ASSOCIATION BETWEEN INHERITED THROMBOPHILIA IN PREGNANCY AND MICRONUCLEUS FREQUENCY IN PERIPHERAL BLOOD LYMPHOCYTES
Šošić GM, Jović N2, Rakić B, Dimitrijević A, Varjačić M
*Corresponding Author: Gordana M. Šošić, B.Sc., Department of Cytogenetic Diagnosis, Obstetrics and Gynecology Clinic, Clinical Center “Kragujevac,” 30 Zmaj Jovina Street, 3400 Kragujevac, Serbia. Tel: +381-63-835-66-24. Fax: +381-34-37-00-73. Email: gordana.sosic.2011.02@gmail.com
page: 5

MATERIALS AND METHODS

Experimental Design and Subjects. The study was designed as a case-control study involving pregnant women admitted to the Department of Obstetrics and Gynecology of our Clinic in 2015. Having been informed, the pregnant women signed the agreement to take part in the study approved by the local Ethics Council (No. 01-12294) and filled in the questionnaire containing basic medical history necessary for research in the field of cytogenetics as well as for evaluations of exposure history. The study included 74 pregnant women of gestational age 11 to 14 weeks. The excluding criteria were the following: exposure to environmental and professional mutagens, exposure to X-ray medical procedures, using oral hormonal contraceptives in the previous year, the presence of other chronic diseases (except thrombophilia), the intake of antibiotics and anti-epileptics during pregnancy and using narcotics. Blood samples from the pregnant women with thrombophilia were taken before starting anticoagulant therapy with LMWH. The pregnant women were grouped according to the value of the MN in the control group [≤4MN/ 1000 binucleated (BN) cells] and the group of cases (>4MN/ 1000BN cells). Blood samples were drawn following the usual procedure and they were kept refrigerated for 24 hours. All the heparinized blood (0.5 mL) was cultured in duplicate in 5 mL complete medium (Gibco® PB-MAX™ Karyotyping Medium; Invitrogen, Carlsbad, CA, USA) and incubated at 37 °C. Cytokinesis-Block Micronucleus Assay. The CBMN assay was performed 44 hours after the cultivation began by adding Cytohalazin B™ (Sigma-Aldrich, St. Louis, MO, USA) to the cultures in final concentration of 4 μg/ mL. After continual incubation of cell cultures for an additional 28 hours, the cells were treated with cold (4 °C) hypotonic (0.56% KC1) solution and fixed three times with fresh Carnoy’s fixative composed of methanol and glacial acetic acid (ZORKA Pharma-HEMIJA d.o.o, Šabac, Serbia) in a ratio of 3:1. The cell material was dripped onto dry and cold microscope slides and the dried slides were colored by 2.0% Giemsa stain solution (BioGnost® d.o.o, Zagreb, Croatia). Micronucleus frequencies were determined by scoring 1000 BN cells per person, according to the criteria previously defined by Fenech et al. [23]. Statistical Data Processing. The entire statistical analysis was performed using the Statistical Package for Social Sciences (SPSS) version 22.0 for Windows software (IBM, Armonk, NY, USA). The results are shown in the tables. To present the results of the categorical variables, absolute values and their percentage distribution were used. The experimental results of continuous numerical values were presented as mean ± standard deviation (SD) values. The χ2 test was applied for determining the differences in the frequency of categorical variables. The significance of the differences between the means of studied variables was tested by the Mann-Whitney and Kruskal-Wallis tests. Binary logistic regression analysis was applied to identify risk factors and to assess the impact of independent variables on the case and control groups. The results are presented as odds ratio (OR) with 95% confidence interval (95% CI) and the p value. Univariate analysis was applied to all the parameters, while multivariate analysis was used for statistically significant parameters. A p value of <0.05 was considered significant.



Number 20
VOL. 20 (2), 2017
Number 20
VOL. 20 (1), 2017
Number 19
VOL. 19 (2), 2016
Number 19
VOL. 19 (1), 2016
Number 18
VOL. 18 (2), 2015
Number 18
VOL. 18 (1), 2015
Number 17
VOL. 17 (2), 2014
Number 17
VOL. 17 (1), 2014
Number 16
VOL. 16 (2), 2013
Number 16
VOL. 16 (1), 2013
Number 15
VOL. 15 (2), 2012
Number 15
VOL. 15, 2012 Supplement
Number 15
Vol. 15 (1), 2012
Number 14
14 - Vol. 14 (2), 2011
Number 14
The 9th Balkan Congress of Medical Genetics
Number 14
14 - Vol. 14 (1), 2011
Number 13
Vol. 13 (2), 2010
Number 13
Vol.13 (1), 2010
Number 12
Vol.12 (2), 2009
Number 12
Vol.12 (1), 2009
Number 11
Vol.11 (2),2008
Number 11
Vol.11 (1),2008
Number 10
Vol.10 (2), 2007
Number 10
10 (1),2007
Number 9
1&2, 2006
Number 9
3&4, 2006
Number 8
1&2, 2005
Number 8
3&4, 2004
Number 7
1&2, 2004
Number 6
3&4, 2003
Number 6
1&2, 2003
Number 5
3&4, 2002
Number 5
1&2, 2002
Number 4
Vol.3 (4), 2000
Number 4
Vol.2 (4), 1999
Number 4
Vol.1 (4), 1998
Number 4
3&4, 2001
Number 4
1&2, 2001
Number 3
Vol.3 (3), 2000
Number 3
Vol.2 (3), 1999
Number 3
Vol.1 (3), 1998
Number 2
Vol.3(2), 2000
Number 2
Vol.1 (2), 1998
Number 2
Vol.2 (2), 1999
Number 1
Vol.3 (1), 2000
Number 1
Vol.2 (1), 1999
Number 1
Vol.1 (1), 1998

 

 


 About the journal ::: Editorial ::: Subscription ::: Information for authors ::: Contact
 Copyright © Balkan Journal of Medical Genetics 2006