Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Archives of Industrial Hygiene and Toxicology

The Journal of Institute for Medical Research and Occupational Health

4 Issues per year

IMPACT FACTOR 2016: 1.395

CiteScore 2016: 1.25

SCImago Journal Rank (SJR) 2016: 0.404
Source Normalized Impact per Paper (SNIP) 2016: 0.721

Open Access
See all formats and pricing
More options …
Volume 63, Issue 3 (Sep 2012)


Chromosomal instability in peripheral blood lymphocytes of patients with reproductive failure assessed by micronucleus assay

Olivera Milošević-Djordjević
  • Faculty of Science, University of Kragujevac, Kragujevac, Serbia
  • Faculty of Medicine, University of Kragujevac, Kragujevac, Serbia
/ Ivana Stošić
  • Faculty of Science, University of Kragujevac, Kragujevac, Serbia
/ Darko Grujičić
  • Faculty of Science, University of Kragujevac, Kragujevac, Serbia
/ Ivanka Zelen
  • Faculty of Medicine, University of Kragujevac, Kragujevac, Serbia
/ Predrag Sazdanović
  • Faculty of Medicine, University of Kragujevac, Kragujevac, Serbia
Published Online: 2012-09-25 | DOI: https://doi.org/10.2478/10004-1254-63-2012-2225

We investigated chromosomal instability in peripheral blood lymphocytes (PBL) of patients with reproductive failure in respect to age, smoking habits, gender, miscarriages, and semen parameters. The study involved 36 individual cases of reproductive failure (18 men and 18 women) attended at the Clinical Centre of Kragujevac, Serbia, and 30 healthy subjects (15 men and 15 women). Micronuclei (MN) frequency was estimated in PBL using the cytokinesis-block micronucleus (CBMN) assay. The baseline MN frequencies were signifi cantly higher (p=0.031; p<0.001) in male [(9.22 ± 4.70) MN per 1000 BN cells] and female patients [(13.50 ± 2.5) MN per 1000 BN cells] than in male and female healthy controls [(6.27 ± 2.66) MN per 1000 BN cells; (6.80 ± 2.98) MN per 1000 BN cells]. The mean baseline MN frequency did not signifi cantly differ between miscarriage groups and between patients with and without normal values of semen parameters. The correlations between poor sperm concentration (<20x106 mL-1), rapid progressive motility (<25 %), normal morphology (<30 %), and MN frequencies were negative, but not statistically signifi cant. We found that only gender signifi cantly infl uenced the MN rates in analysed patients. There were no signifi cant differences between age groups and between smokers and non-smokers in patients and control samples. We conclude that the increase in baseline MN frequency in PBL of patients with reproductive failure corresponds to the increase in chromosomal damage, which occurs as a result of complex events that cause reproductive disorders.

KEYWORDS: age; gender; infertility; micronuclei; semen quality; smoking

  • 1. Layman LC. Human gene mutations causing infertility. J Med Genet 2002;39:153-61.CrossrefPubMedGoogle Scholar

  • 2. Poongothai J, Gopenath TS, Manonayaki S. Genetics of human male infertility. Singapore Med J 2009;50:336-47.PubMedGoogle Scholar

  • 3. Malik A, Jain S, Hakim S, Shukla I, Rizvi M. Chlamydia trachomatis infection & female infertility. Indian J Med Res 2006;123:770-5.Google Scholar

  • 4. Homan GF, Davies M, Norman R. The impact of lifestyle factors on reproductive performance in the general population and those undergoing infertility treatment: a review. Hum Reprod Update 2007;13:209-23.PubMedCrossrefGoogle Scholar

  • 5. Dhont N, van de Wijgert J, Luchters S, Muvunyi C, Vyankandondera J, Temmerman M. Sexual violence, HSV- 2 and HIV are important predictors for infertility in Rwanda. Hum Reprod 2010;25:2507-15.CrossrefWeb of ScienceGoogle Scholar

  • 6. Hristova R, Ko E, Greene C, Rademaker A, Chernos J, Martin R. Chromosome abnormalities in sperm from infertile men with asthenoteratozoospermia. Biol Reprod 2002;66:1781-3.CrossrefPubMedGoogle Scholar

  • 7. Schmid TE, Brinkworth MH, Hill F, Sloter E, Kamischke A, Marchetti F, Nieschlag E, Wyrobek AJ. Detection of structural and numerical chromosomal abnormalities by ACM-FISH analysis in sperm of oligozoospermic infertility patients. Hum Reprod 2004;19:1395-400.CrossrefPubMedGoogle Scholar

  • 8. Nagvenkar P, Desai K, Hinduja I, Zaveri K. Chromosomal studies in infertile men with oligozoospermia & non - obstructive azoospermia. Indian J Med Res 2005;122:34-42.Google Scholar

  • 9. Ceylan GG, Ceylan C, Elyas H. Genetic anomalies in patients with severe oligozoospermia and azoospermia in eastern Turkey: a prospective study. Genet Mol Res 2009;8:915-22.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 10. Schmid TE, Kamischke A, Bollwein H, Nieschlag E, Brinkworth MH. Genetic damage in oligozoospermic patients detected by fluorescence in-situ hybridization, inverse restriction site mutation assay, sperm chromatin structure assay and the Comet assay. Hum Reprod 2003;18:1474-80.CrossrefPubMedGoogle Scholar

  • 11. Shamsi MB, Venkatesh S, Tanwar M, Singh G, Mukherjee S, Malhotra N, Kumar R, Gupta NP, Mittal S, Dada R. Comet assay: a prognostic tool for DNA integrity assessment in infertile men opting for assisted reproduction. Indian J Med Res 2010;131:675-81.Google Scholar

  • 12. Duzcan F, Aybek Z, Tepeli E, Caner V, Cetin GO, Aybek H, Bagci H. Sex chromosome aneuploidy rates in the somatic cells of infertile men. J Reprod Med 2006;51:489-92.PubMedGoogle Scholar

  • 13. Yesilada E, Sahin I, Ozcan H, Yildirim IH, Yologlu S, Taskapan C. Increased micronucleus frequencies in peripheral blood lymphocytes in women with polycystic ovary syndrome. Eur J Endocrinol 2006;154:563-8.CrossrefPubMedGoogle Scholar

  • 14. Moran LJ, Noakes M, Clifton PM, Norman RJ, Fenech MF. Genome instability is increased in lymphocytes of women with polycystic ovary syndrome and is correlated with insulin resistance. Mutat Res 2008;639:55-63.Web of ScienceGoogle Scholar

  • 15. Trkova M, Kapras J, Bobkova K, Stankova J, Mejsnarova B. Increased micronuclei frequencies in couples with reproductive failure. Reprod Toxicol 2000;14:331-5.CrossrefPubMedGoogle Scholar

  • 16. Ishikawa H, Tian Y, Yamauchi T. Infl uence of gender, age and lifestyle factors on micronuclei frequency in healthy Japanese populations. J Occup Health 2003;45:179-81.CrossrefGoogle Scholar

  • 17. Minozzo R, Deimling LI, Gigante LP, Santos-Mello R. Micronuclei in peripheral blood lymphocytes of workers exposed to lead. Mutat Res 2004;565:53-60.Google Scholar

  • 18. Pedersen M, Vinzents P, Petersen JH, Kleinjans JC, Plas G, Kirsch-Volders M, Dostal M, Rossner P, Beskid O, Sram RJ, Merlo DF, Knudsen LE. Cytogenetic effects in children and mothers exposed to air pollution assessed by the frequency of micronuclei and fl uorescence in situ hybridization (FISH): A family study in the Czech Republic. Mutat Res 2006;608:112-20.Google Scholar

  • 19. Bonassi S, Znaor A, Ceppi M, Lando C, Chang WP, Holland N, Kirsch-Volders M, Zeiger E, Ban S, Barale R, Bigatti MP, Bolognesi C, Cebulska-Wasilewska A, Fabianova E, Fucic A, Hagmar L, Joksic G, Martelli A, Migliore L, Mirkova E, Scarfi MR, Zijno A, Norppa H, Fenech M. An increased micronucleus frequency in peripheral blood lymphocytes predicts the risk of cancer in humans. Carcinogenesis 2007;28:625-31.Web of SciencePubMedGoogle Scholar

  • 20. 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.Google Scholar

  • 21. Fenech M. Cytokinesis-block micronucleus cytome assay. Nat Protoc 2007;2:1084-104.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 22. Botto N, Rizza A, Colombo MG, Mazzone AM, Manfredi S, Masetti S, Clerico A, Biagini A, Andreassi MG. Evidence for DNA damage in patients with coronary artery disease. Mutat Res 2001;493:23-30.Google Scholar

  • 23. Petrozzi L, Lucetti C, Scarpato R, Gambaccini G, Trippi F, Bernardini S, Del Dotto P, Migliore L, Bonuccelli U. Cytogenetic alterations in lymphocytes of Alzheimer’s disease and Parkinson’s disease patients. Neurol Sci 2002;23(Suppl 2):S97-8.CrossrefGoogle Scholar

  • 24. Milošević-Djordjević O, Grujičić D, Vasković Z, Marinković D. High micronucleus frequency in peripheral blood lymphocytes of untreated cancer patients irrespective of gender, smoking and cancer sites. Tohoku J Exp Med 2010;220:115-20.Web of ScienceGoogle Scholar

  • 25. Fenech M. Micronuclei and their association with sperm abnormalities, infertility, pregnancy loss, pre-eclampsia and intra-uterine growth restriction in humans. Mutagenesis 2011;26:63-7.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 26. De Palma A, Burrello N, Barone N, D’Agata R, Vicari E, Calogero AE. Patients with abnormal sperm parameters have an increased sex chromosome aneuploidy rate in peripheral leukocytes. Hum Reprod 2005;20:2153-6.CrossrefGoogle Scholar

  • 27. Fenech M. Important variables that influence base-line micronucleus frequency in cytokinesis-blocked lymphocytesa biomarker for DNA damage in human populations. Mutat Res 1998;404:155-65.Google Scholar

  • 28. Mahrous HS. Effects of age, gender, folate and vitamin B12 status on DNA damage. Bull Alex Fac Med 2009;45:195-200.Google Scholar

  • 29. Bolognesi C, Perrone E, Landini E. Micronucleus monitoring of a fl oriculturist population from western Liguria, Italia. Mutagenesis 2002;17:391-7.CrossrefGoogle Scholar

  • 30. Catalan J, Autio K, Kuosma E, Norppa H. Age-dependent inclusion of sex chromosomes in lymphocyte micronuclei of man. Am J Hum Genet 1998;63:1464-72.CrossrefPubMedGoogle Scholar

  • 31. Bonassi S, Neri M, Lando C, Ceppi M, Lin YP, Chang WP, Holland N, Kirsch-Volders M, Zeiger E, Fenech M; HUMN collaborative group. Effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the Human MicroNucleus project. Mutat Res 2003;543:155-66.Google Scholar

  • 32. Hessel H, Radon K, Pethran A, Maisch B, Grobmair S, Sautter I, Fruhmann G. The genotoxic risk of hospital, pharmacy and medical personnel occupationally exposed to cytostatic drugs - evaluation by the micronucleus assay. Mutat Res 2001;497:101-9.Google Scholar

  • 33. Costa C, Teixeira JP, Silva S, Roma-Torres J, Coelho P, Gaspar J, Alves M, Laffon B, Rueff J, Mayan O. Cytogenetic and molecular biomonitoring of a Portuguese population exposed to pesticides. Mutagenesis 2006;21:343-50.CrossrefPubMedGoogle Scholar

  • 34. Hando JC, Tucker JD, Davenport M, Tepperberg J, Nath J. X chromosome inactivation and micronuclei in normal and Turner individuals. Hum Genet 1997;100:624-8.CrossrefPubMedGoogle Scholar

  • 35. Garaj-Vrhovac V, Durinec M, Kopjar N, Oreščanin V. A survey on the cytogenetic status of the Croatian general population by use of the cytokinesis-block micronucleus assay. Mutat Res 2008;649:91-100.Web of ScienceGoogle Scholar

  • 36. Milošević-Djordjević O, Grujičić D, Marinković D, Arsenijević S, Banković S. Effect of various doses of gestogens on micronuclei frequency in human peripheral blood lymphocytes of pregnant women. Hum Reprod 2003;18:433-6.CrossrefGoogle Scholar

  • 37. Grujičić D, Milošević-Djordjević O, Marinković D, Arsenijević S, Banković S, Živanović A, Dimitrijević A. Effect of gestagens on micronucleus frequency in human peripheral blood lymphocytes. Balk J Med Genet 1999;2:23-8.Google Scholar

About the article

Published Online: 2012-09-25

Published in Print: 2012-09-25

Citation Information: Archives of Industrial Hygiene and Toxicology, ISSN (Print) 0004-1254, DOI: https://doi.org/10.2478/10004-1254-63-2012-2225.

Export Citation

This content is open access.

Comments (0)

Please log in or register to comment.
Log in