Sperm Chromatin Dispersion Research Articles

P-008 Microgravity exposure significantly decreases sperm motility and vitality. Can we consider human reproduction outside the Earth?

Abstract Study question Are fresh human sperm samples affected by different gravitational conditions than on Earth? Summary answer Motility and vitality of fresh human sperm samples are significantly decreased under microgravity conditions obtained by parabolic flight. What is known already Microgravity effects on the male reproductive system have mainly been studied in the animal model with diverse results and discouraging extrapolation in humans. While an increased motility was reported in bulls, mice models showed a decrease. Although preliminary data from the Micro-11 experiment presented by NASA reported human sperm alterations after microgravity exposure, our first study failed to show any significant effect of microgravity on “frozen” samples, suggesting that human sperm could be safely shipped outside the earth if important aspects related with cryopreservation were solved. Study design, size, duration Prospective study carried out in collaboration between the ART centre, a Technical University, and an Aviation Club specializing in parabolic flights.Two parabolic flights were conducted between 2020-2021, each consisting of 20 parabolic maneuvers, which means 160 seconds of microgravity exposure per sample. Fifteen sperm samples obtained from healthy men were included in the study in order to analyse the effects of microgravity and compare the results with those obtained in Earth gravity. Participants/materials, setting, methods Fresh sperm samples were checked pre-flight to evaluate vitality, concentration, motility and morphology. Samples were split into two to compare the effects of different gravity exposure: microgravity (flight) and Earth gravity (ground). After the flight, the same analysis were repeated, plus kinematics, DNA fragmentation by sperm chromatin dispersion, apoptosis by magnetic activated cell sorting, and oxidative stress by colorimetric test (Halosperm-Halotech). Computer Aided Semen Analysis (SCA-Scope) was used for cell counting. Main results and the role of chance On comparison of the mean values between fresh samples exposed to microgravity and those maintained on Earth gravity, statistical significant differences (p < 0,05) were found in the following parameters: vitality (69,7 ± 9,9 vs 72,4 ± 9,7 %), motile sperm concentration (23,7 ± 15,3 M/ml vs 31,5 ± 25,1 M/ml), grade “a” sperm concentration (8,7 ± 6,5 vs 11,7 ± 9,9 M/ml), percentage of spermatozoa with progressive motility (30 ± 12,9 vs 36 ± 14,3 %), curvilinear motility-VCL (45,7 ± 12,8 vs 47,7 ± 13,3 μm/s). Under the study conditions, non-statistically significant differences were observed in the other kinematic parameters: Lineal Velocity (VSL), Average Path Velocity (VAP), Straightness (STR), Amplitude of Lateral Head displacement (ALH), Linearity (LIN), Wobble (WOB), Beat-Cross Frequency (BCF), total sperm concentration (81,7 ± 112,1 vs 79,7 ± 89,8 M/ml), morphology (11,3 ± 6,3 vs 10,6 ± 5,3%), DNA fragmentation (14,6 ± 9,6 vs 15,7 ± 9,4), apoptosis (2,8 ± 2,8 vs 3,8 ± 4,4) and oxidative stress, since all samples maintained the same stress level in both splits. Limitations, reasons for caution Parabolic flight is an accepted ground-based method for obtaining microgravity conditions, but provides a short period of elapsed exposure to microgravity. Therefore, the results obtained need to be confirmed by using other platforms that provide a much longer time of exposure. More cases must be analysed to confirm the results. Wider implications of the findings Short exposure to microgravity significantly decreases sperm motility and vitality. Such an effect is likely to be stronger with longer exposure. These findings should be taken into account since this may eventually affect sperm fertilizing capacity and therefore natural conception or ART with fresh/frozen sperm, outside of the Earth. Trial registration number NCT03760783

Comparison of zeta potential and physiological intracytoplasmic sperm injection in obtaining sperms with a lower DNA fragmentation index: A cross-sectional study.

BackgroundThe sperm DNA fragmentation index (DFI) is one of the men's reproductive health criteria that affects assisted reproductive technique outcomes. Efforts in obtaining high-quality mature sperms seem to be necessary. Advanced sperm selection techniques (including physiological intracytoplasmic sperm injection [PICSI], zeta potential, microfluidic, etc.) have gained popularity in this regard.ObjectiveThe study aimed to compare the efficacy of zeta potential and PICSI sperm selection in obtaining sperms with better DNA integrity.Materials and MethodsIn this cross-sectional study, 48 couples were enrolled where the male partner had increased sperm DFI in his ejaculated sample and the female was in normal reproductive health. For each male partner, the semen sample was processed with zeta potential and PICSI techniques, then the sperm DFI of neat semen was compared to zeta and PICSI samples by the sperm chromatin dispersion test.ResultsData showed that both the zeta potential and PICSI technique decreased sperm DFI in comparison with the neat semen sample (p 0.001 for both). In addition, there was a statistically significant difference in sperm DFI between the PICSI and zeta potential samples (p 0.01).ConclusionThe current study showed that both zeta potential and PICSI could result in sperm with a lower DFI. However, PICSI seems to be superior to zeta potential in this regard.

The Role of TNF-α and Its Target HSP-70 in Triggering Apoptosis in Normozoospermic and Non-Normozoospermic Samples.

Objective: Semen analysis is performed as one of the screening tests for infertility, including motility, morphology, and concentration observation. We aimed to investigate the expression rates of tumor necrosis factor-α (TNF-α) and heat shock protein (HSP)-70 as two opposite affectors of apoptosis in men with normal semen parameters and abnormal parameters to find the possible effect of this pathway on sperm parameters. We also aimed to investigate the apoptotic markers (DNA fragmentation and Caspase-3 expression) to observe the correlation of this pathway with apoptosis. Materials and Methods: A total of 32 men who applied for infertility evaluation were included in the study. Semen analysis was performed according to WHO criteria. Liquefaction time, appearance, volume, pH, viscosity, sperm concentration, total motility rate, sperm motility, and percentage of spermatozoa with normal morphology were determined. TNF-α, HSP-70, and Caspase-3 immunolocalization were scored histologically. A sperm chromatin dispersion test was used to observe DNA fragmentation. Results: There was no significant difference in TNF-α protein expression rate (mild level). The HSP-70 expression rate was lower, especially in the head region of normo. Caspase-3 was higher totally in non-normo. DNA fragmentation levels were similar in both the groups. Conclusion: From TNF-α protein expression at the mild level in both the groups, it may be hypothesized that the apoptotic pathway might not be triggered by the extrinsic pathway. We found a negative correlation between HSP-70 and Caspase-3 expressions, providing further evidence that HSP-70 works as an inhibitor to apoptosis. This, particularly on specific points, made us think the communication might begin in the anterior chamber, then flow through the cell body to the tail. HSP-70 expression was lower in normo than in non-normo, indicating the possible role of HSP-70 as an answer to any type of stressor in non-normozoospermic patients. Correspondingly, it may be concluded that HSP has an antiapoptotic effect, causing inhibition in the elimination of abnormal sperm cells impairing sperm parameters.

Evaluation of selected semen parameters and biomarkers of male infertility - preliminary study.

Background: Because the etiopathogenesis of male infertility is multifactorial our study was designed to clarify the relationship between standard semen parameters, testicular volume, levels of reproductive hormones and the fragmentation of sperm nuclear DNA (SDF). Methods: Patients (n = 130) were clustered as subjects: 1) with an abnormal volume (utrasonography) of at least one testis (<12 mL) or with a normal volume of testes and 2) with abnormal levels of at least one of the reproductive hormones (FSH, LH, PRL, TSH, total T - electrochemiluminescence method) or with normal hormonal profiles and 3) with high level of SDF (>30%), moderate (>15-30%) or low (≤15%) (sperm chromatin dispersion test). Results: In subjects with a decreased testicular volume and in subjects with abnormal levels of reproductive hormones, decreased basic semen parameters were found. Participants with abnormal testicular volume had a higher percentage of SDF and a higher level of FSH (Mann-Whitney U test). In turn, men with a high level of SDF had lower testicular volume and conventional sperm parameters than men with a low level of SDF (Kruskal-Wallis test). Conclusions: We showed that spermatogenesis disorders coexisted with decreased testicular volume and increased FSH levels. The disorders of spermatogenesis were manifested by reduced basic sperm characteristics and a high level of sperm nuclear DNA damage.

Evaluation of DNA fragmentation in dog sperm using the sperm chromatin dispersion test.

The study's objective was to adapt the Sperm Chromatin Dispersion (SCD) protocol to evaluate sperm DNA fragmentation and implement a fragmentation control in dogs. Correlation between DNA status and routine sperm parameters was also analysed. To adapt the SCD, two different mercaptoethanol (ME) concentrations were assayed (2.5% and 5%) in fourteen ejaculates from seven dogs and semen incubation with 0.3M NaOH for 15min at room temperature was assayed as a control for sperm DNA fragmentation. Data were analysed using a Mann-Whitney test and either Pearson's or Spearman's correlation. The selected ME concentration to use in the SCD test was 5%, as it produced the largest DNA dispersion halo while preserving the core nucleus structure. Four DNA halo patterns were identified as follows: large dispersion halos, medium halos, small halos and nuclei without halos. Semen incubated with NaOH showed 100% sperm without halos (damaged DNA). A significant positive correlation was observed between sperm with fragmented DNA and sperm with coiled tails. Thus, it was possible to adapt the SCD protocol to evaluate dog sperm DNA fragmentation in raw semen without using a commercial kit and establish incubation with NaOH as a DNA fragmentation control. Only coiled tails showed correlation with DNA fragmentation.

Effects of sperm preparation techniques on sperm survivability and DNA fragmentation.

ObjectiveThis study was performed to determine the effect of swim-up (SU) and density gradient centrifugation (DGC) on sperm survival and DNA fragmentation.MethodsIndividual semen samples were analyzed before each was divided into two aliquots (half for SU and half for DGC) for calculation of sperm survival and the DNA fragmentation index (DFI). Sperm DNA fragmentation was determined using the sperm chromatin dispersion test.ResultsThe DFI of the 63 semen samples processed using both procedures was lower than that of the fresh semen samples. The DFI was significantly lower for samples processed using the SU than DGC method. In the sperm survival test, the SU technique was associated with increased sperm motility and vitality following preparation. After 24 hours, however, the concentration and percentage of surviving sperm were significantly lower in the SU than DGC group.ConclusionsBoth semen preparation techniques help to minimize sperm DNA fragmentation; however, when the DFI is <30%, the SU technique is more appropriate than DGC. While DGC may be superior for intrauterine insemination, the SU method may be preferable for in vitro fertilization or maturation.

Sperm selection with hyaluronic acid improved live birth outcomes among older couples and was connected to sperm DNA quality, potentially affecting all treatment outcomes.

Sperm selection with hyaluronic acid improved live birth outcomes among older couples and was connected to sperm DNA quality, potentially affecting all treatment outcomes.

Dependability of the Sperm Chromatin Dispersion (SCD) Assay for Recurrent Sperm DNA Damage Evaluation of Sperm DNA Fragmentation (SDF) in Men with Presentation of Infertility-A Short Communication

How the male attributes in the generation of a healthy embryo, there exists a need for a sperm that is of good quality with minimum genome injury whose estimationis usually are done by the determination of sperm Deoxyribonucleic Acid (DNA) breakdown or the Sperm DNA Fragmentation (SDF). SDF usually guides with regards to the quality of sperm chromatin in case of men with a presentation of infertility. The causes attributed are the capacity of side actions of escalation of SDF amounts on natural fertility, embryo generation along with implantation, risk of miscarriage along with health of the newborn [1]. Earlier we detailed the role of SDF in Intracytoplasmic Sperm Injection (ICSI) with use of Time Lapse Microscopy (TLM) simultaneously [2]. SDF represents a marker of chromatin injury, takes place at the time of spermiogenesis, halt in the epididymis along withpost ejaculation inview of various factors that are not unique like varicocoele, insufficient life style like toxic exposure secondary to occupational along with experimental factors, aging as well as infections [3]. For the birth of a healthy newborn, genomically healthy sperms are necessary, while enhancement of sperms with injured chromatin in the human ejaculate might result in non stabilization of the genome at the time of fertilization as well as embryogenesis [4]. Thus it has been pointed that the evaluation of sperm chromatin at a molecularlevel might aid in its provision of the paternal factor for infertility, besides point to decide what treatment actions might be feasi ble for enhanced sperm chromatin quality [5].

The Relationship between the Testicular Blood Flow and the Semen Parameters of Rams during the Selected Periods of the Breeding and Non-Breeding Seasons.

Simple SummaryThere is a rising demand for sheep products such as meat, wool, and milk. Within the sheep flock, rams are the most valuable animals as they represent 50% of the genetics of every sheep flock. To improve productivity, monitoring of the rams’ semen quality allows breeders to maximize fertility. This study aimed to introduce the advanced semen evaluation tests and evaluate the correlations between the parameters of the basic and advanced semen evaluation tests and the dynamics of testicular blood flow. Semen was collected from eighteen rams, and a pulse wave Doppler examination was conducted in three routine ram examination periods in Poland: before the breeding season (BBS), during the breeding season (BS), and after the breeding season (ABS). Routine and advanced semen analysis, including computer-assisted sperm analysis (CASA), sperm chromatin dispersion test (SCD), and motile sperm organelle morphology examination (MSOME), were conducted. In Doppler ultrasonography, the peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index (RI), and pulsatility index (PI) were calculated. The high-quality semen was derived during BS, whereas BBS semen demonstrated high sperm abnormalities related to sperm cells DNA fragmentation and vacuolization. The features of higher sperm abnormalities correlated with an increase in RI, PI, and EDV, respectively.The study aimed to conduct advanced semen evaluation tests during routine ram examination periods in the breeding and non-breeding seasons and to investigate their correlation with the dynamics of testicular blood flow. Semen was collected from eighteen rams, and pulse wave Doppler examination before (BBS), during (BS), and after the breeding season (ABS). Routine and advanced semen analysis, including computer-assisted sperm analysis (CASA), sperm chromatin dispersion test (SCD), and motile sperm organelle morphology examination (MSOME), were conducted. In Doppler ultrasonography, the peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index (RI), and pulsatility index (PI) were calculated. In BS period, high sperm concentration (p < 0.0001) and total sperm number/ejaculate (p = 0.008) were noted. During the BBS period, a low percentage of forwarding motility (p = 0.017) and high sperm abnormalities (p = 0.005) were found. Also during this period, both SCD and MSOME revealed high sperm DNA fragmentation (p < 0.0001) and signs of vacuolization (Grade II-IV, p < 0.05). The advanced features of higher sperm abnormalities (Grade IV of MSOME) correlated with an increase RI (ρє <0.60;0.61>) and PI (ρє <0.46;0.52>), whereas the basic percentage of sperm abnormalities correlated with the EDV (ρє <0.44;0.73>) value. One may conclude that the current preliminary study requires further research concerning the monthly examination of a ram to provide full yearly characteristics of the relation between advanced semen evaluation tests and the dynamics of testicular blood flow.

Protective effect of Scrophularia striata combined with trehalose and cysteine added to diluents on cryopreservd goat epididymal sperm

Objective: To evaluate antioxidant effects of Scrophularia (S.) striata ethanol extract, trehalose and cysteine added to diluents on cryopreserved goat epididymal sperms. Methods: Motility and standard motion parameters of sperm were assessed by using computer assisted sperm motility analysis system. Sperm viability was evaluated by eosin-nigrosin staining method. Hypo-osmotic swelling test was used to evaluate membrane health. Thiobarbituric acid testing was used to measure malondialdehyde (MDA) concentrations. To assess DNA fragmentation, sperm chromatin dispersion test was used. In Experiment 1, treatments consisting of basal Tris diluent supplemented with 25, 50 or 100 μg/mL of S. striata ethanol extract gave the best concentration to the freezing diluents. Experiment 2 was carried out to compare the best concentration of S. striata ethanol extract (50 μg/mL) resulting from the first experiment with 150 mM trehalose and/or 5 mM cysteine alone or in combination. Results: S. striata ethanol extract (50 μg/mL) significantly increased sperm viability, motility and progressive motility and at the same time decreased MDA concentration and DNA fragmentation compared to other treatments (P&lt;0.05). In addition, all treatment groups resulted in viability, membrane health, total motility, progressive motility, curvilinear velocity, straight-line velocity higher and MDA lower compared to the control group (P&lt;0.05). Acrosome integrity was significantly higher in 50 μg/mL of S. striata ethanol extract combined with cysteine, trehalose, or cysteine+trehalose groups than those in the control, trehalose, cysteine, and 50 μg/mL of S. striata ethanol extract groups (P&lt;0.05). Regarding DNA, extenders supplemented with 50 μg/mL of S. striata ethanol extract, 50 μg/mL of S. striata ethanol extract+trehalose, and 50 μg/mL of S. striata ethanol extract+trehalose+cysteine were superior to other treatments. Conclusions: Adding 50 μg/mL of S. striata ethanol extract alone or in combination with trehalose and cysteine can improve the quality of cryopreserved epididymal sperms of goats.

Prolonged exposure of human spermatozoa in polyvinylpyrrolidone has detrimental effects on sperm biological characteristics.

Polyvinylpyrrolidone (PVP) has been utilized in intracytoplasmic sperm injection (ICSI) for immobilization and manipulation of spermatozoa. This study aims to determine the suitable time that sperm cells could be safely exposed to PVP during ICSI procedure. Twenty-five normal semen samples were prepared using the swim-up method and then were exposed to 10% PVP at different time intervals (15, 30 and 60min). The effect of PVP on sperm parameters (viability and morphology), DNA fragmentation index (sperm chromatin dispersion test), chromatin quality (aniline blue, toluidine blue and chromomycin A3 staining), acrosome reaction, mitochondrial membrane potential and sperm ultrastructure was assessed at different time intervals. Our results showed that prolonged sperm exposure in PVP for 15, 30 and 60min significantly affects viability and morphology with a concomitant increase in DNA fragmentation and abnormal chromatin structure, while the percentage of acrosome-reacted spermatozoa was additionally increased. In addition, the spermatozoa with high mitochondrial membrane potential were significantly decreased compared to unexposed spermatozoa to PVP. In conclusion, the detrimental effects of PVP were increased significantly following sperm exposure in PVP after 15min. Therefore, the sperm exposure to PVP should be limited to less than 15min during ICSI procedure.

Correlations between abnormalities of morphological details and DNA fragmentation in human sperm.

ObjectiveAs the associations of sperm DNA fragmentation with morphology have not been examined in detail, this study aimed to investigate the relationship between abnormalities of morphological details and DNA integrity in human sperm.MethodsIn this cross-sectional study, men from infertile couples were enrolled at Hue Center for Reproductive Endocrinology and Infertility, Vietnam. Conventional semen parameters, including morphological details, were analyzed following the World Health Organization 2010 criteria. Sperm DNA fragmentation was evaluated using a sperm chromatin dispersion assay. The relationships and correlations between semen parameters, sperm morphology, and the type of halosperm and the DNA fragmentation index (DFI) were analyzed.ResultsAmong 130 men in infertile couples, statistically significant differences were not found in the sperm halo type between the normal and abnormal sperm morphology groups. The percentage of round-head spermatozoa was higher in the DFI >15% group (16.98%±12.50%) than in the DFI ≤15% group (13.13%±8.82%), higher values for amorphous heads were found in the DFI >15% group, and lower values for tapered heads were observed in the DFI ≤15% group; however, these differences were not statistically significant. Small-halo sperm and the DFI were positively correlated with round-head sperm (r=0.243, p=0.005 and r=0.197, p=0.025, respectively). ConclusionThe rate of general sperm morphological abnormalities in semen analysis was not related to sperm DNA integrity. However, round sperm heads were closely associated with sperm DNA fragmentation.

Protective Effects of Trehalose and Pentoxifylline on Goat Sperm Exposed to Chilling-Freezing Process.

Freezing of sperm is known as an important part of assisted reproduction. However, many studies have illustrated that cryopreservation negatively affects the quality and fertility rate of sperm. This study aimed to evaluate the effects of trehalose and pentoxifylline (PTX) in diluents on cooled and frozen-thawed Markhoz goat sperm. Preassessed samples were pooled and diluted with a basic diluent using trehalose and PTX. The cooled sperm showed significant improvement. The motion characteristics of cryopreserved sperm were evaluated based on computer-assisted system analysis. In this study, we investigated the viability, membrane integrity, malondialdehyde concentration, total abnormality, acrosome integrity, and seminal hyaluronidase enzyme. Also, the hypo-osmotic swelling test, mitochondrial activity, apoptotic features, caspase activity, chromatin dispersion test, active mitochondria, and reactive oxygen species (ROS) activity were assessed as complementary parameters. The data illustrate that the total motility, progressive motility, average path velocity (VAP), straight-line velocity (VSL), curvilinear velocity (VCL), and the ratio of sperm chromatin dispersion, viable sperm were improved significantly (p < 0.05) using 3 mM PTX alone or 3 mM PTX plus 50 mM trehalose, while other characteristics indicate significant enhancement by 3 and 6 mM PTX and 50 and 70 mM trehalose alone or in combination, except amplitude of lateral head displacement (ALH), beat/cross frequency (BCF), and intracellular ROS-(O-), which demonstrate no significant difference among treatments. In conclusion, this study indicates that addition of 3 and 6 mM PTX alone or with 50 and 70 mM trehalose seems to reduce the damage caused by cooling and cryopreservation processes.

Incorporating Sperm DNA Fragmentation Index with Computer-Assisted Semen Morphokinematic Parameters as a Better Window to Male Fertility

This study aimed to assess (1) the reproducibility of three sperm chromatin dispersion (SCD) assays for sperm DNA fragmentation, i.e., LensHooke R10® (R10), Halosperm G2® (G2), and BASO® (BA); (2) the correlation between computer-assisted semen analyzer (CASA) morphokinematic parameters and sperm DNA fragmentation index (DFI), and (3) the diagnostic value for male reproduction by combining semen morphokinematic parameters and DFI. Total 50 male participants were recruited, and all collected semen samples underwent semen analyses and SCD assays. Intra- and inter-observer variability of DFI data from different SCD measures was tested. In addition, the predictive ability of CASA parameters and DFI (with different cutoffs, i.e., 15% and 20%) for infertility was assessed using receiver operating characteristic curve analysis. We found that the G2 and R10 produced satisfactory variance coefficients (5.53%, 5.67%) compared to BA (14.8%). The DFI data from the R10 had lower intra-observer variability, in terms of higher intra-class coefficient (0.9615), than that of the G2 (0.8847) or BA (0.8824). Inter-observer variability of three SCD kits in scoring the DFI was comparable and satisfactory (concordance correlation coefficients ranging 0.9895-0.9630). The CASA parameters (i.e., total motility [r = -0.57], progression motility [r = -0.55], and rapidly progressive motility [r = -0.55]) were significantly correlated with DFI (P < 0.001). The predictive ability of the 15%-cutoff DFI data was better than that of the 20%-cutoff or continuous DFI data. The model comprising the CASA parameters, 15%-cutoff DFI, and 4%-cutoff normal morphology had the highest area under curve (0.8125) for infertility. For SCD assay, the R10 was the most reliable SCD assay to detect sperm DNA fragmentation. Combining the sperm DFI with CASA parameters might be a better diagnostic tool for male reproduction.

Sperm DNA Fragmentation Index in Infertility Patients vs Normal Individuals: A Cross-sectional Study

Introduction: Infertility is a major problem among married couples. The male factor contributes to around 30-40% of all cases of infertility. With advancing age, reducing sperm count may magnify the problem. The awareness of qualitative change along with quantity is important for people seeking advice in infertility. Aim: To find the prevalence of sperm DNA fragmentation with Sperm Chromatin Dispersion (SCD) test. Materials and Methods: The present study was prospective cross-sectional study which carried out on semen sample given for routine checkup in department during the period from August 2017 to August 2018. A total of 180 couples, including 88 infertile couples and 92 couples with known fertility were enrolled in the study. Sperm Count and sperm DNA Fragmentation Index (DFI) were calculated and determined by SCD test by kit method. Comparison between groups was done according to distribution of data using Mann-Whitney rank sum (two tailed) statistical test. Results: Among 180 subjects, (88 cases and 92 control) no significant difference in the age was found. The sperm count was not significantly different in cases (group of men of infertile couple) and controls (group of men of proven fertility). The highest value for DFI was 27% in controls and 97% in cases. Statistically significant difference was found in DFI (p=0.008) in both groups. DFI and sperm count showed negative correlation both in cases and controls with a correlation coefficient 0.213 (p-value &lt;0.001) and 0.754 (p-value &lt;0.005) respectively. A significant difference was observed in the median value of sperm DFI of men of infertile couple as compared to men of fertile couple. Conclusion: On comparing, significant difference was observed in the median value of sperm DFI in infertile couple. Sperm DFI at cut-off value 27% has higher diagnostic significance. SCD method is simple, easiest and standard tool to assess DFI.

Correlation of Obesity and Serum Vitamin D Levels with Sperm DNA Integrity, Sperm Quality, and Sperm Viability in Normozoospermia Men.

Obesity, Vitamin D (VD) deficiency, and infertility are important ubiquitous issue; however, the association of obesity and serum VD levels with abnormal sperm is unclear and inconclusive. The current study investigated the correlation of obesity and serum VD levels with sperm DNA integrity and sperm parameters in normozoospermia men. Semen and blood samples from 64 men were divided into two groups: obese and nonobese men based on body mass index (BMI). Sperm motility and viability were determined by computer-aided sperm analysis and eosin-nigrosin staining. DNA fragmentation, determined by sperm chromatin dispersion method. VD concentrations were assessed by the Elisa technique. Serum concentration of VD levels in the obese group was significantly lower than nonobese men (P < 0.05). Sperm motility was significantly reduced in the obese group in comparison to nonobese (P < 0.05). Rapid progressive motility was statistically lower in obese men compared with the nonobese group (P < 0.05). Sperm count and morphology were not statistically significant in both groups. Sperm viability in the nonobese group was significantly decreased in comparison to obese group (P < 0.05). DNA integrity was significantly higher in the obese group as compared with nonobese (P < 0.01). VD deficiency in the obese group showed decreased sperm motility, increased DNA damage, and viability. Adverse consequences of obesity and the possible effect of BMI infertility treatment must be discussed with counseling couples interested in assisted reproductive techniques outcomes, especially in men without any unknown cause.

SPERM CHROMATIN DISPERSION TEST FOR EXAMINATION OF INFERTILE MALE: FROM СLINICAL TRIALS OF FIRST RUSSIAN KIT GEMSTANDART–HALOSPERM L&amp;Q

Objectives. To evaluate the sperm chromatin dispersion in men with infertility using the first Russian kit. Methods. The study was conducted between July and August 2022. Study participants were 492 male volunteers aged 23 to 59 years with infertility which semen samples were examined by GEMSTANDART–HaloSperm L&amp;Q kit so-called Halo test. The outcomes were as a percentage as the sperm DNA Fragmentation Index (DFI), %. The results of Halo test were evaluated according to the following threshold values: in reference range (normal): DFI  15 %, controversial result: from DFI &gt; 15 % to &lt; 25 %, out of reference range (pathological) DFI &gt; 25 %. All of men (n = 492) had the results of a basic semen examination which performed a month before the current clinical trials with a various scenario of pathospermia as well as normospermia. The statistical processed a GraphPad Prism software (version 8.0.1) and Excel statistical package Microsoft® Excel® for Microsoft 365 MSO (verse 2201 16.0.14827.20158). The Pearson’s rank correlation coefficient of parametric measure and descriptive statistics have been applied. Results. Sperm DNA fragmentation (SDF) was not detected in 54.1 % (266/492 = 54.1 %) men whose average age was 36.58 years (DFI  15 %); a poor direct correlation with normal level of DFI and asthenozoospermia (r = 0.20; p &lt; 0.0001), viscosipathy (r = 0.13; p &lt; 0.0001) are presented; a poor inverse correlation with oligoasthenoteratozoospermia (r = – 0.12; p &lt; 0.0001) are presented; the absence of leucospermia was noted. A controversial results for SDF presence was found in 21.3 % (105/492 = 21.3 %) men whose average age was 38.71 years (DFI range from &gt; 15 to &lt; 25 %); a poor direct correlation with oligoasthenoteratozoospermia (r = 0.24; p &lt; 0.0001), oligospermia (r = 0.23; p &lt; 0.0001) and average age of 39 years (r = 0.17; p &lt; 0.0001) was noted; poor inverse correlation with asthenozoospermia (r = – 0.22; p &lt; 0.0001) was presented. SDF was found in 24.6 % (121/492 = 24.6 %) men whose average age was 41.55 (DFI &gt; 25 %); poor direct correlation with a high level of DFI and average age for infertile men in 44 years (r = 0.28; p &lt; 0.0001) was presented. All semen samples (n = 492) presented a poor positive correlation with DFI and age (r = 0.1563; p = 0.0005); oliasthenoterato-zoospermia (r = 0.19; p = 0.0001), leucospemia (r = 0.29; p = 0.0001), and necrozoospermia (r = 0.16; p = 0.0004). Conclusion. The results of the study showed that GEMSTANDART–HaloSperm L&amp;Q kit complements a basic semen examination providing specific information to the clinicians about quality of genetic material in semen samples.

Impact of oxidative stress SNPs on sperm DNA damage and male infertility in a south-east Iranian population.

We examined four single nucleotide polymorphisms in four antioxidant genes (PON1 , CAT , GPx1 and SOD2 ) in 100 infertility cases and 100 controls from an Iranian population-based case-control study to confirm the assumption that polymorphisms in oxidative stress genes increase the risk of sperm DNA damage and idiopathic male infertility. Restriction fragment length polymorphism and tetra-primer amplification refractory mutation system PCR were used to identify genotypes. Sperm DNA damage was assessed using the Sperm Chromatin Dispersion test (Halo Sperm), and the total antioxidant capacity of seminal fluid was determined using the FRAP assay. Our findings demonstrated that alleles Arg-PON1 (rs662) and Ala-MnSOD (rs4880) variant genotypes were considerably linked with a higher risk of male infertility. Linear regression analysis revealed that those with the PON1 Gln192Arg or SOD2 Val16Ala variants have significantly higher levels of sperm DNA fragmentation and lower levels of the total antioxidant capacity in seminal fluid. These findings suggest that genetic differences in antioxidant genes may be linked to oxidative stress, sperm DNA damage, and idiopathic male infertility.

In vitro effects of pentoxifylline and coenzyme Q10 on the sperm of oligoasthenoteratozoospermia patients

The effect of in-vitro sperm incubation with Pentoxifylline (PTX) and Coenzyme Q10 (CoQ10) in Oligoasthenoteratozoospermia (OAT) patients was evaluated. Semen samples were obtained from men with Normozoospermia and men with OAT. Motile sperm from the two groups were subdivided into four subgroups: (i) without incubation with PTX + CoQ10; (ii) incubation with PTX; (iii) Incubation with CoQ10; and (iv) incubation with a combination of PTX + CoQ10. Then, sperm parameters, chromatin, DNA and membrane integrity, protamine deficiency, apoptosis, mitochondrial activity, sperm chromatin dispersion test (SCD), hypo-osmotic swelling test (HOS), chromomycin A3 (CMA3), Terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL), and diaminobenzidine (DAB) assays were evaluated, respectively. Sperm incubated with CoQ10 and a combination of CoQ10 and PTX resulted in a significant increase in the sperm parameters. Also, a significant decrease was noted with a combination of PTX and CoQ10 in normal men. There was a significant difference between CoQ10 treated and CoQ10 + PTX treated groups in comparison with the OAT group in the percentage of the DNA fragmentation, sperm apoptosis, AB+, HOS test + and sperm mitochondrial activity. Incubated sperm with CoQ10, PTX, and in combination with each other can improve sperm parameters in OAT patients.

In Vitro Studying the Effect of Adding Autologous Platelet Rich Plasma (PRP) to the Human Semen on the Sperm DNA Integrity

For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p &lt; 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.