METHODS MONITOR TOXIC SUBSTAN AND/OR INDICATORS OF PRESENCE IN HUMANS&OTHER SPE

监测人类体内有毒物质和/或存在指标的方法

• 批准号: 8362756
• 负责人: BRUCE D HAMMOCK
• 金额: $ 7.02万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362756
• 关键词: Agriculture; Animals; Atrazine; Autoradiography; Binding; Biological; Biological Assay; Biological Markers; Birds; Calibration; Cell Culture Techniques; Chemical Exposure; Chromatography; Coal Mining; Cortisone; Deposition; Development; Dose; Ecosystem; Environmental Exposure; Environmental Impact; Exposure to; Funding; Gel; Goals; Grant; Herbicides; Hormonal; Human; Immunoassay; Isotope Labeling; Label; Laboratory Study; Lung; Mass Spectrum Analysis; Measurement; Measures; Methods; Microbe; Monitor; National Center for Research Resources; Pattern; Plants; Play; Poison; Principal Investigator; Proteins; Quail; Research; Research Infrastructure; Research Personnel; Resource Development; Resources; Respiration; Role; Sampling; Serinus; Soil; Source; Stress; Structure of parenchyma of lung; Superfund; System; Technology; Testosterone; Tissue Model; Toxic Environmental Substances; Toxic effect; Toxicant exposure; Toxin; United States National Institutes of Health; Work; Xenobiotics; accelerator mass spectrometry; anthropogenesis; assay development; base; cost; environmental change; environmental chemical; exposed human population; mercapturate; methyl tert-butyl ether; programs; radiotracer; reproductive; reproductive success; response; two-dimensional; uptake; urinary; wasting

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Anthropogenic environmental toxins, even at low doses, cause some measure of biological change to take place, within plants, animals, microbes, or even humans. The goal of the UC Davis Superfund Program is to discover ways to observe and quantify these biomarkers of environmental impacts, so that the sources and causes of these impacts can be understood, assessed, traced, and remediated. To that end, the Program includes AMS quantitation as one of its competencies in its analytical core. This analytical core serves several of the Program projects, including Soil and Waste Transport, development of Immunochemical Biomarkers, Pulmonary Biomarkers, and Reproductive Biomarkers. Accelerator mass spectrometry (AMS) plays an important role in the assessment of human exposure to toxic substances and in probing the mechanistic basis of toxicity in humans and in other host species. It is a core technology in our program of using biomarkers of environmental exposures to toxic substances from agricultural and industrial activities. We define urinary, pulmonary, reproductive, and circulating biomarkers of specific toxic exposures that are quantifiable using assays such as immunoassays, protein mass spectrometry, chromatography, and direct quantitation of isotope labeled toxins with AMS. AMS also provides calibration of the other assays through correlation of isotope label incorporation from toxins into a host. Quantitation of a derived biomarker is then calibrated by the uptake of toxin indicated by the AMS measurements. In the case of Transport, the investigators are assessing the biological activity of the recently used fuel additive, methyl-tert-butyl-ether (MTBE), which leaked into the ground from fuel depots over the past decade. The binding of 14C-MTBE to mammalian protein is being studied to determine if the compound presents a threat to cellular systems. These laboratory studies are freely done with the levels of 14C needed to interact with cell cultures, but much of the Program is concerned with quantifying biomarkers in natural settings where radiotracer release is not possible. The preferred technology for quantifying recognizable biomarkers is the immunoassay which can eventually be made into field-usable kits. It is important to choose the right target for immunoassay development, such as the most likely metabolite or hormonal response of a chemical exposure. AMS is a particularly valuable technology for the discovery of optimal immunoassay targets because it reveals all metabolites of an isotope-labeled xenobiotic, even at low dose exposures. We found that the di-dealkyl mercapturate metabolites of atrazine were the most prominent lasting biomarkers of this ubiquitous herbicide in humans. Immunoassays are developed for these biomarkers. There are "marker" species in ecosystems which are sensitive to environmental change, much like the canaries of past centuries in coal mines. An increasing number of polutants are being seen as hormonal mimics that act as "poison" to a species by imparing its reproductive success. We are using small quail as one such example and are finding the metabolites of testosterone or cortisone in their fecal droppings, which are used as sample so as to avoid stress effects in a captured bird. The pattern of metabolites will be quantified to find which might be signs of slowly developing environmental stresses. The birds are small, and cannot be heavily dosed, so the sensitivity of AMS is needed. Pulmonary responses to environmental chemicals need to be studied from respiration of environmentally relevant doses. The dose deposition in specific proteins of lung tissue of model animals is poorly quantified by present methods that provide a large exposure followed by protein separation on two dimensional gels followed by long term (1 month) autoradiography. AMS has the sensitivity for appropriate doses and sequential gel separations have been worked out to maximize target protein discovery. The AMS core serves to identify prominent biomarkers of exposure for fieldable assay development and quantifies exposures to labeled compounds for the Program researchers.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 人为的环境毒素，即使在低剂量下，也会在植物，动物，微生物甚至人类中发生某种量度的生物学变化。加州大学戴维斯（UC Davis）超级基金计划的目标是发现观察和量化这些环境影响的这些生物标志物的方法，以便可以理解，评估，追踪和修复这些影响的来源和原因。为此，该计划将AMS定量作为其分析核心的能力之一。该分析核心为几个计划项目提供服务，包括土壤和废物运输，免疫化学生物标志物的发展，肺部生物标志物和生殖生物标志物。 加速器质谱法（AMS）在评估人类暴露于有毒物质的评估以及探测人类和其他宿主物种的毒性基础方面起着重要作用。在我们的计划中，它是使用农业和工业活动对有毒物质的环境暴露的生物标志物的核心技术。我们定义了使用特定有毒暴露的特定毒性暴露的尿，肺，生殖和循环的生物标志物，这些暴露物可使用诸如免疫测定，蛋白质质谱法，色谱法以及与AMS的同位素标记的同位素标记的测定法。 AM还通过将毒素的同位素标签掺入到宿主中的相关性来对其他测定法进行校准。然后，通过AMS测量指示的毒素摄取来校准衍生的生物标志物的定量。 在运输的情况下，研究人员正在评估最近使用的十年中最近使用的燃料添加剂甲基二丁基乙醚（MTBE）的生物学活性，该甲基丁基乙醚（MTBE）在过去十年中从燃料库中泄漏到了地面上。正在研究14C-MTBE与哺乳动物蛋白的结合，以确定该化合物是否对细胞系统构成威胁。这些实验室研究是通过与细胞培养的相互作用所需的14C水平自由完成的，但是大部分程序都涉及在无法释放放射性示波器的自然环境中量化生物标志物。量化可识别生物标志物的首选技术是免疫测定法，最终可以将其制成现场使用的套件。重要的是为免疫测定开发的正确靶标，例如最可能的代谢产物或化学暴露的激素反应。 AMS是发现最佳免疫测定靶标的特别有价值的技术，因为它揭示了同位素标记的异种生物的所有代谢物，即使在低剂量暴露下也是如此。我们发现，阿特拉津的二甲基苏联代谢产物是这种无处不在的除草剂中最突出的持久生物标志物。为这些生物标志物开发了免疫测定。 生态系统中有“标记”物种对环境变化敏感，就像过去几个世纪的煤矿的金丝雀一样。越来越多的政治剂被视为激素模拟物，通过侵犯其生殖成功，将物种“毒药”起来。我们将小鹌鹑作为一个这样的例子，正在其粪便中发现睾丸激素或可的松的代谢产物，它们被用作样品，以避免捕获的鸟类中的压力效应。将量化代谢物的模式，以找到可能会缓慢发展环境应力的迹象。鸟很小，不能大量服用，因此需要AM的灵敏度。 需要根据环境相关剂量的呼吸来研究对环境化学物质的肺反应。动物肺组织特异性蛋白质中的剂量沉积通过当前的方法量化很差，该方法可提供大暴露，然后在二维凝胶上进行蛋白质分离，然后是长期（1个月）放射自显影。 AMS对适当剂量具有敏感性，并且已经制定了顺序的凝胶分离以最大程度地发现靶蛋白。 AMS核心旨在识别用于现场测定开发的突出暴露物的重要生物标志物，并量化对计划研究人员标记化合物的暴露。