Eur J Endocrinol
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

DOI: 10.1530/eje.1.02117
European Journal of Endocrinology, Vol 154, Issue 4, 563-568
Copyright © 2006 by European Society of Endocrinology
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (2)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yesilada, E.
Right arrow Articles by Taskapan, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yesilada, E.
Right arrow Articles by Taskapan, C.
CLINICAL STUDY

Increased micronucleus frequencies in peripheral blood lymphocytes in women with polycystic ovary syndrome

Elif Yesilada1, Ibrahim Sahin2, Hamdi Ozcan3, Ibrahim Halil Yildirim1, Saim Yologlu4 and Cagatay Taskapan5

1 Departments of Medical Biology and Genetics, 2 Endocrinology and Metabolism, 3 Dermatology, 4 Biostatistics and 5 Biochemistry, Faculty of Medicine, Inonu University, 44280, Malatya, Turkey

(Correspondence should be addressed to E Yesilada; Email: eyesilada{at}inonu.edu.tr)


   Abstract
Top
 Abstract
Introduction
 Materials and methods
 Results
 Discussion
 References
 
Objective: We aimed to assess possible genomic instability in women with polycystic ovary syndrome (PCOS).

Design: The frequency of micronuclei in cultured peripheral lymphocytes was used as a biomarker of genomic instability in somatic cells.

Methods: Nineteen women, diagnosed with PCOS and 19 healthy female volunteers of corresponding ages and body-mass index (BMI) were included in the study. Micronuclei frequencies were assessed in cytokinesis-blocked lymphocytes.

Results: The frequency of micronucleated cells (per thousand) was 9.00 (5.00) (interquartile range in parentheses) for patient group and 3.0 (3.0) for the control group (P < 0.0001, Mann-Whitney U-test). The serum levels of follicle-stimulating hormone (FSH), estradiol, prolactin, glucose and dehydroepiandrosterone sulfate (DHEAS) and the homeostasis model of assessment of insulin resistance (HOMA-IR) were not different between the two groups (P > 0.05). Serum total testosterone, luteinizing hormone (LH) and insulin levels and hirsutism score in the PCOS group were significantly (P = 0.007, P < 0.0001, P = 0.009 and P < 0.0001 respectively) higher than those of the control group (2.3 (2.1) nmol/l vs 1.7 (0.4) nmol/l; 8.5 (5.88) mU/ml vs 4.8 (4.4) mU/ml; 6.8 (5.1) µU/ml vs 9.7 (4.2) µU/ml; 19.5 (6.5) vs 4.0 (2.5) respectively). However, the mean level of sex hormone-binding globulin (SHBG) in PCOS group was significantly (P = 0.004) lower than in control group (36.4(22.6) nmol/l vs 48.6(25.2) nmol/l respectively).

Conclusion: These findings suggest that women with PCOS have a high incidence of genomic instability, and this condition is positively correlated with the hirsutism score, BMI, LH and serum total testosterone and insulin levels, and is negatively correlated with SHBG.


   Introduction
Top
 Abstract
 Introduction
Materials and methods
 Results
 Discussion
 References
 
Polycystic ovary syndrome (PCOS), characterized by hyperandrogenism and chronic anovulation, is a common endocrine disorder in women. PCOS, which is associated with polycystic ovaries, hirsutism, obesity and insulin resistance, is a leading cause of female infertility (1). Recently, it has been suggested that women with PCOS show higher levels of leukocyte count, a marker of low-grade inflammation and cardiovascular risk, than controls (2). Epidemiologic studies have demonstrated that the prevalence of PCOS in women is 3.5–10%, depending on the diagnosis criterion (3, 4).

The genetic mechanisms underlying PCOS remain largely unknown (5). Although an autosomal dominant model was proposed (6), later studies did not confirm this. Chromosomal studies in patients with PCOS have produced contradictory findings. There have been reports that X-chromosome aneuploidies and polyploidies (7), as well as other cytogenetic abnormalities (8), are associated with PCOS in a limited number of subjects. One well-characterized case had both endocrine and ultrasound stigmata of PCOS (as well as multiple other anomalies, including heart and facial dysmorphies) and a large deletion on the long arm of the chromosome 11 (9). However, other cytogenetic analyses have failed to identify common karyotypic abnormalities (10).

A number of studies reported that PCOS is an oligogenic disorder, and more studies are necessary to define its genetic basis (5, 11). Different combinations of multiple gene polymorphisms and environmental factors explain the heterogeneity of PCOS (5).

This study aimed to assess, by cytokinesis-blocked micronucleus (CBMN) assay, possible chromosomal instability in women with PCOS. The micronucleus (MN) frequencies in cultured peripheral blood lymphocytes have been used as biomarkers of chromosome damages for many years. MN are cytoplasmic chromatin masses with the appearance of small nuclei that arise from chromosome fragments or intact whole chromosomes lagging behind at the anaphase stage of cell division. Their presence in cells reflects structural and/or numerical chromosomal aberrations arising during mitosis (12–15). The quantification of MN for the evaluation of chromosome damage offers several advantages: the method is simple and fast, and it dose not require the presence of metaphasic cells. Another advantage of the MN assay is the relative ease of scoring and the statistical power obtained from scoring larger numbers of cells than are typically used for meta-phase analysis (14). Fenech and Morley (12) presented a CBMN assay that resolved earlier problems experienced with differences in cell growth by monitoring cell division with the aid of cytochalasin B. This agent allows division of the cell nucleus (karyokinesis), but prevents cell division (cytogenesis), thus resulting in binucleated cells (12–14).


   Materials and methods
Top
 Abstract
 Introduction
 Materials and methods
Results
 Discussion
 References
 
The study comprised 19 women newly diagnosed with PCOS, admitted to endocrinology and metabolism clinics, and 19 control subjects with similar age and body-mass index (BMI). The procedures used were in accordance with the guidelines of the Helsinki Declaration on human experimentation. The study protocol was approved by the ethics committee of Inonu University (Faculty of Medicine). All the human subjects were fully informed and gave written, informed consent.

PCOS was diagnosed by the criteria revised in 2003 (16). Patients who had two or more of the following criteria were defined as having PCOS:

  1. history of oligo- and/or anovulation in reproductive years (<6 menses per year, not in the last 2 years prior to menopause)
  2. clinical and/or biochemical signs of hyperandrogenism: hirsutism score of >6 and/or high total testosterone level
  3. typical ovarian histopathology of polycystic ovaries on ultrasound: multiple follicles in each ovary measuring 2–9 mm in diameter and/or increased ovarian volume (>10 ml).

Other causes of hyperandrogenism, such as hyperprolactinemia, Cushing’s syndrome, androgen-secreting tumors and congenital adrenal hyperplasia, were excluded by past medical history, physical examination and specific laboratory analysis. Moreover, 17-OH progesterone and 17-OH progesterone responses to adrenocorticotropic hormone (ACTH) stimulation test were evaluated in some suspected cases to exclude congenital adrenal hyperplasia.

Hirsutism score was evaluated by a single examiner (dermatologist) by the Ferriman–Gallwey score (FGS), as modified by Hatch et al. (17).

Blood samples were drawn during the midfollicular phase (days 3–7) of the menstrual cycle after overnight fasting. Serum total testosterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, dehydroepiandrosterone sulfate (DHEAS), prolactin, sex-hormone binding globulin (SHBG) and insulin levels were measured by an automated chemiluminescence system (Immulite 2000; Diagnostic Product Cooperation, Los Angeles, CA, USA). Insulin resistance was calculated by the homeostasis method of assessment of insulin resistance (HOMA-IR) (18).

Micronucleus assay

Peripheral blood samples were collected by a heparinized sterile injector. Whole blood (0.5 ml) was added to 5 ml RPMI-1640 medium (Sigma, R0883) with 25% fetal bovine serum (Sigma, F7525), 1% L-glutamine (Sigma, G7513) and 2% phytohemagglutinin (Biological Industries, Kibbutz Beit Haemek, Israel). CBMN assay was carried out by the method of Fenech (13). Cytochalasin B (Serva, 18015, Heidelberg, Germay) was added to the cultures at a final concentration of 6 µg/ml after 44-h incubation stimulation with phytohemagglutinin (14). Cells were harvested after 72-h incubation, and they were treated with a hypotonic solution (0.075 M KCl) for 1 min and fixed in fresh fixative solution (methanol: acetic acid, 3:1). Slides were air-dried and stained with Giemsa. Micro-nucleated cells were analyzed under light-microscopy (x 400) by scoring 1000 binucleated lymphocytes, in which the number of MN was recorded by standard recognition criteria (13). Briefly, MN were 1. morphologically identical to the main nuclei but their diameters were from 1/16 to 1/3 of the mean diameter of the main nuclei; 2. nonrefractile; 3. not linked to the main nuclei.

Subjects with possible confounding factors (such as exposure to physical and chemical mutagens, history of alcohol and coffee consumption or smoking, medication, viral infections suffered in the last 3 months, vaccination, hereditary diseases) that potentially play a role in the induction or expression of MN (7–9) was not included in the study.

Statistical analysis

Statistical analyses were performed with SPSS for Windows, Version 13.0 (SPSS Inc., Chicago, IL, USA). Data are presented as median and interquartile range. All parameters were tested by the Shapiro–Wilks normality test, and its distribution was not normal (P < 0.05). Therefore, statistical differences between groups were determined with the nonparametric Mann–Whitney U-test. Bivariate correlations were performed by calculating the Spearman coefficient. The level of P < 0.05 was considered to be statistically significant (two tails).


   Results
Top
 Abstract
 Introduction
 Materials and methods
 Results
Discussion
 References
 
The results of MN analysis and biochemical characteristics for healthy controls and women with PCOS are given in Table 1Go. There were no statistically significant (P > 0.05) differences between groups, according to anthropometrics parameters (age, BMI and abdominal circumference). As expected, PCOS patients had significantly (P < 0.05) higher hirsutism score. The levels of serum total testosterone, LH and basal insulin were also significantly (P = 0.007, P < 0.001 and P = 0.009 respectively) higher in patients than in controls, whereas the levels of FSH, estradiol, prolactin and glucose were similar (P > 0.05) in the two groups. The level of SHBG was significantly (P = 0.004) lower than control values. HOMA-IR levels were similar (P > 0.05) in patients and controls.


View this table:
[in this window]
[in a new window]
  		Table 1 The frequency of MN in peripheral blood and biochemical characteristics for healthy controls and women with PCOS.
 
Finally, micronucleated cell rates per 1000 binucleated lymphocytes were significantly (P < 0.0001) higher in women with PCOS than in controls. A micro-nucleated lymphocyte is shown in Fig. 1Go. There were positive correlations between micronucleated cell rate and hirsutism score (r = 0.501, P = 0.001), BMI (r = 0.343, P = 0.035), LH (r = 0.501, P < 0.0001) and total testosterone (r = 0.431, P = 0.005). In addition, there was negative correlation between micronucleated cell rate and level of SHBG (r = –0.397, P = 0.014).


Figure 1
View larger version (88K):
[in this window]
[in a new window]
  		Figure 1 (A) Binucleated lymphocyte; (B) micronucleated binucleated lymphocyte. 1000X

 

   Discussion
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
References
 
In the past decade, the association between hyperandrogenism, hyperinsulinemia and PCOS was demonstrated (19–21). Our findings, summarized in Table 1Go, are consistent with those of the previous studies, except for HOMA-IR. Although the HOMA-IR score increased in the PCOS group, this difference was not statistically significant (P = 0.089), possibly because of the small number of individuals investigated.

One of the most promising methods to assess DNA damage is the CBMN assay, which detects both chromosome and genome alterations in binucleated cells (13). Owing to the direct correlation between MN production and genomic damage, the CBMN assay applied to human blood peripheral lymphocytes may be considered a reliable method to quantify genome-induced chromosome damage and/or genomic instability (12–15). To the best of our knowledge, this is the first study to investigate MN frequencies in lymphocytes of woman with PCOS.

Spontaneous or baseline MN frequencies in cultured human lymphocytes and exfoliated cells provide an index of accumulated genetic damage occurring during the life span of these cells. For the application of the MN test as a cytogenetic marker of chromosomal damage, the background level in the ‘normal’ population must be specified. Micronucleated cell levels observed in this study averaged 3.0 (3.0) and are consistent with previous findings that micronucleated cell rates in peripheral lymphocytes vary from 1.56 to 11.10 (22–30). The difference between the lower and upper levels in the control group shows that interindividual variability is observed for MN levels in lymphocytes. Interindividual variation may be explained by lifestyle factors, including environmental exposure, or individual susceptibility factors (31).

MN frequencies in peripheral blood lymphocytes were significantly (P < 0.0001) higher in women with PCOS than in controls. This result agrees with some of the reports dealing with this issue (7–9, 32). These reports suggest that genetic abnormality is present in women with PCOS. In our study, hyperandrogenism and hyperinsulinemia in PCOS patients might also play a central role in increased MN frequencies in peripheral blood lymphocytes. In translating our results into clinical practice, one must keep in mind that our study population consisted of a relatively small number of patients.

Hando et al. (33) showed the X chromosome to be present in 72.2% of the MN that were scored. Although we did not analyze the contents of the MN found in subjects, it has been described that both the X chromosome and autosomes are responsible for the higher rate of MN in peripheral lymphocytes in women (34).

If a subfertile population is reviewed, PCOS is diagnosed in about 75% of patients with anovulatory infertility, and high prevalence rates of PCOS have been reported among women with recurrent miscarriages (35, 36). Balen et al. (37) reported a high prevalence of primary (46%) and secondary (26%) infertility in a large group of PCOS patients. In addition, a higher incidence of chromosomal instability in the infertile population is widely recognized (27). Trkova et al. (27) showed that MN frequencies are increased in couples with infertility or with two or more spontaneous abortions.

Oxidative stress that arises due to an imbalance between generation of reactive oxygen species (ROS) and antioxidant defense has been linked to a number of disease states, including cardiovascular disease, aging and various cancers (38,39). Similarly, increased oxidative stress and decreased antioxidant capacity were reported in women with PCOS (40, 41). Protein, lipids, RNA and DNA all undergo continual damage by ROS, produced either as a result of normal cellular metabolism or as a result of other types of oxidative stresses. DNA damage is generally in the form of mutation or strand breakages (38). In addition, it was documented that oxidative stress induces chromosomal breakage and formation of bone-marrow MN (39). Furthermore, some authors have shown a positive correlation between the extent of lipid peroxidation and genotoxicity reflected by increased MN formation(42–45).

It is generally accepted that PCOS is associated with an increased risk of cardiovascular disease (41, 46, 47). Orio et al. (47) have shown the detrimental effect of PCOS on the cardiovascular system, even in young women asymptomatic for cardiac disease. Another study (28) demonstrated that the frequency of MN in peripheral blood is increased in patients with coronary artery disease.

Women with PCOS are also thought to be at increased risk of cancer (11, 48, 49). It is noteworthy that some investigators have shown that in lymphocytes the MN levels are twofold higher in cancer patients than in corresponding healthy individuals (50, 51). Our finding of increased MN frequency in PCOS may contribute to a better understanding of endogenous mutation process and thus determination of cancer risk in these patients.

In conclusion, this study demonstrated an association between increased micronucleus frequency and PCOS. Our findings suggest that there is genetic instability in peripheral blood lymphocytes in women with PCOS. We suggest that hyperandrogenism, hyperinsulinemia and perhaps oxidative stress are factors contributing to increased MN frequency and chromosomal damage.


   Acknowledgements
 
We thank all participants who volunteered for this study and Dr Hikmet Geckil for his critical reading of the manuscript. This study was presented orally at the European Congress of Endocrinology (2005) in Göteborg, Sweden.


   References
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 References
 

    1. Franks S. Polycystic ovary syndrome. New England Journal of Medicine 1995 333 853–861.[Free Full Text]

    2. Orio F Jr, Palomba S, Cascella T, Di Biase S, Manguso F, Tauchmanova L, Nardo LG, Labella D, Savastano S, Russo T, Zullo F, Colao A & Lombardi G. The increase of leukocytes as a new putative marker of low-grade chronic inflammation and early cardiovascular risk in polycystic ovary syndrome. Journal of Clinical Endocrinology and Metabolism 2005 90 2–5.[Abstract/Free Full Text]

    3. Knochenhauer ES, Key TJ, Kashar-Miller M, Waggoner W, Boots LR & Aziz R. Prevalence of the polycystic ovary syndrome in unselected black and white women of the southeastern United States; a prospective study. Journal of Clinical Endocrinology and Metabolism 1998 83 3078–3082.[Abstract/Free Full Text]

    4. Asuncion M, Calvo RM, San Millan JL, Sancho J, Avila S & Escobar-Morreale HF. A prospective study of the prevalence of the polycystic ovary syndrome in unselected Caucasian women from Spain. Journal of Clinical Endocrinology and Metabolism 2000 85 2434–2438.[Abstract/Free Full Text]

    5. Escobar-Morreale HF, Luque-Ramirez M & San Millan JL. The molecular-genetic basis of functional hyperandrogenism and the polycystic ovary syndrome. Endocrine Reviews 2005 26 251–282.[Abstract/Free Full Text]

    6. Amato P & Simpson JL. The genetics of polycystic ovary syndrome. Best Practice and Research. Clinical Obstetrics and Gynaecology 2004 18 707–718.

    7. Rojanasakul A, Gustavson KH, Lithell H & Nillius SJ. Tetraploidy in two sisters with the polycystic ovary syndrome. Clinical Genetics 1985 27 167–174.[ISI][Medline]

    8. Nur J, Grewal MS, Guron CJ & Buckshee K. C-band polymorphism of chromosome no. 1 in patients with polycystic ovary disease.Asia Oceania Journal of Obstetrics and Gynaecology 1987 13 75–78.

    9. Meyer MF, Gerresheim F, Pfeiffer A, Epplen JT & Schatz H. Association of polycystic ovary syndrome with an interstitial deletion of the long arm of chromosome 11. Experimental and Clinical Endocrinology and Diabetes 2000 108 519–523.

    10. Stenchever MF, Gerresheim F, Pfeffer A, Epplen JT & Schatz H. Cytogenetic evaluation of 41 patients with Stein–Leventhal syndrome. Obstetrics and Gynecology 1968 32 797–801.

    11. Xita N, Georgiou I & Tsatsoulis A. The genetic basis of polycystic ovary syndrome. European Journal of Endocrinology 2002 147 717–725.[Abstract]

    12. Fenech M & Morley AA. Measurement of micronuclei in lymphocytes. Mutation Research 1985 147 29–36.[CrossRef][ISI][Medline]

    13. Fenech M. The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations. Mutation Research 1993 285 35–44.[ISI][Medline]

    14. Fenech M, Holland N, Chang WP, Zeiger E & Bonnasi S. The Human Micronucleus Project – an international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans. Mutation Research 1999 428 271–283.[ISI][Medline]

    15. Kirsch-Volders M, Sofuni T, Aardema M, Albertini S, Eastmond D, Fenech M, Ishidate M Jr, Kirchner S, Lorge E, Morita T, Norppa H, Surralles J, Vanhauwaert A & Wakata A. Report from the in vitro micronucleus assay working group. Mutation Research 2003 540 153–163.[ISI][Medline]

    16. Rotterdam Eshe/ASRM-Sponsored PCOS Consensus Workshop Group. Revised 2003 consensus on diagnostic criteria and long term health risks related to polycystic ovary syndrome. Human Reproduction 2004 19 41–47.[Abstract/Free Full Text]

    17. Hatch R, Rosenfield R, Kim MH & Tredway D. Hirsutism implications, etiology and management. American Journal of Obstetrics and Gynecology 1981 10 815–830.

    18. Matthews DR, Hoske JP, Rudenski AS, Naylor BA, Treacher DF & Turner RC. Homeostasis model of assessment: insulin resistance and cell function from fasting plasma glucose and insulin concentration in man. Diabetologia 1985 28 412–419.[CrossRef][ISI][Medline]

    19. Rosenfield RL. Ovarian and adrenal function in polycystic ovary syndrome. Endocrinology and Metabolism Clinics of North America 1999 28 265–293.[CrossRef][ISI][Medline]

    20. Dunaif A. Insulin resistance and the polycystic ovary syndrome: mechanism and implications for pathogenesis. Endocrine Reviews 1997 18 774–800.[Abstract/Free Full Text]

    21. Sahin I, Serter R, Karakurt F, Demirbas B, Culha C, Taskapan C, Kosar F & Aral Y. Metformin versus flutamide in the treatment of metabolic consequences of non-obese young women with polycystic ovary syndrome: a randomized prospective study. Gynecological Endocrinology 2004 19 115–124.[ISI][Medline]

    22. Nersesyan AK, Vardazaryan NS, Gevorgyan AC & Arutyunyan RM. Micronucleus level in exfoliated buccal mucosa cells of cancer patients. Archive of Oncology 2002 10 35–36.

    23. Diaz S, Fonseca G & Fernandez I. Analysis of lymphocyte and oral mucosa cell micronuclei in Cuban paint industry workers. Hereditas 1990 113 77–80.[ISI][Medline]

    24. Burgaz S, Karahalil B, Bayrak P, Taskin L, Yavuzaslan F, Bokesoy I, Anzion RBM, Bos RP & Platin N. Urinary cyclophosphamide excretion and micronuclei frequencies in peripheral lymphocytes and in exfoliated buccal epithelial cells of nurses handling anti-neoplastics. Mutation Research 1999 439 97–104.[ISI][Medline]

    25. Falck GCM, Hirvonen A, Scarpato R, Saarikoski ST, Migliore L & Norppa H. Micronuclei in blood lymphocytes and genetic polymorphism for GSTM1, GSTT1 and NAT2 in pesticide-exposed greenhouse workers. Mutation Research 1999 441 225–237.[ISI][Medline]

    26. Murrey EB & Edvards JW. Micronuclei in peripheral lymphocytes and exfoliated urothelial cells of workers exposed to 4,4'-methylenebis-(2-chloroaniline) (MOCA). Mutation Research 1999 446 175–180.[ISI][Medline]

    27. Trkova M, Kapras J, Bobkova K, Stankova J & Mejsnarova B. Increased micronuclei frequencies in couples with reproductive failure. Reproductive Toxicology 2000 14 331–335.[CrossRef][ISI][Medline]

    28. Botto N, Rizza A, Colombo MG, Mazzone AM, Manfredi S, Masetti S, Clerco A, Biagini A & Andreassi MG. Evidence for DNA damage in patients with coronary artery disease. Mutation Research 2001 493 23–30.[ISI][Medline]

    29. Baciuchka-Palmaro M, Orsière T, Duffaud F, Sari-Minodier I, Pompili J, Bellon L, De Méo M, Digue L, Favre R & Botta A. Acentromeric micronuclei are increased in peripheral blood lymphocytes of untreated cancer patients. Mutation Research 2002 520 189–198.[ISI][Medline]

    30. Joksic G, Petrovic S & Ilic Z. Age-related changes in radiation-induced micronuclei among healthy adults. Brazilian Journal of Medical and Biological Research 2004 37 1111–1117.

    31. Huber R, Braselmann H & Bauchiner M. Intra-and inter-individual variation of background and radiation-induced micronucleus frequencies in human lymphocytes. International Journal of Radiation Biology 1992 61 655–661.[ISI][Medline]

    32. Parker R, Ming PM, Rajan R, Goodner DM & Reme G. Clinical and cytogenetic studies with polycystic ovary disease. American Journal of Obstetrics and Gynecology 1980 137 656–660.[ISI][Medline]

    33. Hando JC, Nath J & Tucker JD. Sex chromosomes, micronuclei and aging in women. Chromosoma 1994 103 186–192.[ISI][Medline]

    34. Catalan J, Autio K, Wessman M, Lindholm C, Knuutila S, Sorsa M & Norppa H. Age-associated micronuclei containing centromeres and the X chromosome in lymphocytes in women. Cytogenetics and Cell Genetics 1995 68 11–16.[ISI][Medline]

    35. van der Spuy ZM & Dyer SJ. The pathogenesis of infertility and early pregnancy loss in polycystic ovary syndrome. Best Practice and Research. Clinical Obstetrics and Gynaecology 2004 18 755–771.

    36. Kousta E, White DM, Cela E, McCarthy MI & Franks S. The prevalence of polycystic ovaries in women with infertility. Human Reproduction 1999 14 2720–2723.[Abstract/Free Full Text]

    37. Balen AH, Conway GS, Kaltsas G, Techatraisak K, Manning PJ, West C & Jacobs HS. Polycystic ovary syndrome: the spectrum of the disorder in 1741 patients. Human Reproduction 1995 10 2107–2111.[Abstract/Free Full Text]

    38. Ames BN, Gold LS & Willett WC. The causes and prevention of cancer. PNAS 1995 92 5258–5265.[Abstract/Free Full Text]

    39. Simic MG. DNA markers of oxidative processes in vivo: relevance to carcinogenesis and anticarcinogenesis. Cancer Research 1994 54 1918–1923.

    40. Sabuncu T, Vural H, Harma M & Harma M. Oxidative stress in polycystic ovary syndrome and its contribution to the risk of cardiovascular disease. Clinical Biochemistry 2001 34 407–413.[CrossRef][ISI][Medline]

    41. Fenkci V, Fenkci S, Yilmazer M & Serteser M. Decreased total anti-oxidant status and increased oxidative stress in women with polycystic ovary syndrome may contribute to the risk of cardiovascular disease. Fertility and Sterility 2003 80 123–127.[ISI][Medline]

    42. Karmakar R, Banik S, Bandyopadhyay S & Chatterjee M. Cadmium-induced alterations of hepatic lipid peroxidation, glutathione S-transferase activity and reduced glutathione level and their possible correlation with chromosomal aberration in mice: a time course study. Mutation Research 1998 397 183–190.[ISI][Medline]

    43. Mayer C, Schmezer P, Freese R, Mutanen M, Hietanen E, Obe G, Basu S & Bartsch H. Lipid peroxidation status, somatic mutations and micronuclei in peripheral lymphocytes: a case observation on a possible interrelationship. Cancer Letters 2000 152 169–173.[CrossRef][ISI][Medline]

    44. Chandra Mohan KVP & Nagini S. Dose-response effects of tomato lycopene on lipid peroxidation and enzymic antioxidants in the hamster buccal pouch carcinogenesis model. Nutrition Research 2003 23 1403–1416.

    45. Subapriya R, Kumaraguruparan R, Abraham SK & Nagini S. Protective effects of ethanolic neem leaf extract on N-methyl-N-nitro-N-nitrosoguanidine-induced genotoxicity and oxidative stress in mice. Drug and Chemical Toxicology 2004 27 15–26.[ISI][Medline]

    46. Talbott E, Guzick D, Clerici A, Berga S, Detre K, Weimer K & Kuller L. Coronary heart disease risk factors in women with polycystic ovary syndrome. Arteriosclerosis, Thrombosis, and Vascular Biology 1995 15 821–826.[Abstract/Free Full Text]

    47. Orio F Jr, Palomba S, Spinelli L, Palomba S, Tauchmanova L, Zullo F, Lombardi G & Colao A. The cardiovascular risk of young women with polycystic ovary syndrome: an observational, analytical, prospective case-control study. Journal of Clinical Endocrinology and Metabolism 2004 89 3696–3701.[Abstract/Free Full Text]

    48. Hardiman P, Pillay OS & Atiomo W. Polycystic ovary syndrome and endometrial carcinoma. Lancet 2003 361 1810–1812.[CrossRef][ISI][Medline]

    49. Gadducci A, Gargini A, Palla E, Fanucchi A & Genazzani AR. Polycystic ovary syndrome and gynecological cancers: is there a link? Gynecological Endocrinology 2005 20 200–208.[CrossRef][ISI][Medline]

    50. Duffaud E, Orsiere T, Villant P, Pellissier AL, Volot E, Favre R & Botta A. Comparison between micronucleated lymphocyte rates observed in healthy subjects and cancer patients. Mutagenesis 1997 12 227–231.[Abstract/Free Full Text]

    51. Jagetia GC, Jayakrishnan A, Fernandes D & Vidyasagar MS. Evaluation of micronuclei frequency in the cultured peripheral blood lymphocytes of cancer patients before and after radiation treatment. Mutation Research 2001 491 9–16.[ISI][Medline]

Received 19 August 2005
Accepted 12 January 2006




This article has been cited by other articles:


Home page
 MutagenesisHome page
J. M. Battershill, K. Burnett, and S. Bull
Factors affecting the incidence of genotoxicity biomarkers in peripheral blood lymphocytes: impact on design of biomonitoring studies
Mutagenesis, August 4, 2008; (2008) gen040v1.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (2)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yesilada, E.
Right arrow Articles by Taskapan, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yesilada, E.
Right arrow Articles by Taskapan, C.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS