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Title: Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material (DNA). Initial assessment of plant DNA adducts as biomarkers

Abstract

Genetic damage to deoxyribonucleic acid (DNA) has long been suspected of being a fundamental event leading to cancer. A variety of causal factors can result in DNA damage including photodimerization of base pairs, ionizing radiation, specific reaction of DNA with environmental pollutants, and nonspecific oxidative damage caused by the action of highly reactive oxidizing agents produced by metabolism. Because organisms depend on an unadulterated DNA template for reproduction, DNA repair mechanisms are an important defense for maintaining genomic integrity. The objective of this exploratory project was to evaluate the potential for TNT to form DNA adducts in plants. These adducts, if they exist in sufficient quantities, could be potential biomarkers of munitions exposure. The ultimate goal is to develop a simple analytical assay for the determination of blomarkers that is indicative of munitions contamination. DNA repair exists in dynamic equilibrium with DNA damage. Repair mechanisms are capable of keeping DNA damage at remarkably low concentrations provided that the repair capacity is not overwhelmed.

Authors:
; ; ;
Publication Date:
Research Org.:
Pacific Northwest Lab., Richland, WA (United States)
Sponsoring Org.:
Department of Defense, Washington, DC (United States)
OSTI Identifier:
195771
Report Number(s):
PNL-10738
ON: DE96003998
DOE Contract Number:
AC06-76RL01830
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: Aug 1995
Country of Publication:
United States
Language:
English
Subject:
56 BIOLOGY AND MEDICINE, APPLIED STUDIES; 55 BIOLOGY AND MEDICINE, BASIC STUDIES; 45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE; TNT; BIOLOGICAL EFFECTS; METABOLISM; PLANTS; GENETICS; POLLUTION; SOILS

Citation Formats

Harvey, S.D., Clauss, T.W., Fellows, R.J., and Cataldo, D.A. Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material (DNA). Initial assessment of plant DNA adducts as biomarkers. United States: N. p., 1995. Web. doi:10.2172/195771.
Harvey, S.D., Clauss, T.W., Fellows, R.J., & Cataldo, D.A. Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material (DNA). Initial assessment of plant DNA adducts as biomarkers. United States. doi:10.2172/195771.
Harvey, S.D., Clauss, T.W., Fellows, R.J., and Cataldo, D.A. Tue . "Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material (DNA). Initial assessment of plant DNA adducts as biomarkers". United States. doi:10.2172/195771. https://www.osti.gov/servlets/purl/195771.
@article{osti_195771,
title = {Evaluation of the metabolic fate of munitions material (TNT & RDX) in plant systems and initial assessment of material interaction with plant genetic material (DNA). Initial assessment of plant DNA adducts as biomarkers},
author = {Harvey, S.D. and Clauss, T.W. and Fellows, R.J. and Cataldo, D.A.},
abstractNote = {Genetic damage to deoxyribonucleic acid (DNA) has long been suspected of being a fundamental event leading to cancer. A variety of causal factors can result in DNA damage including photodimerization of base pairs, ionizing radiation, specific reaction of DNA with environmental pollutants, and nonspecific oxidative damage caused by the action of highly reactive oxidizing agents produced by metabolism. Because organisms depend on an unadulterated DNA template for reproduction, DNA repair mechanisms are an important defense for maintaining genomic integrity. The objective of this exploratory project was to evaluate the potential for TNT to form DNA adducts in plants. These adducts, if they exist in sufficient quantities, could be potential biomarkers of munitions exposure. The ultimate goal is to develop a simple analytical assay for the determination of blomarkers that is indicative of munitions contamination. DNA repair exists in dynamic equilibrium with DNA damage. Repair mechanisms are capable of keeping DNA damage at remarkably low concentrations provided that the repair capacity is not overwhelmed.},
doi = {10.2172/195771},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Aug 01 00:00:00 EDT 1995},
month = {Tue Aug 01 00:00:00 EDT 1995}
}

Technical Report:

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  • The goals of this effort were to confirm and expand data related to the behavior and impacts of munitions residues upon human food chain components. Plant species employed included corn (Zea mays), alfalfa (Medicago sativa). spinach (Spinacea oleraceae), and carrot (Daucus carota). Plants were grown from seed to maturity (70 to 120 days) in a low-fertility soil (Burbank) amended with either {sup 14}C-TNT or {sup 14}C-RDX at which time they were harvested and analyzed for munitions uptake, partitioning, and chemical form of the munition or munition-metabolite. All four of the plant species used in this study accumulated the {sup 14}C-TNT-more » and RDX-derived label. The carrot, alfalfa, and corn demonstrated a higher percentage of label retained in the roots (62, 73, and 83% respectively). The spinach contained less activity in its root (36%) but also contained the highest TNT specific activity observed (>4600 jig TNT equivalents/g dry wt.). The specific uptake values of RDX for the spinach and alfalfa were comparable to those previously reported for wheat and bean (314 to 590 {mu}g RDX-equivalents/g dry wt. respectively). An exception to this may be the carrot where the specific activity was found to exceed 4200 {mu}g RDX-equivalents/g dry wt. in the shoot. The total accumulation of TNT by the plants ranged from 1.24% for the spinach to 2.34% for the carrot. The RDX plants ranging from 15% for the spinach to 37% for the carrot. There was no identifiable TNT or amino dinitrotoluene (ADNT) isomers present in the plants however, the parent RDX compound was found at significant levels in the shoot of alfalfa (> 1 80 {mu}g/g) and corn (>18 {mu}g/g).« less
  • Munitions material can enter the environment as a result of manufacturing activities and field usage. Predictor methodologies, or biomarkers would enhance evaluation of environmental impacts. The goal of this exploratory study deoxyribonucleic acid (DNA) mutation frequency as a biomarker for munitions exposure. The approach e resolution of an effective repetitive sequence probe for the identification of characteristic mutations, and (2) the development of a testing media [a clonal cell line of carrot (Daucus carota) spension cells]. Commercially available probes demonstrated marginal resolution therefore a low-C{sub o}t library was then constructed. Three colonies from the low-C{sub o}t DNA library were screenedmore » and the DNA isolates sequenced. A suspension culture of carrot (Daucus carota) was developed. A mutation spectra experiment was initiated at a 10-mg TNT/L exposure concentration with the attempt to clone over 1500 single TNT-exposed cells. Over the following six months greater than 98% of the initially isolated cells were unable to survive and produce micro calluses. The remaining calli were too few to be statistically significant and the experiment was terminated. The biomarker concept itself remains to be disproved, but the need for large numbers of uniform clones to differentiate true mutations suggest that more direct techniques using whole tissues need to be developed.« less
  • The objective of the present investigation was to elucidate the environmental behavior and fate of hexahydro-1,3,5-trinitro-1,3,5- triazine (RDX), particularly as related to its transport and chemical form in the food chain. To meet this goal, we needed to adapt and develop suitable analytical methodology to fractionate and characterize both RDX and RDX-derived residues in soil and plant matrices. Using the methodology that we developed, we assessed the chemical and physical fate of RDX in soils and plants. In general, the plant availability and plant mobility of RDX is substantially greater than that previously reported for TNT. 30 refs., 27 figs.,more » 26 tabs.« less
  • The objective of these investigations was to elucidate the environmental behavior and fate of trinitrotoluene (TNT). Emphasis was placed on those chemical transformations occurring in soils and in plant tissues following uptake and on the probable impact of these chemical transformations on the food chain. Analytical methodology was developed to fractionate and characterize both TNT and TNT-derived residues in soil and plant matrices. The procedures developed in this program extend prior art, through the use of matrix-specific extraction and fractionation schemes followed by classical HPLC separations. Methods showed good recovery and reproducibility. 30 refs., 35 figs., 27 tabs.