scholarly journals Flow Cytometry Detection of Sperm DNA Fragmentation and Apoptotic Markers in the Semen of Infertile Males

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Huda Mossa Omran ◽  
Moiz Bakhiet ◽  
Volker Ehemann
Keyword(s):  
Flow Cytometry ◽  
Dna Damage ◽  
Male Infertility ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Chromatin Condensation ◽  
B Cell Lymphoma ◽  
Sperm Dna Fragmentation ◽  
Molecular Defects ◽  
Sperm Dna

The effect of sperm molecular defects on fertilization and pregnancy outcome after assisted reproductive therapy (ART) is widely documented by both research and clinical societies. Sperm DNA fragmentation and abnormal chromatin condensation represent critical causes of male infertility. Advanced androgenic techniques for accurately identifying molecular defects help in selecting an appropriate treatment strategy. Additionally, specific markers of apoptosis are increasingly important in predicting male infertility. The ability of flow cytometry to estimate the quantity of sperm with DNA fragmentation or damage and multifactor measurements in immotile sperm have made this developed technique essential in fertility centers. The study is aimed at assessing the level of DNA fragmentation and apoptosis by measuring flow cytometry using new techniques. Flow cytometry analysis revealed a varying degree of DNA damage. It was able to quantify the degree of impairment even in samples with minimal DNA fragmentation. DNA damage was observed even in samples that were considered normal with a routine semen analysis. Flow cytometry was sensitive to changes in sperm apoptosis. Elevated p53 activity levels were associated with high DNA fragmentation. Meanwhile, B-cell lymphoma 2 (Bcl-2) activities showed a different pattern. In conclusion, flow cytometry for sperm DNA fragmentation and markers of apoptosis can be a valuable tool in assisted reproductive centers.

P–068 CatSper4 cation channel expression is low in male infertility and is affected by cryopreservation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N Kilic ◽  
T İrez ◽  
N Dayiolu
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Middle Part ◽  
Cation Channel ◽  
The Other ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Negative Effects ◽  
Sperm Dna

Abstract Study question Is CatSper4 expression in sperm related to functional parameters and does cryopreservation affect CatSper4 expression? Summary answer In this study, it was aimed to investigate whether CatSper4 has a relationship with sperm parameters and is CatSper 4 affected by cryopreservation. What is known already CatSper membrane channels, known as cation channels, are thought to play an important role in the insufficiency of sperm physiology, acrosome reaction, and chemotaxis movement. There is no study on cation channel distribution in an infertile male patient. In addition, studies conducted in recent years have shown that cryopreservation techniques have negative effects on sperm DNA, but there is no analysis in the literature regarding the effects of cryopreservation on CatSper4 ion channel proteins. Study design, size, duration Samples of the patients who applied to the Andrology laboratory in the Medical Park Hospital IVF unit between March 1 and June 1 in 2020 were included in the study. Also, patients with no family history of no genetic anomalies , no varicocele and azoospermia were included.The study were divided into 4 groups in accordance with the male infertility guideline of the European Association of Urology as normozoospermic (control group), the asthenoteratozoospermia, teratozoospermia, and oligoastenotheratozoospermia. Participants/materials, setting, methods In this prospective study, semen analysis, DNA fragmentation, and CatSper 4 by IHC of control group patients with normospermia (n = 40) and oligospermia(n = 50), asthenospermia(n = 40), and teratozoospermia(n = 38) patients were compared and differences resulting from cryopreservation were evaluated by Wilcoxon signed Ranks Test. Main results and the role of chance It was observed that CatSper4 protein positivity was localized in the middle part of the sperm and it was statistically higher in the normozoospermic patient group compared to the other groups (p = 0,01). When the positivity values of CatSper4 protein before and after freezing were compared in the groups, it was seen that the values decreased (p = 0,001,p=0,01). Sperm DNA fragmentation was found to be lowest in normospermia and statistically significantly higher in other groups. Cryopreservation application increased DNA fragmentation in all groups (p < 0,001 , p < 0,01). Limitations, reasons for caution Unfortunately, embryo screening in patients with low CatSper4 expression is not available in the present study. Soon we plan to screen a broader clinical pregnancy series and present the IVF results associated with CatSper4. Wider implications of the findings: Our study indicated that, CatSper4 expression is quite high in normospermia when compared with the other groups, particularly oligoasthenoteratozoospermia and asthenoteratozoospermia. There are almost no studies on this subject in the literature, and we think that it should be studied in larger patient groups and in unexplained infertile cases. Trial registration number Not applicable

scholarly journals Proteomic Analysis in Seminal Plasma of Fertile Donors and Infertile Patients with Sperm DNA Fragmentation

2020 ◽  
Vol 21 (14) ◽  
pp. 5046
Author(s):  
Alba Fernandez-Encinas ◽  
Agustín García-Peiró ◽  
Javier del Rey ◽  
Jordi Ribas-Maynou ◽  
Carlos Abad ◽  
...  
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Recurrent Miscarriage ◽  
Semen Analysis ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Plasma Samples ◽  
Sperm Dna ◽  
Dispersion Test

Seminal plasma proteomics studies could represent a new approach for the determination of molecular elements driving male infertility, resulting in a better male infertility characterization. The aim of this study is to investigate proteomic differences in seminal plasma samples from fertile and infertile individuals. For that, semen samples were selected according to semen analysis, clinical pathology, and values of sperm DNA fragmentation (alkaline and neutral Comet assay and Sperm Chromatin Dispersion test). A total of 24 seminal plasma samples classified in four groups were processed: fertile donors (FD), recurrent miscarriage patients (RM), asthenoteratozoospermic patients (ATZ), and asthenoteratozoospermic patients with varicocele (ATZ-VAR). Results obtained by 2D-differential gel electrophoresis (2D-DIGE) revealed 26 spots significantly increased in fertile donors when compared to patient groups. Also, eight spots in the ATZ group and two in the ATZ-VAR group were decreased compared to the other groups. Twenty-eight proteins were identified by mass spectrometry (MS), most of them involved in metabolic and cellular processes and with a catalytic or binding function. Protein–protein interactions through Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) tool suggest that a large part of them were associated with each other. Furthermore, most of them were associated with ubiquitin C, indicating that it could play an important regulation role, resulting in a potential male infertility biomarker.

scholarly journals Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Essam-Elden M. Mohammed ◽  
Eman Mosad ◽  
Asmaa M. Zahran ◽  
Diaa A. Hameed ◽  
Emad A. Taha ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Acridine Orange ◽  
Pregnancy Outcome ◽  
Dna Fragmentation ◽  
Chromatin Condensation ◽  
Semen Parameters ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P<0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P<0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

scholarly journals Can Utilization of Sperm DNA Fragmentation (SDA) Tests in Combination with Time Lapse Microscopy Help in Improving ICSI Implantation Rates in Unexplained Recurrent Implantation Failures Utilizing Double Stranded SDA as the Causative Factor for the same

2020 ◽  
Vol 4 (1) ◽  
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Time Lapse ◽  
Sperm Dna Fragmentation ◽  
Time Lapse Microscopy ◽  
Sperm Dna ◽  
Implantation Rates ◽  
Embryo Kinetics

Over 50% of intracytoplasmic sperm injection (ICSI) cycles don’t display implantation. Hence laboratories make their maximum efforts to select the best embryos as far as implantation enhancement is concerned. Further utilization of available technologies like time lapse recording have been made in a large number of artificial reproductive technology (ART) centres. Various studies that utilize embryo kinetics have implicated that time when embryo cleavage may prove to be an important factor that determines the implantation potential of an embryo. With this variety of algorithms mathematic wise have been used to forecast which the best embryos are for transfer. But the efficacy of these might be influenced by multiple confounding factors. Thus work on biomarkers that can forecast good ART warrants newer embryo selection basis. Regarding conventional ICSI, typical standard routine semen analysis involving sperm concentration, motility and morphology does not predict the implantation percentages in an ICSI cycle. Once sperm DNA fragmentation (SDA) methods were inducted they appeared to hold promise in forecasting good ART success. Although certain studies utilizing various techniques like TUNEL. SCSA, SCD proved a relation existed between DNA damage and implantation rates in ICSI but the same was contradicted by others. With this it was thought that bias between evaluation of ejaculate and motile sperm picked up for ICSI, as is known regarding absence of positive association of sperm motility and DNA fragmentation. Thus study by Casanovas et.al., tried to find if there is any correlation of single stranded (ssSDA) and double stranded (dsSDA) sperm DNA damage that might forecast ICSI success and utilizing Neutral Comet Assays along with help of time lapse technology they found that double stranded sperm influenced delay in embryo formation as seen by embryo kinetics and thus interfere with implantation rates. Reproduction of these findings might help in getting a standard for getting best embryos selected in ICSI utilizing SDA and time lapse microscopy.

scholarly journals Correlation of sperm DNA damage with blastocyst formation: systematic review and meta-analysis

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Prashanth K. Adiga ◽  
Srisailesh Vitthala ◽  
Shivaranjeni
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Meta Analysis ◽  
Main Body ◽  
Sperm Dna Fragmentation ◽  
Blastocyst Formation ◽  
Male Factor ◽  
Sperm Dna Damage ◽  
Sperm Dna

Abstract Background The routine semen analysis fails to detect sperm DNA damage which contributes to the majority of male factor infertility. Sperm DNA fragmentation test (DFI) measures the sperm DNA damage. Blastocyst formation is an important step in IVF ± ICSI. At present, the literature lacks any data that correlates DFI and blastocyst formation. Main body of the abstract We searched MEDLINE and other databases till 2020 for the studies that reported on sperm DNA damage and blastocyst formation in assisted reproductive technology (ART). The outcomes analyzed were (1) a comparison of blastulation rates in high DFI and low DFI groups. (2) Comparison of blastulation rates in high DFI and low DFI groups based on (a) different sperm DNA fragmentation assays (COMET, SCD, SCSA, TUNEL), (b) different types of ART (IVF/IVF + ICSI/ICSI). 10 studies were included in this review. A non-significant increase in the blastocyst formation was observed in high DFI group (OR = 0.70; 95% CI = 0.4 to 1.21; P = 0.20) and with SCD and TUNEL assays. Short conclusion Our study emphasizes on sperm DNA fragmentation (sperm DNA damage) as an important marker of blastocyst formation. The results of this meta-analysis suggest that the high sperm DNA fragmentation may not adversely affect the blastocyst formation.

scholarly journals Docosahexaenoic acid in the treatment of male infertility caused by high sperm DNA fragmentation

2021 ◽  
Vol 21 (4) ◽  
pp. 89-97
Author(s):  
I. V. Vinogradov ◽  
A. R. Zhivulko
Keyword(s):  
Dna Damage ◽  
Docosahexaenoic Acid ◽  
Male Infertility ◽  
Dna Fragmentation ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Male Accessory Glands ◽  
Accessory Glands ◽  
Sperm Dna ◽  
High Level

Introduction. Antioxidant supplementation therapy continues to be the main treatment for male infertility associated with high level of sperm DNA damage. Docosahexaenoic acid (DHA) is one of the most promising components of antioxidant supplementation therapy. It also has anti-inflammatory properties that makes it interesting for treatment of patients with high level of sperm DNA damage and inflammation in male accessory glands.Materials and methods.One hundred and seventeen (117) infertile patients with high level of sperm DNA damage were recruited for this randomized, double blind, placebo-controlled study. Semen analysis, MAR-test, SCD test and sperm cryotolerance test were performed to all patients. Subjects were divided into 2 groups with high (>1 mln / ml) and low (<1 mln / ml) semen leucocyte concertation and then randomized into 2 subgroups of active treatment and 2 placebo subgroups. The active treatment subgroups received 1470 mg / day of DHA for 3 months. The placebo group received placebo for the same period. Laboratory tests were repeated after the treatment course had been finished.Results. Statistically significant increase in motility (42 % (25–61 %) vs 25 % (15–47 %), p <0.05), vitality (73 % (63–81 %) vs 41 % (35–64 %), p <0.05), decrease in sperm DNA fragmentation level (21 % (12–28 %) vs 33 % (25–39 %), p <0.05) and leucocyte concentration (1 million / ml (0.7–1.7 million / ml) vs 1,5 million / ml (1.1–2.1 million / ml), p <0.05) were observed in the subgroup with male accessory glands inflammation after treatment. Motility (15 % (8–19 %) vs 8 % (5–11 %), p <0.05) and vitality (37 % (25–46 %) vs 24 % (17–40 %), p <0.05) in this subgroup after a sperm cryotolerance test increased as well. In the subgroup with low semen leucocyte concertation statistically significant increase in motility (43 % (27–63 %) vs 34 % (21–54 %), p <0.05), vitality (77 % (66–85 %) vs 65 % (54.5–76.0 %), p <0.05) and decrease of sperm DNA fragmentation level (9 % (5.5–20.0 %) vs 25 % (18–33 %), p <0.05) were observed. DHA supplementation also resulted in statistically significant increase in motility (17 % (10–23 %) vs 6 % (5.0–10.5 %), p <0.05) and vitality (41 % (32.5–53.0 %) vs 37 % (30–49 %), p <0.05) after a sperm cryotolerance test in that subgroup.Conclusion. DHA supplementation therapy increases motility, vitality, sperm cryotolerance and decreases sperm DNA fragmentation regardless of the presence of an inflammatory process in male accessory glands.

scholarly journals Oxidation of Sperm DNA and Male Infertility

Antioxidants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 97
Author(s):  
Leila Rashki Ghaleno ◽  
AliReza Alizadeh ◽  
Joël R. Drevet ◽  
Abdolhossein Shahverdi ◽  
Mojtaba Rezazadeh Valojerdi
Keyword(s):  
Oxidative Stress ◽  
Male Infertility ◽  
Oxidative Damage ◽  
Dna Fragmentation ◽  
Dna Bases ◽  
De Novo Mutation ◽  
Easy Access ◽  
Sperm Dna Fragmentation ◽  
Dna Base ◽  
Sperm Dna

One important reason for male infertility is oxidative stress and its destructive effects on sperm structures and functions. The particular composition of the sperm membrane, rich in polyunsaturated fatty acids, and the easy access of sperm DNA to oxidative damage due to sperm cell specific cytologic and metabolic features (no cytoplasm left and cells unable to mount stress responses) make it the cell type in metazoans most susceptible to oxidative damage. In particular, oxidative damage to the spermatozoa genome is an important issue and a cause of male infertility, usually associated with single- or double-strand paternal DNA breaks. Various methods of detecting sperm DNA fragmentation have become important diagnostic tools in the prognosis of male infertility and such assays are available in research laboratories and andrology clinics. However, to date, there is not a clear consensus in the community as to their respective prognostic value. Nevertheless, it is important to understand that the effects of oxidative stress on the sperm genome go well beyond DNA fragmentation alone. Oxidation of paternal DNA bases, particularly guanine and adenosine residues, the most sensitive residues to oxidative alteration, is the starting point for DNA damage in spermatozoa but is also a danger for the integrity of the embryo genetic material independently of sperm DNA fragmentation. Due to the lack of a spermatozoa DNA repair system and, if the egg is unable to correct the sperm oxidized bases, the risk of de novo mutation transmission to the embryo exists. These will be carried on to every cell of the future individual and its progeny. Thus, in addition to affecting the viability of the pregnancy itself, oxidation of the DNA bases in sperm could be associated with the development of conditions in young and future adults. Despite these important issues, sperm DNA base oxidation has not attracted much interest among clinicians due to the lack of simple, reliable, rapid and consensual methods of assessing this type of damage to the paternal genome. In addition to these technical issues, another reason explaining why the measurement of sperm DNA oxidation is not included in male fertility is likely to be due to the lack of strong evidence for its role in pregnancy outcome. It is, however, becoming clear that the assessment of DNA base oxidation could improve the efficiency of assisted reproductive technologies and provide important information on embryonic developmental failures and pathologies encountered in the offspring. The objective of this work is to review relevant research that has been carried out in the field of sperm DNA base oxidation and its associated genetic and epigenetic consequences.

Do serum vitamin D levels correlate with semen analysis parameters and sperm DNA fragmentation?

2012 ◽  
Vol 98 (3) ◽  
pp. S47-S48
Author(s):  
L. Rubal ◽  
A.M. Hernandez ◽  
S. Ingles ◽  
M. Scrooc ◽  
K. Bendikson
Keyword(s):  
Vitamin D ◽  
Dna Fragmentation ◽  
Semen Analysis ◽  
Serum Vitamin ◽  
Sperm Dna Fragmentation ◽  
Serum Vitamin D ◽  
Sperm Dna ◽  
Vitamin D Levels

scholarly journals 86 THE CLINICAL IMPORTANCE OF SPERM DNA FRAGMENTATION ASSAY IN THE ASSESSMENT OF MALE INFERTILITY

2010 ◽  
Vol 20 ◽  
pp. S34-S35
Author(s):  
S. Venkatesh ◽  
A. Singh ◽  
M.B. Shamsi ◽  
R. Kumar ◽  
D.N. Mitra ◽  
...  
Keyword(s):  
Male Infertility ◽  
Dna Fragmentation ◽  
Clinical Importance ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

The assessment of sperm DNA fragmentation in the saltwater crocodile (Crocodylus porosus)

2017 ◽  
Vol 29 (3) ◽  
pp. 630 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
J. L. Fernández ◽  
J. Gosálvez
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Saltwater Crocodile ◽  
Crocodylus Porosus ◽  
Dispersion Test ◽  
Sperm Nuclei

Herein we report a method of assessing DNA fragmentation in the saltwater crocodile using the sperm chromatin dispersion test (SCDt) after including frozen–thawed spermatozoa in a microgel (Halomax; Halotech DNA, Madrid, Spain). Following controlled protein depletion, which included a reducing agent, sperm nuclei with fragmented DNA showed a homogeneous and larger halo of chromatin dispersion with a corresponding reduced nucleoid core compared with sperm with non-fragmented DNA. The presence of DNA damage was confirmed directly by incorporation of modified nucleotides using in situ nick translation (ISNT) and indirectly by studying the correlation of the SCDt with the results of DNA damage visualisation using a two-tailed comet assay (r = 0.90; P = 0.037). Results of the SCDt immediately following thawing and after 5 h incubation at 37°C in order to induce a range of DNA damage revealed individual crocodile differences in both the baseline level of DNA damage and DNA longevity.

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