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Purpose: To compare the effects of microfluidic sperm sorting, density gradient and swim-up methods on the oxidative reduction potential (ORP) of split semen samples from a single patient population., Materials and Methods: A prospective controlled study was conducted to compare the effects of three different semen processing methods using split semen samples from the same population of infertile men. The primary outcome was the ORP. Secondary outcomes were the sperm concentration, progressive motility rate and total sperm motility., Results: A total of 57 split semen samples were included in this study. The ORP was significantly lower in the microfluidic group compared to the density gradient and swim-up groups (p < 0.05). The ORP/sperm concentration ratio was significantly lower in the microfluidic and density gradient groups compared to the swim-up group (p < 0.05).Total sperm concentration was significantly higher in the density gradient group than the microfluidic and swim-up groups (p < 0.05). Motility was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05). The progressive motile sperm rate was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05)., Conclusions: Microfluidic sperm sorting was better for selecting highly motile sperm and yielded a lower ORP than conventional sperm preparation methods.

Objective: To compare the effect of microfluiding sperm sorting chip and density gradient methods on ongoing pregnancy rates (PRs) of patients undergoing IUI., Design: Retrospective cohort study., Setting: Hospital IVF unit., Patient(s): Couples with infertility undergoing IUI cycles between 2017 and 2018., Intervention(s): Not applicable., Main Outcome Measure(s): Ongoing PRs., Result(s): A total of 265 patients were included in the study. Microfluid sperm sorting and density gradient were used to prepare sperm in 133 and 132 patients, respectively. Baseline spermiogram parameters, including volume, concentration, motility, and morphology, were similar between the two groups. Total motile sperm count was lower in the microfluiding sperm sorting group at baseline (35.96 ± 37.69 vs. 70.66 ± 61.65). After sperm preparation sperm motility was higher in the microfluid group (96.34 ± 7.29 vs. 84.42 ± 10.87). Pregnancy rates were 18.04% in the microfluid group and 15.15% in the density gradient group, and ongoing PRs were 15.03% and 9.09%, respectively. After using multivariable logistic regression and controling for confounding factors, there was a significant increase in ongoing PRs in the microfluid sperm sorting group. The adjusted odds ratio for ongoing pregnancy in the microfluid group compared with the density gradient group was 3.49 (95% confidence interval 1.12-10.89)., Conclusion(s): The microfluid sperm sorting method significantly increased the ongoing PRs compared with the density gradient group in IUI cycles., (Copyright © 2019 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The aim of this study is to evaluate the impact of ovarian reserve and age of women on early morphokinetic parameters of embryos with a time-lapse monitoring system. In total, 197 infertile couples with poor ovarian reserve (Group 1, n = 41), normal ovarian reserve (Group 2, n = 59), or polycystic ovaries (Group 3, n = 97) were included. The time from insemination to the following events were analyzed: pronuclear fading (Pnf) and cleavage to 2, 3, 4 and 5 cells. The optimal ranges for morphokinetic parameters of t5, s2 and cc2 in each group were also evaluated. In total, 1144 embryos were evaluated. Morphokinetic parameters did not differ statistically between the groups. Data were analyzed according to different age groups (20-30, 30-40, >40). The morphokinetic parameters did not differ statistically in Group 1 and 3. In Group 2, the times from insemination to tPnf, t2, t3, t4 were significantly shorter in the younger age group than the older age group (p < 0.05). The percentages of optimal embryos, according to t5, s2 and cc2, did not differ statistically between the groups. In conclusion, ovarian reserve did not seem to affect the morphokinetic parameters of embryos.

Purpose: The aim of this study is to evaluate expression of deoxyribonucleic acid (DNA) synthesis and repair markers in testicular tissues of azoospermic men in whom sperm retrieval could and could not be achieved as a result of microdissection testicular sperm extraction (micro-TESE) procedure., Materials and Methods: In this prospective cohort study, testicular tissues were retrieved from 60 Non-obstructive Azoospermia (NOA) patients who underwent micro-TESE procedure. These patients were divided into twogroups: micro-TESE positive group, which included 30 NOA patients from whom sperm could be extracted via micro-TESE procedure; and micro-TESE negative group, which included 30 NOA patients from whom sperm retrieval could not be achieved via micro-TESE procedure. Expression and distribution patterns of poly(ADP-ribose) polymerase-1 (PARP-1) and proliferative cell nuclear antigen (PCNA) in extracted tissues were assessedby immunohistochemical staining to reveal any differences in DNA synthesis and repair between the two groups., Results: Micro-TESE positive group exhibited significantly stronger immunoreactivity for both PCNA and PARP-1 (P = .001 and P = .001 respectively). The results of this study reveal that both DNA synthesis and repair markers were expressed strongly in patients who experienced successful micro-TESE procedure., Conclusion: Although further studies are needed to support these findings, PARP-1 and PCNA expression in testicular tissues of NOA patients could be promising predictive factors for micro-TESE procedure success.

Purpose: This study seeks to evaluate the association between follicular fluid (FF) coenzyme Q10 (CoQ10) levels, embryo morphokinetics, and pregnancy rate., Methods: Sixty infertile patients who underwent intracytoplasmic sperm injection (ICSI) cycles were included in the study. For each patient, CoQ10 level of the follicular fluid was measured by high-performance liquid chromatography system. After the ICSI of each oocyte, the relationship between the level of CoQ10 content of each follicular fluid, the subsequent embryo quality, and embryo morphokinetics was investigated. The relationship between the level of CoQ10 content of each follicle and optimal time-lapse parameters for the embryos of these follicles including t5, s2, and cc2 was also analyzed. The embryos were further classified into four categories, namely, grades A, B, C, and D, according to morphokinetic parameters using t5-t2 and t5-t3 (cc3). Each follicular fluid analysis was performed for a single oocyte of a single embryo which was transferred to the patients. Additionally, follicular fluid CoQ10 levels and pregnancy rates were evaluated., Results: Follicular fluid CoQ10 levels were significantly higher in grades A and B than grades C and D embryos (p < 0.05). The concentration of CoQ10 levels was significantly higher in the pregnant group (p < 0.05). There was no significant correlation between optimal t5 and s2 morphokinetic parameters and CoQ10 levels. However, CoQ10 levels were significantly higher in follicular fluid of embryos which had optimal cc2 (p < 0.05)., Conclusion: High follicular fluid CoQ10 level is associated with optimal embryo morphokinetic parameters and higher pregnancy rates.

Ovarian hyperstimulation syndrome (OHSS) is the most serious iatrogenic complication of IVF cycles. Although the development of effective treatment strategies for this syndrome is important, preventing OHSS is more crucial. Triggering ovulation with a gonadotrophin-releasing hormone (GnRH) agonist is one method used to avoid OHSS. In this paper, three patients who developed severe OHSS after undergoing GnRH agonist triggering and freezing of all embryos in a GnRH antagonist protocol are described. A review of the literature is also provided. This report highlights the ongoing risk of severe OHSS even after GnRH agonist triggering combined with freezing all embryos in GnRH antagonist cycles. Other prevention strategies might be considered for extreme hyper-responders., (Copyright © 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)

This study was designed to evaluate the effect of luteal-phase administration of single-dose gonadotrophin-releasing hormone (GnRH) agonist on pregnancy, implantation and live birth rates in patients who received GnRH antagonist for pituitary suppression. The study population consisted of 164 patients who underwent intracytoplasmic sperm injection (ICSI) after ovulation induction by gonadotrophins and GnRH antagonist for the prevention of a premature LH surge. For luteal-phase support, all the cases received intravaginal 600 mg micronized progesterone. In this prospective study, patients were randomly assigned to two groups. In one group, patients received an additional single dose of GnRH agonist (0.5 mg leuprolide acetate) subcutaneously on day 6 after ICSI, whereas the patients in the other group did not. Although the number of embryos transferred and the grade of the embryos were similar in the two groups, the patients in the luteal-phase agonist group had significantly higher rates of implantation and clinical pregnancy rates ( P < 0.05). When the two groups were compared, there were also statistically significant differences in multiple pregnancy and live birth rates ( P < 0.05). Administration of single-dose GnRH agonist as a luteal-phase support in ovarian stimulation-GnRH antagonist cycles in addition to standard luteal support seems to be effective in all cycle outcome parameters.

The purpose of this study was to evaluate the effectiveness of combined approach on the prevention of severe ovarian hyperstimulation syndrome (OHSS) in high risk patients undergoing controlled ovarian hyperstimulation for IVF. The combined approach consisted of: (1) step-down administration of gonadotropins; (2) lowering the dose of human chorionic gonadotropin; (3) intravenous albumin infusion at the time of oocyte retrieval and (4) progesterone use for luteal support. Total of 87 high risk patients with a serum estradiol level >11,010 pmol/l or 3000 pg/ml on HCG day were managed by this combined approach and their results were compared with 274 low risk patients. In all high risk patients, the gonadotrophin dose were decreased starting as early as on day 4 of ovarian stimulation as necessary, ovulation was triggered by a decreased HCG dose of 5000-7000 IU according to the level of estradiol, intravenous infusion of 20% human albumin, 50-100ml were given just 1h before the oocyte retrieval and luteal support was provided either by 50mg progesterone in oil, IM or 600 mg micronized progesterone orally or vaginally until the day of beta-HCG determination. All patients were followed by serial ultrasonographic examinations and complete blood count analysis after embryo transfer to detect the early signs of OHSS and to allow early intervention. Age and duration of infertility were similar in both groups. Although the number of gonadotrophin ampoules used (22.7 +/- 4.7 versus 27.8+/-3.7; P<0.05) was significantly lower, estradiol levels (16,764 +/- 6936 pmol/l versus 8870 +/- 2456 pmol/l; P<0.05) and mean number of oocytes (18.3 +/- 5.9 versus 10.6+/-5.4; P<0.05) were significantly higher in study group. There was no significant difference between groups in terms of the mean number of transferred embryos (3.2 +/- 1.1 versus 3.4+/-1.1) and rate of pregnancies (50.5% versus 40.1%). There was only one moderate and no severe OHSS case in the high risk group, while five moderate and one severe OHSS cases developed in the control group consisting of low risk patients. In conclusion, intravenous albumin combined with low dose HCG, early step-down administration of gonadotropins and progesterone use for luteal support, so called combined approach, proved to be effective in the prevention of severe ovarian hyperstimulation syndrome in documented high risk patients.

Purpose: Our purpose was to evaluate the progression of embryos derived from round spermatid injection to the blastocyst stage and compare the results with those obtained by the use of testicular or epididymal spermatozoa., Methods: Thirty-eight patients with azoospermia enrolled in this study. In 29 patients with obstructive or nonobstructive azoospermia, spermatozoa were recovered from epididymis or testis. In the remaining nine cases with nonobstructive azoospermia, only round spermatids were found in seven, whereas in two of the patients, there were no elongated or round spermatids. Six of these cases underwent round spermatid injection., Results: Twenty-one of 29 patients with injection of spermatozoa underwent embryo transfer on day 3, and 10 pregnancies (47.6%) were obtained. In eight cycles, embryos were further cultured for delayed transfer. In six cases undergoing round spermatid injection, no transfer was performed on day 3 and extended culture with delayed embryo transfer was applied. The mean number of fertilized oocytes and mean number of embryos on day 3 and also the fertilization rate and mean number of good-quality embryos on day 3, mainly grade 1 or 2, were statistically significantly higher in the spermatozoa group than the round spermatid injection group. Compared to the spermatozoa group, the number of arrested embryos was significantly higher and the number of blastocyst-stage embryos and number of good-quality blastocysts were significantly lower in the spermatid injection group. No blastocysts developed in two spermatid cycles and embryo transfer was not possible, and in the remaining four cycles, after at least one blastocyst transfer, no pregnancies were achieved. However, in eight cycles with extended culture in the spermatozoa group, embryo transfers were achieved in all and three pregnancies, for a pregnancy rate of 37.5%, were obtained after blastocyst transfer., Conclusions: Our preliminary results showed that round spermatid injection was associated with a significantly lower fertilization and embryo development rate and a significantly higher developmental arrest rate compared with the injection of spermatozoa. Extended culture and delayed embryo transfer did not improve the clinical outcome after round spermatid injection, and these results suggested a developmental failure in embryos preventing successful implantation after round spermatid injection.

Purpose: In this prospective randomized study the effects of enzymatic treatment of zona pellucida of blastocysts on implantation and pregnancy rates were evaluated in a group of patients who had more than five embryos on day 3., Methods: Forty-six patients with a mean age of 29.8 +/- 4.5 years and mean duration of infertility of 6.72 +/- 0.63 years had blastocyst stage transfers, with a mean number of 2.9 +/- 0.1 embryos replaced per patient. Patients were randomly divided into two groups. The first group consisted of patients (n = 22) who had zona intact blastocyst stage transfers and the second group consisted of patients (n = 24) who had zona manipulated (enzymatic treatment) blastocyst stage transfers. Patient and cycle characteristics were similar in both groups. A commercial cell and serum-free sequential culture system was used for all embryos., Results: Overall blastocyst formation rate was 50.3%. Transfer could be done in all patients. The positive beta-human chorionic gonadotropin rate in the zona intact group was 50% (11/22) and in the zona-manipulated group was 70.8% (17/24). Clinical pregnancy, ongoing pregnancy, and implantation rates in zona intact and manipulated groups were 45.5%, 27.3%, and 19%, and 62.5%, 45.8%, and 24% respectively. Although implantation and pregnancy rates in the zona-manipulated group were higher, there were no statistically significant differences in terms of these variables between two groups. No triplet pregnancy was obtained in either group, and the twin pregnancy rate was 20% (2/10) in the zona intact group and 13.3% (2/15) in the zona-manipulated group., Conclusions: With further improvements in the embryo culture systems it will become possible in the near future to achieve high implantation rates even with single blastocyst transfers. Enzymatic treatment of the zona pellucida seems that it does not alter the pregnancy and implantation rates, but further studies with larger group of patients are needed to clarify the real effect of this zona manipulation on pregnancy outcome.

Objective: To assess the effects of cysts developed prior to the commencement of luteal phase gonadotropin-releasing-hormone-agonist (GnRH-a) in IVF cycles., Design: Retrospective analysis., Setting: In vitro fertilization program in a tertiary hospital infertility clinic., Patients: Women stimulated for IVF-ET. All stimulations were down-regulated with GnRH-a commenced on day 21 in a long protocol before gonadotropin stimulation., Main Outcome Measure: Pregnancy rate., Results: Twenty-five of 121 cycles had cysts >15 mm (20.6%); these were classified as baseline (nonfunctional) (n = 12) or corpus luteum (n = 13) cysts. They had significantly longer suppression periods and lower peak estradiol levels, and used more gonadotropins during stimulation. Cysts had no impact on the number of oocytes collected or fertilization and pregnancy rates. Patients with baseline cysts had a greater duration of suppression and required more gonadotropin than those with corpus luteum cysts or those without cysts. The cycle outcomes were similar between the groups, but baseline cyst formation is likely to increase the cost of IVF., Conclusion: These findings suggest that baseline cyst formation causes longer suppression duration and greater gonadotropin utilization, although an impact on pregnancy rates is unlikely.

Objective: To assess the effectiveness of intravenous administration of albumin in prevention of ovarian hyperstimulation syndrome (OHSS) in patients of an in-vitro fertilization program., Study Design: Prospective randomized study. Patients with hCG day E2 levels are 11010 pmol/l (3000 pg/ml) or more were recruited into two groups. Group A (n = 27) received 10 g 20%, 50 ml human albumin infusion before oocyte pick-up and no medication was administered in Group B (n = 28)., Results: Patients were similar in terms of cycle characteristics. No moderate-severe OHSS developed in Group A whereas one severe OHSS case and four moderate OHSS cases developed in Group B. Statistical analysis revealed a significant (P < 0.05) protection in albumin treated group., Conclusion: Human albumin proves effectiveness in prevention of moderate-severe OHSS.

We analyzed whether patent but anatomically deranged tubes were more commonly encountered in infertile women than in fertile women. The fertile group consisted of 100 women who were apparently normal with respect to fertility and the infertile group consisted of 100 infertile women with no certain etiology identified. The individual tubal structure was recorded at laparoscopy and the results of the two groups were then compared. Tubal pathology was found to be significantly higher in the infertile group; however, this marked difference arose from the contribution of a significantly higher frequency of peritubal pelvic adhesions because no difference was found between the two groups regarding anatomic variations of the fallopian tube. We conclude that tubal derangement which could only be identified with laparoscopy significantly contributes to infertility.