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This retrospective cohort study determined the relative efficacy of blastocyst and cleavage-stage transfers in patients with differing numbers of zygotes. A total of 1116 women whose embryo transfers were planned independently of patient characteristics were included. Cleavage-stage (D3) and blastocyst-stage (D5) transfer outcomes were analyzed per number of zygotes. The D5 group included transfer cancellations as the intention-to-treat population. The effect of the embryo transfer date on the clinical outcomes (clinical pregnancy and implantation rates) was analyzed using multivariate logistic regression. Among the patients, 584 and 532 underwent D3 and D5 embryo transfers, respectively. The clinical pregnancy rates were significantly higher in D5 patients with ≥ 6 zygotes (25.7% vs 48.3%). The multivariate logistic regression analysis for clinical pregnancy did not show significant differences between the blastocyst and cleavage-stage transfers in patients with ≤ 5 zygotes (0.874 [0.635-1.204]). Compared to the cleavage-stage, blastocyst-stage transfers for patients with ≥ 6 zygotes resulted in a three-fold increase in clinical pregnancy rates (3.122 [1.797-5.425]). Blastocyst transfers were not inferior to cleavage-stage embryo transfers among patients with few zygotes and were preferable for patients with several zygotes.

Assisted reproduction is presumed to increase monozygotic twin rates, with the possible contribution of laboratory and medical interventions. Monozygotic dichorionic gestations are supposed to originate from the splitting of an embryo during the first four days of development, before blastocyst formation. Single embryo transfers could result in dichorionic pregnancies, currently explained by embryo splitting as described in the worldwide used medical textbooks, or concomitant conception. However, such splitting has never been observed in human in vitro fertilization, and downregulated frozen cycles could also produce multiple gestations. Several models of the possible origins of dichorionicity have been suggested. However, some possible underlying mechanisms observed from assisted reproduction seem to have been overlooked. In this review, we aimed to document the current knowledge, criticize the accepted dogma, and propose new insights into the origin of zygosity and chorionicity.

Purpose This retrospective cohort study determined the relative efficacy of blastocyst and cleavage-stage transfers in patients with differing numbers of zygotes. Methods A total of 1116 women whose embryo transfers were planned independently of patient characteristics were included. Cleavage-stage (D3) and blastocyst-stage (D5) transfer outcomes were analyzed per number of zygotes. The D5 group included transfer cancellations as the intention-to-treat population. The effect of the embryo transfer date on the clinical outcomes (clinical pregnancy and implantation rates) was analyzed using multivariate logistic regression. Results Among the patients, 584 and 532 underwent D3 and D5 embryo transfers, respectively. The clinical pregnancy rates were significantly higher in D5 patients with ≥ 6 zygotes (25.7% vs 48.3%). The multivariate logistic regression analysis for clinical pregnancy did not show significant differences between the blastocyst and cleavage-stage transfers in patients with ≤ 5 zygotes (0.874). Compared to the cleavage-stage, blastocyst-stage transfers for patients with ≥ 6 zygotes resulted in a three-fold increase in clinical pregnancy rates (3.122). Conclusion Blastocyst transfers were not inferior to cleavage-stage embryo transfers among patients with few zygotes and were preferable for patients with several zygotes.

Objective The primary aim of this study is to investigate whether there are any minor sonographic uterine findings, not typical for adenomyosis, in endometriosis patients. The secondary objective is to determine the prevalence of sonographic features of adenomyosis in an infertile population with endometriosis. Study design The investigation was of 291 infertile women with endometriosis, either manifesting endometrioma (OMA) or diagnosed through laparoscopy, who were investigated for two‐dimensional transvaginal sonographic (2D-TVS) features of adenomyosis. These patients were grouped as either having endometriosis with adenomyosis (EwA,n = 121) or without adenomyosis (EwoA, n = 170). Additionally, patients without both endometriosis and 2D-TVS features of adenomyosis constituted the control group (n = 170). Results At least one 2D-TVS feature of adenomyosis was detected in 41.6 % (n = 121) of women with endometriosis. Asymmetrical myometrial thickening of uterine walls (57.9 %), hyperechogenic islands (47.1 %), and fan-shaped shadowing (46.9 %) were relatively more prevalent 2D-TVS findings among EwA patients. Multiple OMA (p = 0.038), OMA ≥ 4 cm (p = 0.034), and total OMA volumes were found to be higher (p = 0.004) in the EwA group. Additionally, uterine volumes were found to be 96.7 cm3, 73.0 cm3, and 64.2 cm3 in the EwA, EwoA, and control groups, respectively (EwA vs EwoA, p Conclusion A stepwise and statistically significant volume increase from the control group to the EwoA and then to the EwA group may reflect a spectrum of uterine involvement in endometriosis. This might indicate that many uterine endometriosis cases are still hidden from view, possibly demonstrating an "iceberg phenomenon".

GİRİŞ ve AMAÇ: Oosit toplama günü semen örneklerinde rastlantısal olarak izole lökospermi saptanan olguların in vitro fertilizasyon / intrasitoplasmik sperm enjeksiyonu (İVF/İCSİ) tedavi sonuçlarını değerlendirmek. YÖNTEM ve GEREÇLER: Akdeniz Üniversitesi Tıp Fakültesi Üremeye Yardımcı Tedavi Merkezi’nde, Ocak 2015 ve Mart 2016 tarihleri arasında, İVF/İCSİ tedavisi uygulanmış hastalara ait bilgiler elektronik veri tabanı kullanılarak gözden geçirildi. Oosit toplama günü yapılan incelemede izole lökospermi (≥1x10(6)/mL) saptanan hastaların (n=70) belirlenmesinin ardından, indeks olguya en yakın dönemde oosit toplama işlemi gerçekleştirilmiş; kadın yaşı, metafaz 2 (M2) oosit ve önceki tedavi sayıları eşleştirilmiş ancak lökospermi saptanmayan (˂1x10(6)/mL) hastalar (n=70) seçilerek kontrol grubu oluşturuldu. Fertilizasyon oranları, embriyo kaliteleri ve gebelik sonuçları gruplar arasında karşılaştırıldı. BULGULAR: Gruplar arasında M2 oosit ve 2-pronuclei ortalamaları benzer bulundu (p>0.05). Ancak fertilizasyon oranının lökospermi grubunda daha düşük olduğu saptandı (%61.4 ve %69.7, p=0.012). Klivaj evresi (2-3. gün) yüksek kalite embriyo sayıları ve fertilize oositten yüksek kalitede klivaj embriyo gelişim oranı benzer bulundu (p>0.05). Hasta başına yüksek kalitede blastokist sayısı ve yüksek kaliteli klivaj evresindeki embriyolardan yine yüksek kaliteli blastokist gelişim oranında istatistiksel farklılık saptanmadı (p>0.05). Buna karşılık, total fertilizasyon başarısızlığının (%14.3 ve %2.9, p=0.016) ve siklus iptallerinin (%34.3 ve %17.1, p=0.020) lökospermi grubunda daha sık olduğu saptandı. Gebelik sonuçları incelendiğinde, kontrol grubunda siklus başına pozitif beta-insan koryonik gonadotropin (β-hCG) oranı daha yüksek bulunmasına karşın (p=0.033), embriyo transferi başına pozitif β-hCG ve klinik gebelik oranlarında gruplar arasında istatistiksel olarak anlamlı fark izlenmedi (p>0.05).TARTIŞMA ve SONUÇ: Lökospermi fertilizasyonu etkileyebileceğinden total fertilizasyon başarısızlığına ve siklus iptallerine neden olabilir. Ancak, fertilizasyonun sağlıklı şekilde gerçekleşmesi halinde embriyo kalitesinin ve gebelik sonuçlarının olumsuz etkilenmediği söylenebilir., INTRODUCTION: To investigate the effect of isolated leukocytospermia detected incidentally at the day of oocyte retrieval on in vitro fertilization / intracytoplasmic sperm injection (IVF/ICSI) treatment outcomes.METHODS: This retrospective study included patients who underwent IVF/ICSI treatment at the Akdeniz University Reproductive Centre between January 2015 and March 2016. The electronical database was screened to identify the patients with isolated leukospermia detected at the day of oocyte retrieval. Seventy patients whose semen samples had leukospermia (≥1x10(6)/mL) were retrospectively matched for women’s age, metaphase 2 (M2) oocytes and previous unsuccessful attempts at IVF with 70 patients without leukospermia (˂1x10(6)/mL). Fertilization rates, embryo qualities and pregnancy outcomes were compared between groups.RESULTS: The number of M2 oocytes and 2-pronuclei were similar between groups (p>0.05). However, fertilization rate was found to be significantly lower in leukospermia group (%61.4 vs. %69.7, p=0.012). The number of high quality embryos as well as developmental rate of high quality embryos from fertilized oocytes and high quality blastocysts from high quality cleavage stage embryos were comparable between groups (p>0.05). Leukospermia group had increased number of patients with total fertilization failure (%14.3 vs. %2.9, p=0.016) and cycle cancelation (%34.3 vs. %17.1, p=0.020) rates. Although positive beta-human chorionic gonadotropin (β-hCG) rate per cycle was higher in the control group (p=0.033), positive β-hCG and clinical pregnancy rates per embryo transfer were found to be comparable between the groups (p>0.05). DISCUSSION AND CONCLUSION: Leukospermia might affect the fertilization and therefore increase total fertilization failures and cycle cancelations. However, if fertilization occurs successfully, this situation might not have adverse effects on embryo development and pregnancy outcomes.

Amaç: Ardışık kültür ortamı ile kesintisiz tek basamaklı kültür ortamının erken dönemde embriyo kaliteleri üzerine olan etkilerini araştırarak, farklı kültür ortamlarında geliştirilmiş embriyoların olası implantasyon potansiyellerini incelemek. Gereç ve yöntem: Akdeniz Üniversitesi Tıp Fakültesi Üremeye Yardımcı Tedavi Merkezi’nde, Aralık 2014 ve Aralık 2015 tarihleri arasında, in vitro fertilizasyon tedavisi uygulanmış hastalara ait bilgiler elektronik veri tabanı kullanılarak incelendi. Ardışık basamaklı kültür medyumu uygulanan hastaların (n=144) belirlenmesinin ardından, her hasta için indeks olguya en yakın dönemde oosit toplama işlemi gerçekleştirilmiş; yaş ve metafaz-2 oosit sayıları eşleşen ancak tek basamaklı embriyo kültür medyumunun kullanıldığı hastalar (n=144) seçildi. Embriyo kaliteleri ve gebelik sonuçları gruplar arasında karşılaştırıldı. Bulgular:Gruplar arasında fertilize oosit, gelişimin 2-3. gününde yüksek kalite embriyo sayıları ile fertilize oositten yüksek kalitede klivaj embriyo gelişiminde istatistiksel olarak anlamlı fark saptanmadı (p>0.05). Ancak, hasta başına yüksek kalitede blastokist sayısının (p=0.048) ve yüksek kaliteli klivaj evresindeki embriyolardan yine yüksek kaliteli blastokist gelişim oranının (p=0.043) tek basamaklı kültür ortamında daha yüksek olduğu bulundu. Benzer şekilde, blastokist transferi yapılan olgular istatistiksel olarak tek basamaklı kültür grubunda daha fazla idi (p˂0.001). Gebelik sonuçları değerlendirildiğinde, embriyo transferi yapılan hastalarda klinik gebelik (p=0.038) oranı tek basamaklı kültür ortamında daha yüksek bulundu. Ancak implantasyon oranında gruplar arasında istatistiksel olarak anlamlı fark izlenmedi (p>0.05). Sonuç: Çalışmamız tek basamaklı medyum uygulamalarıyla ardışık medyum ile benzer oranlarda yüksek kalitede embriyo gelişimi sağlanabileceğini göstermektedir. Ayrıca, blastokist gelişim oranı artarak olası gebelik sonuçlarında iyileşme sağlanabilir. Pam Tıp Derg 2016;9(3):212-218, Purpose: The aim of this study was to investigate the effect of sequential vs. single-step media on early human embryo development and the embryos’ implantation potential. Materials and methods:This retrospective study included patients who underwent in vitro fertilization (IVF) treatment at the Akdeniz University Reproductive Centre between December 2014 and December 2015. The electronical database was screened to identify the type of culture media used for IVF. A hundred forty four women whose embryos were cultured in sequential media were retrospectively matched for age and metaphase-2 oocytes with 144 women whose embryos were cultured in single-step media. Embryo quality and pregnancy outcomes were compared between sequential and single-step media groups. Results: The number of fertilized oocytes, high quality embryos on day 2-3 and developmental rate of high quality embryos from fertilized oocytes were found to be comparable between the groups (p>0.05). However, the number of high quality blastocysts (p=0.048) and developmental rate of high quality blastocysts from high quality cleavage stage embryos (p=0.043) were significantly higher in single-step media group. Subsequently, singlestep media group had more embryos transferred at blastocyst stage (p˂0.001). Dealing with the pregnancy outcomes, clinical pregnancy per embryo transfer was found to be higher in single-step media group (p=0.038). There was no significant difference in implantation rates between the groups (p>0.05). Conclusion: We have shown that a single step culture media supports high quality embryo development equivalently to established sequential media. Moreover, higher number of blastocysts available for transfer with single step media, might result in a concurrent improve in pregnancy rates. Pam Med J 2016;9(3):212-218

Aim: To evaluate the effect of instant urinary catheterization on pregnancy rates in women who are on bed rest after embryo transfer (ET). Methods: This retrospective study included patients who underwent intracytoplasmic sperm injection (ICSI) treatment at the Akdeniz University Fertility Clinic between January 2015 and March 2016. All patients were advised bed rest for an hour after ET. The clinic's electronic database was screened to identify the uterine anatomy of the patients, transferred embryo quality, and urinary catheterization status. Only women with an anteverted and anteflexed uterus and cycles that resulted in high-quality blastocyst transfers were included in this analysis. Finally, 71 cycles with urinary catheterization and 81 cycles with no catheterization were analyzed. Results: The clinical (53.5 vs. 40.7%, p = 0.115) and ongoing pregnancy (43.7 vs. 35.8%, p = 0.323) rates were comparable between urinary catheterization and non-catheterization groups. Multivariate analysis by logistic regression including confounding factors revealed that urinary catheterization was independently associated with clinical pregnancy (OR 2.06, 95% CI 1.03-4.13, p = 0.041). However, no significant association was detected between catheterization and the clinically more relevant, ongoing pregnancy (OR 1.65, 95% CI 0.82-3.30, p = 0.160). Conclusion: Our preliminary findings on the use of instant urinary catheterization after ET are promising and justify further investigation in larger prospective studies.

Ozet : Gonadotropin serbestlestirici hormon (GnRH) agonisti ile oosit maturasyonunun tetiklendigi ve sonrasinda tum embriyolarin donduruldugu 27 yasindaki olguda luteal destek olmaksizin olusan spontan klinik gebelik sunulmustur. Polikistik over sendromu (PKOS) tanisiyla, oncesinde 3 sefer kontrollu ovaryan stimulasyon ve intrauterin inseminasyon uygulanmis olan hastaya GnRH antagonist protokolu ile intrasitoplasmik sperm enjeksiyonu tedavisi planlandi. Stimulasyonun 11. gununde ≥11 mm 25 adet folikul saptanmasi uzerine GnRH agonisti uygulanarak final oosit maturasyonu saglandi. Blastokist evresine kadar gelisim gosteren yuksek kalitedeki embriyolarin tumu vitrifiye edildi. Taze embriyo transferi planlanmamasi dolayisiyla hastaya luteal faz destegi verilmeyerek yedi gun sureyle cinsel iliski onerilmedi. Ancak, oosit toplanmasinin 5 hafta sonrasinda amenore yakinmasiyla tarafimiza tekrar basvuran hastaya yapilan transvajinal ultrasonografide fetal kalp atimi muspet spontan tekiz gebelik izlendi. Gunumuzde, GnRH agonistlerle tetikleme sonrasi luteal fazin nasil desteklenmesi gerektigine dair yogun tartismalar devam etmektedir. Bu bulgular, ovaryan stimulasyon sonrasi suprafizyolojik steroid seviyelerine ragmen, her hastanin korpus luteum fonksiyonlarinin devaminin farklilik gosterebilecegine isaret etmektedir. Dolayisiyla, PKOS ve beraberinde endojen luteinizan hormon aktivitesinin yuksek oldugu hastalar, korpus luteumun sinirli ancak luteal faz icin yeterli aktivitesinin devam edebilecegi bir alt grubu olusturuyor olabilir. Anahtar kelimeler: GnRH agonist tetikleme, luteal faz destegi, spontan gebelik SPONTANEOUS CLINICAL PREGNANCY FOLLOWING GNRH AGONIST TRIGGERING AND FREEZE ALL POLICY Abstract: We describe a 27 year-old patient undergoing follicle stimulation with gonadotrophin releasing hormone (GnRH) agonist and freeze-all approach, presented with spontaneous clinical pregnancy without supplementation of luteal phase. The patient had three attempts with controlled ovarian hyperstimulation and intrauterine insemination due to polycystic ovary syndrome (PCOS) before referring to our clinic for intracytoplasmic sperm injection. A GnRH antagonist protocol was assigned and ovarian stimulation commenced. After the detection of 25 follicles ≥11 mm, final oocyte maturation was triggered with GnRH agonist on the eleventh day of stimulation. Since freeze all approach was taken, no luteal phase support was administered. High quality blastocysts were vitrified. The couple was advised to refrain from sexual intercourse for seven days. The patient, however, presented with amenorrhea five weeks after the day of oocyte retrieval. A clinical pregnancy with fetal cardiac activity was determined on ultrasound examination. Currently, there is an ongoing debate on how best to counteract the negative impact of agonist triggering on the luteal phase. Our case demonstrates that corpus luteum functions might diverse despite supra-physiological sex steroids resulting from ovarian stimulation. Therefore, in case of PCOS consisting high endogenous LH activity, corpus luteum might be rescued and luteolysis might limit itself in such cycles shortly after implantation. Keywords: GnRH agonist trigger, luteal phase support, spontaneous conception

OZET: Hucre siklusunun farkli asamalarinda oosit maturasyon aresti ile karsilasilabilir. Uremeye yardimci tedavi uygulamalarinda tum oositlerin mayozu tamamlayamamalari nadir olarak karsilasilan bir durumdur. Sundugumuz olguda, onceki tedavisinde matur oosit elde edemedigimiz bir vakada, GnRH agonisti (Triptorelin asetat 0.2 mg) ve standart doz hCG’yi (hCG 5000 IU) beraber uygulanarak matur oosit elde etmeye calistik. Buna karsin, “ikili tetikleme” ile yine oosit maturasyonunun metafaz-1 asamasinda duraksadigini saptadik. Her ne kadar olgumuzda “ikili tetikleme” stratejisi etkin olmamissa da, oosit maturasyon aresti farkli nedenlere bagli olabileceginden sonuclarimizin genellenmemesi daha uygun olabilir. Bu hususta, oosit maturasyon aresti ile karsilasilan her olgunun literaturde yer almasinin, yontemler basarisiz olsa dahi, uzun vadede problemin cozumune yonelik katki saglayabilecegini dusunmekteyiz. ANAHTAR KELIMELER: oosit maturasyon aresti, gonadotropin serbestlestirici hormon agonisti, ikili tetikleme DUAL TRIGGERING (GNRH AGONIST AND HCG) IN REPEATED TOTAL OOCYTE MATURATION ARREST ABSTRACT: Maturation arrest of human oocytes might occur at various stages of the cell cycle. A total failure of human oocytes to complete meiosis is rarely observed during assisted conception cycles. We describe here an infertile couple for whom all oocytes failed to mature during their previous in vitro fertilization treatment. Even with the use of dual triggering concept (Triptorelin asetat 0.2 mg plus hCG 5000 IU) oocyte maturation was again arrested at metaphase 1 stage in their next cycle. Since failure to resume meiosis might arise from different etiologies, we cannot conclude that “dual trigger” is not an alternative treatment option for the patients with history of recurrent oocyte maturation arrest. However, we do believe that our observation will guide future researchers. KEYWORDS: oocyte maturation arrest, gonadotrophin releasing hormone agonist, dual trigger

Dichlorvos is an organophosphorus insecticide that is used worldwide for pest control in agriculture and household use. Vitamins C and E are potential antioxidants protecting cells from oxidative stress. Vitamin C+ vitamin E, dichlorvos, a combination of vitamin C+ vitamin E + dichlorvos, or corn oil (control) were given to rats via oral gavage for 7 weeks. Body and testis weights, sperm parameters, hormone levels, histo- and cytopathological changes in testes were investigated at the end of 24 h and the 4th and 7th weeks comparatively with the control group. Body and testis weights, sperm morphology, FSH, LH, and testosterone levels were decreased significantly at the end of 4th and 7th weeks in the dichlorvosand vitamins + dichlorvos-treated groups. A statistically significant decline in sperm motility and testosterone levels occurred by the end of 7th week in the dichlorvos- and vitamins + dichlorvos-treated groups. Light and electron microscopy revealed necrosis, edema and cellular damage in testicular tissues of the dichlorvos- and vitamins + dichlorvos-treated rats at the end of 4th and 7th weeks. In conclusion, dichlorvos caused subacute and subchronic reproductive toxicity, but vitamins did not confer protection. (C) 2011 Elsevier GmbH. All rights reserved.

Magnetic cell sorting allows large numbers of viable cells to be isolated with high purity and yield. It is accepted that the process does not cause physical damage to the cell. Today, magnetic nanoparticles are widely used in separation of macromolecules or cells from various cell suspensions. Studies have shown that nonapoptotic sperm fractions selected by MACS technique have morphologically superior quality, higher percentage of motility, viability, improved apoptosis indices and routine sperm parameters. Reduction of apoptotic spermatozoa within the ejaculate by means of the MACS system results in a distinct reduction of spermatozoa with DNA fragmentation. The initial clinical trials of MACS of human spermatozoa in assisted reproduction showed a statistically significant improvement in the embryo cleavage rates.

Magnetic activated cell sorting (MACS) eliminates apoptotic spermatozoa based on the presence of externalized phosphatidylserine residues. We evaluated the outcome of male fertility treatment when intracytoplasmic sperm injection (ICSI) into human oocytes was performed with non-apoptotic MACS-selected spermatozoa.196 couples were treated by ICSI following spermatozoa preparation by MACS (study group; 122 couples) or density gradient centrifugation (DGC) (control group; 74 couples). Fertilization, cleavage, pregnancy, and implantation rates were analyzed.The percentage of sperm with normal morphology after MACS selection was improved. Cleavage and pregnancy rates were higher, respectively, in the study group than in control. A slightly higher implantation rate was also observed in the study group.MACS selection of human spermatozoa increased cleavage and pregnancy rates in oligoasthenozoospermic ART cases. This novel method for selecting non-apoptotic spermatozoa for ICSI is safe and reliable, and may improve the assisted reproduction outcome.