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: 7977077
• 负责人: BRUCE D HAMMOCK
• 金额: $ 3万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-04 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7977077
• 关键词: Agriculture; Animals; Atrazine; Autoradiography; Binding; Biological; Biological Assay; Biological Markers; Birds; Calibration; Cell Culture Techniques; Chemical Exposure; Chromatography; Coal Mining; Computer Retrieval of Information on Scientific Projects Database; Cortisone; Deposition; Development; Dose; Ecosystem; Environmental Exposure; Environmental Impact; Exposure to; Funding; Gel; Goals; Grant; Herbicides; Hormonal; Human; Immunoassay; Institution; Isotope Labeling; Label; Laboratory Study; Lung; Mass Spectrum Analysis; Measurement; Measures; Methods; Microbe; Monitor; Pattern; Plants; Play; Poison; Proteins; Quail; Research; 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; 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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Project Description 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 项目描述 人为环境毒素，即使是低剂量，也会导致植物、动物、微生物甚至人类发生某种程度的生物变化。加州大学戴维斯分校超级基金计划的目标是发现观察和量化这些环境影响生物标志物的方法，以便了解、评估、追踪和补救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。该分析核心服务于该计划的多个项目，包括土壤和废物运输、免疫化学生物标记物、肺部生物标记物和生殖生物标记物的开发。 加速器质谱 (AMS) 在评估人类接触有毒物质以及探索人类和其他宿主物种毒性的机制基础方面发挥着重要作用。它是我们利用农业和工业活动中有毒物质环境暴露的生物标志物计划的核心技术。我们定义了特定毒性暴露的尿液、肺部、生殖和循环生物标志物，这些生物标志物可以使用免疫测定、蛋白质质谱、色谱法和 AMS 直接定量同位素标记毒素等测定进行量化。 AMS 还通过毒素与宿主的同位素标记掺入相关性来校准其他测定。然后通过 AMS 测量指示的毒素吸收来校准衍生生物标志物的定量。 就运输业而言，研究人员正在评估最近使用的燃料添加剂甲基叔丁基醚（MTBE）的生物活性，该添加剂在过去十年中从燃料库泄漏到地下。正在研究 14C-MTBE 与哺乳动物蛋白质的结合，以确定该化合物是否对细胞系统构成威胁。这些实验室研究是根据与细胞培养物相互作用所需的 14C 水平自由完成的，但该计划的大部分内容涉及在不可能释放放射性示踪剂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，该技术最终可以制成现场可用的试剂盒。为免疫测定开发选择正确的靶标非常重要，例如化学暴露最可能的代谢物或激素反应。 AMS 是发现最佳免疫分析靶标的一项特别有价值的技术，因为它可以揭示同位素标记的异生素的所有代谢物，即使在低剂量暴露下也是如此。我们发现莠去津的二脱烷基硫醇酸酯代谢物是这种人类普遍存在的除草剂最显着的持久生物标志物。针对这些生物标志物开发了免疫测定法。 生态系统中存在对环境变化敏感的“标记”物种，就像过去几个世纪煤矿里的金丝雀一样。越来越多的污染物被视为激素模仿物，通过损害物种的繁殖成功率对物种起到“毒药”作用。我们以小鹌鹑为例，在其粪便中发现了睾酮或可的松的代谢物，将其用作样本，以避免对捕获的鸟类产生压力影响。代谢物的模式将被量化，以发现哪些可能是环境压力缓慢发展的迹象。家禽体型较小，不能大量给药，因此需要 AMS 的灵敏度。 肺部对环境化学品的反应需要通过环境相关剂量的呼吸来研究。目前的方法很难量化模型动物肺组织特定蛋白质中的剂量沉积，这些方法提供大量暴露，然后在二维凝胶上进行蛋白质分离，然后进行长期（1个月）放射自显影。 AMS 具有对适当剂量的敏感性，并且已制定出顺序凝胶分离以最大限度地发现目标蛋白。 AMS 核心用于识别显着的暴露生物标志物，以进行可现场分析开发，并为项目研究人员量化标记化合物的暴露情况。