Project 4: Measurement and Engineering Solutions to Detect and Prevent N-Nitrosamine Exposure

项目摘要/摘要--项目4 N-亚硝基(-N-N=O)化合物包括一些已知和包含的最强的化学诱变剂 具有数百种不同结构的化学物质家族。然而，只有少数N-亚硝胺是 通过EPA标准方法进行常规测量，这些方法在预期的低水平下无效 对健康造成影响。为了克服这一挑战并减少人类接触，项目4试图测量 并在家庭中使用N-亚硝胺时销毁一套N-亚硝胺，这些家庭最需要它们来保护公众 健康和环境。首先，具体目标1是开发方法来检测一套已建立的和 新型N-亚硝胺的多维气相色谱-质谱联用(For 鉴定)和火焰电离检测(用真实标准或通过有效碳进行定量 当真实的标准既不能合成也不能购买时，数字概念)。这项技术使 对近十几个关键环境参数的估计，这样在复合体中检测到的新分子 混合物将拥有随时可用的物理化学指标，以支持环境命运模型和预测暴露。 将与社区参与核心合作，使用传统的 收集方法以及新的识别-固相萃取(R-SPE)材料，浓缩N- 亚硝胺(将在项目3中开发)和提取物的部分将提供给项目2以进行诱变 评估。关于NDMA水平的知识将向项目1提供信息，以便与环境相关的水平 可以使用对NDMA致敏的新型基因工程小鼠来研究它们的生物学影响。 此外，数据管理和分析核心将“什么”和“多少”的数据与 生物影响告知风险。有了N-亚硝胺环境水平的知识，就有针对性 二是开发基于电化学过程的新型家庭水过滤技术和 用酶破坏进行分子识别。后者依赖于基因工程微生物， 作为酶的非生物支架；因此，它克服了生物反应器面临的许多经典挑战 并为利用生物技术的进步为期待已久的水革命提供了巨大的可能性 治疗。特殊目标3将测试降解产物，以确保它们不再具有遗传毒性 通过与Project 1合作，使用“化学感应”细胞作为DNA损伤的动画探测器。 受训者将推动研究，并将得到研究经验和培训协调的支持 核心以确保最佳的专业发展。此外，项目4将受益于由 行政核心，确保与其他项目和核心以及 向利益相关者，包括居住在奥林化学超级基金附近的人提供研究转化的机会 Site和PassamAquoddy部落。通过相互作用和相互依赖的系统方法，项目4 将有助于保护人类健康免受危险物质的伤害。

PROJECT SUMMARY/ABSTRACT – PROJECT 4 N-nitroso (-N-N=O) compounds include some of the most powerful chemical mutagens known and encompass a family of chemicals with hundreds of different structures. However, only a handful of N-nitrosamines are routinely measured via EPA Standard Methods, and these methods are not effective at the low levels anticipated to cause health impacts. To overcome this challenge and reduce human exposure, Project 4 seeks to measure and destroy a suite of N-nitrosamines at the point of use in homes, where they are needed most to protect public health and the environment. First, Specific Aim 1 is to develop methods to detect a suite of established and novel N-nitrosamines using multi-dimensional gas chromatography paired with both mass spectrometry (for identification) and flame ionization detection (for quantification with authentic standards or via an effective carbon number concept when authentic standards can be neither synthesized nor purchased). This technique enables estimation of nearly a dozen critical environmental parameters, such that novel molecules detected in complex mixtures will have at-the-ready physicochemical metrics to feed environmental fate models and predict exposure. Water samples will be collected in collaboration with the Community Engagement Core using traditional collection methods as well as novel Recognition-Solid Phase Extraction (R-SPEs) materials that concentrate N- nitrosamines (to be developed in Project 3), and fractions of extracts will be provided to Project 2 for mutagenicity assessment. Knowledge about the levels of NDMA will inform Project 1 so that environmentally relevant levels can be studied for their biological impacts using novel genetically engineered mice that are sensitized to NDMA. In addition, the Data Management and Analysis Core will integrate data for “what” and “how much” with data on biological impact to inform risk. Armed with knowledge of environmental levels of N-nitrosamines, Specific Aim 2 is to develop novel in-home water filtration technologies based on electrochemical processes and molecular recognition with enzymatic destruction. The latter relies on genetically engineered microbes that act as non-living scaffolds for enzymes; as such, it overcomes many classical challenges faced by bioreactors and opens up vast possibilities to leverage advances in biotechnology for long-awaited revolutions in water treatment. Specific Aim 3 will test the degradation products to ensure that they are no longer genotoxic by collaborating with Project 1 to use “Chem-Sense” cells that act as animate detectors of DNA damage. Trainees will drive the research and will be supported by the Research Experience and Training Coordination Core to ensure optimal professional development. In addition, Project 4 will benefit from activities organized by the Administrative Core that ensure close collaborations with other Projects and with the Cores, as well as opportunities for research translation to stakeholders, including people living near the Olin Chemical Superfund Site and the Passamaquoddy Tribe. Via a Systems Approach of interactions and interdependencies, Project 4 will contribute to the protection of human health from hazardous substances.