Detection of oxidized and glycated proteins in clinical samples using mass spectrometry — A user's perspective

Background

Proteins in human tissues and body fluids continually undergo spontaneous oxidation and glycation reactions forming low levels of oxidation and glycation adduct residues. Proteolysis of oxidised and glycated proteins releases oxidised and glycated amino acids which, if they cannot be repaired, are excreted in urine.

Scope of Review

In this review we give a brief background to the classification, formation and processing of oxidised and glycated proteins in the clinical setting. We then describe the application of stable isotopic dilution analysis liquid chromatography-tandem mass spectrometry (LC-MS/MS) for measurement of oxidative and glycation damage to proteins in clinical studies, sources of error in pre-analytic processing, corroboration with other techniques – including how this may be improved – and a systems approach to protein damage analysis for improved surety of analyte estimations.

Major conclusions

Stable isotopic dilution analysis LC-MS/MS provides a robust reference method for measurement of protein oxidation and glycation adducts. Optimised pre-analytic processing of samples and LC-MS/MS analysis procedures are required to achieve this.

General significance

Quantitative measurement of protein oxidation and glycation adducts provides information on level of exposure to potentially damaging protein modifications, protein inactivation in ageing and disease, metabolic control, protein turnover, renal function and other aspects of body function. Reliable and clinically assessable analysis is required for translation of measurement to clinical diagnostic use. Stable isotopic dilution analysis LC-MS/MS provides a “gold standard” approach and reference methodology to which other higher throughput methods such as immunoassay and indirect methods are preferably corroborated by researchers and those commercialising diagnostic kits and reagents. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.     

Low-dimensional compounds containing cyano groups. XIX. Crystal structure, spectroscopic, thermal and magnetic properties of {[Cu(tn)2]3[Pt(CN)4]2}[Pt(CN)4] (tn = 1,3-diaminopropane) complex

Violet prismatic crystals of {[Cu(tn)₂]₃[Pt(CN)₄]₂}[Pt(CN)₄] (tn = 1,3-diaminopropane) were crystallized from the water-methanol solution containing CuCl₂·2H₂O, tn and K₂[Pt(CN)₄]·3H₂O. Prepared complex was characterized using elemental analysis, infrared and UV-Vis spectroscopy, magnetic measurement and thermal analysis. X-ray analysis revealed an ionic character of the complex containing mononuclear square planar [Pt(CN)₄]²⁻ complex anions and penta-nuclear [Cu(tn)₂-Pt(CN)₄-Cu(tn)₂-Pt(CN)₄-Cu(tn)₂]²⁺ complex cations. The inner Cu(II) atom of the complex cation is hexa-coordinated, whereas two crystallographically equivalent peripheral Cu(II) atoms are penta-coordinated in the shape of a deformed square pyramid. Four v(CN) absorption bands observed in the IR spectrum are in agreement with the higher number of crystallographically different cyano groups and a broad highly asymmetric band observed in the reflectance UV-Vis spectrum is consistent with the presence of both hexa- and penta-coordinated Cu(II) atoms in the structure. The temperature dependence of the inverse susceptibility suggests the presence of a weak antiferromagnetic exchange coupling between Cu(II) ions. The complex is stable up to 210 °C when its two-stage thermal decomposition starts.     

Structural determination Vitex cymosa Bertero active principle: Diastereoselective synthesis of (±)-trans-4-hydroxy-6-propyl-1-oxocyclohexan-2-one and its antinociceptive activity

The diastereoselective synthesis of (±)-trans-4-hydroxy-6-propyl-1-oxocyclohexan-2-one, as a mixture trans:cis (3:1), was accomplished using a protocol that combine the Prins cyclization and RuO₄ oxidation. The synthesis this lactone allowed the elucidation of the correct structure of the substance isolated from the barks of Vitex cymosa. The δ-lactones mixture showed significant antinociceptive properties in preliminary tests using the tail flick model assay.     

黄土高原植被演替对土壤碳库及δ~(13)C的影响

采集宁夏云雾山草原植被演替阶段的土壤和植物样品,测定土壤容重(BD)、土壤含水量(WC)、土壤有机碳(SOC)含量、以及土壤和植物优势种稳定碳同位素组成(δ13C),并比较了它们之间的差异,分析其反映的环境信息。结果表明:在表层0~10 cm深度,灌木地SOC含量为71.7 g·kg-1,显著高于草地的54.6 g·kg-1;灌木地0~20 cm深度SOC含量占1 m深SOC含量的58.1%,而草地在0~40 cm深度的SOC含量占1 m深的34.1%;短期植被演替(约10 a)对土壤剖面0~20 cm层的δ13C值有显著影响,其中对0~10 cm层δ13C影响最大;草地演替为灌木地后δ13C偏负1.4‰,30 cm以下土壤δ13C值对短期植被变化不敏感;通过δ13C在C3植被的短期演替过程中的响应关系发现,δ13C值作为土壤碳库更替和碳循环的研究工具有很好的辨识力;灌木的δ13C值从叶子到根系存在明显差异,逐渐偏正,变化为2.2‰,草地为1.9‰;通过对0~40 cm深度植物根系δ13C值的测定发现,灌木地由表层的-28.15‰变为-26.11‰,偏正了2.04‰,草地根系δ13C值从表层的-27.08‰变为-27.57‰,偏负了0.49‰。     

沙棘属植物叶片碳稳定同位素含量与气候的关系

本试验以131个沙棘属植物种群为研究对象,通过测定其叶片碳稳定同位素(δ13C)值,分析了碳稳定同位素特征与环境因子之间的关系.结果 表明:沙棘属植物叶片的δ13C值介于-24.65‰～-29.11‰‰,平均值为-26.97‰,属于C3植物,叶片δ13C值变异系数为种内大于种间,表明环境因子是影响沙棘属植物叶片δ13C...     

应用稳定同位素研究广西东方洞食物网结构和营养级关系

稳定性同位素技术已经成为研究生态系统食物网结构和营养级关系及其动态变化的重要手段。应用稳定同位素技术(δ13C和δ15N)研究了东风洞的食物网结构和营养级关系。研究结果表明,东风洞主要为腐食食物链,由于在洞穴内黑暗带中无光照也无植物生长,所以碳源主要为东风洞中的土壤有机质,作为第一营养级;主要摄食关系为马陆与土壤有机物质,螺类与土壤有机物质等;在该洞生态系统中,将3种马陆直接作为初级消费者,以隅蛛和其它马陆平均值之间的差值(2.04‰)作为东风洞食物网稳定氮同位素的富集因子。根据营养级模型可知,马陆类群、细长钻螺等土壤动物,裸灶螽和涂闪夜蛾形成第二营养级,即初级消费者;第三营养级包括蜘蛛类、盲蛛、地蜈蚣以及黑眶蟾蜍,为次级消费者。     

荒漠植物梭梭和沙拐枣的花环结构及C4光合特征

 为了探讨梭梭（Haloxylon ammodendron）和沙拐枣（Calligonum mongolicum）适应高温强光荒漠环境的光合作用机构及特征,通过对其同化枝的解剖结构观察,δ13C值分析,以及气体交换测定表明：二者均具有花环结构(Kranz anatomy),肉细胞（Mesophyll cell）呈栅栏状,其内侧是维管束鞘细胞（Bundle sheath cell）,小维管束与维管束鞘细胞相接。在栅栏组织和贮水组织中,梭梭具有形状巨大的含晶细胞;沙拐枣具有大量的粘液细胞。梭梭和沙拐枣同化枝的δ13C值分别为-14.3‰和-14.8‰,在不同生长季节和土壤水分条件下,二者的δ13C值变化在-14‰到-16‰之间。梭梭和沙拐枣的CO2补偿点分别为2 μmol•mol－1和4 μmol•mol－1,光饱和点分别为1 660和1 756 μmol•m－2•s－1,表观光合量子效率分别为0.044和0.057 mol CO2•mol－1 photons。这表明,广泛分布于我国荒漠地区的木本植物梭梭和沙拐枣为C4植物,其光合途径不随生长季节和水分条件的变化而改变。     

MrgprA3-expressing pruriceptors drive pruritogen-induced alloknesis through mechanosensitive Piezo2 channel

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A sesquiterpene δ-lactone from Zinnia juniperifolia

Juniperin, a constituent of Zinnia juniperifolia, is a sesquiterpene δ-lactone with an elermane skeleton. Its structure and stereochemistry are based on chemical and spectroscopic studies.