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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 人为的环境毒素，即使是低剂量的，也会引起某种程度的生物变化，在 植物、动物、微生物，甚至人类。加州大学戴维斯分校超级基金项目的目标是找到观察 并量化这些环境影响的生物标记物，以便这些影响的来源和原因 了解、评估、跟踪和补救。为此，该计划包括加速器质谱仪(AMS) 量化是其分析核心的能力之一。该分析核心服务于该计划的几个项目， 包括土壤和废物运输、免疫化学生物标记物的开发、肺生物标记物和 生殖生物标志物。 AMS在评估人类对有毒物质的暴露和探讨其机制方面发挥着重要的作用。 对人类和其他寄主物种的毒性基础。这是我们使用生物标记物的计划的核心技术 环境暴露于农业和工业活动中的有毒物质。我们定义了尿路，肺， 特定毒性暴露的生殖和循环生物标志物，可使用以下分析方法进行量化 免疫分析、蛋白质质谱仪、层析和AMS直接定量同位素标记毒素。 AMS还通过将毒素中的同位素标记掺入到 主持人。然后，通过AMS测量所指示的毒素摄取量来校准衍生生物标志物的定量。 在运输的案例中，调查人员正在评估最近使用的燃料添加剂甲基的生物活性- 叔丁基醚(MTBE)，过去十年从燃料库泄漏到地下。~(14)C-MTBE的结合 正在对哺乳动物的蛋白质进行研究，以确定该化合物是否对细胞系统构成威胁。这些 实验室研究是免费进行的，需要与细胞培养物相互作用的14C水平，但该计划的大部分 致力于在不可能释放放射性示踪剂的自然环境中对生物标志物进行量化。首选的 用于量化可识别生物标志物的技术是最终可用于现场的免疫分析 工具包。重要的是为免疫分析发展选择正确的靶点，例如最有可能的代谢物或 接触化学物质后的荷尔蒙反应。AMS是一种特别有价值的发现 免疫分析之所以成为目标，是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 发现阿特拉津的二脱烷基硫代谢物是最显著的持久生物标志物。 人类中普遍存在的除草剂。针对这些生物标记物开发了免疫分析方法。 生态系统中有一些对环境变化敏感的“标志性”物种，就像过去的金丝雀一样。 在煤矿里呆了几百年。越来越多的变种人被视为荷尔蒙模拟物，对 通过赋予其繁殖成功来保护物种。我们使用小鹌鹑作为一个这样的例子，并正在寻找 粪便中睾酮或可的松的代谢物，用作样本以避免应激影响 在一只被捕获的鸟身上。代谢物的模式将被量化，以找出哪些可能是缓慢发展的迹象 环境压力。这些鸟体型较小，不能大量服用，因此需要AMS的敏感性。 肺对环境化学品的反应需要从与环境相关的呼吸来研究 剂量。目前的方法对模型动物肺组织特定蛋白的剂量沉积很难进行定量。 这提供了大量的暴露，然后在二维凝胶上分离蛋白质，然后长期(1个月) 放射自显影。AMS对适当剂量具有敏感性，已制定出顺序凝胶分离方法 最大限度地发现目标蛋白质。 AMS核心用于识别暴露的显著生物标记物，用于现场测试开发并量化 计划研究人员暴露于有标签的化合物。

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 accelerator mass spectrometry (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. 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.