Grantee Research Project Results
2000 Progress Report: Influence of Genotype on Reproductive Effects of Air Pollution in Young Men
EPA Grant Number: R827019Title: Influence of Genotype on Reproductive Effects of Air Pollution in Young Men
Investigators: Evenson, Donald P. , Marshall, Don , Rubes, Jiri , Sram, Radim
Institution: South Dakota State University , Veterinary Research Institute
Current Institution: South Dakota State University
EPA Project Officer: Aja, Hayley
Project Period: September 1, 1998 through September 30, 2000 (Extended to November 30, 2003)
Project Period Covered by this Report: September 1, 1999 through September 30, 2000
Project Amount: $274,670
RFA: Interindividual Variation in Human Susceptibility to Environmentally-caused Disease (1998) RFA Text | Recipients Lists
Research Category: Human Health
Objective:
This research project is testing the hypothesis that men with commonly occurring genetic mutations for enzymes involved in metabolizing reactive intermediates of polycyclic aromatic hydrocarbon (PAH) components of air pollution will be more susceptible to DNA damage and/or aneuploidy induced in their germ cells. Genotypes for GSTM1, GSTT1 and NAT2 will be determined from archived blood samples. Sperm DNA damage will be evaluated in archived semen samples by the following methods (some in the developmental stage): (1) the Sperm Chromatin Structure Assay (SCSA); (2) DNA strand breaks as measured by a DNA end labeling technique (TUNEL) and single cell gel electrophoresis (COMET assay); and (3) sperm aneuploidy (hyperhaploidy and diploidy) measured using 3-color fluorescence in situ hybridization (FISH).Progress Summary:
Samples Collected. Sperm were obtained by masturbation from 15 normal healthy donors, 20 years old at the initiation of the study. All men lived in the Teplice district of the Czech Republic. Consent, medical questionnaires and lifestyle features were obtained from each. Seven semen samples were obtained from each donor over a 2-year period within 1 week's time in the late winter and early autumn.
Semen Processing. Semen was allowed to liquefy at room temperature and then analyzed according to standard procedures. Semen volume, sperm concentration, total sperm number, the percent of motile and morphologically normal sperm were assessed. For FISH, smears were made onto clean microscopic slides after thawing archived samples at room temperature, and air-dried. The remaining semen was aliquoted and frozen at -80?C without cryopreservation.
FISH Data. The fish data were previously reported; significant interindividual differences were observed for all categories of disomes and diploidies (p<0.001). These differences were persistent over the 2-year sampling period.
Genotype vs. SCSA Data. Men in this study have been genotyped; however, due to the lack of high pollution periods, the SCSA data were not different and thus no correlation could be made with the genotype data.
COMET and TUNEL Data. Progress is being made on the analysis of these samples by the COMET and TUNEL assays. We expect to be finished by the end of our one-year extended project period. The major delay has been to develop and interpret the COMET data with our new Vis-COMET image analysis system.
SCSA Data on the Czech Men. The SCSA data are similar to data collected from hundreds of other semen samples. It is of great interest however, to note that the percent of abnormal sperm in the young Czech men was higher than from men in previous studies done in the United States. From semen samples obtained from the entire study (266 samples) 46 percent of the men had a probability of high fertility (<15 % with damaged DNA) in comparison with 68 percent of U.S. men, 35 percent fell in the intermediate to poor fertility (15 % to <30 % damaged DNA) in comparison with 25 percent of U.S. men, and 19 percent were likely infertile (>30 % damaged DNA) in comparison with 6 percent of U.S. men. It is not clear whether this poorer quality sperm chromatin is due to the younger age of the men (18-20), or more likely, that the baseline of damage is due to a generally poorer environment.
Of significance was the very important observation that the SCSA data were correlated with the level of air pollution. Specifically, no difference was noted between low and medium levels of pollution, but there was an approximately 50 percent increase in the percent of sperm with damage DNA in the high pollution group (p<0.05) by the Kuskal-Wallis test.
Future Activities:
The major efforts of this coming period are to finish the COMET and TUNEL data. In particular, we will first finish those sets of about six longitudinal samples obtained from single donors. This allows for an internal control over time as related to environmental exposure. Furthermore, we expect that these data sets will provide valuable information as to the nature of the sperm nuclear defects and how they may relate to each other.
We also are doing a study on the effects of age on SCSA data to hopefully shed some light on the important question of why the young men had poorer quality sperm chromatin than the studies previously done in the United States.
Journal Articles on this Report : 8 Displayed | Download in RIS Format
| Other project views: | All 31 publications | 21 publications in selected types | All 16 journal articles |
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Alvarez JG, Sharma RK, Ollero M, Saleh RA, Lopez MC, Thomas AJ, Evenson DP, Agarwal A. Increased DNA damage in sperm from leukocylospermic semen samples as determined by the sperm chromatin structure assay. Fertility and Sterility 2002;78(2):319-329 |
R827019 (2000) R827019 (Final) |
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Evenson DP, Jost LK, Corzett M, Balhorn R. Characteristics of human sperm chromatin structure following an episode of influenza and high fever: a case study. Journal of Andrology 2000;21:739-746. |
R827019 (2000) R827019 (Final) |
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Changes in susceptibility of bovine sperm to in situ DNA denaturation during prolonged storage at ambient temperature under conditions of exposure to reactive oxygen species and nuclease inhibitor. Reproduction, Fertility and Development. |
R827019 (2000) |
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Larson KL, DeJonge CJ, Barnes AM, Jost LK, Evenson DP. Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Human Reproduction 2000;15:1717-1722. |
R827019 (2000) R827019 (Final) |
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Larson KL, Brannian JD, Singh NP, Burbach JA, Jost LK, Hansen KP, Kreger DO, Evenson DP. Chromatin structure in globozoospermia: a case report. Journal of Andrology 2001;22(3):424-431. |
R827019 (2000) R827019 (Final) |
not available |
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Ollero M, Gil-Guzman E, Lopez MC, Sharma RK, Agarwal A, Larson K, Evenson D, Thomas Jr AJ, Alvarez JG. Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility. Human Reproduction 2001;16(9):1912-1921. |
R827019 (2000) R827019 (Final) |
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Perreault SD, Rubes J, Robbins WA, Evenson DP, Selevan SG. Evaluation of aneuploidy and DNA damage in human spermatozoa: applications in field studies. Andrologia 2000;32(4-5):247-254. |
R827019 (2000) R827019 (Final) |
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Selevan SG, Borkovec L, Slott VL, Zudova Z, Rubes J, Evenson DP, Perreault SD. Semen quality and reproductive health of young Czech men exposed to seasonal air pollution. Environmental Health Perspectives 2000;108(9):887-894. |
R827019 (2000) R827019 (Final) |
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Supplemental Keywords:
human health, atmosphere, exposure, health effects, cellular, susceptibility, genetics, industry., RFA, Health, Air, Scientific Discipline, ENVIRONMENTAL MANAGEMENT, Susceptibility/Sensitive Population/Genetic Susceptibility, Health Risk Assessment, Risk Assessments, Biology, genetic susceptability, Environmental Chemistry, Endocrine Disruptors - Human Health, Children's Health, endocrine disruptors, Biochemistry, Endocrine Disruptors - Environmental Exposure & Risk, air toxics, Genetics, Risk Assessment, genetic predisposition, health risks, interindividual variation, human exposure, sensitive subpopulations, sperm, environmentally-caused disease, aromatic hydrocarbons, Czech Republic, developmental processes, endocrine disrupting chemicals, exposure, gender, genotype, reproductive effects, reproductive processes, biological effects, reproductive, toxicants, young men, air pollution, human health risk, PAH, reproductive health, pollution exposure, DNA damage, ethnic, exposure assessment, genetic diversity, race ethnicityProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.