Roger W. Giese, Project Leader, Northeastern University
See Project 1 news here.
The long-term 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. Much of the analysis will be based on mass spectrometry (MS); three types of MS will be employed: High performance liquid chromatography matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry [(HPLC)MALDI-TOF/TOF-MS], Gas chromatography electron impact (electron capture) time-of-flight mass spectrometry [GC-EI(EC)-TOF-MS], and High performance liquid chromatography electrospray ionization time-of-flight mass spectrometry [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 three kinds of assays will be applied to each of these types of samples:
- Direct Detection, the xenobiotic (or a laboratory derivative thereof) is directly detected in a mass spectrometer;
- DNA Adduct, the xenobiotic forms a DNA adduct in vivo that is detected by Mass Tag Profiling;
- Pro-oxidant, the xenobiotic is detected after metabolism-like oxidation in a Nucleotide Exposure Assay;
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 three 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 PROTECT.
- Accu-Seal Corporation, San Marcos, CA, has provided a medical-grade bag sealer which we are using for sample preparation.
- Sigma Aldrich Corporation, St. Louis, MO, is providing a diversity of chromatographic packings, also for use in sample preparation.
- Thermo-Fisher, Pittsburgh, PA, is providing a discounted reagent.
- Yankee Containers, North Haven, CT, donated transport vessels for samples between Boston and Puerto Rico.
- Industrial Fabrics Corm, Minneapolis, MN, donated nylon netting.
- Affymetrix, Santa Clara, CA, donated a modified nucleotide standard.