scholarly journals Analysis of sperm DNA fragmentationin infertility patients

2012 ◽  
Vol 61 (3) ◽  
pp. 141-147 ◽  
Author(s):  
Evgeniya Michailovna Shilnikova ◽  
Irina Dmitrievna Fedorova ◽  
Alexander Mkrtichevich Gzgzyan

Using Terminal Deoxynucleotidyl Transferase dUTP Nick End Labelling (TUNEL) technique the assessment of the frequency of the spermatozoa with fragmented DNA in the ejaculate from sperm donors and men with infertility was analyzed. The DNA fragmentation rate was higher in spermatozoa of carriers of a chromosomal structural abnormality compared with the control group. There was no correlation between the sperm DNA fragmentation rate and the parameters of semen analysis. The direct linear correlation between the frequency of the spermatozoa with fragmented DNA and vacuole sperm head was found. The DNA fragmentation rate was not correlated to the frequency of the spermatozoa with bulb, amorphous heads or spermatozoa with abnormal acrosome

2019 ◽  
Vol 20 (4) ◽  
pp. 39-44
Author(s):  
S. Sh. Khayat ◽  
E. E. Bragina ◽  
E. A. Arifulin ◽  
E. M. Lazareva ◽  
T. M. Sorokina ◽  
...  

The study objective is to analyze the content of spermatozoa with single and double-stranded DNA breaks in different age groups.Materials and methods. The level of DNA fragmentation was studied in 300 ejaculate samples obtained from 266 sub- or infertile men. The group 1 included 150 samples obtained from 131 patients under the age of 45 (21–44 years), the group 2 included 150 samples obtained from 135 patients above the age of 45 (45–68 years). Mean ages were 34.8 ± 3.9 and 48.6 ± 3.1 years, respectively. The number of sperm with fragmented DNA was evaluated using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method on ejaculate smears. The number of spermatozoa with >15 % of fragmented DNA was considered elevated. Standard semen analysis was performed in 117 and 97 men from the groups 1 and 2, respectively.Results. The number of sperm with fragmented DNA varied in ejaculated samples from 1.5 to 64.5 %. Mean number of sperm with DNA breaks in the group 1 (12.0 ± 6.0 %) was significantly lower than in the group 2 (16.1 ± 8.3 %, p <0.05). Mean sperm count in the ejaculate of the group 1 (267.0 ± 198.7 million) was significantly higher than in the group 2 (201.0 ± 162.9 million, p = 0.02).Conclusion. We revealed that in men over the age of 45 years, the percentage of spermatozoa with DNA fragmentation is higher than in men under 45 years of age, it may indirectly indicate an increased level of reactive oxygen species in the seminal plasma in older patients. 


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N Kilic ◽  
T İrez ◽  
N Dayiolu

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 &lt; 0,001 , p &lt; 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


2019 ◽  
Vol 91 (2) ◽  
Author(s):  
Viktor Alargkof ◽  
Larissa Kersten ◽  
Romil Stanislavov ◽  
Zdravko Kamenov ◽  
Panagiotis Nikolinakos

Objective: This exploratory retrospective study aimed to compare the level of Sperm DNA Fragmentation (SDF) and investigate its association with bulk semen parameters, for the first time in Bulgarian patients with varicocele, using a distinct methodology. Material and methods: Standard semen analysis was performed according to the 2010 criteria of the European Society of Human Reproduction and Embryology - Nordic Association for Andrology (ESHRE-NAFA-2010) and DNA fragmentation was assessed using the Halosperm® kit. The total sample included 28 males: the control group consisted of men with normal genital examination and unknown fertility (n = 10), group one consisted of men with varicocele, normozoospermia and DNA fragmentation > 15% (n = 9) and group two consisted of men with varicocele, abnormal sperm parameters and DNA fragmentation > 15% (n = 9). Results: DNA fragmentation was found to be higher in patients with abnormal sperm parameters (43.78 ± 30.78) compared to the normozoospermic group (21.22 ± 3.93) (p = 0.008). In normozoospermic patients, no statistically significant correlations were observed between SDF and bulk semen parameters. In patients with abnormal sperm parameters, DNA fragmentation exhibited significant very strong negative association with motility (a+b), vitality and typical morphology (p < 0.001). Conclusions: DNA integrity assays could be used for a better evaluation and management of male infertility, particularly in normozoospermic varicocele patients.


2020 ◽  
Vol 47 (4) ◽  
Author(s):  
L. L. Voroncova ◽  
S. O. Kenijz ◽  
V. A. Kovalenko

Abstract Purpose of the study. To study the features of changes of spermiologic values and degree of spermatozoa DNA fragmentation depending on the presence/absence of toxocariasis invasion. Materials and methods. 89 men aged 20 to 45 years were examined, which were divided into 5 groups. The first (control) group was 12 fertile men; the second group (comparison group) – 27 infertility patients with normal level of DNA fragmentation of sperm and without antibodies to toxocariasis; The third group – 20 infertility men with normal level of DNA fragmentation of sperm and presence of antibodies to toxocariasis. By the fourth and fifth groups included 15 infertility men with high levels of DNA fragmentation of sperm and the presence of antibodies to toxocariasis and lack of respectively. A comprehensive research was conducted for all men that included analysis of spermogram according to WHO recommendations, determination of sperm DNA fragmentation and presence of toxocariasis invasion. Results. As a result of spermograms in the studied groups of men were identified the violations of ejaculate fertility (asteno-, teratoand oligozoospermia). It has been established that the degree disturbance of spermatogenesis (including significant oligo- and teratozoospermia) is depending on the fragmentation of DNA in male gametes and the presence of antibodies to toxocara. At the same time more severe patozoospermia observed in the presence of elevated levels of fragmented sperms and the absence of antibodies to toxocara antigens, which is the basis for further research. Conclusions. Thus, the determination of sperm DNA fragmentation and availability of toxocariasis invasion in men with reproductive disorders is a necessary component of a comprehensive examination, which may allow to improve the diagnostics of male infertility and to promote the development of the right tactics and optimal treatment regimens. Keywords: male infertility, spermogram, sperm DNA fragmentation, toxocariasis invasion


2012 ◽  
Vol 98 (3) ◽  
pp. S47-S48
Author(s):  
L. Rubal ◽  
A.M. Hernandez ◽  
S. Ingles ◽  
M. Scrooc ◽  
K. Bendikson

2017 ◽  
Vol 29 (3) ◽  
pp. 630 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
J. L. Fernández ◽  
J. Gosálvez

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.


2020 ◽  
Vol 9 (12) ◽  
pp. 3976
Author(s):  
Alberto Pacheco ◽  
Arancha Blanco ◽  
Fernando Bronet ◽  
María Cruz ◽  
Jaime García-Fernández ◽  
...  

Magnetic-activated cell sorting (MACS) can be used to separate apoptotic sperm with high proportions of fragmented DNA from the rest, thus improving the overall quality of the seminal sample. Therefore, the aim of this retrospective study was to investigate the efficiency of the MACS technique to increase reproductive outcomes in patients with high levels of sperm DNA fragmentation (SDF) undergoing intracytoplasmic sperm-injection (ICSI) cycles. In this study, we analyzed a total of 724 assisted-reproduction-technique (ART) cycles that were divided into two groups: the study group (n = 366) in which the MACS selection technique was performed after density-gradient centrifugation (DGC), and the control group (n = 358) in which only DGC was used for sperm selection. Reproductive outcomes were analyzed in both groups according to three different ART procedures: preimplantation genetic testing for aneuploidy (PGT-A), and autologous and oocyte-donation cycles. The MACS group showed significantly lower miscarriage rates in autologous ICSI cycles, higher pregnancy rates in oocyte-donation cycles, and a significant increase in live-birth rates in both autologous and oocyte-donation cycles. Overall, these results suggested that the MACS technique can be effectively used to eliminate sperm with high SDF levels, and therefore may help to improve reproductive outcomes in couples undergoing ART.


2019 ◽  
Vol 26 (12) ◽  
pp. 1575-1581 ◽  
Author(s):  
Senay Cankut ◽  
Turgay Dinc ◽  
Mehmet Cincik ◽  
Guler Ozturk ◽  
Belgin Selam

Aim: Human sperm DNA fragmentation is one of the factors suggested for male infertility. The ratio of sperm DNA damage in semen may adversely affect both the fertilization rate and the embryo development of in vitro fertilization/ intracytoplasmic sperm injection cycles. Sperm cryopreservation both increases the success rates in assisted reproductive techniques (ARTs) and contributes to the preservation of fertility before testis surgery, chemotherapy, and radiotherapy. The aim of the current study is to determine sperm DNA fragmentation, following cryopreservation. Methods: A cross-sectional, observational study was conducted at a university hospital infertility clinic. One hundred (n = 100) volunteer fertile men (ages between 21 and 39 years) with normozoospermic sperm parameters were involved in the current study. Sperm DNA damage was evaluated with the Halosperm technique and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Fresh samples were studied in liquid form. The remaining samples were kept frozen and then thawed after 1 month and reevaluated with the Halosperm technique and TUNEL assay. Results were then compared between the fresh and frozen samples. Results: Sperm DNA fragmentation results with the Halosperm technique both before and after cryopreservation were 25% (5%-65%) and 40% (6%-89%), respectively, with a statistically significant increase (15%; P < .001). Sperm DNA fragmentation results by TUNEL assay before and after cryopreservation were 17% (3%-43%) and 36% (7%-94%), respectively, with a statistically significant increase (19%; P <.001). Conclusion: The current data demonstrate increased sperm DNA damage after cryopreservation. Further studies may contribute to development of less harmful techniques and cryoprotectants in order to improve the results of ART.


2016 ◽  
Vol 44 (6) ◽  
pp. 1283-1291 ◽  
Author(s):  
Lin-Tao Xue ◽  
Rui-Xue Wang ◽  
Bing He ◽  
Wei-Ying Mo ◽  
Li Huang ◽  
...  

Objective To investigate the effect of sperm DNA fragmentation on the fertilization rate, embryo development and pregnancy outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in a cohort of Chinese couples. Methods Infertile couples that had undergone assisted reproductive technology at our centre between January 2011 and December 2013 were included in this retrospective study. Fractions of prepared sperm samples were evaluated for sperm DNA fragmentation on the day of oocyte recovery. Results Of the 550 couples selected, 415 had undergone IVF and 135 ICSI. Sperm DNA fragmentation rate was significantly negatively correlated with the fertilization rate in the ICSI cycles but not the IVF cycles. No association was found between sperm DNA fragmentation and cleavage rate or good quality embryo formation rates in IVF or ICSI cycles. Receiver operating characteristic (ROC) curve analysis showed that the sperm DNA fragmentation rate was a statistically significant prognostic indicator of the clinical fertilization rate in ICSI cycles; a rate > 22.3% was associated with a lower fertilization rate following ICSI compared with a rate ≤ 22.3%. Conclusions High values of sperm DNA fragmentation were associated with a low fertilization rate following ICSI but were not associated with alterations in pregnancy or live birth rates in either ICSI or IVF in this cohort of Chinese couples.


2005 ◽  
Vol 17 (2) ◽  
pp. 282
Author(s):  
D. Evenson

Sperm DNA integrity is obviously important for normal embryo development and pregnancy outcome. Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. The Sperm Chromatin Structure Assay (SCSA) treats sperm with low pH to denature DNA at the sites of DNA strand breaks, followed by acridine orange (AO) staining of green for native DNA and red for denatured DNA, as measured by flow cytometry (FCM), as well as % sperm with high DNA stainability (HDS: immature sperm with intact DNA related to decreased fertilization rates). FCM-sorted sperm from each SCSA-defined population (normal, moderate, and high DNA fragmentation and HDS sperm) show that the moderate DNA fragmentation index (DFI) population has the same image analysis characteristics as normal sperm without significant comets. Thus, an ICSI technician is not likely to differentiate between a normal and a moderate DFI sperm. The TUNEL assay uses an enzyme to add a fluorochrome-labeled base to a 3′-OH broken DNA strand. Both light microscopy and flow cytometry are used for measuring the % and extent of DNA fragmentation but cannot measure the level of HDS. For the COMET assay, sperm are suspended in an electrophoretic gel, placed on a glass microscope slide, digested with proteases and RNAse, subjected to an electric field, and then stained with a DNA dye. The % of comet positive sperm is scored, but the extent of fragmentation is difficult to define and the % HDS cannot be determined. Small pieces of fragmented DNA migrate in the gel forming a “comet.” All three methods have been used for both research and clinical diagnosis and as prognosis for livestock (bulls, boars, rams, stallions) and humans. Light microscope techniques suffer from a lack of statistical soundness needed for clinical decisions as well as present a potential bias in selection of sperm for measurements. Due to the thousands of sperm randomly selected for flow cytometry measurements, the data are statistically robust. Data from all three kinds of measurements in over a hundred manuscripts clearly show that sperm DNA fragmentation has a negative impact on embryo growth and pregnancy. Infertile animals may have nearly all of the sperm with fragmented DNA. Fertility ratings in bulls and boars are clearly related to the percent and extent of DNA fragmentation. Threshold levels for fertile/sub fertile/infertile differ for different species. Likewise different methods/laboratories have suggested various threshold levels to characterize a man with a highly fertile to low/very poor potential. The range of sperm with fragmented DNA is from ∼2% to 100%. The SCSA method has defined a 27–30% DFI as the point in which a man is placed into a statistical category of taking a longer time to achieve in vivo pregnancy, more intrauterine insemination and routine IVF cycles, or no pregnancy. Current data suggest that ICSI may help overcome the diminished pregnancy prognosis with high DFI over the other ART or natural methods.


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