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• W2169459352 abstract "The aim of this study was to investigate the fertilization and pregnancy rates of artificial reproductive technologies using semen samples without (<1 × 106/mL) and with (≥1 × 106/mL) leukocytospermia. The overall fertilization rate was 63.4% (range, 44.4–87.5%) in nonleukocytospermic couples and 64.3% (range, 45.3–100.0%) in leukocytospermic couples, whereas the corresponding pregnancy rates were 34.5% and 50%, respectively. These results show that leukocytospermia may not necessarily have a negative effect on outcome after either in vitro fertilization or intracytoplasmic sperm injection. The aim of this study was to investigate the fertilization and pregnancy rates of artificial reproductive technologies using semen samples without (<1 × 106/mL) and with (≥1 × 106/mL) leukocytospermia. The overall fertilization rate was 63.4% (range, 44.4–87.5%) in nonleukocytospermic couples and 64.3% (range, 45.3–100.0%) in leukocytospermic couples, whereas the corresponding pregnancy rates were 34.5% and 50%, respectively. These results show that leukocytospermia may not necessarily have a negative effect on outcome after either in vitro fertilization or intracytoplasmic sperm injection. When evaluating an infertile couple, the World Health Organization (WHO) recommends performing a semen analysis on the male partner (1World Health Organization WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction.4th ed. Cambridge University Press, Cambridge1999Google Scholar). The WHO classifies a leukocyte count ≥1 × 106/mL in the semen sample as indicative of leukocytospermia. Reports on the influence of leukocytospermia on semen quality are contradictory. Leukocytes produce reactive oxygen species (ROS), which are harmful to spermatozoa (ROS have been reported to damage sperm morphology and motility as well as DNA integrity; (2Alvarez J.G. Sharma R.K. Ollero M. Saleh R.A. Lopez M.C. Thomas Jr., A.J. et al.Increased DNA damage in sperm from leukocytospermia semen samples as determined by the sperm chromatin structure assay.Fertil Steril. 2002; 78: 319-329Abstract Full Text Full Text PDF PubMed Scopus (124) Google Scholar, 3Repping S. van Weert J.M. Mol B.W. de Vries J.W. van der Veen F. Use of the total motile sperm count to predict total fertilization failure in in vitro fertilization.Fertil Steril. 2002; 78: 22-28Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar)), and recent reports indicate that leukocytospermia has a negative impact on semen quality as a result of ROS (4Rodin D.M. Larone D. Goldstein M. Relationship between semen cultures, leukospermia, and semen analysis in men undergoing fertility evaluation.Fertil Steril. 2003; 79: 1555-1558Abstract Full Text Full Text PDF PubMed Scopus (63) Google Scholar, 5Kiessling A.A. Lamparelli N. Yin H.Z. Seibel M.M. Eyre R.C. Semen leukocytes: friends or foes?.Fertil Steril. 1995; 64: 196-198Abstract Full Text PDF PubMed Google Scholar, 6Tomlinson M.J. White A. Barratt C.L. Bolton A.E. Cooke I.D. The removal of morphologically abnormal sperm forms by phagocytes: a positive role for seminal leukocytes?.Hum Reprod. 1992; 7: 517-522Crossref PubMed Scopus (149) Google Scholar, 7Ford W.C. Regulation of sperm function by reactive oxygen species.Hum Reprod Update. 2004; 10: 387-399Crossref PubMed Scopus (220) Google Scholar, 8Agarwal A. Saleh R.A. Bedaiwy M.A. Role of reactive oxygen species in the pathophysiology of human reproduction.Fertil Steril. 2003; 79: 829-843Abstract Full Text Full Text PDF PubMed Scopus (1069) Google Scholar, 9Henkel R. Kierspel E. Stalf T. Mehnert C. Menkveld R. Tinneberg H.R. et al.Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients.Fertil Steril. 2005; 83: 635-642Abstract Full Text Full Text PDF PubMed Scopus (224) Google Scholar, 10Yilmaz S. Koyuturk M. Kilic G. Alpak O. Aytoz A. Effects of leukocytospermia on semen parameters and outcomes of intracytoplasmatic sperm injection.Int J Androl. 2005; 28: 337-342Crossref PubMed Scopus (22) Google Scholar). This may include a negative impact on fertilization and pregnancy rates. With the possibility of a treatment for leukocytospermia, it is important that the effect of leukocytes on assisted reproductive technologies (ART) outcomes should be investigated in detail (11Lackner J.E. Herwig R. Schmidbauer J. Schatzl G. Kratzik C. Marberger M. Correlation of leukocytospermia with clinical infection and the positive effect of anti-inflammatory treatment on semen quality.Fertil Steril. 2006; 86: 601-605Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 12Sigman M. Glass S. Campagnone J. Pryor J.L. Carnitine for the treatment of idiopathic asthenospermia: a randomized, double-blind, placebo-controlled trial.Fertil Steril. 2006; 85: 1409-1414Abstract Full Text Full Text PDF PubMed Scopus (107) Google Scholar). The aim of this pilot study was to compare the effect of leukocytospermia (≥1 × 106/mL) on the outcomes (fertilization and pregnancy rates) of IVF and intracytoplasmic sperm injection (ICSI) in couples with and without leukocytospermia. A total of 75 couples who were treated for infertility at the Department of Endocrinology and Reproduction at the Medical University of Vienna between January and June 2006 were investigated. All patients signed informed consent. Couples included individuals with female factor infertility and male factor infertility. The limited number of patients did not allow any subgroup analysis according to any specific female or male infertility reason. Leukocytospermia was the factor being studied, therefore, only the presence of leukocytospermia was a group characteristic. Semen analysis was carried out according to WHO procedures (1World Health Organization WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction.4th ed. Cambridge University Press, Cambridge1999Google Scholar). Sperm concentration, progressive forward motility (motility grade a), normal sperm morphology, and leukocyte count were recorded. The determination of leukocytes was carried out using peroxidase staining in two steps as described in the WHO laboratory manual. Patients were divided into two groups according to the presence or absence of leukocytospermia: group1, no leukocytospermia (leukocytes <1 × 106/mL); group 2, leukocytospermia (leukocytes ≥1 × 106/mL). IVF and ICSI outcomes in nonleukocytospermic and leukocytospermic couples were then compared by analyzing fertilization and pregnancy rates. All female partners received midluteal, long-protocol down-regulation with a gonadotropin-releasing hormone antagonist (decapeptyl 0.1 mg; Ferring, Vienna) 0.1 mg/day administered SC, followed by SC recombinant FSH (150–200 IU/day; Puregon, Organon, Oss, Netherlands; 150–225 IU/day; Gonal-F, Merck-Serono, Munich, Germany). IM hCG (10,000 IU) was administered when two or more follicles reached a diameter of ≥17 mm, which approximately corresponded to days 9–13 of stimulation. Oocyte retrieval was performed 34–36 hours after hCG injection. IVF involved a conventional insemination method when the semen sample was normal and ICSI in the case of pathological semen findings. ET was performed on the third day after oocyte retrieval. Pregnancy was determined by urine hCG 14 days after ET. Fertilization was evaluated 16–18 hours after injection or insemination. Statistical analysis was carried out using the Statistical Package for the Social Sciences (ver. 10.0.7; SPSS Inc., Chicago) software. All demographic data are presented as the median and 25th and 75th quartiles. Statistical differences were assessed using the Mann-Whitney U-test. It was assumed that nonleukocytospermia would result in higher pregnancy rates, therefore the odds ratio (OR) of obtaining a pregnancy in those who were nonleukocytospermic were calculated in comparison with those who were leukocytospermic. Overall, 75 couples underwent ART: 44 underwent IVF, and 31 underwent ICSI. Ten semen samples in the IVF group and 10 in the ICSI group showed leukocytospermia. The median age of the women in the IVF group was 33.5 years (range, 31.0–38.8 years) and did not differ significantly from that in the ICSI group (35.0 years; range, 31.0–38.0 years; P=.501). Table 1 details nonleukocytospermic and leukocytospermic semen parameters and corresponding fertilization and pregnancy rates. Using the spearman rho correlation, no significant influence of leukocytes on any semen parameter could be detected. The women's age showed a statistically significant impact on pregnancy rates only in the nonleukocytospermic IVF group (r = 0.431, P=.011) and in the nonleukocytospermic ICSI group (r = 0.715; P<.001); women's age did not have a statistically significant impact on fertilization rates. This finding on women's age can be explained by the similar ages within the groups, as described above.Table 1Nonleukocytospermic (n = 55) and leukocytospermic (n = 20) semen parameters and fertilization and pregnancy rates for the overall ART group.NonleukocytospermiaLeukocytospermiaPaMann-Whitney test; χ2 used to compare pregnancy rates.Sperm concentration, ×106/mL (range)31.0 (9.0–83.9)35.5 (9.1–74.5).905Progressive forward motility rate, % (range)17.0 (3.0–30.0)13.0 (4.5–29.5).976Normal sperm morphology, % (range)30.0 (19.0–49.0)35.0 (16.5–55.5).353Leukocyte count, ×106/mL (range)0.0 (0.0–0.3)1.5 (1.0–2.4).001Fertilization rate, % (range)63.4 (44.4–87.5)64.3 (45.3–100.0).777Pregnancy rate, %34.550.0.224Note: Data are given as medians and interquartile ranges.a Mann-Whitney test; χ2 used to compare pregnancy rates. Open table in a new tab Note: Data are given as medians and interquartile ranges. Calculation of ORs revealed that nonleukocytospermia in the ICSI group had the greatest impact on fertilization, with this group having the best chance of becoming pregnant, with an OR of 0.952. Neither of the ORs for the overall ART (0.691) or IVF groups (0.539) provided evidence that nonleukocytospermia conferred any advantage on the chances of becoming pregnant. With regard to the reported damage by leukocytospermia on spermatozoa, the results of this study are surprising and are in contrast to a report by Yilmaz et al. (10Yilmaz S. Koyuturk M. Kilic G. Alpak O. Aytoz A. Effects of leukocytospermia on semen parameters and outcomes of intracytoplasmatic sperm injection.Int J Androl. 2005; 28: 337-342Crossref PubMed Scopus (22) Google Scholar), who compared 34 leukocytospermic semen samples with 36 nonleukocytospermic semen samples and subsequent ICSI outcomes. Fertilization rates from leukocytospermic samples were lower than those from nonleukocytospermic semen samples, whereas pregnancy rates were similar (30.0% and 30.6%, respectively). One reason for these findings could be that the median leukocyte concentration in leukocytospermic semen samples in the study by Yilmaz et al. was 2.68 (range, 1.5–6.1) × 106/mL, which is markedly higher than the median leukocyte concentration (1.5 [range, 1.0–2.4] ×106/mL) found in our study. These findings suggest, however, that the effects of leukocytes might be dependent on their concentration; studies on this hypothesis are still lacking. Another reason for the similar fertilization and pregnancy rates from samples with and without leukocytospermia found in our study could be the observed lack of a negative influence of leukocytes on sperm motility. In the leukocytospermic semen samples, there was no statistically significant correlation between leukocyte concentration and sperm motility (progressive forward motility: r = −0.174, P=.464; total motility: r = −0.075, P=.792). However, 10 of 20 couples with leukocytospermia underwent ICSI in which sperm motility is bypassed by the procedure itself. Additionally, in the ICSI group, in which leukocytospermia was seen to have had the highest levels (1.5 × 106/mL vs. 1.47 × 106/mL; P= .203), male factor infertility was present in 74.2% of the samples. These findings suggest an association between leukocytospermia and male factor infertility (although not statistically significant in our study), but that outcome after ART is not impaired by use of leukocytospermic samples. On the basis of the premise that leukocytes should harm sperm via ROS, Zorn et al. (13Zorn B. Vidmar G. Meden-Vrtovec H. Seminal reactive oxygen species as predictors of fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization and intracytoplasmatic sperm injection.Int J Androl. 2003; 26: 279-285Crossref PubMed Scopus (111) Google Scholar) published a report in which they compared IVF and ICSI outcomes in correlation with ROS levels. Although the investigators found a negative association between ROS levels and IVF outcome, seminal ROS did not affect ICSI outcome. They also assumed that the ICSI procedure itself protected sperm against any additional harm of ROS, as contact with leukocytes might be reduced. However, the investigators did not report detailed information on semen analysis, especially leukocyte concentration, despite the fact that leukocytes are thought to be the major source of ROS (2Alvarez J.G. Sharma R.K. Ollero M. Saleh R.A. Lopez M.C. Thomas Jr., A.J. et al.Increased DNA damage in sperm from leukocytospermia semen samples as determined by the sperm chromatin structure assay.Fertil Steril. 2002; 78: 319-329Abstract Full Text Full Text PDF PubMed Scopus (124) Google Scholar, 9Henkel R. Kierspel E. Stalf T. Mehnert C. Menkveld R. Tinneberg H.R. et al.Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients.Fertil Steril. 2005; 83: 635-642Abstract Full Text Full Text PDF PubMed Scopus (224) Google Scholar). Overall, Zorn et al. (11Lackner J.E. Herwig R. Schmidbauer J. Schatzl G. Kratzik C. Marberger M. Correlation of leukocytospermia with clinical infection and the positive effect of anti-inflammatory treatment on semen quality.Fertil Steril. 2006; 86: 601-605Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar) found ROS to have a greater influence on IVF outcome than on ICSI outcome. One possible explanation for these observations might be that ROS are essential for capacitation and fertilization (7Ford W.C. Regulation of sperm function by reactive oxygen species.Hum Reprod Update. 2004; 10: 387-399Crossref PubMed Scopus (220) Google Scholar, 14De Jonge C. Biological basis for human capacitation.Hum Reprod Update. 2005; 11: 205-214Crossref PubMed Scopus (131) Google Scholar). Reports on ROS in the female reproductive tract during or after fertilization support these assumptions (8Agarwal A. Saleh R.A. Bedaiwy M.A. Role of reactive oxygen species in the pathophysiology of human reproduction.Fertil Steril. 2003; 79: 829-843Abstract Full Text Full Text PDF PubMed Scopus (1069) Google Scholar, 15Agarwal A. Said T.M. Bedaiwy M.A. Banerjee J. Alvarez J.G. Oxidative stress in an assisted reproductive techniques setting.Fertil Steril. 2006; 86: 503-512Abstract Full Text Full Text PDF PubMed Scopus (243) Google Scholar). Attaran et al. (16Attaran M. Pasqualotto E. Falcone T. Goldberg J.M. Miller K.F. Agarwal A. et al.The effect of follicular fluid reactive oxygen species on the outcome of in vitro fertilization.Int J Fertil Womens Med. 2002; 45: 314-320Google Scholar) investigated the follicular fluid of 53 women who underwent IVF for ROS levels. Women who became pregnant had significantly higher ROS levels than those who did not become pregnant. Thus, Attaran et al. (16Attaran M. Pasqualotto E. Falcone T. Goldberg J.M. Miller K.F. Agarwal A. et al.The effect of follicular fluid reactive oxygen species on the outcome of in vitro fertilization.Int J Fertil Womens Med. 2002; 45: 314-320Google Scholar) concluded that ROS at low concentrations could be a potential marker for the success of IVF. This again underlines the double-edged effect of ROS on fertilization. Therefore, the impact of ROS on fertility may be a question of degree rather than of the presence or absence of a detectable pathology. Taking into account that leukocytes are the primary source of ROS, this could also be true for leukocytes in semen samples. This suggestion could explain the multiple reports in the literature that show a poor correlation between leukocytospermia and semen quality (4Rodin D.M. Larone D. Goldstein M. Relationship between semen cultures, leukospermia, and semen analysis in men undergoing fertility evaluation.Fertil Steril. 2003; 79: 1555-1558Abstract Full Text Full Text PDF PubMed Scopus (63) Google Scholar). Leukocytes, in any concentration, are quite common in semen samples, but their correlation with bacteriospermia, signs of clinical infections in asymptomatic males, or reduced semen quality is poor (11Lackner J.E. Herwig R. Schmidbauer J. Schatzl G. Kratzik C. Marberger M. Correlation of leukocytospermia with clinical infection and the positive effect of anti-inflammatory treatment on semen quality.Fertil Steril. 2006; 86: 601-605Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 17Lackner J. Schatzl G. Horvath S. Kratzik C. Marberger M. Value of counting white blood cells (WBC) in semen samples to predict the presence of bacteria.Eur Urol. 2006; 49: 148-153Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar). Tomlinson et al. (6Tomlinson M.J. White A. Barratt C.L. Bolton A.E. Cooke I.D. The removal of morphologically abnormal sperm forms by phagocytes: a positive role for seminal leukocytes?.Hum Reprod. 1992; 7: 517-522Crossref PubMed Scopus (149) Google Scholar) described the phagocytosis of abnormal sperm through leukocytes, which is thus a positive effect of leukocytospermia. Lackner et al. (11Lackner J.E. Herwig R. Schmidbauer J. Schatzl G. Kratzik C. Marberger M. Correlation of leukocytospermia with clinical infection and the positive effect of anti-inflammatory treatment on semen quality.Fertil Steril. 2006; 86: 601-605Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar) showed that the WHO definition of male accessory gland infection does not categorically represent any clinical symptomatic infection and that there was no correlation between leukocytospermia and a reduction in semen quality. Therefore, it would appear that leukocytes do not always have negative effects on semen quality. Here again, as previously suggested, the same may be true for leukocytes as for ROS, that is, that it is not the actual presence of leukocytes that results in damage but, for example, a specific threshold. In summary, to evaluate an influence of leukocytes on semen quality or ART outcome, a future aim should be to define specific cut-off levels for leukocytes at which semen quality or fertilization and pregnancy rates are impaired. In conclusion, leukocytospermia was found to have no negative influence on fertilization or pregnancy rates after either IVF or ICSI; however, one limiting factor of this study is the small number of leukocytospermic semen samples in both the IVF and ICSI groups." @default.
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• W2169459352 title "Effect of leukocytospermia on fertilization and pregnancy rates of artificial reproductive technologies" @default.
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