Percentage Of Sperm DNA Fragmentation Research Articles

Analyzing and Examining Smoking Effects and Oxidative Stress Markers in Seminal Plasma in Iraqi Infertility Men

The oxidative stress (OS) and sperm DNA fragmentation have been associated with sub-infertility in males and infertility due to a lack of means of protection and defense (antioxidants) for spermatozoa and an increase in reactive oxygen spices (ROS) related to wrong lifestyle (cigarettes and hookah) or what also known as Shisha. The study included (90) men between the ages of (23-40) years, (60), infertile men who smoked cigarettes and hookahs. They were divided into two groups, the first oligozoospermia and the second asthenozoospermia, according to the guidelines of the World Health Organization for seminal fluid analysis (SFA), 5 th edition, WHO(2010). Each group included (30) smoking infertile males, with a control group included (30) non-smoking fertile males. The study aimed to estimate the concentration level of Ascorbic Acid (AA) in the seminal plasma of two groups of infertile men smoking cigarettes and hookahs using enzyme-linked immunosorbent assay (ELISA) method, with calculating the concentration of Malondialdehyde (MDA) as well as estimating the percentage of sperm DNA fragmentation (SDF) using sperm chromatin dispersion (SCD), Halo view. Compared with healthy fertile men of non-smokers of cigarettes and hookahs as one of the biomarkers of oxidative stress in infertile men in Nineveh Governorate-Iraq. The results indicated that there were significant differences at the probability level (p<0.01) for the studied parameters (AA, MDA, SDF) compared to the control group (fertile men). The results showed an increase in the percentage of sperm DNA fragmentation with increasing the concentration of MDA in seminal plasma and decreasing the concentration of AA. Cigarette and hookah smoking increased the generation of ROS with a decrease in the water-soluble non-enzymatic antioxidant Ascorbic acid (AA) and thus increased the oxidative processes of the plasma membranes of spermatozoa and the occurrence of oxidative stress and impaired fertility and caused infertility. Keywords: male infertility, oxidative stress, sperm DNA fragmentation, Ascorbic acid, MDA. Bionatura http://dx.doi.org/10.21931/RB/CSS/2023

Male reproductive health after 3 months from SARS-CoV-2 infection: a multicentric study

PurposeWhile SARS-CoV-2 infection appears not to be clinically evident in the testes, indirect inflammatory effects and fever may impair testicular function. To date, few long-term data of semen parameters impairment after recovery and comprehensive andrological evaluation of recovered patients has been published. The purpose of this study was to investigate whether SARS-CoV-2 infection affect male reproductive health.MethodsEighty patients were recruited three months after COVID-19 recovery. They performed physical examination, testicular ultrasound, semen analysis, sperm DNA integrity evaluation (TUNEL), anti-sperm antibodies (ASA) testing, sex hormone profile evaluation (Total testosterone, LH, FSH). In addition, all patients were administered International Index of Erectile Function questionnaire (IIEF-15). Sperm parameters were compared with two age-matched healthy pre-COVID-19 control groups of normozoospermic (CTR1) and primary infertile (CTR2) subjects.ResultsMedian values of semen parameters from recovered SARS-CoV-2 subjects were within WHO 2010 fifth percentile. Mean percentage of sperm DNA fragmentation (%SDF) was 14.1 ± 7.0%. Gelatin Agglutination Test (GAT) was positive in 3.9% of blood serum samples, but no positive semen plasma sample was found. Only five subjects (6.2%) had total testosterone levels below the laboratory reference range. Mean bilateral testicular volume was 31.5 ± 9.6 ml. Erectile dysfunction was detected in 30% of subjects.ConclusionOur data remark that COVID-19 does not seem to cause direct damage to the testicular function, while indirect damage appears to be transient. It is possible to counsel infertile couples to postpone the research of parenthood or ART procedures around three months after recovery from the infection.

Effect of Sperm Selection by Magnetic-Activated Cell Sorting in D-IUI: A Randomized Control Trial.

Clinical outcome in assisted reproduction techniques (ARTs) is mainly influenced by the quality of gametes used. It is known that a high percentage of sperm DNA fragmentation (DNAf) decreases the success of ART clinical results. Therefore, techniques such as magnetic-activated cell sorting (MACS) help to improve results in cases of patients with a high percentage of DNAf. Cryopreservation of sperm in donor intrauterine insemination (D-IUI) treatments increases sperm DNAf, so patients using these sperm samples can benefit from using this technique. This prospective randomized national multicenter study analyzed clinical outcomes of 181 D-IUI treatments. MACS was performed after density gradient centrifugation (DGC) in 90 thawed semen donor samples (MACSG), whereas only DGC was performed in 91 thawed semen donor samples (CG). To our knowledge, this is the first study analyzing the effect of MACS on D-IUI cycles. Our results show no significant differences in gestation, live birth, or miscarriage rates between the two groups. We believe that further studies with a larger sample size are needed to evaluate the application of MACS in combination with standard IUI donor sperm preparations in fertility clinics.

Chlamydia trachomatis Infection: its relation to semen parameters and sperm DNA integrity

The present study was conducted to determine the effect of Chlamydia trachomatis (CT) infection on semen quality and sperm DNA integrity. The study included 60 infertile male patients with CT infection and 25 age matched controls. Diagnosis of patients was based on detection of CT IgA by ELISA in seminal plasma and CT plasmid DNA in the semen sediment. All patients and controls were subjected to the following investigations: history taking, conventional semen analysis, detection of CT IgA, Plasmid DNA in semen samples, reactive oxygen species (ROS) and percentage of DNA fragmentation. There was significant increase in semen ROS levels and the percentage of sperm DNA fragmentation in the CT patient group when compared to the control group (P<0.05) and in those with leukocytospermia when compared to those without leukocytospermia (P<0.05). In the patient group with CT infection there was a positive correlation between the percentage of DNA fragmentation, ROS (r = 0.82 with P<0.0001) and pus cell count. (r = 0.7 with P<0.0001). In patients with leukocytospermia, there was a positive correlation between the percentage of DNA fragmentation, ROS (r = 0.9 with P<0.0001) and pus cell count (r = 0.83 with P<0.0001). In conclusion, sperm concentration, mobility, and viability, are significantly decreased in patients with CT compared to controls. ROS levels and the percentage of sperm DNA fragmentation significantly increased in CT patients especially patients with leukocytospermia.

Retrospective Monocentric Clinical Study on Male Infertility: Comparison between Two Different Therapeutic Schemes Using Follicle-Stimulating Hormone.

Follicle-stimulating hormone (FSH) is a therapeutic option in patients with idiopathic oligozoospermia and normal FSH serum levels. However, few studies have evaluated which dose of FSH is more effective. The aim of this study was to compare the clinical efficacy of the two most frequently used FSH treatment regimens: 75 IU daily vs. 150 IU three times a week. Patients were retrospectively assigned to two groups. The first group (n = 24) was prescribed highly purified FSH (hpFSH) 75 IU/daily (Group A), and the second group (n = 24) was prescribed hpFSH 150 IU three times a week (Group B) for three months. Before and after treatment, each patient underwent semen analysis, evaluation of the percentage of DNA-fragmented spermatozoa, assessment of testicular volume (by ultrasonography), and measurement of FSH and total testosterone (TT) serum levels. Treatment with hpFSH significantly improved conventional sperm parameters. In detail, sperm concentration increased significantly after treatment only in Group A, whereas total sperm count, percentage of spermatozoa with progressive motility, normal morphology, or alive improved significantly in both groups. Interestingly, the percentage of sperm DNA fragmentation decreased significantly in both groups after treatment with hpFSH. FSH serum levels were expectably higher at the end of the treatment than before hpFSH was administered to both groups. Remarkably, TT serum levels only increased significantly in Group A. Finally, testicular volume was significantly higher in Group A after treatment, while it did not change significantly compared to baseline in Group B. The percentage of FSH responders did not differ significantly between the two groups (8/24 vs. 6/24). The daily administration of hpFSH 75 IU seems more effective than using 150 IU three times a week. However, this therapeutic scheme implies a higher number of injections and slightly higher costs.

The impact of varicocelectomy on sperm DNA fragmentation and pregnancy rate in subfertile men with normal semen parameters: A pilot study

ABSTRACT Objective: To evaluate the outcome of microscopic subinguinal varicocelectomy on sperm DNA fragmentation (SDF) and pregnancy rate in men with normal semen parameters. Patients and methods: A pilot study that included male patients with a minimum of a 1-year history of male subfertility, normal semen parameters, a high percentage of SDF, and clinically palpable varicoceles. Microscopic subinguinal varicocelectomy was carried out for 45 patients (study group), while 40 patients had no intervention (control group). Semen analysis and SDF were measured before and at 6 months after the varicocelectomy. The pregnancy rate was assessed at the 6- and 12-month follow-ups. Results: Between July 2014 and January 2019, 85 subfertile men were included in the study and completed 12 months of follow-up. The two groups were comparable in terms of their age, body mass index, infertility duration, infertility type, varicoceles laterality, and varicoceles grade (P values = 0.84, 0.34. 0.35, 1, 0.39, and 0.46, respectively). At 6 months after varicocelectomy, the mean SDF was reduced in both groups, and this reduction was statistically higher in the varicocelectomy group (P < 0.001). After 1-year, spontaneous pregnancy was achieved in 62% of the patients in the varicocelectomy group compared to 30% in the control group (P = 0.009). Conclusion: Varicocelectomy has a positive impact on SDF and spontaneous pregnancy in infertile men with clinically palpable varicoceles and normal semen parameters. Abbreviations: BMI: body mass index; DFI: DNA fragmentation Index; SDF: sperm DNA fragmentation

Exogenous melatonin advances the ram breeding season and increases testicular function

Governed by melatonin, ovine reproductive seasonality limits production outcomes due to periods of decreased reproductive efficiency. Though it is established that slow-release melatonin implants improve out of season reproductive performance in the ewe, the comprehensive effects of exogenous melatonin in the ram remain inconclusive. This study aimed to ultimately clarify the ability of exogenous melatonin to alter ram reproductive function during the non-breeding season and the subsequent breeding season. Hence, we investigated the effect of exogenous melatonin on reproductive endocrinology, semen quality and production, testicular size and libido in Merino and Poll Dorset rams (n = 31, using a subset of 18 rams for analysis of semen production and quality). Melatonin treatment resulted in elevation of melatonin in seminal plasma from 1–8 weeks post-implantation and in blood plasma at 6 weeks post-implantation. The blood plasma testosterone of implanted rams was greater than controls at both 6 weeks post-implantation and during the following breeding season. Implanted rams exhibited increased testicular size and number of sperm per ejaculate from 3–12 weeks post-implantation but did not demonstrate any change in sperm motility or morphology in response to treatment. Compared to their control counterparts, melatonin-treated Poll Dorset rams exhibited a lower percentage of sperm DNA fragmentation during several weeks of the non-breeding season. Though melatonin increased the likelihood of ejaculate collection in Poll Dorset rams (P < 0.05), libido was otherwise unaffected by treatment. Melatonin did not alter seminal plasma concentrations of inhibin A or Anti-Mullerian hormone, however, for the first time in the ram we have shown Anti-Mullerian hormone to be positively correlated with the number of sperm per ejaculate and sperm motility (r = 0.464 and 0.3242 respectively, P < 0.001), and inhibin A to be correlated to the number of sperm per ejaculate (r = 0.1786, P = 0.0135). These results indicate that melatonin is able to both systemically upregulate reproduction and act directly upon testicular function in the ram.

Predictive value of oxidative stress testing in semen for sperm DNA fragmentation assessed by sperm chromatin dispersion test.

Although standard semen analysis is a cornerstone of male infertility investiation, it has significant limitations. Seminal oxidative stress and DNA damage have shown potential to better predict male fertility potential and provide improved diagnostic and management strategies. The aim of this study was to investigate whether seminal oxidation-reduction potential (ORP), a new parameter to measure oxidative stress directly, can accurately predict the percentage of sperm DNA fragmentation (SDF) and thereby serve as a surrogate marker in the evaluation of male infertility. ORP was evaluated in 3968 patients using the MiOXSYS system and SDF in 1147 patients using the Halosperm G2 test kit. Both parameters were analyzed in 1068 patients, along with seminal analysis, according to WHO guidelines 2010 (5th edition). SDF correlated positively with seminal ORP normalized for sperm concentration (n=1068; r=.218; P<.0001) as well as with ORP normalized for the motile sperm concentration (motORP; n=1068; r=.387; P<.0001). MotORP can significantly (P<.0001) better predict SDF than ORP normalized for the sperm concentration (area under the curve (AUC): 0.719 vs 0.623) (specificity: 71.5%; sensitivity: 61.9%; PPV: 47.1%; NPV: 82.1%). Moreover, motORP can significantly (P<.0001) better predict (AUC: 0.826 vs 0.771) normozoospermia with high sensitivity (82.75%) and specificity (68.5%). Unlike other oxidative stress (OS) markers, ORP provides a global vision of the redox balance in semen. Moreover, SDF can also be induced by mechanisms different from OS, which could explain why its predictive power was low. The higher predictive power of motORP reflects the impact of seminal ROS on motility rather than inherent DNA breaks present in immature and aborted apoptotic spermatozoa. The evaluation of motORP seems a more promising parameter than ORP normalized for sperm concentration for the prediction of SDF and normozoospermia. However, even if ORP and SDF are inter-related, they measure independent sperm functions and one test cannot replace the other in the evaluation of sperm function defects.

Codeine-induced sperm DNA damage is mediated predominantly by oxidative stress rather than apoptosis

ABSTRACT Background: Opioids have been implicated to induce infertility. Although codeine remains the most used opioid for recreational purpose, no study has documented its effect on sperm quality. Elucidating the effect of codeine on sperm cells and the associated mechanisms may provide an insight into preventing drug-induced sperm damage. Twenty-one New Zealand white rabbits were randomized into three groups; control and codeine-treated. The codeine-treated groups received either 4 or 10mg/kg b.w of codeine for six weeks. Results: Codeine treatment led to significant decrease in sperm count, motility, viability, normal morphology, and sperm membrane integrity. This was associated with significant rise in sperm DNA fragmentation, oxidative damage, and caspase 3 activity. The percentage of sperm DNA fragmentation correlates positively with 8-hydroxy-2'-deoxyguanosine, a biomarker of oxidative DNA damage, and caspase 3 activity, a biomarker of apoptosis. The observed correlation was stronger between sperm DNA fragmentation and oxidative DNA damage than sperm DNA fragmentation and caspase 3 activity. Conclusion: This study revealed that chronic codeine exposure causes sperm DNA fragmentation and poor sperm quality primarily via oxidative stress rather than activation of caspase 3-dependent apoptosis. Findings of the present study may explain drug-induced male factor infertility, particularly, those associated with opioid use.

The relationship between sperm DNA fragmentation, free radicals and antioxidant capacity with idiopathic repeated pregnancy loss

More than two consecutive miscarriages in less than 20 weeks of gestation is defined as recurrent spontaneous miscarriage. Various causes such as uterine anatomical anomalies, genetic factors, and infectious and endocrine disorders have been reported for RPL. However, approximately 50% of the causes are unknown, which can be due to male factors. Several studies have been done on semen parameters to determine the unknown causes and risk factors for miscarriages, however, only studying common semen parameters have not been sufficient. In this study, the relationship between sperm DNA fragmentation, the amount of free radicals, and total antioxidant capacity (TAC) in semen have been considered as a risk factor for spontaneous miscarriage. Semen samples were collected from 42 men whose partners had a history of spontaneous miscarriage and 42 fertile men as the control group. Volume, pH, viscosity, concentration, and motility of semen, as well as sperm morphology were measured. Sperm DNA fragmentation was analyzed by the sperm chromatin structure assay (SCSA) and TUNEL methods, the amount of sperm free radicals was measured by the luminescence method and the total amount of semen antioxidant was measured using the TAC kit. The results have shown that sperm motility in the experimental group was significantly less than the control group (P = 0.001). The percentage of sperm DNA fragmentation and the amount of free radicals in the experimental group were significantly higher than the control group (P < 0.001). The total amount of antioxidant was lower in the experimental group compared to the control. Spouses of men with lower sperm motility and higher DNA fragmentation had a higher chance of spontaneous miscarriage when compared to the control group. The results of this study support the hypothesis that sperm DNA fragmentation is a major contributor to spontaneous miscarriage.The relationship between SDF, ROS and TAC with RPL.

Nosema ceranae changes semen characteristics and damages sperm DNA in honeybee drones.

In this study, we aimed to determine how infection with Nosema ceranae spores affected semen volume, sperm concentration, and fragmentation of sperm DNA in honeybees. A total of 120 one-dayold drones were marked with queen bee marking numbers, and equally divided into two groups. Drones in the infected group were individually fed with honey syrup containing 200 000 N. ceranae spores per 1 µL, while those in the uninfected (control) group were fed honey syrup without spores. The groups were then placed in two separate colonies. Fourteen days later, ejaculate was collected from the drones and was analyzed for semen volume, sperm concentration per 1 µL semen, and sperm DNA fragmentation. Compared to uninfected controls, the N. ceranae spore-infected drones showed significantly decreased semen volume and sperm concentration, as well as a higher percentage of sperm DNA fragmentation.

Impact of a mild scrotal heating on sperm chromosomal abnormality, acrosin activity and seminal alpha-glucosidase in human fertile males.

The aim of this study was to observe sperm aneuploidy, DNA integrity, seminal alpha-glucosidase (NAG) and acrosin activity (AA) under testicular heat stress (SH). Spermatozoa were obtained from 30 healthy adult volunteers subjected to scrotal warming at 43°C for 30-40min on two successive days per week for 3months between February 2012 and September 2016. Aniline blue (AB), acridine orange (AO) staining, TUNEL assay and FISH analysis to evaluate sperm function, sperm DNA integrity and chromosomal abnormalities were carried on before, during and after SH. Sperm AA and NAG was measured by microplate reader. The mean parameters of sperm parameters, AA and NAG were significantly decreased. In contrast, the mean percentage of sperm DNA fragmentation and the proportion of aneuploidy of chromosomes 13, 18, 21, X and Y were significantly increased for spermatozoa collected during SH versus before SH (p<.01-.001). After stopping scrotal heating for 3months, most parameters were completely restored to pre-SH levels. Sperm parameters, sperm DNA integrity, chromosomes, AA and NAG are affected by scrotal exposure to constant SH temperatures several degrees over normal physiological temperature, and after treatment, these parameters were reversibly restored to the level before SH in adult men.

Sperm DNA fragmentation: An early and reliable marker of air pollution

Environmental factors could have a key role in the continuous and remarkable decline of sperm quality observed in the last decades. This study compared the seminal parameters and sperm DFI in men living in areas with different levels of air pollution. Results demonstrate that both steel plants workers and patients living in a high polluted area show a mean percentage of sperm DNA fragmentation above 30%, highlighting a clear sperm damage. In this work, two different techniques were used to measure sperm DNA damage in patients’ groups, finding in both cases a high sperm DFI in patients living in polluted areas. We candidate sperm DNA fragmentation as a valuable early marker of the presence and harmful effects of pollution. We suggest that sperm DNA evaluation could be both an indicator of individual health and reproductive capacity, and a suitable datum to connect the surrounding environment with its effects.

Effect of Lepidium meyenii (maca) on testicular function of mice with chemically and physically induced subfertility.

The aim of this study was to evaluate the effect of Lepidium meyenii (maca) in chemically and physically subfertile mice. After 35days, the following groups of mice were evaluated: control, sham, chemical subfertility, chemical subfertility-maca-supplemented, physical subfertility, physical subfertility-maca-supplemented and maca-supplemented only. Motility (32.36%±5.34%) and sperm count (44.4±5.37×10(6) /ml) in the chemically and physically subfertile mice (11.81%±4.06%, 17.34±13.07×10(6) /ml) decreased compared to the control (75.53%±2.97% and 57.4±19.6 10(6) /ml) and sham (53.5%±7.86% and 58.4±14.10 10(6) /ml). Maca was able to reverse the deleterious effect of motility (76.36±1.97) as well as sperm count (53.5±9.18×10(6) /ml) on chemical subfertility. In contrast, maca did not reverse the effects of induced physical subfertility nor motility (18.78%±14.41%) or sperm count (20.17±11.20×10(6) /ml). The percentage of sperm DNA fragmentation in the physically subfertile mice increased (11.1%±19.29%) compared to the control (0.84%±0.85%). However, in the physically subfertile group, maca decreased sperm DNA fragmentation (2.29%±2.30%) closer to the sham (1.04%±0.62%) and the control (0.84%±0.85%). The group supplemented only with maca showed 0.54%±0.50% of spermatozoa with DNA fragmentation. Yet, the differences observed were statistically not significant. In conclusion, it appears that maca activates the cytochrome P450 system after chemically induced subfertility. However, it does not reverse the low mitochondrial membrane potential in spermatozoa compromised in the physical subfertility group.

Infertile men older than 40years are at higher risk of sperm DNA damage.

BackgroundThe effect of paternal age on semen quality is controversial. In this retrospective study, the aim was to investigate the effects of advancing age on sperm parameters including reactive oxygen species (ROS), total antioxidant capacity (TAC) and sperm DNA damage in infertile men. We also examined whether paternal age >40 y is associated with higher risk of sperm DNA damage.MethodsA total of 472 infertile men presenting for infertility were divided into 4 age groups: group A: patients ≤ 30 y; group B: patients 31- 40 y, group C: ≤ 40 y and group D: patients >40 y. The following tests were performed - semen analysis according to WHO 2010 criteria, seminal ROS by chemiluminescence, TAC by colorimetric assay and sperm DNA damage by TUNEL assay - and the results were compared amongst the 4 age groups.ResultsThere was no statistical difference in conventional semen parameters, TAC and ROS with advancing paternal age as well as between different age groups. However, a significant negative association was noted between sperm DNA damage and advancing paternal age. Men >40 y showed higher levels of sperm DNA damage (24.4 ± 18.5%) compared to younger men (<30 y; 16.7 ± 11.2%; p <0.05).ConclusionsInfertile men over the age of 40 y have a greater percentage of sperm DNA fragmentation compared to infertile men aged 40 y and below. Advanced paternal age (>40 y) may increase the risk of sperm DNA damage in infertile men.

The semen quality of the mobile phone users.

The increased use of mobile phones, the media's attention for general health, and the increase of idiopathic male infertility suggest to investigate the possible consequences of an excessive use of mobile phones on semen quality. To evaluate the conventional and some of the main biofunctional sperm parameters in healthy men according to the different use of the mobile phone. All the enrolled subjects in this study were divided into four groups according to their active cell phone use: group A= no use (no.=10 subjects); group B= <2 h/day (no.=16); group C= 2-4 h/day (no.=17); and group D= >4 h/day (no.=20). Among the subjects of the group D (>4 h/day), a further evaluation was made between the "trousers users"(no.=12) and "shirt users"(no.=8), and they underwent semen collection to evaluate conventional and biofunctional sperm parameters (density, total count, morphology, progressive motility, apoptosis, mithocondrial membrane potential, chromatin compaction, DNA fragmentation). None of the conventional sperm parameters examined were significantly altered. However, the group D and the trousers users showed a higher percentage of sperm DNA fragmentation compared to other groups. These results suggest that the sperm DNA fragmentation could represent the only parameter significantly altered in the subjects who use the mobile phone for more than 4 h/day and in particular for those who use the device in the pocket of the trousers.

Correlation between neutral alpha-glucosidase activity and sperm DNA fragmentation

To evaluate the association between neutral alpha-glucosidase (NAG) activity and sperm DNA fragmentation (DFI), ejaculates from 24 men undergoing evaluation for sperm DNA damage as a part of infertility assessment were analysed. The mean +/- SD and range for the semen quality of the 24 ejaculates are as follows: volume (3.1 +/- 1.3, 1.8-6.0 ml); sperm concentration (45.6 +/- 41.1, 1.3-151.2 x 10(6) ml(-1)); sperm motility (52.8 +/- 28.8, 1-95%); sperm with fragmented DNA (17.6 +/- 15.4, 1.7-56.0%); sperm with immature chromatin (9.6 +/- 3.8, 2.5-19.1%); NAG activity (37.9 +/- 18.3, 4.4-75.3 mU ml(-1)). The only sperm parameter significantly correlated with neutral alpha-glucosidase is the percentage of sperm DFI [correlation coefficient (r) = 0.4376, P = 0.03].

Comparative study on determination of sperm DNA integrity by sperm chromatin dispersion test and by acridine orange staining test

Objective To analyze and compare the accuracy and utility of determining sperm DNA integrity by sperm chromatin dispersion (SCD) test and acridine orange staining (AO) test.Methods The level of DNA fragmentation was determined by SCD test and AO test in 32 adult healthy fertile men and 27 idiopathic oligozoospermia (IO) patients.Sperm nuclei with large DNA dispersion halos or with mediumsized halos were normal and nuclei with small-sized halos or no halo were abnormal.The normal sperm DNA was double strands and stained by AO as green.The damaged sperm DNA was single strand,which were stained as green for native DNA and red for denatured DNA.Results The percentage of sperm nuclei with large halos,medium-sized halos,small-sized halos and no halo evaluated bv SCD test in IO patients were (49.9±13.8)%,(11.5±5.4)%,(11.9±6.1)%and(26.7±10.0)%,and they were(73.2±6.2)%,(14.7±6.3)%,(6.8±2.9)%and(5.3±2.2)%in healthy control group,respectively.There was significant difference between two groups(t=8.576,P＜0.01;t=2.083,P＜0.05;t=4.284,P＜0.01;t=11.823,P＜0.01).The percentage of sperm DNA fragmentation was (38.6±12.1)%in IO patients and was (12.1±5.2)% in fertile men,respectively.A statistically significant difference was found between IO patients and healthy control group under SCD test (t=11.995,P＜0.01).AO test showed no significant differences between IO patients and healthy control group (t=1.626,P＞0.05).The percentage of sperm DNA fragmentation evaluated by AO test in IO patients was (45.5 ±13.8)%,and it was (39.8±13.3)%in healthy control group.Conclusions Sperm DNA fragmentation may lead to male infertility.The SCD is effective in testing the sperm DNA fragmentation as a screening procedure to determine semen quality during basic infertility investigation for clinical use. Key words: Oligospermia; Spermatozoa; DNA damage; Staining and labeling; Genetic techniques

Sperm deoxyribonucleic acid fragmentation as a prognostic indicator of assisted reproductive technology outcome

To examine sperm DNA fragmentation in semen used for assisted reproduction procedures to establish this factor's prognostic role in fertilization rate, embryo development, pregnancy rate, and outcome. Prospective study. Department of Medicine and Biology of Reproduction of the Edouard Herriot Hospital in Lyon, France. 322 couples, divided into 88 cycles of in vitro fertilization (IVF) or 234 cycles of intracytoplasmic sperm injection (ICSI). Sperm DNA fragmentation was detected in sperm obtained 2 to 5 months before the ART procedure. Sperm DNA fragmentation was measured with the terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) technique. There was a negative statistical correlation between the rate of fragmentation and the semen characteristics. A statistically significant negative relationship was found for sperm DNA fragmentation and fertilization when ICSI and IVF were compared. With ICSI, a statistically significant negative relationship was found between fertilization rate and percentage of sperm DNA fragmentation (DNA fragmentation index, or DFI). The risk of nontransfer due to blocked embryo development increased when the DFI exceeded 15% (18.2% for ICSI vs 4.2% for IVF) with an odds ratio of 5.05. The miscarriage risk increased fourfold when the DFI exceeded 15% (37.5% for ICSI vs 8.8% for IVF). Sperm DNA fragmentation measured 2 to 5 months before the assisted reproduction procedure was a prognostic indicator of the fertilization, pregnancy, and miscarriage rates and the pregnancy outcome.

Longitudinal study of sperm DNA fragmentation as measured by terminal uridine nick end-labelling assay

One major limitation in the use of sperm DNA fragmentation as measured by the TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay is the paucity of solid data on the stability of this parameter. The objective of our study was to evaluate variations in the degree of sperm DNA fragmentation, as measured by the TUNEL assay, over a 6 month period. Five donors provided semen samples (total 107) on the average three times per month, and 10 infertility patients provided semen samples every 4 weeks (total 58). The mean percentage of sperm DNA fragmentation for donors was 13.18%, the within-donor standard deviation (SD(W) = 3.79%) was small compared to between-donor (SD(B) = 17.56%). For the group of patients, the mean percentage of sperm DNA fragmentation was 22.44%, with SD(W) of 4.43% within patients and SD(B) of 29.48% between patients. No seasonal rhythm was observed during the study. The intra-class correlation coefficient for all subjects combined was 0.83. Compared to sperm concentration, individual coefficients of variation for sperm DNA fragmentation indicated less variability in four subjects, but were similar in the others. This longitudinal study shows that sperm DNA fragmentation is a parameter with good stability (repeatability) over time; it can be taken as a baseline both in healthy fertile men and in patients from infertility couples.