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      Oxidation–reduction potential and sperm DNA fragmentation, and their associations with sperm morphological anomalies amongst fertile and infertile men

      research-article
      a , * , a , a , b , a , a , c , d , e , f , a , a
      Arab Journal of Urology
      Elsevier
      ART, assisted reproductive techniques, AUC, area under the curve, ICSI, intracytoplasmic sperm injection, IUI, intrauterine insemination, IVF, in vitro fertilisation, NPV, negative predictive value, ORP, oxidation–reduction potential, OS, oxidative stress, PPV, positive predictive value, ROC, receiver operating characteristic, ROS, reactive oxygen species, SCD, sperm chromatin dispersion, Male infertility, Oxidation-reduction potential, Sperm, Sperm DNA fragmentation, Sperm morphology

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          Abstract

          Objective

          To assess seminal oxidation–reduction potential (ORP) and sperm DNA fragmentation (SDF) in male infertility and their relationships with sperm morphology in fertile and infertile men.

          Patients and methods

          Prospective case-control study comparing the findings of infertile men ( n = 1168) to those of men with confirmed fertility ( n = 100) regarding demographics and semen characteristics (conventional and advanced semen tests). Spearman rank correlation assessed the correlation between ORP, SDF, and different morphological indices. Means of ORP and SDF were assessed in variable levels of normal sperm morphology amongst all participants.

          Results

          Infertile patients had a significantly lower mean sperm count (32.7 vs 58.7 × 10 6 sperm/mL), total motility (50.1% vs 60.4%), and normal morphology (5.7% vs 9.9%). Conversely, infertile patients had significantly higher mean head defects (54% vs 48%), and higher ORP and SDF values than fertile controls. ORP and SDF showed significant positive correlations and significant negative correlations with sperm head defects and normal morphology in infertile patients, respectively. ORP and SDF were significantly inversely associated with the level of normal sperm morphology. Using receiver operating characteristic curve analysis, ORP and SDF threshold values of 1.73 mV/10 6 sperm/mL and 25.5%, respectively, were associated with 76% and 56% sensitivity and 72% and 72.2% specificity, respectively, in differentiating <4% from ≥4% normal morphology.

          Conclusion

          A direct inverse relationship exists between seminal ORP and SDF with various levels of normal sperm morphology. Using ORP and SDF measures in conjunction with standard semen morphology analysis could validate the result of the fertility status of patients.

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          Most cited references26

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          Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios

          Sperm DNA fragmentation (SDF) has been generally acknowledged as a valuable tool for male fertility evaluation. While its detrimental implications on sperm function were extensively investigated, little is known about the actual indications for performing SDF analysis. This review delivers practice based recommendations on commonly encountered scenarios in the clinic. An illustrative description of the different SDF measurement techniques is presented. SDF testing is recommended in patients with clinical varicocele and borderline to normal semen parameters as it can better select varicocelectomy candidates. High SDF is also linked with recurrent spontaneous abortion (RSA) and can influence outcomes of different assisted reproductive techniques. Several studies have shown some benefit in using testicular sperm rather than ejaculated sperm in men with high SDF, oligozoospermia or recurrent in vitro fertilization (IVF) failure. Infertile men with evidence of exposure to pollutants can benefit from sperm DNA testing as it can help reinforce the importance of lifestyle modification (e.g., cessation of cigarette smoking, antioxidant therapy), predict fertility and monitor the patient’s response to intervention.
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            The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation.

            Sperm DNA fragmentation is being increasingly recognized as an important cause of infertility. We herein describe the Sperm Chromatin Dispersion (SCD) test, a novel assay for sperm DNA fragmentation in semen. The SCD test is based on the principle that sperm with fragmented DNA fail to produce the characteristic halo of dispersed DNA loops that is observed in sperm with non-fragmented DNA, following acid denaturation and removal of nuclear proteins. This was confirmed by the analysis of DNA fragmentation using the specific DNA Breakage Detection-Fluorescence In Situ Hybridization (DBD-FISH) assay, which allows the detection of DNA breaks in lysed sperm nuclei. Sperm suspensions either prepared from semen or isolated from semen by gradient centrifugation were embedded in an agarose microgel on slides and treated with 0.08 N HCl and lysing solutions containing 0.8 M dithiothreitol (DTT), 1% sodium dodecyl sulfate (SDS), and 2 M NaCl. Then, the slides were sequentially stained with DAPI (4',6-diamidino-2-phenylindole) and/or the Diff-Quik reagent, and the percentages of sperm with nondispersed and dispersed chromatin loops were monitored by fluorescence and brightfield microscopy, respectively. The results indicate that all sperm with nondispersed chromatin displayed DNA fragmentation, as measured by DBD-FISH. Conversely, all sperm with dispersed chromatin had very low to undetectable DBD-FISH labeling. SCD test values were significantly higher in patients being screened for infertility than in normozoospermic sperm donors who had participated in a donor insemination program. The coefficient of variation obtained using 2 different observers, either by digital image analysis (DIA) or by brightfield microscopy scoring, was less than 3%. In conclusion, the SCD test is a simple, accurate, highly reproducible, and inexpensive method for the analysis of sperm DNA fragmentation in semen and processed sperm. Therefore, the SCD test could potentially be used as a routine test for the screening of sperm DNA fragmentation in the andrology laboratory.
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              Role of reactive oxygen species in male infertility.

              Human spermatozoa exhibit a capacity to generate ROS and initiate peroxidation of the unsaturated fatty acids in the sperm plasma membrane, which plays a key role in the etiology of male infertility. The short half-life and limited diffusion of these molecules is consistent with their physiologic role in key biological events such as acrosome reaction and hyperactivation. The intrinsic reactivity of these metabolites in peroxidative damage induced by ROS, particularly H2O2 and the superoxide anion, has been proposed as a major cause of defective sperm function in cases of male infertility. The number of antioxidants known to attack different stages of peroxidative damage is growing, and it will be of interest to compare alpha-tocopherol and ascorbic acid with these for their therapeutic potential in vitro and in vivo. Both spermatozoa and leukocytes generate ROS, although leukocytes produce much higher levels. The clinical significance of leukocyte presence in semen is controversial. Seminal plasma confers some protection against ROS damage because it contains enzymes that scavenge ROS, such as catalase and superoxide dismutase. A variety of defense mechanisms comprising a number of anti-oxidants can be employed to reduce or overcome oxidative stress caused by excessive ROS. Determination of male infertility etiology is important, as it will help us develop effective therapies to overcome excessive ROS generation. ROS can have both beneficial and detrimental effects on the spermatozoa and the balancing between the amounts of ROS produced and the amounts scavenged at any moment will determine whether a given sperm function will be promoted or jeopardized. Accurate assessment of ROS levels and, subsequently, OS is vital, as this will help clinicians both elucidate the fertility status and identify the subgroups of patients that respond or do not respond to these therapeutic strategies. The overt commercial claims of antioxidant benefits and supplements for fertility purposes must be cautiously looked into, until proper multicentered clinical trials are studied. From the current data it appears that no single adjuvant will be able to enhance the fertilizing capacity of sperm in infertile men, and a combination of the possible strategies that are not toxic at the dosage used would be a feasible approach.
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                Author and article information

                Contributors
                Journal
                Arab J Urol
                Arab J Urol
                Arab Journal of Urology
                Elsevier
                2090-598X
                2090-5998
                01 February 2018
                March 2018
                01 February 2018
                : 16
                : 1
                : 87-95
                Affiliations
                [a ]Department of Urology, Hamad Medical Corporation, Doha, Qatar
                [b ]Department of Urology, Cleveland Clinic Foundation, Cleveland, USA
                [c ]Department of Surgery, Hamad General Hospital, Hamad Medical Corporation, Doha, Qatar
                [d ]College of Medicine, Qatar University, Doha, Qatar
                [e ]School of Health and Education, University of Skövde, Skövde, Sweden
                [f ]Department of Obstetrics and Gynecology, Women’s Hospital, Hamad Medical Corporation, Doha, Qatar
                Author notes
                [* ]Corresponding author at: Department of Urology, Hamad Medical Corporation, PO Box 3050, Doha, Qatar. amajzoub@ 123456hamad.qa
                Article
                S2090-598X(17)30148-1
                10.1016/j.aju.2017.11.014
                5922185
                29713539
                17d44a46-6eb6-4f2e-b465-5aaa56dc6a4b
                © 2017 Production and hosting by Elsevier B.V. on behalf of Arab Association of Urology.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 14 September 2017
                : 11 November 2017
                : 21 November 2017
                Categories
                Diagnosis

                art, assisted reproductive techniques,auc, area under the curve,icsi, intracytoplasmic sperm injection,iui, intrauterine insemination,ivf, in vitro fertilisation,npv, negative predictive value,orp, oxidation–reduction potential,os, oxidative stress,ppv, positive predictive value,roc, receiver operating characteristic,ros, reactive oxygen species,scd, sperm chromatin dispersion,male infertility,oxidation-reduction potential,sperm,sperm dna fragmentation,sperm morphology

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