Department of Genetic Ecotoxicology
Head: Radim J. Sram, M.D., D.Sc.
Scientists:
Miroslav Dostal, M.D., D.Sc.
Bozena Novotna, Ph.D.
Pavel Rössner, M.D., D.Sc.
Pavel Rössner, Jr., Ph.D.
Jana Schmuczerova, Ph.D.
Jan Topinka, D.Sc.
Ph.D. Students:
Mgr. Olena Beskid
Oksana Sevastyanova, M.D.
Mgr. Vlasta Svecova
Technical Assistants:
Dr. Irena Chvatalova
Dr. Eva Dejmkova
Mrs. Lenka Dostalova
Mrs. Jana Herlova
Ing. Zdenka Lnenickova
Mgr. Alena Milcova
Mgr. Zuzana Novakova
Mgr. Andrea Rössnerova
Ing. Ivo Solansky
Mrs. Olga Stverakova
Mrs. Jolana Vankova
Address:
Videnska 1083, 142 20 Prague 4
Phone: (420) 241 062 596
Fax: (420) 241 062 785
E-mail: sram@biomed.cas.cz
The Department of Genetic Ecotoxicology (DGE) is a joint venture of the Health Institute of Central Bohemia
and the Institute of Experimental Medicine.
The research of the DGE is concerned with the genetic damage caused by genotoxic and carcinogenic compounds such as PAHs, alkenes, etc. The
effects of genotoxic xenobiotics are studied in vitro using acelullar assays and cell cultures, such as human diploid lung fibroblasts, as well as in vivo by
carrying out human transitional molecular epidemiologic studies and observational epidemiologic studies. The research is based on the application of advanced
scientific technologies such as the 32P-postlabelling assay for DNA adduct analysis in WBC, lymphocytes and placentas, HPLC analysis of 32P-labeled DNA
adducts, the expression of p53 and p21WAF1 proteins in plasma and lymphocytes, the comet assay to assess DNA damage in lymphocytes, cytogenetic
analysis of chromosomal aberrations by conventional method as well as by fluorescence in situ hybridization (FISH), radioimmunological methods, flow cytometry
for the phenotyping of lymphocyte subpopulations and methods for the analysis of genetic polymorphisms CYP1A1, GSTM1, GSTT1, GSTP1, NAT2, EPHX, XPD,
XRCC1, hOGG1, p53, MS, MTHFR and APOE (PCR, RLFP techniques).
The DGE has been responsible for the scientific management of the Program "AIR POLLUTION AND HEALTH", which consists of several projects studying
the impact of environmental pollution on the health of the population in North Bohemia and in the capital, Prague.
Current research at the Department focuses on the following main topics:
- Molecular epidemiology studies: biomarkers of exposure to mutagens and carcinogens
- Pregnancy outcome
- Immunity and childhood morbidity
- In vitro studies
- Embryotoxicity of genotoxic compounds
- Apoptosis in developing systems
Molecular epidemiology studies: biomarkers of exposure to mutagens and carcinogens
The objectives of these studies are to simultaneously evaluate personal exposure, internal measure of
exposure, biologically effective dose of exposure, early biological effect and metabolic susceptibility using different biomarkers as mentioned above.
Cotinine in urine, as determined by an RIA assay, as a marker of exposure to tobacco smoke and the levels of vitamin A, C, E and folic
acid in plasma are measured as possible confounding factors.
Fig. 1 Evaluation of chromosomal aberrations by fluorescent in situ
hybridization (FISH) method (reciprocal translocation of chromosome 1).
In the project "Exposure of model population" (Program AIR POLLUTION AND HEALTH) the impact of air pollution on city policemen
working outdoors in downtown Prague for more than 8 h/day (EXP) was studied.Volunteers from a suburban area working indoors were selected as controls (CON).
The results of multivariate regression analysis indicate that smoking, vitamin C, polymorphisms of GSTM1, XPD repair gene in exon 23, and epoxide hydrolase
gene (EPHX) in exon 4 were significant predictors for total DNA adduct levels. Smoking, exposure to ambient air pollution and polymorphism of XPD repair
gene in exon 6 were significant predictors for "like" B[a]P-DNA adduct. Using FISH analysis a significant increase in all cytogenetic endpoints was observed
in the EXP group, due to a significant increase in the genomic frequency of translocations in EXP vs. CON groups. The frequencies of chromosomal aberrations
determined by FISH were affected by GSTP1, hOGG1 and XPD polymorphisms. Ambient air exposure to PAHs significantly increased FISH cytogenetic endpoints in
nonsmoking policemen working in downtown Prague. The project "Atherosclerotic changes and DNA adducts" (IGA MZ - grant NM-10-3) was based on the
idea that atherosclerosis and carcinogenesis may share some common mechanisms of the genotoxic action of exogenous compounds such as polycyclic aromatic
hydrocarbons (PAHs). The main objective of this study was to test the hypothesis that "bulky" aromatic DNA adducts in the smooth muscle cells (SMC) of thoracic
aortas taken at autopsy from sudden and accidental death male subjects are associated with the stage of atherosclerosis. We found significantly higher DNA adduct
levels in "Cases" as compared with "Controls". "Cases" also had significantly greater age, elevated heart weight and plasma cholesterol levels, and a higher frequency of
overweight subjects as compared to "Controls". Multivariate logistic regression revealed that the "bulky" aromatic DNA adducts, which are most likely related to
environmental exposure to genotoxic chemicals, remain a statistically and biologically significant predictor of the stage of atherosclerosis even after adjustment
for all other significant predictors in the model. Our results showed that the risk of atherosclerosis is also associated with some of genetic susceptibility markers
(GSTT1 and CYP1A1-MspI polymorphisms). The fact that "bulky" aromatic DNA adduct levels predict the progression of atherosclerosis independently of smoking
indicates that the formation of atherosclerotic plaques may also be initiated by environmental exposures other than tobacco smoke.
The DGE has also participated in several EC projects: "Air pollution distribution of adult population in Europe", "Biomarkers of genotoxicity
of urban air pollution: A dose-response study", "Biomonitoring of occupationally exposed population to acrylonitrile" and "Effects of PAHs in environmental pollution
on exogeneous and endogeneous DNA damage" as well as the HEI project "Transitional epidemiology study for 1,3-butadiene biomarkers".
Pregnancy outcome
Several recent studies suggested that the increased risk of adverse human reproductive effects in the District
of Teplice could be related to the high level of air pollution in this region. A population-based project "Pregnancy outcome" was carried out to test this
presumption. A long-term project, the major aim of which is to evaluate the impact of urban air pollution on reproduction quality in the capital city of Prague
(with heavy traffic pollution), has been recently initiated. The major objective of this project is a comparison of the reproductive effects of air pollution
in different regions.
Exposure is estimated from air pollution monitoring, questionnaire information and the assessment of selected biomarkers. Based on statistical
power analysis, low birthweight, premature birth and intrauterine growth retardation (IUGR) were chosen as the main reproductive outcomes. Two basic approaches were
combined: In a prospective cohort study of all pregnancies in Teplice and Prachatice, the effect of the simultaneous action of air pollution and many other factors
on pregnancy outcomes was analyzed using methods of multivariate logistic regression analysis. The statistical models accounted for possible critical exposure
windows for different outcomes. In a nested case control study, a sample of cases (adverse outcomes) and suitable controls were studied. Selected biomarkers were
determined in venous blood, cord blood and placenta (DNA adducts assayed by 32P-postlabelling, comet assay, cell immunity markers and levels of vitamins A, E, C and
folic acid) with the aim of evaluating the exposure, response and state of mother or child. Metabolic genotypes (GSTM1, GSTT1, NAT2, CYP1A1 and EPHX) were also
determined to investigate the effect of genetically determined susceptibility on the level of internal exposure.
A considerably higher prevalence of all adverse outcomes was found in the highly polluted district of Teplice compared with Prachatice.
A significant and dose-related relationship between the risk of fetal growth retardation and the levels of fine particles (PM10, PM2.5) was observed in the Teplice
region. No similar association was observed in Prachatice - a region with considerably lower levels of particles. On the other hand, a strong dose-related association
between the risk of IUGR and the concentration of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) was found in both Teplice and Prachatice.
Fig. 2: Intrauterine growth retardation (IUGR) by carcinogenic PAHs in Teplice. Adjusted Odds Ratios for IUGR were significantly
increased for medium and high concentrations of c-PAHs during the first month of gestation. Concentration intervals for c-PAHs were:
Low = < 15 ng/m3 (reference level), Med = 15 to 30ng/m3 and
High = 30 ng/m3 or higher.
PAHs are mostly adsorbed on the surfaces of fine particles. Therefore, based on the above results, it was hypothesized
that c-PAHs, rather than particles, adversely affect fetal growth. Carcinogenic PAHs are known to inhibit epidermal growth factor receptors and to alter early
trophoblast proliferation. In this way the fetoplacental exchange of oxygen and nutrients may be reduced to a sufficient extent to affect fetal growth.
The results concerning the impact of air pollution on pregnancy outcome for the first time revealed that air pollution may influence human
reproductive processes, such as fetal growth or fecundability. The induced changes may include not only morphologic birth defects but also some subtle functional
changes affecting their carriers throughout their lives. Children are thought to be very sensitive to xenobiotics from conception. Their development is complex
and is influenced by various environmental factors as well as by their mother´s deleterious lifestyle such as cigarette smoking or poor diet.
Immunity and childhood morbidity
A study of the morbidity of children aged between 0-3 years has been carried out in the families enrolled in
the Pregnancy Outcome Study. Immune biomarkers in cord and maternal blood were determined in study participants at delivery. Comparisons were made between
children living in the district Teplice, known for its high air pollution, and children living in Prachatice, a district with
consistently lower particulate and SO2 exposures. Children born in Teplice experienced a significantly higher rate of otitis media (and otalgia), gastrointestinal
infections, upper respiratory infections, and pneumonia, but did not differ in their risk of bronchitis or viral infections such as varicella. These findings
remained significant even when multiple linear regression models accounted for education, maternal age, maternal smoking, and other smokers in the household,
breastfeeding, and attendance at day care. The results suggest that air pollution may alter early childhood susceptibility to infections. This stage of the study
was supported by the Ministry of Environment of the Czech Republic in the period 1997-1999. The study now continues (2001-2004) under the sponsorship of the Health
Effects Institute (Boston, MA) to follow-up children born in 1997-1999.
In vitro studies
These studies are concerned with the investigation of the DNA binding activities of single carcinogenic chemicals,
their environmental mixtures, and different chemical compounds associated with PM10 air particles. DNA adduct formation is studied using an acelullar system,
human diploid lung fibroblasts, NCI-H322 and Hep G2 cell lines with the aim to better understand the mechanism of action of these complex mixtures. Western
blotting and ELISA are used to examine the induction of p53 and p21WAF1 protein levels following treatment of these cells with PAH compounds and environmental mixtures.
Both these proteins are involved in cell cycle control; cell cycle alteration has been studied as well.
Embryotoxicity of genotoxic compounds
The significance of the genotoxic effects of xenobiotics during the embryonic period of life is analyzed
using a chick embryo model. DNA and chromosome damage is related to (i) the kinetics of the tested compound in embryonic and extraembryonic tissues , (ii) cell
proliferation and cell death and (iii) final morphological and functional development of particular morphogenetic systems and the whole embryo. Special attention is
devoted to hematopoietic cells and the embryonic kidney. Image analysis is used to quantify the differences between control and exposed individuals.
Fig.3 Erythroid cells of the chick embryo exhibiting a different degree of nuclear DNA fragmentation.
(a) healthy cell - all DNA in the nucleus ("head of comet"),
(b) cell with the low level of DNA damage (DNA fragments in the "tail of comet"),
(c) cell with the high degree of DNA damage (minimum of DNA in the "head of comet").
Apoptosis in developing systems
Apoptosis represents a mechanism to eliminate mutant cells from an organism. Using single cell gel
electrophoresis, the extent of apoptotic DNA fragmentation is measured in bone marrow aspirates of patients with myelodysplastic syndromes with the aims
of (i) precise diagnosis, (ii) a comparison of the impairment of particular cell lineages, (iii) a follow-up of disease progression and the efficiency of therapy.
Besides DNA fragmentation, membrane markers of cell differentiation and cell death, involving mapping the levels of sialylation of membrane glycoproteins, are
investigated.
The role of apoptosis in the development of genetically determined male sterility is studied in rats with the Hd mutation.
The levels of apoptosis in particular stages of spermatogenesis in control and mutant rats are correlated with histological findings.
Relevant publications before 1998
1. Sram, R.J., Zudova, Z., Kuleshov, N.P. (1980) Cytogenetic analysis of peripheral lymphocytes in workers occupatinally exposed to epichlorohydrin.
Mutation Res. 70: 115-120
2. Sram, R.J., Kuleshov, N.P. (1980) Monitoring the occupational exposure to mutagens by the cytogenetic analysis of human peripheral lymphocytes.
Arch. Toxicol. Suppl. 4: 11-20
3. Sram, R.J., Tomatis, L., Clemmesen, J., Bridges, B.A. (1981) An evaluation of the genetic toxicity of epichlorhydrin.
Mutation Res. 87: 299-319
4. Sram, R.J., Dobias, L., Pastorkova, A., Rössner, P., Janca, L. (1983) Effect of ascorbic acid prophylaxis on
the frequency of chromosome abberations in the peripheral lymphocytes of coal-tar workers. Mutation
Res. 120: 181-186
5. Sram, R.J., Hola, N., Kotesovec, F., Vavra, R. (1985) Chromosomal abnormalities in soft coal open-cast
mining workers. Mutation Res. 144: 271-275
6. Kocisova, J., Rössner, P., Binkova, B., Bavorova, H., Sram, R.J. (1988) Mutagenicity studies on paracetamol in human volunteers.
I. Cytogenetic analysis of peripheral lymphocytes and lipid peroxidation in plasma. Mutation Res. 209: 161-165
7. Topinka, J., Binkova, B., Sram, R.J. (1991) DNA-repair capacity and lipid peroxidation in chronic alcoholics.
Mutation Res. 263: 133-136
8. Sram, R.J., Binkova, B., Dobias, L., Rössner, P., Topinka, J.,Vesela, D., Vesely, D., Stejskalova, J.,
Bavorova, H., Rericha,V. (1993) Monitoring genotoxic exposure in uranium miners. Environ. Health Perspect. 99: 303-305
9. Binkova, B., Dobias, L., Wolff, T., Sram, R.J. (1994) 32P-postlabelling analysis of DNA adducts in tissues of
rats exposed to coke-oven emissions. Mutation Res. 307: 355-363
10. Sorsa, M., Autio, K., Demopoulos, N.A., Järventaus, H., Rössner, P., Sram, R.J, Stephanou, G.,
Vlachodimitropoulos, D. (1994) Human biomonitoring of occupational exposure to 1,3-butadiene. Mutation Res. 309: 321-326
11. Dobias, L., Cerna, M., Rössner, P., Sram, R.J. (1994) Genotoxicity and carcinogenicity of metronidazole.
Mutation Res. 317: 177-194
12. Binkova, B., Lewtas, J., Miskova, I., Lenicek, J., Sram, R.J. (1995) DNA adducts and personal air monitoring of
carcinogenic polycyclic aromatic hydrocarbons in an environmentally exposed population. Carcinogenesis 16: 1037-1046
13. Binkova, B., Lewtas, J., Miskova, I., Rössner, P., Cerna, M., Mrackova, G., Peterkova, K., Mumford, J., Meyer,
S., Sram, (1996) R.J. Biomarker studies in Northern Bohemia. Environ. Health Perspect. 104 Suppl. 3: 591-597
14. Sram, R.J., Benes, I., Binkova, B., Dejmek, J., Horstman, D., Kotesovec, F., Otto, D., Perreault, S.D., Rubes, J.,
Selevan, S.G., Skalik, I., Stevens, R.K., Lewtas, J. (1996) Teplice Program - the impact of air pollution on human health. Environ. Health Perspect.
104 Suppl. 4: 699-714
15. Topinka, J., Binkova, B., Mrackova, G., Stavkova, Z., Peterka, V., Benes, I., Dejmek, J., Lenicek, J., Pilcik, T.,
Sram, R.J. (1997) Influence of GSTM1 and NAT2 genotypes on placental DNA adducts in an environmentally
exposed population. Environ. Mol. Mutagenesis 30: 184-195
Publications 1998-2005
1. Binkova, B., Lenicek, J., Benes, I., Vidova, P., Gajdos, O., Fried, M., Sram, R.J. (1998) Genotoxicity of coke
oven and urban air particulate matter in in vitro assays acellular assay coupled with 32P-postlabeling and HPLC analysis of DNA adducts. Mutation Res. 414:77-94
2. Binkova, B., Topinka, J., Mrackova, G., Gajdosova, D., Vidova, P., Stavkova, Z., Peterka, V., Pilcik, T., Rimar,
V., Dobias, L., Farmer, P.B., Sram, R.J. (1998) Coke oven workers study: The effect of exposure and GSTM1
and NAT2 genotypes on DNA adducts in white blood cells and lymphocytes as determined by 32P-postlabeling. Mutation Res. 416: 67-84
3. Costa, D.J., Slott, V., Binkova, B., Myers, S.R., Lewtas, J. (1998) Influence of GSTM1 and NAT2 genotypes on
the relationship between personal exposure to PAH and biomarkers of internal dose. Biomarkers 3: 411-424
4. Heringova, L., Dostal, M., Jelinek, R. (1998) Cell cycle alteration within chick embryonic anlagen after
cyclophosphamide treatment. Teratogenesis, Carcinogenesis, Mutagenesis 14: 63-72
5. Kalina, I., Brezani, P., Gajdosova, D., Binkova, B., Salagovic, J., Habalova, M., Mrackova, G., Dobias, L., Sram,
R.J. (1998) Cytogenetic monitoring in coke oven workers. Mutation Res. 417: 9-17
6. Rössner, P., Sram, R.J., Bavorova, H., Ocadlikova, D., Cerna, M., Svandova, E. (1998) Spontaneous level of
chromosomal aberrations in peripheral blood lymphocytes of control individuals of the Czech Republic population. Toxicol.Lett. 96,97: 137-142
7. Sram, R.J., Podrazilova, K., Dejmek, J., Mrackova, G., Pilcik, T. (1998) Single cell gel electrophoresis assay:
sensitivity of peripheral white blood cells in human population studies. Mutagenesis 13: 99-103
8. Sram, R.J., Rossner, P., Peltonen, K., Podrazilova, K., Mrackova, G., Demopoulos, N.A., Stephanou, G.,
Vladimiropoulos, D., Darroudi, F., Tates, A.D. (1998) Chromosomal aberrations, sister-chromatid exchanges,
cells with high frequency of SCE, micronuclei and comet assay parameters in 1,3-butadiene exposed workers. Mutation Res. 419: 145-154
9. Topinka, J., Schwarz, L., Kiefer, F.,Wiebel, F.J., Gajdos, O., Vidova, P., Dobias, L., Fried, M., Wolff, T., Sram,
R.J. (1998) DNA adducts formation in mammalian cell cultures by polycyclic aromatic hydrocarbons (PAH)
and nitro-PAH in coke oven emissions in cell extract. Mutation Res. 419: 91-106
10. Novotna, B., Kubinova, L., Vankova, J., Jelinkova, L. (1998) Quantitation of cell death and DNA cleavage
during chick embryo limb development. In: Fluorescence Microscopy and Fluorescence Probes (ed. J.Slavik), New York:Plenum Press, pp.255-260
11. Novotna, B., Sedmera, D., Dusek, Z., Holejsovska, I., Bila, V., Kren, V. (1998) Apoptotic fragmentation of DNA
during limb morphogenesis of rats with polydactyly-luxate syndrome. BEMS Journal 1: 40-47
12. Sedmera, D., Novotna, B., Bila, V., Kren, V. (1998) The role of cell death in limb development of rats
manifesting Lx allele on different genetic backgrounds. Eur. J. Morphol. 36: 173-181
13. Sram, R.J., Binkova, B., Rössner, P., Rubes, J., Topinka, J., Dejmek, J. (1999) Adverse reproductive outcomes
from exposure to environmental mutagens. Mutation Res. 428: 203-215
14. Leonard, A., Gerber, G.B., Stecca, C., Rueff, J., Borba, H., Farmer, P.B., Sram, R.J., Czeizel, A.E., Kalina, I.
(1999) Mutagenicity, carcinogenicity, and teratogenicity of acrylonitrile. Mutation Res. 436: 263-283
15. Binkova, B., Vesely, D., Vesela, D., Jelinek, R., Sram, R.J. (1999) Genotoxicity and embryotoxicity of urban air
particulate matter collected during winter and summer period in two different districts of the Czech Republic. Mutation Res. 440: 45-58
16. Dobias, L., Kusova, J., Gajdos, O., Vidova, P., Gajdosova, D., Havrankova, J., Fried, M., Binkova, B., Topinka,
J. (1999) Bioassay-directed chemical analysis and detection of mutagenicity in ambient air of the coke oven.
Mutation Res., 445:285-293
17. Dejmek, J., Selevan, S.G., Benes, I., Solansky, I., Sram, R.J. (1999) Fetal growth and maternal exposure to particulate
matter during pregnancy. Environ. Health Perspect. 107: 475-480
18. Sram, R.J. (1999) Impact of air pollution on reproductive health. (Editorial.) Environ. Health Perspect. 107: A542-A543
19. Phillips, D.H., Castegnaro, M. on behalf of the trial participant (Binkova, B.,Topinka, J.) (1999) Standardization
and validation of DNA adduct postlabelling methods: report of interlaboratory trials and production of recommended protocols. Mutagenesis 14: 301-315
20. Machala, M., Vondracek, J., Ulrich, R., Rubes, J., Sram, R.J. (1999) Evidence for estrogenic and TCDD-like
activities in extracts of blood and semen of men. Organohalogen Compounds 42: 83-86
21. Binkova, B., Strejc, P., Boubelik, O., Stavkova, Z., Chvatalova, I., Sram, R.J. (1999) DNA adducts and human
atherosclerotic lesions. Homeostasis 39: 168-173
22. Novotna, B., Ooms, D., Vilhelmova, M., Vankova, J., Hooghe, R. (1999) DNA fragmentation and changes of
membrane glycosylation in blood cells of chick embryos treated with 5-bromo-2-deoxyuridine during organogenesis. Neoplasma 46: 53-54
23. Zhao, Ch., Vodicka, P., Sram, R.J., Hemminki, K. (2000) Human DNA adducts of 1,3-butadiene, an important
environmental carcinogen. Carcinogenesis 21: 107-111
24. Epstein, H., Dejmek, J., Subrt, P., Vitnerova, N., Sram, R.J. (2000) Trace metals and pregnancy outcome in the
Czech Republic. Environ. Epidemiol. Toxicol. 2: 13-19
25. Machala, M., Ulrich, R., Vondracek, J., Rubes, J., Sram, R.J. (2000) Presence of environmental toxicants in
semen and blood of young men from the Teplice Region, Czech Republic. Environ. Epidemiol. Toxicol. 2: 24-31
26. Sram, R.J., Binkova, B. (2000) Molecular epidemiology studies on occupational and environmental exposure to
mutagens and carcinogens (1997-1999). Environ. Health Perspect. 108 Suppl. 1: 57-70
27. Dejmek, J., Jelinek, R., Solansky, I., Benes, I., Sram, R.J. (2000) Fecundability and parental exposure to
ambient sulfur dioxide. Environ. Health Perspect. 108 Suppl. 1: 647-654
28. Dejmek, J., Solansky, I., Benes, I., Lenicek, J., Sram, R.J. (2000) The impact of polycyclic aromatic
hydrocarbons and fine particles on pregnancy outcome. Environ. Health Perspect. 108: 1159-1164
29. Cerna, M., Pochmanova, D., Pastorkova, A., Benes, I., Lenicek, J., Topinka, J., Binkova, B. (2000) Genotoxicity
of urban air pollutants in the Czech Republic. Part I. Bacterial mutagenic potencies of organic compounds adsorbed on PM10 particulates. Mutation Res. 469: 71-82
30. Topinka, J., Schwarz, L.R., Wiebel, F.J., Cerna, M., Wolff, T. (2000) Genotoxicity of urban air pollutants in the
Czech Republic. Part II. DNA adduct formation in mammalian cell by extractable organic matter. Mutation Res. 469: 83-93
31. Binkova, B., Giguere, Y., Rössner, P.Jr., Dostal, M., Sram, R.J. (2000) The effect of dibenzo[a]pyrene on
human diploid lung fibroblasts: The induction of DNA adducts, expression of p53 and p21WAF1 proteins and cell cycle distribution. Mutation Res. 471: 57-70
32. Begemann, P., Sram, R.J., Neumann, H.G. (2001) Hemoglobin adducts of epoxybutene in workers occupationally
exposed to 1,3-butadiene. Archives Toxicol. 74: 680-687
33. Zhao, Ch., Vodicka, P., Sram, R.J., Hemminki, K. (2001) DNA adducts of 1,3-butadiene in humans: relationships
to exposure, GST genotypes, single-strand breaks, and cytogenetic end points. Environ. Mol. Mutagenesis 37: 226-230
34. Dostal, M., Giguere, Y., Fait, T., Zivny, J., Sram, R.J. (2001) The distribution of major lymphocyte subsets in
cord blood is associated with its pH. Clin.Biochem. 34: 119-124
35. Binkova, B., Strejc, P., Boubelik, O., Stavkova, Z., Chvatalova, I., Sram, R.J. (2001) DNA adducts and human
atherosclerotic lesions. Int. J. Hyg. Occup.Health 204: 49-54
36. Albertini, R.J., Sram, R.J., Vacek, P.M., Lynch, J., Wright, M., Nicklas, J.A., Boogaard, P.J., Henderson, R.F.,
Swenberg, J.A., Tates, A.D., Ward, J.B. (2001) Biomarkers for assessing occupational exposures to 1,3-butadiene. Chem. Biol.
Interact. 135-136: 429-453
37. Sram, R.J. (ed.) (2001) Teplice Program: Impact of Air Pollution on Human Health. Praha: Academia, 318 p.
38. Georgiadis, P., Stoikidou, M., Topinka, J., Kaila, S., Gioka, M., Katsouyanni, K., Sram, R.J., Kyrtopoulos, S.A.
(2001) Personal exposures to PM 2.5 and polycyclic aromatic hydrocarbons and their relationship to environmental tobacco smoke
at two locations in Greece. J.Expos.Anal.Environ.Epidemiol. 11: 169-183
39. Georgiadis, P., Topinka, J., Stoikidou, M., Kaila, S., Gioka, M., Katsouyanni, L., Sram, R.J., Autrup, H.,
Kyrtopoulos, S.A. (2001) Biomarkers of genotoxicity of air pollution (the AULIS project): bulky DNA adducts in
subjects with moderate to low exposures to airborne polycyclic aromatic hydrocarbons and their relationship to
environmental tobacco smoke and other parameters. Carcinogenesis 22: 1447-57
40. Kyrtopoulos, S.A., Georgiadis, P., Autrup, H., Demopoulos, N., Farmer, P., Haugen, A., Katsouyanni, K.,
Lambert, B., Ovrebo, S., Sram, R.J., Stefanou, G., Topinka, J. (2001) Biomarkers of genotoxicity of urban air
pollution. Overview and descriptive data from a molecular epidemiology study on populations exposed to
moderate-to-low levels of polycyclic aromatic hydrocarbons: the AULIS project. Mutation Res. 496: 207-228
41. Dusek, Z., Holejsovska, I., Novotna, B., Zemanova, Z. (2001) Embryotoxicity of 1,2-dibromoethane in chick
embryos in ovo: early and late effects. Eur. J. Morphol. 39/2: 105-112
42. Rubes, J., Vozdova, M., Rezacova, O., Robbins, W.A., Perreault, S.D., Wyrobek, A. (2002) Stable variants of
sperm aneuploidy among healthy men show associations between germinal and somatic aneuploidy. Am. J. Hum. Genet. 70: 1507-1519
43. Dejmek, J., Ginter, E., Solansky, I., Podrazilova, K., Stavkova, Z., Benes, I., Sram, R.J. (2002) Vitamin C, E and
A levels in maternal and fetal blood for Czech and Gypsy ethnic groups in the Czech Republic. Int. J. Vit. Nutr. Res. 72: 183-190
44. Dejmek, J., Solansky, I., Podrazilova, K., Sram, R.J. (2002) The exposure of nonsmoking and smoking mothers
to environmental tobacco smoke during different gestational phases and fetal growth. Environ. Health Perspect. 110: 601-606
45. Hertz-Picciotto, I., Dostal, M., Dejmek, J., Selevan, S., Wegienka, G., Gomez-Caminero, A., Sram, R.J. (2002)
Air pollution and distribution of lymphocyte immunophenotypes in cord and maternal blood at delivery. Epidemiology 13: 172-183
46. Rössner, P. Jr., Binkova, B., Chvatalova, I., Sram, R.J. (2002) Acrylonitrile exposure: the effect on p53 and
p21WAF1 protein levels in the blood plasma of occupationally exposed workers and in vitro in human diploid lung fibroblasts. Mutation Res. 517: 239-250
47. Biros, E., Kalina, I., Kohut, A., Bogyiova, E., Salagovic, J., Sulla, I. (2002) Allelic and haplotype frequencies of
the p53 polymorphisms in brain tumor patients. Phys. Res. 51: 59-64.
48. Biros, E., Kohut, A., Biros, I., Kalina, I., Bogyiova, E., Stubna, J. (2002) A link between the p53 germ line
polymorphisms and white blood cells apoptosis in lung cancer patients. Lung Cancer 35: 231-235
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