Are ICSI results affected by months and seasons? A retrospective cohort study of fresh and frozen cycles.

In present study, we aimed to investigate the effect of seasonal variation on ICSI results both in fresh and frozen embryo transfer cycles. Between January 2007 and December 2019, a total of 4039 ICSI cycles (3227 fresh and 812 frozen ET) which resulted in embryo transfer were included in the study. We found no significant difference in the pregnancy outcome among different seasons and months. The best results were obtained for spring (41.0%) and the least for winter (37.1%) in fresh cycles and the best for spring (42.5%) and the least for winter (29.7%) in FET cycles. In monthly distribution, the best results were obtained for September (57.0%) and the least for November (24.1%) in fresh cycles and the best for October (49.3%) and the least for December (20.2%) in FET cycles. Our study did not show any significant influence of the months or seasons on clinical pregnancy rates in either fresh and frozen embryo transfers. However, the difference among months in frozen cycles was notable although it did not reach statistical significance. It can be suggested that the strict control of reproductive hormones especially in fresh cycles as well as the standardized laboratory temperature minimize the influence of seasonal effect on ICSI outcome.

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.

Factors related to follicular oxidative stress in intracytoplasmic sperm injection cycles and its effects on granulosa cells.

The aim of the present study was to investigate several common conditions that may potentially be correlated with follicular oxidative status during an intracytoplasmic sperm injection (ICSI) cycle and that include the serum oestrogen level on the day of oocyte pick-up, maternal age and pregnancy outcome. Patients that were enrolled in the study were classified randomly into three groups using their numerical order. The first group were classified based on maternal age (<35 and ≥35 years) (n = 398), the second group on the serum oestradiol (E2) level on the day of human chorionic gonadotropin (hCG) administration (levels >90th percentile and ≤ 90th percentile) (n = 491) and the third group on pregnancy outcome (positive/negative) (n = 376). The groups were matched for the other variables (stimulation protocol, dose of gonadotropin, duration of stimulation, antral follicle count, body mass index, basal follicle stimulating hormone (FSH), and E2 levels and day of hCG trigger) to prevent the possible contribution of those parameters to the results. Each group was matched for other variables (stimulation protocol, dose of gonadotrophin, duration of stimulation, antral follicle count, body mass index, basal FSH and E2 levels and day of hCG trigger) that may have affected the outcome, except for the parameter under investigation. Maternal age (P = 0.044,168 r = 0.418), oestrogen level on day of hCG administration (P = 0.001, r = 0.436) and pregnancy outcome (AUC = 0.65, P = 0.071) were found to be correlated with follicular oxidative status. The results obtained will help us to shield patients from possible situations that may cause oxidative stress and therefore adverse outcomes of an ICSI cycle.

Effects o follicular fluid oxidative status on human mural granulosa cells, oocyte competency and ICSI parameters.

PURPOSE: The aim of the present study was to understand the molecular and genetic alterations involved in follicular fluid oxidative process by investigating human mural granulosa cells and to find possible biomarkers for oocyte competency and ICSI outcome measures. METHODS: A total of 166 patients were included in the study. Total antioxidant and oxidant levels of follicular fluids were measured on the day of oocyte pick-up and oxidative status were calculated. Expression profiles of three potential target proteins in cases of oxidative stress (Hsp70, Tgf-ß, Notch1), DNA status and chromatin integrity of mural granulosa cells were analyzed. RESULTS: TAS levels were positively correlated with the Hsp70 and Tgf-ß expression patterns of mural granulosa cells. Mature oocyte rate and fertilization rates were affected negatively by the presence of oxidative stress and a significant positive correlation was found with the oxidative status and the fertilization rate, whereas no correlation with the remaining ICSI parameters in the overall group. CONCLUSIONS: Oxidative stress detected in follicular fluid adversely affects fertilization rates post-ICSI however no effect on the remaining parameters including embryo quality, pregnancy, and implantation rates. DNA damage, chromatin integrity were increased, whereas Hsp70 and Tgf-ß were decreased in mural granulosa cells in cases of oxidative stress which may indirectly reflect the oocyte competency and may be used as biomarkers for ICSI outcome measures.

Effects of human sperm cryopreservation on apoptotic markers in normozoospermic and non-normozoospermic patients.

SummaryThe negative effects of cryopreservation on sperm parameters are well documented but little information is known about molecular basis of the process. The aim of the present study was to investigate the possible effects of sperm cryopreservation on main apoptotic signs including DNA fragmentation and caspase-3 activation and to determine if these effects vary according to sperm parameters. Sperm samples of 72 patients were cryopreserved. The patients were sub-grouped as normozoospermic or non-normozoospermic patients according to their semen parameters. DNA fragmentation rates and caspase-3 activation levels were analyzed before and after cryopreservation in both groups. Mean DNA fragmentation rate was increased significantly from 23.98% in neat semen samples to 27.34% after cryopreservation (P = 0.03). DNA fragmentation rates were slightly higher in non-normozoospermic patients compared with the normozoospermic patients in both the neat semen and after cryopreservation (23.25 and 24.71% vs. 26.32 and 28.36%, respectively) although the difference obtained were not statistically significant. An increasing trend for caspase-3 activations (0.093 vs. 0.116) was observed after cryopreservation but the differences were not statistically significant. Caspase-3 activation was found to be slightly higher in non-normozoospermic patients both in the neat semen and after cryopreservation compared with the normozoospermic patients but the differences were not statistically significant. Caspase-3 expression was also shown using immunocytochemistry in both fresh ejaculated sperm and thawed sperm after cryopreservation but at different localizations. The cryopreservation process had detrimental effects on sperm quality but the quality of the sperm samples was not adversely effective for the apoptotic markers including DNA fragmentation and caspase-3 activation patterns. In fact, it was the cryopreservation process itself that adversely effected the above apoptotic markers and apoptosis. It was concluded therefore that sperm cell cryopreservation triggers apoptosis after thawing and this process adversely affects semen parameters.

Male infertility, azoozpermia and cryptozoospermia incidence among three infertility clinics in Turkey.

OBJECTIVE: Semen parameters are directly correlated with the infertility of the male. Incidence rates of male factor infertility, azoospermia and cryptozoospermia differ according to many factors such as geographic region, age, occupation and body weight. The aim of the present study is to determine the incidence of male factor infertility, azoospermia and cryptozoospermia among patients who have been admitted to three separate infertility clinics in Turkey for infertility investigation and analyze the outcomes of these patients. MATERIAL AND METHODS: A total of 9733 men, who have been admitted to 3 infertility clinics in Turkey due to infertility between March 2011 and October 2016, were included in the study. Male infertility, azoozpermia and cryptozoospermia incidence were calculated according to WHO criteria. RESULTS: Male factor infertility was determined in 3114 (32%) of the patients including cases with azoospermia and cryptozoospermia. Azoospermia cases were observed in 570 (5.85%) and cryptozoospermia in 850 (8.73%) men. Azoospermic, and cryptozoospermic patients constitute 18.3%, and 27.2% of the male infertility cases. Sperm retrieval rates in diagnostic or oocyte pick-up plus testicular sperm extraction groups were found to be comparable (16.39%, and 41.3%, respectively). CONCLUSION: The data obtained may help to estimate the number of in vitro fertilization cycles and testicular sperm extraction cases, to determine social security policies, and reproductive potential, and in the light of these data to establish social insurance policies. These data will help patients to decide on treatment alternatives, and guide the urologists about the issue.

Laser assisted zona thinning technique has no beneficial effect on the ART outcomes of two different maternal age groups.

PURPOSE: Laser assisted zona thinning is a technique used for facilitating embryo's hatching process and is commonly used for the embryos of poor prognosis patients with advanced maternal age, repeated implantation failures, poor embryo quality or thick zona pellucida. The aim of this study was to investigate the effect of zona thinning on both good or poor prognosis patients of two different age groups. METHODS: We investigated two different age groups (group 1: <35 years and group 2: ≥35 years ) and compared the effect of assisted zona thinning (groups 1A and 2A) versus not-thinned controls (groups 1B and 2B) in both groups. RESULTS: The clinical pregnancy rates were 57% and 56% in groups 1A and 1B (p = 0.86), 43% and 38% in groups 2A and 2B (p = 0.59) respectively. CONCLUSIONS: Our results suggest that laser assisted zona thinning of day 3 embryos has no beneficial effect on clinical pregnancy and implantation outcomes.