scholarly journals Induced lipid peroxidation in ram sperm: semen profile, DNA fragmentation and antioxidant status

Reproduction ◽  
2016 ◽  
Vol 151 (4) ◽  
pp. 379-390 ◽  
Author(s):  
Thais Rose dos Santos Hamilton ◽  
Letícia Signori de Castro ◽  
Juliana de Carvalho Delgado ◽  
Patrícia Monken de Assis ◽  
Adriano Felipe Perez Siqueira ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Dna Fragmentation ◽  
Sperm Quality ◽  
Ovis Aries ◽  
Sperm Dna Fragmentation ◽  
Mitochondrial Potential ◽  
Sperm Analysis ◽  
Sperm Dna ◽  
Significant Difference ◽  
Ram Sperm

Action of reactive oxygen species, protamination failures and apoptosis are considered the most important etiologies of sperm DNA fragmentation. This study evaluated the effects of induced lipid peroxidation susceptibility on native semen profile and identified the mechanisms involved in sperm DNA fragmentation and testicular antioxidant defense on Santa Ines ram sperm samples. Semen was collected from 12 adult rams (Ovis aries) performed weekly over a 9-week period. Sperm analysis (motility, mass motility, abnormalities, membrane and acrosome status, mitochondrial potential, DNA fragmentation, lipid peroxidation and intracellular free radicals production); protamine deficiency; PRM1, TNP1 and TNP2 gene expression; and determination of glutathione peroxidase (GPx), glutathione reductase, catalase (CAT) and superoxide dismutase activity and immunodetection in seminal plasma were performed. Samples were distributed into four groups according to the sperm susceptibility to lipid peroxidation after induction with ascorbate and ferrous sulfate (low, medium, high and very high). The results were analyzed by GLM test and post hoc least significant difference. We observed an increase in native GPx activity and CAT immunodetection in groups with high susceptibility to induced lipid peroxidation. We also found an increase in total sperm defects, acrosome and membrane damages in the group with the highest susceptibility to induced lipid peroxidation. Additionally, the low mitochondrial membrane potential, susceptible to chromatin fragmentation and the PRM1 mRNA were increased in the group showing higher susceptibility to lipid peroxidation. Ram sperm susceptibility to lipid peroxidation may compromise sperm quality and interfere with the oxidative homeostasis by oxidative stress, which may be the main cause of chromatin damage in ram sperm.

P–025 Sperm selection using a modified “swim up” technique in absence of sperm centrifugation improve sperm DNA fragmentation and decreases miscarriage rate

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Cabell. Vives ◽  
P Belchin ◽  
C Lopez-Fernandez ◽  
M Fernandez-Rubio ◽  
J Guerrero-Sanchez ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
P Value ◽  
Sperm Dna Fragmentation ◽  
Sperm Selection ◽  
Male Factor ◽  
Miscarriage Rate ◽  
Sperm Dna ◽  
Group 2

Abstract Study question Is it useful to avoid sperm centrifugation in laboratory routine work to improve sperm quality and reproductive outcome in Assisted Reproduction Techniques (ART)? Summary answer Exclusion of sperm centrifugation for sperm selection using neat sperm samples (IO-lix), increases sperm quality in the collected subpopulation decreasing miscarriage rate after using ICSI. What is known already Inclusion of sperm centrifugation in ART is an aggressive intervention for sperm selection with ineludible production iatrogenic damage affecting sperm integrity. The application of IMSI, PICSI or microfluidic devices avoid sperm centrifugation and may improve the quality of the subsample obtained. However, these methodologies may result time consuming, expensive or producing poor results when the quality of the sperm is limited. We have already shown that a modified swim-up avoiding centrifugation (called IO-lix) is a low-cost and efficient alternative to microfluidic devices, recovers 100 times more concentration and reduces sperm DNA fragmentation with no significant differences to other methodologies. Study design, size, duration This is a retrospective study from 2018 to 2020 which includes patients with an average of age of 38.2 years using their own oocytes with ICSI as fertilization technique. Two aleatory groups of patients were made: Group 1: 88 cycles with 503 fertilized oocytes and 206 blastocysts were obtained with sperm samples processed by IO-lix and Group 2: 303 cycles, 1451 fertilized oocytes and 591 blastocysts using a standard “swim up” technique to process sperm. Participants/materials, setting, methods A total of 391 ICSI cycles were included in this retrospective study. The male factor was similar in both groups and they showed altered SDF previously to the cycle. We compared data of the motility and SDF of sperm samples before and after applying IO-lix and we analyzed by X2 contingence test differences on miscarriage rates between groups 1 and 2. Main results and the role of chance General sperm parameter changes after IO-lix showed that averaged sperm concentration observed in neat ejaculated samples was 62M/SD=46.4. Values obtained after IO-lix in the same samples were 12.3M/SD8.0. Averaged sperm motility in neat samples was 54%/SD=9.3 and 70.9%/SD=13.2 after IO-lix. Finally, sperm DNA fragmentation in neat samples was 35.8%/SD17.3, while these values decreased to 9.2%/SD=3.9 after IO-lix. About reproductive outcome results, significant differences were not obtained on the development to blastocyst stage rate comparing both groups (X2=0.003; p value = 0.954; Alpha 0.05). In the case of IO-lix processed samples, the pregnancy rate was 59.42% in Group 1 and 44.72% in Group 2 (X2=0.651; p value =0.419; Alpha 0.05). A total of 9 miscarriages of 41 clinical pregnancies (21.95%) were observed after IO-lix, while this number increases to 59 out of 123 clinical pregnancies, which means the 47.96% of the embryo transfers, when “swim-up” was used. In this case significant differences were obtained (X2=3.935; p value = 0.0.047; Alpha 0.05). Limitations, reasons for caution Being a pilot study aimed to understand the results of IO-lix in ART, correlations have not been stablished between the levels of sperm improvement after IO-lix and paired results of ART. This study would be necessary, specially to identify the possible origin of miscarriage associated to the male factor. Wider implications of the findings: Elimination of sperm centrifugation using a combined strategy of gradients and “swim-up” for sperm isolation, reduce miscarriage rate and produce equivalent results of blastocyst development to those obtained with “swim-up”. Being a cost-effective and improving laboratory workload, its use for sperm selection is recommended. Trial registration number Not applicable

scholarly journals Obesity as a factor in spermatogenesis disorders (experimental study)

2020 ◽  
Vol 21 (2) ◽  
pp. 36-43
Author(s):  
A. A. Artamonov ◽  
S. V. Bogolyubov ◽  
T. I. Eliseeva ◽  
O. B. Pozdnyakov ◽  
A. V. Astakhova
Keyword(s):  
Total Cholesterol ◽  
Dna Fragmentation ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Study Objective ◽  
Diet Induced Obesity ◽  
White Rats ◽  
Sperm Dna ◽  
Significant Difference

Introduction. In recent years, the effects of obesity on male fertility have been extensively investigated. The results of existing studies are extremely contradictory.The study objective was to determine the effect of obesity on the male reproductive system using the biological model of laboratory rats as an example.Materials and methods. In vivo modeling of diet-induced obesity. The study was conducted on 22 laboratory sexually mature white rats weighing 140–160 g. The animals were divided into two groups: 1 control (10 animals) and 2 rats with diet-induced obesity (12 animals). After 12 weeks, the animals were removed from the experiment. All rats underwent: calculation of the Lee index (body mass index in rats), determination of the concentration and viability of spermatozoa in a suspension of sperm from the epididymis, determination of glucose level of total cholesterol and triglycerides in the blood, study of sperm DNA fragmentation, histological examination testis: calculating the crosssectional area of the seminiferous tubule; determination of the number of non-functioning tubules and tubules with desquamated spermiogenic epithelium; determination of the average spermatogenesis index.Results. In the study groups there were no differences in glucose and total cholesterol levels. However, a statistically significant, significant difference in the level of triglycerides in the blood was revealed. The concentration of sperm and their viability in the studied groups did not differ. The level of sperm DNA fragmentation in the experimental group is significantly higher than in the control group (31.5 ± 10.1 and ± 1.4 %, respectively, p <0.05). Morphometric evaluation of histological preparations did not establish differences in the cross-sectional area of the seminiferous tubules and the average spermatogenesis index in the studied groups. In rats with obesity, compared with the control group, significantly more non-functioning tubules (2.9 ± 0.3 and 8.4 ± 0.3; p <0.05) and tubules with desquamated spermatogenic epithelium (1.8 ± 0.3 and 8.8 ± 0.5; p <0.05).Conclusion. Diet-induced obesity causes impaired spermatogenesis, and damage to the sperm genetic material in male white rats.

P–091 Magnetic Activated Cell Sorting (MACS) improves euploid blastocysts rate in pre-implantation genetic testing cycles with high levels of sperm DNA fragmentation and advanced paternal age

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Scarselli ◽  
E Cursio ◽  
A Colasante ◽  
V Zazzaro ◽  
P Andrea ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Dna Fragmentation ◽  
Sperm Quality ◽  
Annexin V ◽  
Paternal Age ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Number Of Patients ◽  
Blastocyst Rate

Abstract Study question Can MACS increase euploid blastocyst rate in Pre-implantation Genetic Testing (PGT) cycles for AMA-APA (Advanced Maternal-Paternal Age) in patients with high sperm DNA fragmentation (SDF)? Summary answer A slight increase in euploid blastocyst rate was found using MACS in infertile patients with high SDF undergoing PGT cycles compared to the control group. What is known already Many authors have shown a close correlation between the presence of apoptotic markers on spermatozoa and the failure of assisted reproduction treatments. In normal physiological conditions, apoptotic spermatozoa with phosphatidylserine (PS) residues externalized on the plasma membrane, are eliminated along female genital tract, preventing oocyte fertilization. MACS eliminates apoptotic sperm whit PS residues using superparamagnetic microbeads conjugated with annexin V. This technique reduces the proportion of sperm with high rates of SDF and can be used to maximize ART procedures results. MACS application improves sperm quality, fertilization, cleavage and pregnancy rates reducing miscarriage rate. Study design, size, duration From June to November 2020, 10 couples in which MACS was applied to select non-apoptotic spermatozoa, were randomly enrolled in our study (MACS group) and 8 couples without MACS were considered as controls (No-MACS Group). All couples in both groups underwent a PGT cycle and had high sperm DNA Fragmentation (&gt; 20%). A higher rate of euploid and diploid-euploid mosaic blastocysts were obtained in the MACS group compared to the control group. Participants/materials, setting, methods Patients with severe oligoastenoteratozoospermia were excluded. MACS protocol was performed as follows: semen sample was analyzed (WHO 2010) and washed with buffered medium; pellet was removed and a swim-up was performed. Retrieved spermatozoa were washed with a binding buffer (Miltenyi Biotec), centrifuged (400 g x 4 minutes) and supernatant discarded. Pellet was covered with Annexin-V and re-suspended. After 15 minutes incubation at room temperature, the sample was eluted through the column and collected for ICSI. Main results and the role of chance In MACS group, female and male mean age ± SD were 41.6 ± 2.1 and 43.5 ± 7.3, respectively. Female and male mean age ± SD were 41.7 ± 2.8 and 44.6 ± 8.1 in the No-MACS group, respectively. In MACS and No-MACS groups, injected oocytes were 44 and 35, fertilized oocytes were 32 (72.3%) and 27 (77.1%) (NS), blastocyst formation rates were 71.8% (23/32) and 48.1% (13/27) (NS), respectively. In No-MACS group, only 1 euploid and 1 diploid-euploid mosaic blastocysts were obtained (1/13 = 8%) (NS). In MACS group, 4 euploid blastocysts were formed (4/23 = 17.4%) whereas mosaic diploid-euploid blastocysts were 3/23 (13.0%) (NS). Aneuploid blastocysts were 16/23 (69.6%) in MACS group and 11/13 (84.6%) in No-MACS group (NS). Limitations, reasons for caution AMA and APA of couples enrolled should be considered as a limit of the study. A larger number of patients and biopsied blastocysts are needed to analyze clinical results and perform a robust statistical analysis establishing if MACS is useful to improve transferable blastocyst rate in patients with high SDF. Wider implications of the findings: MACS is useful to select non apoptotic sperms; although fertilization, cleavage and blastocyst rates are not improved, aneuploid blastocysts rate slightly decreases using MACS. It I possible that, selecting spermatozoa free from PS residues, MACS allows to choose spermatozoa with a better DNA packaging, thus affecting the embryo ploidy. Trial registration number non applicable

scholarly journals The correlation between male age, sperm quality and sperm DNA fragmentation in 320 men attending a fertility center

2008 ◽  
Vol 26 (1) ◽  
pp. 41-46 ◽  
Author(s):  
Thomas Winkle ◽  
Bernd Rosenbusch ◽  
Friedrich Gagsteiger ◽  
Thomas Paiss ◽  
Nicole Zoller
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Sperm Dna Fragmentation ◽  
Male Age ◽  
Sperm Dna

scholarly journals Is there a connection with basic sperm parameters and age?

2020 ◽  
Vol 13 (4) ◽  
pp. 58-64
Author(s):  
A.I. Ryzhkov ◽  
◽  
I.S. Shormanov ◽  
S.Yu. Sokolova ◽  
◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Male Fertility ◽  
Reproductive Technologies ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Who Guidelines ◽  
Negative Relation ◽  
Sperm Analysis ◽  
Mobile Forms ◽  
Sperm Dna

Introduction. TStandard sperm examination, performed according to WHO guidelines, remains the main method of assessment of male fertility. At the same time, this research method has a number of significant limitations, including a low predictive value regarding the outcomes of assisted reproductive technologies (ART) programs and the outcome of naturally occurring pregnancies. The limitations of standard sperm examination dictate the need for additional methods for assessing male fertility. The most promising and widely used test is the assessment of the level of sperm DNA fragmentation. Aim. To evaluate the associations between the levels of sperm DNA fragmentation and the age of the patients along with the following parameters used as a part of standard sperm analysis: semen volume, total number of spermatozoa, percentages of progressively-mobile, non-progressively-mobile and immobile forms, percentages of morphologically normal forms and the forms bearing head defects within the structure of total number of morphological anomalies, as well as semen leukocyte count. Materials and methods. Study materials were the examination results from 121 males aged from 21 to 53 years old (mean age 32.7±4.5 years old), undergoing an examination within the Clinical Institution «Mother and Child – Yaroslavl» during a time period from January 2019 until April 2020. Standard sperm analysis procedures were carried out according to latest edition of WHO Guidelines (2010). The determinations of germ cell DNA fragmentation levels were performed using the TUNEL method. Results. The test results have revealed a weak negative relation between the sperm DNA fragmentation (%) and the percentage of progressively-mobile forms (%) – r = -0.26 (p <0.01). The correlation of sperm DNA fragmentation with age and other parameters was considered statistically insignificant. The second stage of the analysis have demonstrated that the increased degree sperm DNA fragmentation is 1.8-fold more often found in the patients having signs of astenozoospermia (23.6%) comparing to the patients showing normal degree of spermatozoa mobility (13.1%), (p <0.05). Conclusions. The level of sperm DNA fragmentation correlates with the percentage of progressively-mobile forms of spermatozoa (negative relation) and no correlations were found with other semen parameters and with the age. The rates of increased levels of sperm DNA fragmentation are 1.8-fold more often found in astenozoospermia patients comparing to the patients showing normal degrees of spermatozoa mobility.

scholarly journals Antioxidant treatment of increased sperm DNA fragmentation: Complex combinations are not more successful

2020 ◽  
Vol 92 (4) ◽  
Author(s):  
Cevahir Ozer
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Single Agent ◽  
Included Study ◽  
Sperm Dna Fragmentation ◽  
Antioxidant Treatment ◽  
Sperm Dna Damage ◽  
Treatment Groups ◽  
Sperm Dna ◽  
Significant Difference

Objective: Oral antioxidant supplementation is part of the treatment of infertility associated with oxidative stress-related sperm damage. It is possible to assume that the combined use of antioxidants will be better than single agent use. The purpose of this study was to compare the effectiveness of different antioxidant combinations in infertile men with increased sperm DNA fragmentation. Materials and methods: We retrospectively reviewed the records of 637 patients who underwent antioxidant support therapy for increased sperm DNA damage between 2014 and 2019. Patients with DNA damage of 30% or more were included study. Result: A total of 163 patients with follow-up data and who fulfilled the study criteria were included in the study. There were four different treatment groups. No statistically significant differences were found between the groups. After 3 months of antioxidant treatment, there was a statistically significant decrease in sperm DNA damage in all treatment groups. However, there was no statistically significant difference between the treatment groups. Conclusions: The complexity of the antioxidant combination may not contribute to the success of the treatment or may cause possible side effects, increase the cost of treatment and decrease patient compliance.

scholarly journals Short communication. Stallion sperm quality after combined ejaculate fractionation and colloidal centrifugation

2015 ◽  
Vol 13 (4) ◽  
pp. e04SC02 ◽  
Author(s):  
Francisco Crespo ◽  
Jaime Gosálvez ◽  
Stephen D. Johnston ◽  
Joaquina De la Torre
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Rate Of Increase ◽  
Sperm Sample ◽  
Artificial Vagina ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

<p>This study investigated the possible additive benefit of ejaculate fractionation and colloidal centrifugation on stallion sperm quality. Using an open-end artificial vagina, the sperm-rich fraction (FRAC-1) was separated from the rest of the ejaculate (FRAC-2) and a third sperm sample representing the combined ejaculate was reconstituted post-ejaculation (RAW). Each semen sample was processed for colloidal centrifugation. The percentage of abnormal spermatozoa was 17.8 ± 7.0% in RAW and 14.6 ± 9.5% in FRAC-1 but decreased to 11.4 ± 4.7% and 9.6 ± 6.9% respectively, after colloidal centrifugation. A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation. The rate of increase in sperm DNA fragmentation during the first 6 h of incubation at 37 ºC was 1.8 ± 0.9% per hour in RAW semen and 2.0 ± 2.0% per hour in FRAC-1 but this significantly decreased to 1.3 ± 1.4% and 0.9 ± 0.8% respectively after colloidal centrifugation. While stallion seminal characteristics can be improved using colloidal centrifugation, further enhancement is possible if the ejaculate is initially fractionated.</p>

scholarly journals Effect of bacterial infection on sperm quality and DNA fragmentation in subfertile men with Leukocytospermia

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fatemeh Eini ◽  
Maryam Azizi Kutenaei ◽  
Fayegheh Zareei ◽  
Zeinolabedin Sharifian Dastjerdi ◽  
Maryam Hosseinzadeh Shirzeyli ◽  
...  
Keyword(s):  
Bacterial Infection ◽  
Dna Fragmentation ◽  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Seminal Fluid ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

Abstract Background Although bacterial infections have been recognized as a possible cause of male infertility, the effect of bacterial infections on sperm quality and sperm DNA fragmentation remains controversial. The current study aimed to investigate the prevalence rate of bacterial infection in subfertile men and its effect on semen quality. Seminal fluid was collected from 172 male members of infertile couples attending the andrology infertility center and a group of 35 fertile subjects as a control. Sperm parameters and DNA fragmentation were evaluated based on the type of bacteria in all ejaculates. Results From the 172 patients investigated for infertility, 60 (34.88%) patients had a positive culture for pathogenic bacteria of different species. Leukocytospermia was significantly higher in infected samples in comparison with non-infected samples (p < 0.05). Sperm concentration and motility and morphology were significantly lower in infected than non-infected samples. Moreover, sperm DNA fragmentation was significantly higher in infected than non-infected samples. Besides, our results showed that sperm DNA fragmentation was correlated significantly with leukocytospermia (R: 0.22, p < 0.01). Conclusion The present study suggested that bacterial infection significantly correlated with leukocytospermia could impair male fertility potential through decreasing sperm motility, morphology, and DNA integrity.

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