Batuhan Özmen

Three years of routine vitrification of human zygotes: is it still fair to advocate slow-rate freezing?

Cryopreservation of human oocytes and embryos is a necessary tool in assisted reproduction treatment that leads to an increased cumulative outcome while decreasing costs. Vitrification is a cryopreservation technique that leads to a glass-like solidification, with rapid cooling of cells or tissues. Nowadays vitrification is claimed to be the future of cryopreservation of human embryos due to improved survival rates and clinical outcomes. This study was conducted at a university clinic to assess the safety and efficiency of vitrification of human zygotes as a routine procedure. A total of 849 pronuclear-stage (PN) zygotes were vitrified between March 2004 and July 2006. During this period, 103 cycles of cryopreserved embryo transfer were completed. In total, 339 PN zygotes were thawed resulting in an 89% survival rate (302 PN zygotes). The mean number of embryos per transfer was 2.2. The pregnancy rate obtained was three times higher (36.9%) than that obtained with the slow-rate freezing method (10.2%) used previously in the same centre. In conclusion, vitrification of human zygotes at the pronuclear stage seems to be a successful and reliable method with favourable outcomes and can be recommended as a routine technique for cryopreservation of human embryos.

Current aspects of blastocyst cryopreservation

Cryopreservation of human gametes and embryos has become an essential part of assisted reproduction. Successful cryopreservation of human blastocysts is increasingly relevant as extended in-vitro culture of human embryos becomes more common, permitting routine use of blastocyst transfer in IVF programmes. This reduces the number of embryos transferred, thereby reducing multiple pregnancies and maximizing cumulative pregnancy rates per oocyte retrieval. The superiority of blastocyst freezing over earlier stage freezing in terms of implantation per thawed embryo transferred improves overall expectations for the cryopreservation programme. Therefore, a reliable procedure for the cryopreservation of blastocysts is needed because, after transfer, only a small number of supernumerary blastocysts are likely to be available for cryopreservation. Since the early 1980s, two common techniques have been used in cryopreservation: the conventional slow cooling method and the more recent rapid procedure known as vitrification. Vitrification has become an attractive alternative to slow freezing, since it appears to result in significantly higher survival and pregnancy rates. The aim of this review is to focus on the cryopreservation of human blastocysts using slow and rapid protocols and to assess the impact of the crypreservation protocol used on the survival, implantation and pregnancy rates.

DNA damage of human spermatozoa in assisted reproduction: origins, diagnosis, impacts and safety

Sperm DNA contributes half the offsprings genomic material and abnormal DNA can lead to derangements in the reproductive process. Normal sperm genetic material is required for successful fertilization, as well as for further embryo and fetal development that will result in a healthy child. Thus, the damage to sperm DNA is critical in assisted reproductive techniques which are increasingly used to treat infertile couples. There has been improving data about the effects of human sperm DNA damage or fragmentation. As well, increasing knowledge concerning the effects of DNA damage on embryo and fetal development has been attained. This review aims to summarize the present knowledge on the impact of human sperm cell DNA damage on male infertility and outcome in the context of safety.

Relationship between sperm DNA damage, induced acrosome reaction and viability in ICSI patients.

The DNA damage in human spermatozoa is a relevant predictor of prognosis in male infertility, whereby increased sperm DNA damage impairs the outcomes of artificial reproduction. Theoretically, DNA damage should alter the special cellular functions of human spermatozoa, and lead to diminished acrosome reaction with reduced fertilization rates. Nevertheless, intracytoplasmic sperm injection (ICSI) has been reported to alleviate such negative outcomes due to DNA damage. This study investigated the relationship between DNA fragmentation and acrosome reaction as well as viability in ICSI patients. The study enrolled 42 men undergoing ICSI due to poor sperm parameters. The DNA fragmentation indexes (DFI) were 4-10% in 38% of the cases, and > or = 10% in 19% of the cases. The results of both acrosome reaction and viability assays showed negative correlations with DFI values in all cases and especially in cases with fertilization rates <60% (P < 0.05). However, such correlations were not found in cases with fertilization rates >60%. There were no live deliveries in patients with high DFI levels (>10%). In conclusion, negative correlations were identified between increased DNA damage, and acrosome reaction and/or viability of human spermatozoa, especially in cases with reduced fertilization rates.

Ovarian aging and premature ovarian failure

Physiological reproductive aging occurs as a result of a decrease in the number and quality of oocytes in ovarian cortex follicles. Although the reason for the decrease in the quality of the pool and follicular oocytes is not fully understood, endocrine, paracrine, genetic, and metabolic factors are thought to be effective. Nowadays, in order to understand the mechanisms of ovarian aging, genomic research has gained importance. The effect of co-factors, such as telomerase and ceramide, in the ovarian aging process is only getting ascertained with new research studies. The most important tests in the assessment of ovarian aging are antral follicle count and anti-Mullerian hormone.

The effects of subchorionic hematoma on pregnancy outcome in patients with threatened abortion.

OBJECTIVE To assess the effects of ultrasonographically detected subchorionic hematomas on pregnancy outcomes in patients with vaginal bleeding within the first half of pregnancy. MATERIAL AND METHODS Patients diagnosed with threatened abortion due to painless vaginal bleeding and who were followed up in an in-patient service during the first vaginal bleeding between January 2009 and December 2010 were included in this retrospective cohort study. Patients were divided into two groups according to the presence of subchorionic hematoma. Miscarriage rates and pregnancy outcomes of ongoing pregnancies were compared between the groups. RESULTS There were no statistically significant differences between the groups regarding demographic parameters, including age, parity, previous miscarriage history, and gestational age at first vaginal bleeding. While 13 of 44 pregnancies (29.5%) with subchorionic hematoma resulted in miscarriage, 25 of 198 pregnancies (12.6%) without subchorionic hematoma resulted in miscarriage (p=.010). The gestational age at miscarriage and the duration between first vaginal bleeding and miscarriage were similar between the groups. The outcome measures of ongoing pregnancies, such as gestational week at delivery, birth weight, and delivery route, were also similar between the groups. CONCLUSION Ultrasonographically detected subchorionic hematoma increases the risk of miscarriage in patients with vaginal bleeding and threatened abortion during the first 20 weeks of gestation. However, it does not affect the pregnancy outcome measures of ongoing pregnancies.

Is omitting the 3rd hour measurement in the 100 g oral glucose tolerance test feasible

Abstract Background: To evaluate the diagnostic value of the 3rd hour plasma glucose level in the 100 g oral glucose tolerance test (OGTT). Methods: Records of all pregnant patients with abnormal 50 g glucose challenge test (GCT) between January 2005 and December 2013 were reviewed (n=1963). The 100 g OGTT results were analyzed separately for both Carpenter & Couston (CC) and National Diabetes Data Group (NDDG) criteria. Results: The number of patients diagnosed with gestational diabetes mellitus (GDM) was 297 (15.1%) according to CC criteria and 166 (8.4%) according to NDDG criteria. The 1st hour plasma glucose level showed the highest correlation with GDM diagnosis (ρ=0.595 for CC and ρ=0.567 for NDDG). However, the 3rd hour plasma glucose level showed the weakest correlation with GDM diagnosis (ρ=0.216 for CC and ρ=0.213 for NDDG). The 3rd hour value of 100 g OGTT was one of the two elevated measurements in 10.8% of patients when CC criteria are used and in 13.8% of patients when NDDG criteria are used. Conclusion: Omitting 3rd hour plasma glucose measurement in 100 g OGTT results in unacceptable rates of underdiagnosed patients.

Signet-ring stromal tumor of the ovary: an extremely rare neoplasm.

Signet-ring stromal tumor (SRST) of the ovary is an extremely rare neoplasm. Herein, we present a SRST of the ovary, which is the twelfth report in the literature. A 44 year-old, G4P2 patient was admitted with the complaint of polymenorrhea. She was operated on for persistent semisolid ovarian mass measuring 5 cm in diameter. The pathological examination confirmed the diagnosis of a benign ovarian SRST. In conclusion, SRST is an extremely rare benign ovarian tumor with good prognosis according to the current literature. Although the recurrence rate or malignant transformation potential of these tumors are not yet known, close follow-up in the post-operative period may be beneficial.

A retrospective comparative study of prednisolone use in antagonist co-treated assisted reproductive technology cycles for patients with good prognosis

Objective: To investigate the impact of peri-implantation prednisolone use and its duration in antagonist co-treated assisted reproductive technology (ART) cycles of patients with good prognosis. Materials and Methods: Infertile patients treated with gonadotropin-releasing hormone antagonist protocol between January 2010 and June 2013 were included. The patients in group A (n=196) received no prednisolone. The patients in groups B (n=397) and C (n=371) received 5 mg oral prednisolone daily, for 4 and 12 days following embryo transfer, respectively. The main outcome parameter was live birth rate. Results: The ages of the groups were 30.1±4.6, 31.5±4.5, and 30.9±4.7 years, respectively (p=0.163). There was no statistically significant difference between the groups regarding cycle characteristics. Implantation rates were 20.7%, 24.6%, and 23.8%, respectively (p=0.163). Miscarriage rates were 1.5%, 3.5%, and 3.2%, respectively (p=0.859). Live birth rates were 28.7%, 29.3%, and 32.8%, respectively (p=0.482). Conclusion: Empiric prednisolone administration during the peri-implantation period does not seem to have beneficial effects in ART cycles of patients with good prognosis.

A live birth after transfer of a day 2 embryo derived from frozen-thawed zygotes that had undergone polar body biopsy: a case report

OBJECTIVE To present a live birth after freezing and thawing of biopsied oocytes. DESIGN Case report. SETTING Artificial reproduction unit of a university hospital. PATIENT(S) A primary infertile couple with asthenoteratozoospermia and repeated failures of intracytoplasmic sperm injection (ICSI). INTERVENTION(S) Screening of aneuploidy during the fourth ICSI cycle with polar body biopsy (PB) for repeated failures of artificial reproductive techniques and a transfer of a cryopreserved day 2 embryo derived from cryopreserved zygotes with slow-rate freezing after PB. MAIN OUTCOMES MEASURE(S) Live birth, viability, and survival. RESULT(S) A successful pregnancy and a live birth were presented after a transfer of day 2 embryos derived from oocytes that underwent PB and subsequent cryopreservation. CONCLUSION(S) Pregnancy can be obtained subsequent to cryopreservation and thawing after PB.