透势, 维持较高的细胞膨压, 减缓PEG渗透胁迫对光合系统的破坏作用, 保证相对较高的光合速率和生长速度, 从而有效增强了三角叶滨藜对渗透胁迫的适应能力。     

我国珍稀濒危植物保护红线的划定

本文依据《国家生态保护红线——生态功能基线划定技术指南(试行)》原则和植物保护的具体情况, 探讨了我国珍稀濒危植物保护红线划定的原则与方法。研究选取《国家重点保护野生植物名录》中收录的所有物种作为研究对象。将其定义为红线保护植物, 其中的I级保护植物定义为红线关键植物, 并基于文献资料及标本记录等数据建立了我国植物的属性数据库和地理分布数据库。在GIS支持下, 以建立的数据库为基础研究了我国红线保护植物的地理分布特征; 基于保护生物学理论, 结合我国自然保护区的就地保护现状进行了分析和评价; 以热点地区和GAP分析为理论基础, 在其分布地中识别具有代表性的热点区域以及不同属性植物分布的重点区域, 进而结合土地利用和人类干扰因素, 划定我国珍稀濒危植物生境整体保护的红线。通过在全国尺度上的分析, 得出我国珍稀濒危植物的整体保护红线面积为71.63万km², 占陆域国土面积的7.45%。植物生境保护红线划定的原则、方法和划定方案的研究对于更准确地划定生态安全预警红线具有重要意义, 同时可以为我国国土生态安全格局的构建提供依据。     

水稻雄配子体发育过程中tRNA片段的生物信息学分析

tRNA片段(tRF)是tRNA通过非随机剪切产生的RNA片段, 其产生和功能机制尚不明确; 而在水稻(Oryza sativa)雄配子体发育过程中, 人们对tRNA更是知之甚少。通过高通量测序, 在水稻雄配子体发育过程中发现了长度范围较大的tRFs; 进一步采用logo对tRFs两端的序列进行分析, 发现了4个有序列特征(其中3个未见报道)和1个无序列特征的酶切位点; 通过NCBI Blast预测了tRF靶基因, 发现其大多靶向转座因子。研究结果对揭示tRF产生机制以及水稻雄配子体发育研究有一定的参考价值。     

A nonlinear optimization model for fishing vessel evacuation during typhoon emergencies

We present an optimization model for fishing vessel evacuation during typhoon emergencies. Typhoon forecasts often do not accurately predict the typhoon path, so a risk assessment method is developed to evaluate the risk of fishing vessel evacuation processes. Risk assessment is the objective function of the fishing vessel evacuation optimization model. The model used is a many-to-many nonlinear assignment problem. A hybrid closed loop algorithm is used and its efficiency exceeds the traditional algorithm. The model is tested on a typhoon scenario based on harbors' distribution in Zhejiang Province, China.     

Plant regeneration through direct somatic embryogenesis,antioxidant properties,and metabolite profiles of Swertia corymbosa (Griseb.) Wight ex C.B. Clarke

A protocol for induction of direct somatic embryogenesis and subsequent plant regeneration for the medicinally important and endangered plant of Swertia corymbosa has been developed for the first time. In the present study, in vitro derived leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) individually and in combination with cytokinins for its effectiveness to induce the differentiation of somatic embryos. The antioxidant activities of methanolic extracts from non-embryonic callus (NEC), pre-embryoid masses (PEM), somatic embryos at globular stage (SEG), somatic embryos at heart-shaped stage (SEHS), and cotyledonary embryos (SEC) of S. corymbosa were evaluated using three in vitro assays: scavenging of free radicals (DPPH and ABTS) and ferric reducing antioxidant test. In all cases, the methanolic extract from SEG demonstrated better antioxidant activity than those taken from other tested samples. Higher amounts of swertianin (1), methylswertianin (2), and 1,2- dihydroxy-6-methoxyxanthone-8-O-β-D-xylopyranosyl (3) were found in SEG stage followed by SEHS and PEM when compared to NEC and SEC.