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: 7359008
• 负责人: BRUCE D HAMMOCK
• 金额: $ 2.79万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7359008
• 关键词: METHODS; MONITOR; TOXIC; SUBSTAN; INDICATORS

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. 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 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NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。人为环境毒素，即使在低剂量下，也会在植物、动物、微生物甚至人类体内引起某种程度的生物变化。加州大学戴维斯分校超级基金项目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。该分析核心服务于几个计划项目，包括土壤和废物运输，免疫化学生物标志物，肺生物标志物和生殖生物标志物的开发。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项目的核心技术。我们定义了特定有毒物质暴露的尿液、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、色谱法和用AMS直接定量同位素标记毒素等测定方法进行量化。AMS还通过从毒素到宿主的同位素标记掺入的相关性来提供其他测定的校准。然后通过AMS测量所指示的毒素摄取来校准导出的生物标志物的定量。 在运输方面，调查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14 C-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。这些实验室研究是自由进行的，需要与细胞培养物相互作用的14 C水平，但该计划的大部分内容涉及在自然环境中定量生物标志物，其中放射性示踪剂释放是不可能的。用于定量可识别的生物标志物的优选技术是免疫测定法，其最终可以制成现场可用的试剂盒。重要的是要选择正确的免疫检测目标，如最可能的代谢物或激素反应的化学品暴露。AMS是发现最佳免疫测定靶点的特别有价值的技术，因为它揭示了同位素标记的异生素的所有代谢物，即使在低剂量暴露下。我们发现阿特拉津的二脱烷基巯基尿酸代谢物是这种普遍存在于人类中的除草剂的最突出的持久生物标志物。针对这些生物标志物开发了免疫测定。 生态系统中有一些对环境变化敏感的“标志”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拟物，通过损害物种的繁殖成功来充当物种的“毒药”。我们正在以小鹌鹑为例，在它们的粪便中发现睾丸激素或可的松的代谢物，这些粪便被用作样本，以避免对捕获的鸟类产生压力效应。代谢物的模式将被量化，以发现可能是缓慢发展的环境压力的迹象。鸟类很小，不能被大量使用，因此需要AMS的灵敏度。 肺对环境化学品的反应需要从环境相关剂量的呼吸来研究。目前的方法对模型动物肺组织中特定蛋白质的剂量沉积定量较差，这些方法提供了大量暴露，然后在二维凝胶上分离蛋白质，然后进行长期（1个月）放射自显影。AMS对适当的剂量具有灵敏度，并且已经制定了连续的凝胶分离以最大限度地发现靶蛋白。 AMS核心用于识别暴露的突出生物标志物，用于现场检测开发，并为项目研究人员量化标记化合物的暴露。