Scavenging Activity of Azelaic Acid on Hydroxyl Radicals in Vitro

Azelaic acid is an aliphatic dicarboxylic acid (HOOC-(CH₂)₇-COOH) which has recently been shown to have some practical therapeutic applications in skin diseases of different etiologies. It possesses diverse biological activities and its mechanisms of action are still under investigation. Azelaic acid, as disodium salt (C₉2Na), at concentrations from 0.05 mM to 1.0 mM is capable of inhibiting significantly the hydroxylation of l-tyrosine to l-DOPA due to hydroxylradicals (HO) produced by Fenton reaction. Similarly C₉,2Na significantly inhibits the heterogeneous photocatalytic oxidation of toluene to cresols, and the peroxidation of arachidonic acid (C20: 4, n6), due to HO formed by dissolved oxygen in the presence of UV-irradiated semiconductor TiO₂ (photo-Fenton type reaction). C₉2Na decomposition and its by-products formation are quantifiable only at high HO concentrations. On the contrary, C₉2Na is not a scavenger of O₂ generated by xanthine-xanthine oxidase system. Under the same experimental conditions, mannitol behaves like C₉2Na. These data indicate that HO scavenging capacity of C₉2Na in vitro and represent a useful tool for further investigations on the mechanisms of action of azelaic acid in biological systems.     

黑籽南瓜NBS类基因CfRFN2的克隆及VIGS验证

黑籽南瓜(Cucurbita ficifolia)是云南特有的具有高抗枯萎病遗传性状的瓜类种质资源。为鉴定黑籽南瓜中NBS-LRR类基因的抗病功能,该研究从其叶片中克隆了NBS类基因CfRFN2 (GenBank ID:MK618462),测序全长为4 303 bp,完整的编码框长度为4 092 bp,编码1 363个氨基酸残基,该基因注释为拟南芥抗病蛋白At4g27190类转录体X1的同源基因,含有1个NB-ARC和2个LRR结构域,属于具有信号肽的可溶性蛋白。核苷酸相似性分析显示,CfRFN2与其他瓜类NBS类基因相似性在87%～98%之间;系统进化树分析表明,CfRFN2蛋白和瓜类的其他NBS类抗病蛋白聚为一个分支,其中CfRFN2蛋白与中国南瓜和美洲南瓜的RPS2、印度南瓜的RPS2-like亲缘关系最近,其次是黄瓜的At4g27190和苦瓜的At4g27220,与甜瓜的Atg27190亲缘关系相对较远;组织表达特性分析表明,CfRFN2基因在黑籽南瓜叶片中表达量最高,其次是茎,而在果皮和根中表达量较低。该研究采用烟草脆裂病毒载体系统,构建了黑籽南瓜VIGS沉默载体pTRV2-CfRFN2,含沉默载体的农杆菌侵染黑籽南瓜幼苗后接种枯萎病菌,qRT-PCR检测表明,接种后2 d和4 d的转pTRV2-CfRFN2沉默组植株的CfRFN2基因表达量比接种后同时期的野生型植株显著降低(分别下降34.75%和98.27%),病情指数增加为野生型的1.32倍,初步证明黑籽南瓜CfRFN2基因具有抗枯萎病的功能,推测该基因可能在黑籽南瓜抗枯萎病防御过程中发挥着重要作用。该研究中NBS类基因CfRFN2的克隆和VIGS验证为黑籽南瓜更多优异基因的克隆和功能验证奠定了前期基础,也为发掘黑籽南瓜优异抗病基因和开展瓜类分子育种提供新信息。     

Deciphering Spatial Protein–Protein Interactions in Brain Using Proximity Labeling

Cellular biomolecular complexes including protein–protein, protein–RNA, and protein–DNA interactions regulate and execute most biological functions. In particular in brain, protein–protein interactions (PPIs) mediate or regulate virtually all nerve cell functions, such as neurotransmission, cell–cell communication, neurogenesis, synaptogenesis, and synaptic plasticity. Perturbations of PPIs in specific subsets of neurons and glia are thought to underly a majority of neurobiological disorders. Therefore, understanding biological functions at a cellular level requires a reasonably complete catalog of all physical interactions between proteins. An enzyme-catalyzed method to biotinylate proximal interacting proteins within 10 to 300 nm of each other is being increasingly used to characterize the spatiotemporal features of complex PPIs in brain. Thus, proximity labeling has emerged recently as a powerful tool to identify proteomes in distinct cell types in brain as well as proteomes and PPIs in structures difficult to isolate, such as the synaptic cleft, axonal projections, or astrocyte–neuron junctions. In this review, we summarize recent advances in proximity labeling methods and their application to neurobiology.     

基于集合经验模态分解去趋势的水分利用效率对气候变化响应的高程分异——以“21世纪海上丝绸之路”沿线省份为例

水分利用效率(WUE)是表征陆地碳-水循环耦合关系的重要指标，但其对气候变化响应的高程分异仍不清楚。通过集合经验模态分解(EEMD)去趋势和偏相关方法，以"21世纪海上丝绸之路"沿线省份为研究区，揭示WUE对气候变化的响应及其随高程的分异。研究结果表明：(1)研究区内WUE多年均值由中心向南北递减。不同植被类型的WUE多年均值由高到低依次为：常绿针叶林、混交林、常绿阔叶林、稀树灌木草地、耕地和城市建设用地。(2)51.11%的区域表现出均温与WUE的正相关；而81.46%地区表明温差的扩大会使得WUE增加；有近一半的研究区表明最高温的升高有利于提高WUE，而最低温的作用则相反；有67.99%的区域表明降水增多反而会导致WUE的减少。(3)在大多数土地覆盖类型，日温差和最低温主要与WUE呈正相关，而最高温和降水主要与WUE呈负相关。在常绿针叶林、耕地和城市建设用地，日均温与WUE呈负相关。在其他三种植被类型下则呈正相关。(4)在低海拔地区，均温与WUE呈负相关而在中高海拔地区则转变为正相关关系。而最高温则正好相反。降水与WUE的负相关关系系数随高度的增加而不断加强，而温差和最低温与WUE的正相关关系也随高度的增加而剧烈波动增强。     

Arabica coffee and olive oils mitigate malathion-induced nephrotoxicity in rat: In silico,immunohistochemical and biochemical evaluation

Malathion (MAL) is an organophosphate insecticide that disrupts the body's antioxidant system; it is one of the earliest organophosphate insecticides extensively used as dust, emulsion, and vapor control a wide variety of insect pests under different conditions. This experimentation aims to evaluate the influence of Arabica coffee oil and olive oil on MAL-induced nephrotoxicity in male rat. 6 sets bearing the same number of animals were applied to this experiment. Each set comprised 10 rats. The first set of rats was used as the control group; rats in the second set were exposed to MAL measured at 100 mg/kg body weight for 7 weeks. Animals in the third and fourth set were treated with 400 mg/kg body weight of Arabica coffee oil and olive oil, and 100 mg/kg body weight of MAL. The fifth, together with the sixth set, were fed with a similar proportion of Arabica coffee oil and olive oil as administered to the third set of rats. After the experimental duration, rats of group 2 showed severe biochemical alterations, including significant increases of creatinine, uric acids, and urea nitrogen (BUN), resulting in marked decreases in serum albumin values and total protein (TP). Severe histopathological and immunohistochemical alterations of kidney tissues were observed in exposed MAL-intoxicated rats. Administration of these oils reduced the detected biochemical, histopathological modifications caused by MAL intoxication. Two active ingredients in Arabica coffee oil (oleic acid) and olive oil (hydroxytyrosol) showed good cyclooxygenase-2 (COX 2) interaction. Moreover, oleic acid from coffee oil and olive oil exhibited impressive association with xanthine oxidase (XO). The current finding showed that coffee oil and olive oil could be appraised as possible and a likely deterrence component against nephrotoxicity brought about by MAL.     

Systems ecological accounting for wastewater treatment engineering: Method,indicator and application

Wastewater treatment facility is vital for sustainable urban development. In the course of removing contaminants and discharging ready-for-reuse water, wastewater treatment consumes resources and triggers environmental emission during its lifetime. A comprehensive framework to analyze the embodied ecological elements as natural resources and environmental emissions of wastewater treatment is presented in this work. The systems method as a combination of process and input–output analyses is applied and a set of indicators are accordingly devised. Two representative ecological elements, i.e., greenhouse gases emissions and solar emergy of alternative wastewater treatment systems, i.e., a traditional activated sludge wastewater treatment plant and a constructed wetland have been taken into consideration. For each ecological element, five indicators have been calculated and compared to assess the impact on climate change and resources utilizing style of the case systems. The framework raised in this paper is fully supportive for optimal decision-making among different wastewater treatment technologies, and could be transplanted to be applied to systems ecological accounting for other production systems.     

浙江省生境质量时空演变与生态红线评估

生境质量是反映生物多样性状况与局地生态功能的重要指标,在高质量发展背景下研究区域生境质量的时空演变具有重要意义。以浙江省为研究区,基于InVEST模型、热点分析及地理探测器模型探究生境质量的时空演变与影响因素,并利用生境质量结果对浙江省生态红线开展了定量评估。结果表明:(1)2000-2015年,浙江省生境质量均值呈减速下降趋势,空间上形成了西北、西南、中东高和东北、中部低的分布格局;生境退化度呈现"中心-外围"的圈层辐射结构。(2)热点分析显示,生境质量与生境退化度在乡镇尺度上集聚特征相似、冷热点空间分布趋势相反。(3)地理探测分析发现,地形(高程、坡度)是影响浙江省生境质量的主要因素,植被因素(NPP、NDVI)的贡献度随时间推移逐渐增大;浙江省生境质量空间分异受到自然因子与社会经济因子的协同作用。(4)浙江省生态红线的生境质量整体较高且稳定,不同红线类型的生境质量存在差异;高生境质量区与生态红线的错位区域主要分布在浙西南、西北部山区,而北部、中部以及东部相对较少。基于此,对生态红线调整、区域生态功能区划提出对应的策略,以期提升浙江省生态空间管控。     

Anti-salmonella properties of kefir yeast isolates: An in vitro screening for potential infection control

The rise of antibiotic resistance has increased the need for alternative ways of preventing and treating enteropathogenic bacterial infection. Various probiotic bacteria have been used in animal and human. However, Saccharomyces boulardii is the only yeast currently used in humans as probiotic. There is scarce research conducted on yeast species commonly found in kefir despite its claimed potential preventative and curative effects. This work focused on adhesion properties, and antibacterial metabolites produced by Kluyveromyces lactis and Saccharomyces unisporus isolated from traditional kefir grains compared to Saccharomyces boulardii strains. Adhesion and sedimentation assay, slide agglutination, microscopy and turbidimetry assay were used to analyze adhesion of Salmonella Arizonae and Salmonella Typhimurium onto yeast cells. Salmonella growth inhibition due to the antimicrobial metabolites produced by yeasts in killer toxin medium was analyzed by slab on the lawn, turbidimetry, tube dilution and solid agar plating assays. Alcohol and antimicrobial proteins production by yeasts in killer toxin medium were analyzed using gas chromatography and shotgun proteomics, respectively. Salmonella adhered onto viable and non-viable yeast isolates cell wall. Adhesion was visualized using scanning electron microscope. Yeasts-fermented killer toxin medium showed Salmonella growth inhibition. The highest alcohol concentration detected was 1.55%, and proteins with known antimicrobial properties including cathelicidin, xanthine dehydrogenase, mucin-1, lactadherin, lactoperoxidase, serum amyloid A protein and lactotransferrin were detected in yeasts fermented killer medium. These proteins are suggested to be responsible for the observed growth inhibition effect of yeasts-fermented killer toxin medium. Kluyveromyces lactis and Saccharomyces unisporus have anti-salmonella effect comparable to Saccharomyces boulardii strains, and therefore have potential to control Salmonella infection.     

Critical role of bioanalytical strategies in investigation of clinical PK observations,a Phase I case study

RG7652 is a human immunoglobulin 1 (IgG1) monoclonal antibody (mAb) targeting proprotein convertase subtilisin/kexin type 9 (PCSK9) and is designed for the treatment of hypercholesterolemia. A target-binding enzyme-linked immunosorbent assay (ELISA) was developed to measure RG7652 levels in human serum in a Phase I study. Although target-binding assay formats are generally used to quantify free therapeutic, the actual therapeutic species being measured are affected by assay conditions, such as sample dilution and incubation time, and levels of soluble target in the samples. Therefore, in the presence of high concentrations of circulating target, the choice of reagents and assay conditions can have a significant effect on the observed pharmacokinetic (PK) profiles. Phase I RG7652 PK analysis using the ELISA data resulted in a nonlinear dose normalized exposure. An investigation was conducted to characterize the ELISA to determine whether the assay format and reagents may have contributed to the PK observation. In addition, to confirm the ELISA results, a second orthogonal method, liquid chromatography tandem mass spectrometry (LC-MS/MS) using a signature peptide as surrogate, was developed and implemented. A subset of PK samples, randomly selected from half of the subjects in the 6 single ascending dose (SAD) cohorts in the Phase I clinical study, was analyzed with the LC-MS/MS assay, and the data were found to be comparable to the ELISA data. This paper illustrates the importance of reagent characterization, as well as the benefits of using an orthogonal approach to eliminate bioanalytical contributions when encountering unexpected observations.     

我国三大城市群地区城市和农业用地地表热环境效应对比研究

以大中小城市协同发展为特征的城市群已成为我国城市化发展的主要形式，城市化和农业作为城市群地区最主要的土地利用活动，其气候效应是国际研究的热点。然而过去研究多关注大城市的热岛效应，对更为普遍的农业活动以及中小城市城市化的气候效应认识十分薄弱。基于MODIS地表温度数据，以自然林地为参照，提出了一种可逐像元估算土地利用地表热环境效应的新方法，进而对比分析了我国三大城市群地区（京津冀、长三角和珠三角）城市和农业用地地表热环境效应及其驱动因子差异。结果表明各城市群白天城市热岛效应明显，地级以上城市年平均热岛强度达3.2℃以上，但最强热岛均未发生在核心城市。夜晚热岛效应明显减弱，京津冀和长三角部分城市甚至出现冷岛效应。农业用地在白天亦表现出明显的增温效应，特别是在京津冀地区，而在夜晚除珠三角城市群外，降温效应明显，京津冀和长三角地区平均降温2.3℃和0.7℃。虽然城市用地平均增温强度大于农业用地，但农业用地因面积优势对区域温度变化起控制作用。白天城市和农业用地整体导致各城市群温度明显增加，京津冀增温最高（4.0℃），夜晚二者导致长三角和京津冀地区平均温度降低。研究还发现各城市群城市和农业用地地表热环境效应时空异质性极大，主要受植被、地表反照率、气候背景和人口密度控制。本文对制定缓解气候变化的土地利用策略具有重要的指导意义。