Genotoxic assessment in peripheral blood lymphocytes of post-polio individuals using sister chromatid exchange analysis and micronucleus assay

Abstract

Environmental pollution is a complex issue because of the diversity of anthropogenic agents, both chemical and physical, that have been detected and catalogued. The consequences to biota from exposure to genotoxic agents present an additional problem because of the potential for these agents to produce adverse change at the cellular and organism levels. Past studies in virus have focused on structural damage to the DNA of environmental species that may occur after exposure to genotoxic agents and the use of this information to document exposure and to monitor remediation. In an effort to predict effects at the population, community and ecosystem levels, in the present study, we attempt to characterize damage occurring through genotoxic agents like 5-bromo-2-deoxyuridine, BrdU, using sister chromatid exchange technique and the formation of micronuclei (MN) in the peripheral lymphocytes of the post-polio syndrome sequelae affected by poliovirus. Analysis of structural chromosomal aberrations (CAs) and involvement of the specific chromosome break were pursued in this study. They revealed a significantly higher incidence of CAs (chromatid and chromosome breaks) in patients compared with controls, where the specific chromosome break has emerged as specific. Also, the maximum numbers of breaks were found to be in chromosome 1 at the position 1p36.1. The results also suggest a correlation between CAs and content of MN.

Keywords

post-polio syndrome chromosomal aberrations sister chromatid exchange micronucleus cytochalasin B 5-bromo-2-deoxyuridine

References

Harnden DG Cytogenetic studies on patients with virus infections and subjects vaccinated against yellow fever. Am J Hum Genet 1964; 16: 204-213.

Fortunato EA , Dell’Aquila ML , and Spector DH Specific chromosome 1 breaks induced by human cytomegalovirus. Proc Natl Acad Sci USA 2000; 97: 853-858.

Carrano AV , Minkler JL , Stetka DG , and Moore DH Variation in the baseline sister chromatid exchange frequency in human lymphocytes . Environ Mutagen 1980; 2: 325-337.

Glenn GM , Ricciardi RP Adenovirus 5 early region 1A host range mutants hr3, hr4 and hr5 contain point mutations which generate single amino acid substitution. J Virol 1985; 56: 66-74.

Ragonese P. , Fierro B. , and Salemi G. Prevalence and risk factors of post-polio syndrome in a cohort of polio survivors . J Neurol Sci 2005; 236: 31-35.

Bhatnagar A. , Rani R. , and Ghosh PK Chromosome aberrations and sister-chromatid exchanges (SCEs) in peripheral blood lymphocytes of patients suffering from poliomyelitis. Mutat Res 1984; 141: 55-58.

Latt SA Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc Natl Acad Sci USA 1973; 70: 3395-3399.

Lin MS , Comings DE , and Alfi OS Optical studies of the interaction of 4’-6-diamidino 2-phenylindole with DNA and metaphase chromosomes. Chromosoma 1977; 60: 15-25.

Kato H. Spontaneous sister chromatid exchange s detected by BudR-labelling method . Nature 1974; 251: 70-72.

10.

Huang CC Induction of a high incidence of damage to the X chromosomes of Rattus (Mastomys) natalensis by base analogues, viruses & carcinogens. Chromosoma 1967; 23: 162-179.

11.

Zakharov AF , Egolina NA Differential spiralization among mammalian mitotic chromosomes. I. BUdRrevealed differentiation in Chinese hamster chromosomes . Chromosoma 1972; 38: 341-355.

12.

Kim MA Chromatid Austausch und Heterochromatin Veranderungen menschlicher Chromosomen nach BrdU-Markierang. Humangenetik 1974; 25: 179-188.

13.

Goto K. , Akematsu T. , Shimazu H. , and Sugiyama T. Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining. Chromosoma 1975; 53: 223-230.

14.

Goto K. , Maeda S. , Kano Y. , and Sugiyama T. Factors involved in differential Giemsa staining of sister chromatids. Chromosoma 1978; 66: 351-359.

15.

Scheres JJ , Hustinx T. , Ruttem FJ , and Merkx G. ‘‘Reverse’’ differential staining of sister chromatids . Exp Cell Res 1977; 109: 466-468.

16.

Burkholder GD Reciprocal Giemsa staining of late DNA replicating regions produced by low and high pH sodium phosphate. Exp Cell Res 1978; 111: 489-492.

17.

Sakanishi S. , Takayama S. Reverse differential staining of sister chromatid after substitution with BUdR and incubation in sodium phosphate solution. Exp Cell Res 1978; 115: 448-450.

18.

Takayama S. , Sakanishi S. Differential Giemsa staining of sister chromatids after extraction with acids. Chromosoma 1977 ; 64: 109-115.

19.

Peace BE , Succop P.

Spontaneous micronucleus frequency and age: what are normal values?

Mutat Res 1999; 425: 225-230.

20.

Countryman PI , Heddle JA The production of micro-nuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res 1976; 41: 321-332.

21.

Perry P. , Wolff S. New Giemsa method for differential staining of sister chromatids. Nature 1974; 261: 156-158.

22.

S. Karger. An international system for Human Cytogenetic Nomenclature , Mitelman F. ed. Basel : ISCN 1995.

23.

Mariani T. Fragile sites and statistics. Hum Genet 1989 ; 81: 319-322.

24.

Fenech M. The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations. Mutat Res 1993; 285: 35-44.

25.

Eastmond DA , Tucker JD Identification of aneuploidyinducing agents using cytokinesis-blocked human lymphocytes and an antikinetochore antibody. Environ Mol Mutagen 1989; 13: 34-43.

26.

Kligerman AD , Doerr CL , Tennant AH , and Zucker RM Cytogenetic studies of three triazine herbicides. I. In vitro studies. Mutat Res 2000; 465: 53-59.

27.

Jubelt B. , Agre JC Characteristics and management of post polio Syndrome. JAMA 2000; 284: 412-414.

28.

Aya T. , Makino S. Notes on chromosome abnormalities in cultured leucocytes from serum hepatitis patients. Proc Jpn Acad 1966; 42: 270-274.

29.

Bartsch HD , Habermehl KO , and Diefenthal W. Chromosomal damage after infection with poliomyelitis virus. Exp Cell Res 1967; 48: 671-675.

30.

Korenberg JR , Freedlender E. Giemsa technique for the detection of sister chromatid exchanges. Chromosoma 1974; 483: 55-360.

31.

Gratzner H. , Pollack A. , Ingram D. , and Lief R. Deoxyribonucleic acid replication in single cells and chromosomes by immunologic techniques. J Histochem Cytochem 1976; 24: 34-39.