The Association of ICSI Outcomes with Semen and Blood Bisphenol A Concentrations of the Male Partner.

This study investigated the association of blood and semen Bisphenol A (BPA) levels of the male partner on the reproductive outcome in intracytoplasmic sperm injection (ICSI) treatment cycles. For this prospective study (ClinicalTrials.gov identifier: NCT02703584), blood and semen samples of the male partner of the 75 women who had ICSI were analyzed. The study group consisted of men who had ICSI for male factor infertility other than azoospermia, while men with normal spermiogram whose partners underwent ICSI due to tubal factor infertility were taken as the study group. Habitual consumption of drinking water from plastic carboys/bottles (PBW) at home was also questioned in both groups as it was considered as chronic BPA exposure. The association of ICSI outcome with blood BPA (bBPA) and semen BPA (sBPA) levels was analyzed in both groups. No significant correlation was found between sperm parameters and bBPA levels in both groups. A negative correlation was found between sBPA levels and total sperm count and progressive sperm motility in men who consumed PBW. Embryo development arrest was found to be significantly higher in patients who have high sBPA levels. Although sBPA levels were not different in PBW consumers, bBPA levels were found to be significantly lower in those who consumed tap water (TW) than those who used PBW. Elevated bBPA were associated with a significant decrease in clinical pregnancy rate. Considering the widespread human exposure to BPA, the effect of BPA on the male reproductive system needs to be further examined.

Evaluation of Clinical Outcomes after Poor-Quality Embryo Transfer and Prognostic Parameters.

We aimed to investigate the clinical results following poor-quality embryo transfer and the parameters to foresee the prognosis. In this study, 2123 cycles that had day 3 and day 5 single-fresh embryo with poor-quality embryo transfers and good-quality embryo transfers were compared. The cycles according to transfer day were evaluated by conducting a subgroup analysis. The correlation between all the obtained demographic characteristics, controlled ovarian stimulation parameters, and cycle results were analysed. Clinical pregnancy was established in 53 patients that underwent transfer in the poor-quality embryo group (14.9%). Of these patients, 36 had live birth (live birth rate per clinical pregnancy 67.9%). In cleavage-stage embryos, live birth rates per clinical pregnancy were higher in poor-quality blastocyst transfer. When analysing the factors affecting live births in the poor-quality embryo group, as the total gonadotropin dose increases, the probability of live birth decreases, as in the probability of hCG positivity. In conclusion, although the probability of pregnancy is low, when clinical pregnancy is established, there is a high chance of having a live birth after poor-quality embryo transfers. This could be regarded as an acceptable option in cycles when only poor-quality embryos are available.

Therapeutic effects of melatonin in long-term exposure to 2100MHz radiofrequency radiation on rat sperm characteristics.

INTRODUCTION: Radiofrequency electromagnetic fields (RF-EMFs) are one of the risk factors for male reproductive health and melatonin can be an ideal candidate for therapeutic development against RF-induced male fertility problems due to its antioxidant properties. The possible therapeutic role of melatonin in the destructive effects of 2100MHz RF radiation on rat sperm characteristics is investigated in the present study. METHODS: Wistar albino rats were divided into four groups and the experiment continued for ninety consecutive days; Control, Melatonin (10mg/kg, subcutaneously), RF (2100MHz, thirty minutes per day, whole-body), and RF+Melatonin groups. Left caudal epididymis and ductus deferens tissues were placed in sperm wash solution (at 37°C) and dissected. The sperms were counted and stained. Measurements of the perinuclear ring of the manchette and posterior portion of the nucleus (ARC) were performed and the sperms were examined at an ultrastructural level. All of the parameters were evaluated statistically. RESULTS: The percentages of abnormal sperm morphology were significantly increased with RF exposure, while the total sperm count was significantly decreased. RF exposure also showed harmful effects on acrosome, axoneme, mitochondrial sheath, and outer dense fibers at the ultrastructural level. The number of total sperms, sperms with normal morphology increased, and ultrastructural appearance returned to normal by melatonin administration. DISCUSSION: The data showed that melatonin may be a beneficial therapeutic agent for long-term exposure of 2100MHz RF radiation-related reproductive impairments.

Clinical significance of serum and follicular fluid ceramide levels in women with low ovarian reserve.

Objective: Ceramide (CER) is a bioactive component of the mitochondrial membrane. In this study, we will investigate the clinical importance of serum CER (sCER) and follicular fluid CER (ffCER) levels in the lipid synthesis pathway and their effect on poor oocyte quality and in vitro fertilization (IVF) outcome. Materials and Methods: This cross-sectional, case-control study was conducted in the IVF unit of a maternity hospital in the capital of Turkey, Ankara. A total of 88 women undergoing their first IVF cycle were included in this study patients were divided into 2 groups according to current diagnostic criteria for their ovarian reserves. Baseline sCER levels, and ffCER concentrations retrieved on the oocyte pickup day were measured. Results: The mean age, body mass index, and infertility duration of the patients was similar between the groups (all p>0.05). There was also no significant difference in the clinical pregnancy rates (38.6% vs. 47.7%, p=0.127). sCER (15.6±6.5 vs. 23.5±8.9) and ffCER (82.5±34.3 vs. 116.4±46.5) levels were statistically significantly lower in the low ovarian reserve (LOR) group (both p<0.001). The performed receiver operating characteristic curve analysis revealed that sCER and ffCER levels could predict both LOR and pregnancy. Conclusion: This is the first study evaluating the sCER and ffCER levels of patients undergoing IVF treatment. CER may be used as an ovarian reserve markers and a biomarker capable of predicting IVF outcomes.

Comparison of the impact of laser-assisted hatching on fresh cleavage and blastocyst embryo transfer and association with pregnancy outcomes.

Objective: Assisted hatching (AH) techniques can improve live birth (LB) and clinical pregnancy (CP) rates. Since there are limited data regarding this subject, we investigated the impact of laser-assisted hatching (LAH) on fresh embryo transfer (ET) and association with pregnancy outcomes in unselected patient population. Materials and Methods: This retrospective study included the fresh ETs performed at our center between April 2010 and April 2019. Among 3.782 fresh ETs, 3.286 underwent LAH (n=1.583 at cleavage stage and n=1.703 at blastocyst stage) while 496 underwent non-assisted hatching (NAH) (n=213 at cleavage stage and n=283 at blastocyst stage). The ETs were performed at the blastocyst or cleavage stages, and single or double embryos were transferred. LB rate was the primary outcome, while secondary outcomes were the pregnancy test, monozygotic twinning (MZT), and CP rates. Results: The LAH and NAH groups showed similar LB, pregnancy test, CP, and MZT rates at cleavage and blastocyst stages. On the other hand, LAH significantly affected LB rates at the blastocyst stage (20.6% at blastocyst stage vs. 16% at the cleavage stage, p=0.001). Conclusion: In conclusion, LAH does not improve reproductive outcomes of fresh blastocyst-stage and cleavage-stage ETs. However, LAH significant impacts LB rates in the blastocyst stage than the cleavage stage.

The relationship between good quality embryo rates and IVF outcomes/embryo transfer policies in extended embryo culture.

This study aims to investigate which parameters affect the change in good quality embryo rates during the cleavage stage and whether they have any effect on embryo transfer policies and IVF results. We analysed changes in good quality embryo (grades 1 and 2) rates during the period on days 2, 3 and 5; patients with five or fewer embryos (group 1), 6-10 embryos (group 2) and more than 10 embryos (group 3). The good quality embryo rates decreased in all groups on day 5. When the infertility reasons are studied among all of the groups, ovulatory dysfunction is found to be significantly higher in group 2 compared to group 1 and unexplained infertility was found to be significantly higher in group 2 compared to group 1 and group 3. Total antral follicle, mature oocyte and total oocyte counts were found to be significantly lower in group 1. However, there is no significant difference found among all of the groups for ß-HCG levels and clinical pregnancies. Changes in good quality embryo rates at the cleavage stage in extended embryo culture do not have an impact on IVF results.IMPACT STATEMENTWhat is already known on this subject? The number and quality of embryos in the cleavage stage are important parameters affecting the embryo transfer decision on day 5. There is still insufficient knowledge concerning changes in the percentage of increased good quality embryo transfers associated with IVF outcomes during the second to the third day, and the third to the fifth day.What do the results of this study add? Day 5 embryo transfer is possible in patients with a low number of embryos, according to our results. The good quality embryo rates of patients with a low number of embryos at the cleavage stage are more promising compared to patients having more than five embryos.What are the implications of these findings for clinical practice and/or further research? An extended embryo culture option can be used on patients with a low number of embryos for clinical practice.

Evaluation of embryo transfer through a scoring system: ETSS (Embryo transfer scoring system).

INTRODUCTION: To create a scoring system by including all of the factors that are recommended for an ideal ET and to investigate its correlation with the Β-HCG results. MATERIALS AND METHODS: This study was conducted as a retrospective trial between January 2009 and December 2018. Women who had a single ET between the specified dates were included in the study. The embryo grade, ET day, distance between the fundus to embryo transfer site measured via ultrasonography, endometrial thickness on ET day, and presence of mucus and blood in the catheter after transfer were the variables evaluated. Each one of the five variables that constituted the scoring system were rated separately. RESULTS: Overall, 1652 patients participated in this research. Antral follicle count (13,3 ± 8 vs. 14,6 ± 8,2, p: 0,001), endometrial thickness on the ET day (9.9 ± 2 vs.10.3 ± 2, p = 0.006) and number of mature oocytes (8.6 ± 6 vs. 9 ± 5.1, p: 0.003) were significantly higher in patients with positive Β-HCG values. The total score in the Β-HCG positive group was 9.8 ± 1.4 versus 8.9 ± 1.4 in the Β-HCG negative group (p < 0.001). The best ETSS cut-off value for predicting Β-HCG positivity was 9.5, with 82% sensitivity and 67% specificity (AUC:0.808). CONCLUSION: Our scoring system is an important step toward standardization, as it offers a new, practical, cost-free, and applicable scoring system based on pre- and post-ET measurements and laboratory data.

Phospholipase C-zeta levels are not correlated with fertilisation rates in infertile couples.

In mammals, 'oocyte activation' is triggered by certain proteins, one of which is phospholipase C-zeta. Recent evidence suggests that low expression of phospholipase C-zeta might be associated with male infertility, while a limited number of studies claimed the opposite. This study was designed to test whether quantity of phospholipase C-zeta and in vitro fertilisation rates are correlated or not, assessed by flow cytometry. Semen samples from 43 infertile couples were analysed for the percentage and mean fluorescent intensity (MFI) of phospholipase C-zeta protein. Results were confirmed by immunofluorescent labelling. Patients with a fertilisation rate of 40% or lower were involved in the low fertilisation group, while the high fertilization group consisted of patients with a fertilisation rate of 60% and higher. Quantitative analyses by flow cytometry showed no significant difference among the low fertilisation and high fertilisation groups when phospholipase C-zeta ratio or MFI was considered. No correlation was found between pregnancy rates and phospholipase C-zeta quantity. None of the total fertilisation failure cases were lack of phospholipase C-zeta. In fact, fertilisation was possible even when phospholipase C-zeta levels were very low. Thus, we concluded that phospholipase C-zeta quantity cannot be considered as a diagnostic tool for male infertility.

The effect of plastic bottled water consumption on outcomes of ICSI cycles undertaken for unexplained infertility.

RESEARCH QUESTION: Do bisphenol A (BPA) levels in maternal urine, serum and follicular fluid affect embryo quality and intracytoplasmic sperm hinjection (ICSI) cycle outcomes in women with unexplained infertility? DESIGN: Prospective study conducted between 1 April 2019 and 30 September 2019. The study cohort consisted of 82 women aged between 23 and 33 years who underwent intracytoplasmic sperm injection owing to unexplained infertility and provided urine, blood and follicular fluid samples on the day of oocyte retrieval. Consumption of drinking water from plastic carboys or bottles at home were considered as chronic BPA exposure. Demographic features and IVF outcomes of the patients were collected. RESULTS: Among the 82 women with unexplained infertility, clinical pregnancy was achieved in 22 (26.8%) patients after the IVF and embryo transfer cycle. The patients who consumed tap water had statistically significantly lower BPA values in three body fluids compared with patients who consumed plastic bottled water (all P < 0.001). Women who had grade 1 embryos transferred had lower serum BPA values than women who had grade 2 embryos transferred (10.8 ± 5.2 versus 26.9 ± 22 ng/ml, P = 0.003). Serum and follicular fluid BPA levels were statistically significantly higher in women who failed to achieve clinical pregnancy (P < 0.001, P = 0.006, respectively) and obtain a live birth (both P = 0.007). CONCLUSIONS: A negative relationship was found between serum and follicular fluid BPA levels and embryo quality, clinical pregnancy and live birth in these women. In addition, the BPA levels of women who consume tap water at home were lower than those who use plastic bottled water.

Effect of Antagonist Start Day on Cycle Outcomes in Poor Responders.

BACKGROUND: Despite the great advances in Assisted Reproductive Technologies (ART), management of poor responders has remained a great challenge. Gonadotropin releasing hormone antagonist (GnRH-ant) has been offered as a patient friendly protocol. In the literature, conflicting data exists about the effect of the GnRH-ant starting day on cycle outcomes. AIM: The aim of this study is to evaluate the effect of GnRH-ant starting day on cycle outcomes of patients with poor ovarian response defined by Bologna criteria. SETTING AND DESIGN: This retrospective cohort study was conducted at an ART clinic of a tertiary hospital. MATERIALS AND METHODS: A total of 361 cycles using flexible GnRH-ant, 195 in Group A (GnRH-ant administered before day 6 of stimulation) and 166 cycles in Group B (GnRH-ant started on or after day 6), were selected retrospectively for the study. STATISTICAL ANALYSIS: Statistical analysis of data was carried out using using IBM SPSS Statistics Software (20.0, SPSS Inc., Chicago, IL, USA). Independent samples t-test and Mann-Whitney U test were used to analyze the variables. RESULTS: Total antral follicle count was significantly higher in Group A compared to Group B (P = 0.009). Duration of stimulation was significantly shorter (P < 0.01) and total dose of gonadotropin used was lower in Group A when compared to Group B (P < 0.01). While higher number of oocytes was retrieved from Group A (P = 0.037), no between-group differences were observed in number of mature oocytes, fertilized oocytes, clinical pregnancy rate or ongoing pregnancy rate (OPR) per embryo transfer (P > 0.05). CONCLUSION: Early GnRH-ant start may point out a favourable response to ovarian stimulation in poor responders. However, clinical or OPRs were not different from the late GnRH-ant start group.

Impact of Transferring a Poor Quality Embryo Along with a Good Quality Embryo on Pregnancy Outcomes in IVF/ICSI Cycles: a Retrospective Study.

Background The number and the quality of embryos transferred are important predictors of success in in vitro fertilization (IVF) cycles. In the presence of more than one good quality embryo on the transfer day, double-embryo transfer (DET) can be performed with these embryos, but generally, different quality embryos are present in the available transfer cohort. We aimed to investigate the effect of transferring a poor quality embryo along with a good quality embryo on IVF outcomes. Methods In this study, 2298 fresh IVF/intracytoplasmic sperm injection (ICSI) cycles with two good quality embryos (group A), one good and one poor quality embryo (group B), and single good quality embryo (group C) transfers were examined. All groups were divided into two subgroups according to the transfer day as cleavage or blastocyst stage. Clinical pregnancy and live birth rates were the primary outcomes. Results In the cleavage stage transfer subgroups, the clinical pregnancy rates were lower in the single-embryo transfer (SET) subgroup compared with DET subgroups, but the difference was not statistically significant compared with DET with mixed quality embryos. The live birth rates were comparable between the three groups. In the blastocyst transfer subgroups, the clinical pregnancy and live birth rates were significantly higher in DET with two good quality embryos than DET with mixed quality embryos and SET groups. Multiple pregnancy rates were higher in both DET groups in terms of transfer day (p = 0.001). Conclusion DET with mixed quality embryos results with lower clinical pregnancy and live birth rates compared with DET with two good quality embryos at the blastocyst stage. At cleavage stage transfer, there is no difference in live birth rates between the two groups.

Is leptin receptor expression triggered in the case of embryo transfer to endometrium coculture?

Background/aim: A synchronized dialogue between maternal and embryonic tissues is required for successful implantation. Low uterine receptivity is responsible for two-thirds of implantation failures and leptin is effective in the physiology of reproduction by binding to specific receptors. In this study, we investigateleptin receptor expression in cases of embryo transfer to endometrial coculture. Materials and methods: Biopsy materials were taken from 20 females with indication for coculture application and were cultured in an appropriate medium after the epithelial cells were isolated. The grown cells were cultured in chamber slides as the first group. For the second group, day 3 embryo was added to chamber slides and the development was observed. The embryo was transferred 1 or 2 days later and other cells (after the transfer process) were used to form the second group. After fixation, immunohistochemical staining was performed with anti-leptin primary antibody. Results: Regarding the coculture without embryo transfer, moderate leptin receptor immunoreactivity was seen in the perinuclear region and the cell membrane. Also, regarding the coculture with embryo transfer, moderate leptin receptor immunoreactivity was seen in the cytoplasm and strong leptin receptor immunoreactivity was seen in the cell membrane. Conclusion: Embryo transfer to endometrium coculture triggers leptin receptor expression

Value of Post-transfer Day-12 Beta Human Chorionic Gonadotropin Levels for Pregnancy Outcome Prediction of Intracytoplasmic Sperm Injection Cycles.

BACKGROUND: Several markers were studied previously in order to predict the pregnancy outcome of assisted reproductive techniques; however, serum beta human chorionic gonadotropin was found to be the most predictive marker. AIMS: To evaluate the value of serum beta human chorionic gonadotropin levels in discriminating biochemical and clinical pregnancies 12 days after embryo transfer, while determining the factors predicting ongoing pregnancy was established as the secondary aim. STUDY DESIGN: Retrospective cross-sectional study. METHODS: A total of 445 pregnant cycles were retrospectively analysed in 2359 embryo transfer cycles. Patients were divided into two groups according to the outcome of pregnancy: biochemical and clinical. RESULTS: The cut-off value of beta human chorionic gonadotropin levels on day 12 in predicting clinical pregnancies was 86.8 IU/mL with 65.1% sensitivity and 74.7% specificity [CI: 0.76 (0.71-0.81). Receiver operating characteristic curve analysis revealed different cut-off values for embryo transfer days (57 mIU/mL for day 3 embryo transfer CI: 0.59-0.79 and 87 mIU/mL for day 5 embryo transfer, CI: 0.74-0.86). Subgroup analysis of clinical pregnancies revealed a significant difference between ongoing pregnancies and early fetal losses regarding duration of infertility (81.3±54.4 vs. 100.2±62.2 months), serum oestradiol on hCG day (2667.4±1276.4 vs. 2094.6±1260.5 pg/mL), number of transferred embryos (1.9±0.8 vs. 1.5±0.7) and the prevalence of diminished ovarian reserve as an indication (2.3% vs 12.2%). CONCLUSION: Beta human chorionic gonadotropin levels on day 12 following embryo transfer provide an important parameter for the prediction of clinical pregnancy; however, other stimulation parameters are indicated in the prediction of ongoing pregnancies.

Does combining magnetic-activated cell sorting with density gradient or swim-up improve sperm selection?

PURPOSE: The present study aimed to evaluate whether combining the magnetic-activated cell sorting (MACS) with density-gradient (DG) or swim-up (SU) sperm separation techniques can improve sperm selection to obtain higher quality spermatozoa. METHODS: Two commonly used sperm selection techniques, SU and DG, were compared to MACS combined with either SU or DG. Spermatozoa obtained from normozoospermic (n = 10) and oligozoospermic (n = 10) cases were grouped as SU, DG, SU+MACS, and DG+MACS followed by the analysis of sperm morphology, motility, DNA integrity, and the levels of Izumo-1 and PLCZ proteins. RESULTS: Although spermatozoa obtained by SU or DG when combined with MACS have improved aspects when compared to SU or DG alone, results did not reach a statistically significant level. Moreover, separation with MACS caused a significant loss in the numbers of total and rapid progressive spermatozoa. CONCLUSIONS: Considering the cost/benefit ratio, MACS application together with traditional techniques may only be preferred in certain cases having higher concentrations of spermatozoa, but it does not seem to be an ideal and practical sperm selection technique for routine use.

Investigation of functional sperm membrane integrity after consequence of anti-hypoosmotic swelling test technique: an electron microscopic study.

OBJECTIVE: Infertility is described as the inability of a couple to conceive after 1 year of unprotected intercourse. Male factor is assumed to be responsible for 50% of cases of infertile couples. It is a common clinical problem, affecting approximately 13-15% of couples worldwide. In this study we aimed to investigate the effects of hypoosmotic swelling (HOS) and anti-HOS tests on sperm membrane integrity at the ultrastructural level. STUDY DESIGN: Twenty-nine infertile and 10 fertile men were included in this study. The fertile and infertile subjects were classified according to Kruger and WHO criterion. All semen samples were examined and ana-yzed based on WHO guidelines. Sperm viability was determined by using the eosin Y staining method. After HOS and anti-HOS tests wore applied, the samples were evaluated at the ultrastructural level. RESULTS: Normal structural features of all regions of sperm were observed in sections of sham normospermia. Some histopathological changes wore seen in HOS and anti-HOS group sections. CONCLUSION: The HOS procedure was found not to cause degenerative changes in the sperm ultrastructure. The anti-HOS procedure can be applied in normospermic and oligospermic groups.

Obesity is not associated with the poor pregnancy outcome following intracytoplasmic sperm injection in women with polycystic ovary syndrome.

OBJECTIVE: To determine if body mass index has an effect on the outcome of in vitro fertilization in patients with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation. MATERIAL AND METHODS: The study included 337 cycles. Patients were stratified into the following 3 groups: normal weight, overweight, and obese. The primary outcome measures were response to ovarian hyperstimulation, the fertilization rate, the implantation rate, and the clinical and ongoing pregnancy rates. RESULTS: Total gonadotropin consumption increased, and the number of retrieved oocytes decreased as the body mass index increased. The implantation rate and clinical pregnancy rate were similar in all 3 groups. In response to the mid-luteal long protocol, the cycle cancellation rate was lower and the number of retrieved oocytes was higher in the overweight and obese groups, as compared to the antagonist protocol. CONCLUSION: The body mass index did not affect the outcome of in vitro fertilization in women with polycystic ovary syndrome. Additional research is required to better understand the role of stimulation protocols on the cycle outcome.

Effects of atorvastatin on smoking-induced alveolar injury in rat lungs.

BACKGROUND: Smoking is one of the most serious health care issues worldwide, as one third to one half of all people who smoke eventually use tobacco habitually. Chronic smoke exposure causes airway and lung parenchymal inflammation and the destruction of alveolar cell walls. Statins may have anti-inflammatory effects that would play a role in preventing the cellular damage associated with smoking. OBJECTIVE: The aim of this study was to investigate whether atorvastatin protects against smoking-induced inflammation in alveolar epithelial type I (ATI) and type II (ATII) cells in the lungs of rats. METHODS: Adult male albino Wistar rats (200-250 g) were randomly divided into 3 groups and exposed to cigarette smoke 8 hours per day for 15 days. During that 15-day period, the 2 treatment groups received atorvastatin 0.5 or 1.0 mg/kg/d in 2 mL of methyl cellulose solution and the control group received 2 mL of methyl cellulose solution alone, all via nasogastric catheter. After the 15 days, the lungs were excised and the tissues were examined by transmission electron microscopy. RESULTS: Thirty rats were divided into 3 groups of 10 rats each. All rats survived the 15 days. In the atorvastatin 0.5-mg group, no changes were found in the ATI cells or in the blood-air barrier. In the atorvastatin 1.0-mg group, we observed hyperplasia in the common basal membranes. Hypertrophy, mitochondrial crystolysis (MC), and intracytoplasmic edema (ICE) were detected in the ATI cells in the 1.0-mg group, while chromatin condensation, atrophic appearance, cell shrinkage, and cyto-plasmic vacuolization were observed in the ATII cells. The rough endoplasmic reticulum (rER) tubules of the ATII cells appeared spiral-shaped. In the control group, minimal ICE was detected in the ATI cells. However, microvillus deformation, pseu-dopod formation, edema, mitochondrial swelling, and MC were observed in the ATII cells. We also observed MC, several pinocytic vesicles, and normal rER tubules in the endothelial cells of the control group. CONCLUSIONS: The administration of atorvastatin 0.5 mg/kg/d was associated with some attenuation of lung injury caused by smoke inhalation in these rat lungs. However, atorvastatin 1.0 mg/kg/d was associated with lung damage. Future studies are needed to evaluate the dose-response relationship of atorvastatin to smoking-induced alveolar damage.