Project 1: Discovery of Xenobiotics Associated with Preterm Birth

早产是一个发病率不断增加的主要医学问题，异物可能起到了作用。该项目的长期目标是发现导致早产的外源物质。将对环境和生物样本进行检测，这些样本将主要来自波多黎各，因为那里的早产发生率异常高。虽然将进行非目标(筛选)分析，但主要关注Superfund和相关污染物，因为波多黎各有许多Superfund站点。大部分分析将基于质谱仪(MS)；将使用三种类型的MS：(HPLC)MALDI-TOF/TOF-MS、GC-EI(EC)-TOF-MS和HPLC-ESI-TOF-MS。我们将寻求发现“推定的PTB-外源生物”，即符合以下两个标准之一的外源生物： 它们与早产有关，或者与早产无关，但它们是常见的污染物，具有可能导致这种情况的生物活性。环境样品将是自来水和地下水，包括经过电解修复(解毒)的样品。生物样本将是妊娠组织(孕膜、胎盘)、细胞培养、外周血白细胞和尿液。妊娠组织将来自大鼠和人类，白细胞和尿液将来自人类。 四种分析方法中的一种或多种将被应用于这些类型的样品中的每一种：1.直接检测，在质谱仪中直接检测外源生物(或其实验室衍生物)；2.DNA加合物，外源生物在体内形成DNA加合物，由质量标签分析检测；3.促氧化剂，外源生物在核苷酸暴露分析中代谢样氧化后被检测到；4.生物检测：在对来自人类胎盘的培养细胞进行的生物检测中，首先检测细胞毒性(初级筛选)，然后检测细胞凋亡、遗传毒性、炎症和氧化应激(次级筛选，反映可能导致早产的机制)。将开发新的或改进的分析方法，并将在该项目中研究新的方法学组合，以促进通过这四种战略发现假定的PTB外源化合物。因此，该项目将在复杂样品中发现有毒异物的整个领域取得普遍适用的进展。通过样本交换、协同测试和数据协同分析， 无论是直接还是通过核心，该项目都与Protect中的所有其他项目集成在一起 程序。

Preterm birth is a major medical problem of increasing incidence, and xenobiotics may contribute. The longterm goal of this project is to discover xenobiotics that contribute to preterm birth. Environmental and biological samples will be tested, and these samples will mostly come from Puerto Rico because of the unusually high incidence of preterm birth there. While nontargeted (screening) analysis will be conducted, primary attention will be given to Superfund and related contaminants, since there are many Superfund sites in Puerto Rico. Much of the analysis will be based on mass spectrometry (MS); three types of MS will be employed: (HPLC)MALDI-TOF/TOF-MS, GC-EI(EC)-TOF-MS, and HPLC-ESI-TOF-MS. We will seek to discover "putative PTB-xenobiotics," defined as xenobiotics that meet either one of the following two criteria: they correlate with preterm birth, or they do not but are common as contaminants and have a bioactivity that could contribute to this condition. The environmental samples will be tap water and ground water, including samples that have been remediated (detoxified) by electrolysis. The biological samples will be pregnancy tissues (gestational membrane, placenta), cell cultures, peripheral blood leukocytes and urine. The pregnancy tissues will come from rats and humans, and the leukocytes and urine will come from humans. One or more of four kinds of assays will be applied to each of these types of samples: 1. Direct Detection, the xenobiotic (or a laboratory derivative thereof) is directly detected in a mass spectrometer; 2. DNA Adduct, the xenobiotic forms a DNA adduct in vivo that is detected by Mass Tag Profiling; 3. Pro-oxidant, the xenobiotic is detected after metabolism-like oxidation in a Nucleotide Exposure Assay; 4. Bioassay, the xenobiotic is detected in a bioassay with cultured cells derived from human placenta¿based initially on monitoring for cytotoxicity (primary screen) and subsequently for apoptosis, genotoxicity, inflammation, and oxidative stress (secondary screen, reflecting mechanisms that could contribute to preterm birth). New or improved analytical methodology will be developed, and new combinations of methodology will be studied in this project to enhance the discovery of putative PTB-xenobiotics by these four strategies. The project, thereby, will result in generally-applicable advances in the overall field of discovering toxic xenobiotics in complex samples. Through exchange of samples, collaborative testing, and collaborative analysis of data, either directly or through the cores, this project integrates with all of the other projects in the PRoTECT program.