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Öğe A histopathological evaluation of ovarian hyperstimulation syndrome on reproductive and vital organs and the role of the VEGF-PKA pathway in a mouse model(Karger, 2021) Karabulut, Seda; Korkmaz, Oya; Erdem Altun, Ceren; Keskin, İlknurOvarian hyperstimulation syndrome (OHSS) is one of the most common and iatrogenic complications of in vitro fertilization therapy, which is an exaggerated response to excess hormones resulting in the development of a large number of maturing follicles. Although the complications of and reasons for the condition are well known, the overall histopathological effects on systemic organs and the extent of the damage have not been fully elucidated. Besides, the mechanism that underlies the situation is not very well known. The aim of the present work was to analyse the histopathological effects of OHSS on reproductive (uterus and ovary) and vital organs (liver and kidney) and the possible role of the VEGF-PKA pathway in triggering the condition. Balb/c mice were used to establish an OHSS model. The OHSS group were injected with overdose PMSG while the normal responder group were injected with an optimal dose. Histopathological evaluation was utilised in the liver, kidney, ovary, and uterus stained with hematoxylin and eosin, Masson's trichrome, and periodic acid-Schiff stain. The expression profiles of VEGF (vascular endothelial growth factor), PKA (protein kinase A), and p-PKA (an activated form of PKA) were detected with immunohistochemistry and Western blotting. OHSS was demonstrated to have a negative histopathological effect on all of the organs analysed. These effects were associated with an overall increase in the expression levels of VEGF, PKA, and p-PKA. OHSS has a serious histopathological negative effect on the systemic and reproductive organs and is proven to affect overall health, and thus should be considered a dangerous complication during ART techniques. The activation of the VEGF-PKA pathway, which is indicated by the expression levels of VEGF, PKA, and p-PKA, is demonstrated to accompany this complication, which should be further elucidated to understand the mechanisms underlying the condition.Öğe Administration of autologous platelet rich plasma and the impact on outcomes of assisted reproduction treatment in infertile women(IMR Press, 2021) Kutlu, Pelin; Karabulut, Seda; Korkmaz, OyaObjective: There are diverse findings concerning the use of platelet-rich plasma (PRP) in assisted reproduction treatment (ART) cycles of infertile women with low ovarian reserve, ovarian failure or implantation failure due to endometrial problems. A debate continues regarding the benefit of this technique because of the low number of evidence-based studies. Mechanism: Different PRP preparation methods have been described in the literature, aiming to obtain the highest number of platelets following centrifugation steps. Findings in brief : Research on the use of PRP in female infertility aims to improve the deteriorated hormonal profile, gamete production and implantation of the embryo into the endometrium. Conclusions: This paper reviewed literature evaluating the impact of PRP on the outcomes of subsequent ART cycles in infertile women. PRP is a safe and easy-to-apply procedure and can be used as an 'add-on' therapy in patients with reduced ovarian reserve, ovarian failure or implantation failure prior to the in vitro fertilization (IVF) cycle, although it is still regarded as an empirical treatment method. Further studies should be conducted to enlighten the subject.Öğe Anticancer properties of phenylboronic acid in androgen-dependent (lncap) and androgen-independent (pc3) prostate cancer cells via map kinases by 2d and 3d culture methods(2025) Gürsoy Gürgen, Duygu; Güneş, Arzu; Köse, Oğuzhan; Kaplan, Arife Ahsen; Karabulut, Seda; Tunalı, Muzaffer Başak; Keskin, İlknurObjective: This study utilized three cell lines: normal prostate epithelial RWPE-1, androgen-dependent LNCaP, and androgen-independent PC3. We investigated the inhibitory effects of phenylboronic acid (PBA)’s inhibitory effect on cellular proliferation due to its ability to disrupt microtubule formation in prostate cancer cell lines. Additionally, this study aimed to assess the cytotoxic effects of PBA on prostate cancer cells using two-dimensional (2D) and three-dimensional (3D) cell culture models. Methods: The IC50 values of PBA and colchicine were determined through viability assays in 2D and 3D models. Colony formation, proliferation, and migration assays were conducted. Immunofluorescence intensity analysis of MAPKKK proteins (ERK, JNK, p38) was performed to explore the mechanism of cellular response to PBA. Results: The IC50 values were determined for each treatment group. After 48-hour of PBA treatment, migration was inhibited more effectively than with colchicine in both cancer cell lines. After 24-hour, PBA reduced colony formation and proliferation. PBA treatment for 24-hour decreased JNK expression in PC3 and LNCaP cells in 2D models. Both PBA and colchicine increased p38 expression in PC3 spheroids. PBA’s effects on cell deformation were visualized in semi-thin sections, marking the first ultrastructural observation of PBA-induced morphological defects in cancer cells. Conclusion: PBA exerts antimitotic effects by inhibiting proliferation and migration and triggers diverse metabolic responses across different cell lines. Furthermore, PBA’s low toxicity on RWPE-1 cells suggests its potential as a promising chemotherapeutic agent for future studies.Öğe Artificial oocyte activation with calcium ionophore for frozen sperm cycles(Taylor and Francis Ltd., 2018) Karabulut, Seda; Aksünger, Özlem; Ata, Can; Sağıroğlu, Yusuf; Keskin, İlknurFertilization problems are the major problems that may be faced in 30-55% of the patients during an intracytoplasmic sperm injection (ICSI) cycle. A successful oocyte activation depends on factors related to both sperm and oocyte, and one of the important factors that mediates the process is Ca2+ concentration within the oocyte. Artificial oocyte activation (AOA) is a method used for fertilization problems that commonly involve the usage of Ca2+ ionophores and is usually used in problems such as total fertilization failure (TFF) and globozoospermia. The aim of the present study was to investigate possible effects of AOA for different groups of patients with fertilization failure. Four groups of patients (previous TFF, low oocyte number, severe sperm quality, and frozen sperm (FS) group) that underwent ICSI with AOA were included in the study. All groups had similar control groups with same indications except TFF, where AOA was not performed. Fertilization rates were significantly higher in the TFF group than those observed in other AOA groups. Fertilization rates and quality of embryos observed in the remaining AOA groups were higher than those of the controls, which were statistically insignificant. Prgenancy rates were higher in all AOA groups compared to the controls, although the differences were significant in FS group only. Quality of embryos and pregnancy rates were lower in the TFF group compared to the remaining AOA groups indicating possible concomitant problems. Fertilization rates, quality of embryos and pregnancy rates seemed to be increased in all indication groups suggesting that not only TFF patients but also a wide variety of patients with different indications may benefit from AOA.Abbreviations: ICSI: Intracytoplasmic sperm injection; ARTs: Assisted reproductive techniques; Ca: Calcium; AOA: Artificial oocyte activation; TFF: Total fertilization failures; OAT: Oligoasthenoteratozoospemia; IVF: In vitro fertilization; SOAT: Severe OAT; LON: Low ooctye number; FS: Frozen sperm; hCG: human chorionic gonadotrophin; PVP: polyvinylpyrrolidone; HSA: human serum albuminÖğe Assessing the impact of the novel sperm selection technique 'annexin-v coated polystyrene bead technique' on mouse assisted reproductive techniques outcomes: preliminary findings(2024) Karabulut, Seda; Yalım Camcı, İrem; Erdem Altun, Ceren; Usta, Melek; Yiğit, PakizeICSI is one of the most commonly used techniques to treat infertility. The sperm selection for the procedure is done ‘randomly’ by the embryologist according to the motility and morphology parameters which is known not to reflect the potential of a sperm for fertilization, pregnancy and a healthy childbearing. Since the apoptosis rate is higher in sperm cells of infertile patients, it is more likely to choose an apoptotic sperm by the 'random selection method'. We recently introduced a novel sperm selection technique namely ‘Annexin-V coated polystrene bead technique’(APB-Tech), for the selection of non-apoptotic sperm cells. The principal of the technique is based on the binding affinity of an apoptotic sperm to ‘Annexin-V covered beads’ enabling to distinguish a viable and a healthy sperm by light microscopy. The aim of the present study was to observe the effects of this technique on ICSI outcomes in mice. Sibling-oocyte trial was conducted and the outcome measures were compared with the results of traditional sperm selection method. Embryo and blastocyst qualities and blastocyst development rates were significantly increased in APB-Tech group, while the other parameters were not affected. Promising results obtained from the technique reflect its promising potential as a new and powerful tool for sperm selection and thus infertility techniques.Öğe Chronic unpredictable stress disturbs the blood-testis barrier affecting sperm parameters in mice(Elsevier Ltd, 2021) Kolbaşı, Bircan; Bülbül, Muhammet Volkan; Karabulut, Seda; Erdem Altun, Ceren; Çakıcı, Çağrı; Ülfer, Gözde; Müdok, Tangül; Keskin, İlknurResearch question: Does chronic stress affect the key proteins and sperm parameters of the blood & ndash;testis barrier (BTB)? Design: C57Bl/6 mice were divided into two groups: a non-treated control group and a chronic unpredictable stress (CUS) applied group. The stress status of the animals was confirmed with behavioural tests. Histopathologic evaluation was conducted by haematoxylin and eosin staining and electron microscope. Malondialdehyde, corticosterone and testosterone levels were evaluated in peripheral blood. Expression levels of BTB proteins, namely zonula occludens-1 (ZO-1), claudin-11 (CLDN11) and clathrin in Sertoli cells, were assessed by Western blotting and immunofluorescence techniques. Sperm samples were collected from cauda epididymis, and sperm parameters analysed. Results: The stress model was confirmed by behavioural tests. Histopathological evaluation of the testes demonstrated a mild degeneration in seminiferous tubules. Malondialdehyde (P = 0.008) and corticosterone levels increased (P = 0.004) and testosterone levels decreased (P = 0.005) in the CUS group. Electron microscopic evaluation confirmed the damage in BTB integrity in the CUS group. Western blot analysis showed that ZO-1 and CLDN11 levels were significantly decreased, although clathrin levels were unchanged. Although sperm concentration and total motility rate were not significantly different between the groups, progressive motility (P = 0.03), normal sperm morphology (P = 0.04), chromatin integrity (toluidine blue) (P = 0.002) and the acrosomal reaction rate (P = 0.002) were significantly decreased, and acrosomal abnormality rate was dramatically increased (P = 0.04) in the CUS group. Conclusions: In mice, CUS disrupted BTB integrity and impaired sperm parameters. A decrease in ZO-1 and CLDN11 expression levels may be proposed as the causative factor.Öğe Comparison of machine learning classification techniques to predict implantation success in an IVF treatment cycle(Elsevier Science Ltd, 2022) Yiğit, Pakize; Bener, Abdulbari; Karabulut, SedaResearch question: Which machine learning model predicts the implantation outcome better in an IVF cycle? What is the importance of each variable in predicting the implantation outcome in an IVF cycle?Design: Retrospective cohort study comprising 939 transferred embryos between 2014 and 2018 in an IVF centre in Turkey with 17 selected features. The algorithms were Logistic Regression (LR), Decision Tree (DT), Naive Bayes (NB), Random Forest (RF), Support Vector Machine (SVM), Neural Network (Nnet), Gradient Boost Decision Tree (GBDT), eXtreme Gradient Boosting (XGBoost) and Super Learner (SL). The results were evaluated with performance metrics (F1 score, specificity, accuracy and area under the receiver operating characteristic curve [AUROC]) with 10-fold cross-validation repeated ten times.Results: RF and SL models achieved the highest performance and showed F1 scores of 74% and 73%, specificity of 94%, an accuracy of 89%, and AUROC of 83%. In addition, the model identified the top features as maternal age, embryo transfer day, total gonadotrophin dose and oestradiol concentration.Conclusions: The present study revealed that machine learning algorithms successfully predicted implantation rates in an IVF attempt. In addition, maternal age is by far the most important predictor of IVF success when compared with other variables.Öğe Detailed morphological analysis of axolotl sperm(Korean Society of Veterinary Science, 2021) Keskin, İlknur; Gürsoy Gürgen, Duygu; Avinca, Didem; Özdemir, Ekrem Musa; Keskin, Suat Utku; Karabulut, SedaThe axolotl has extraordinary regeneration capacity compared to other vertebrates. This remarkable potential has been attributed to its life-long neoteny, characterized by the exhibition of embryonic characteristics at the adult stage. A recent study provided a detailed morphological analysis of the sperm morphology of the Ambystoma mexicanum using routine and detailed histological techniques. The primary purpose of the present study is to describe a simple and inexpensive method for evaluating the morphology of axolotl sperm. In this study, spermatophore structures were collected and spread on slides and air-dried. The slides were stained with periodic acid Schiff, toluidine blue, Masson's trichrome, Giemsa, Spermac, and Diff-Quik dye for a morphological examination. The slides were coated with gold/palladium for a scanning electron microscopy examination. The sperm of the axolotl consisted of an elongated head, a neck, and a flagellum covered with an undulating membrane. The lengths of the midpiece, tail, and head were 8.575 µm, 356.544 µm, and 103.661 µm, respectively. In the flagellum part, the wavy membrane structure, whose function has not been explained, surrounds the tail. The data obtained from this study will constitute an important step in designing future research on the reproductive and regeneration capacity of the axolotl.Öğe Does a microfluidic chip for sperm sorting have a positive add-on effect on laboratory and clinical outcomes of intracytoplasmic sperm injection cycles? A sibling oocyte study(Wiley, 2019) Yalçınkaya Kalyan, Ender; Can Çelik, Seren; Okan, Özlem; Akdeniz, Gülden; Karabulut, Seda; Çalışkan, ErayThe most recent technologies for sperm sorting involve microfluidics. However, the most important question whether their use is of any advantage in terms of laboratory and clinical IVF/ICSI outcomes still remains controversy. In this study, we aimed to investigate whether a microfluidic sperm sorting device (Fertile Plus (R)) has a positive add-on effect on laboratory and clinical outcomes. Sibling oocytes of 81 patients were assigned to two sperm sorting groups including swim up and Fertile Plus (R). All embryos were cultured until day 5/6. Fertilisation, embryo quality and blastocyst development were assessed as primary outcomes among 81 patients; clinical pregnancy, implantation and live birth rates were analysed as secondary outcomes as a subgroup analysis due to transfer cancellations. No statistically significant differences were found between groups in terms of all outcomes analysed in laboratory and clinical terms (p > .05 for all). The results of this study suggest that sorting spermatozoa through Fertile chip does not improve laboratory outcomes significantly and does not seem to have a positive contribution to clinical outcomes.Öğe Dondurulmuş ejekülat ve dondurulmuş testiküler spermin sonuçlarının intrasitoplazmik sperm enjeksiyonu (ICSI) parametrelerine etkisinin değerlendirilmesi(Avrasya Üroonkoloji Derneği, 2019) Karabulut, Seda; Korkmaz, Oya; Sağıroğlu, Yusuf; Keskin, İlknurAmaç: Çalışmamızda, dondurulmuş ejekülat ve dondurulmuş testiküler spermin İntrasitoplaz - mik Sperm Enjeksiyonu (ICSI) sonuçları üzerine etkisinin araştırılması amaçlanmıştır. Gereç ve Yöntemler: Çalışmamızda 11.10.2016- 12.09.2017 tarihleri arasında infertilite nedeniyle ICSI tedavisine alınan ve spermleri dondurulan 36 çift çalışmaya alınmıştır. Hastaların 14 tanesine ejekülat sperm kriyoprezervasyonu, 22 tanesine de micro testiküler sperm ekstraksiyonu (TESE) sperm kriyoprezervasyonu gerçekleştirilmiştir. İki grubun ICSI sonuçları (fertilizasyon oranı, embriyo gelişme oranı ve gebelik oranları) karşılaştırılmıştır. Bulgular: Sperm parametreleri değerlendirildi- ğinde ejekülat ve TESE spermlerinde dondurma ön- cesi ve dondurma sonrası toplam motilite ve canlılık oranları arasında istatistiksel bir fark belirlenme - miştir (>0.05). Fertilizasyon oranları, implantasyon oranları ve embriyo gelişme oranları arasında ista - tistiksel olarak anlamlı bir fark gözlemlenmemiştir (>0.05). Gebelik oranları ise dondurulmuş ejekülat grubunda dondurulmuş testiküler sperm grubuna oranla istatistiksel olarak anlamlı derecede yüksek bulunmuştur (<0.05). Sonuç: Sperm kriyoprezervasyonun uzun yıl- lardır ICSI tedavilerinde kullanılmasına rağmen yöntemin etkinliği üzerine araştırmalar devam etmektedir. Çalışmamızın bulguları doğrultusun - da spermin ICSI sürecini olumsuz ekilememesine rağmen gebelik oranlarında anlamlı düşüşe neden olduğu sonucuna varılmıştır. Bunun, implantasyon sürecinde spermin oynadığı rol nedeniyle olabilece- ği düşünülmektedir. Bu etkinin ortaya koyulabilme- si ve implantasyon sürecinde görev alan mekaniz - maların aydınlatılabilmesi için, daha büyük hasta gruplarıyla yapılmış moleküler seviyede çalışmalara ihtiyaç duyulmaktadır.Öğe Effects o follicular fluid oxidative status on human mural granulosa cells, oocyte competency and ICSI parameters(Elsevier, 2020) Karabulut, Seda; Korkmaz, Oya; Kutlu, Pelin; Eren Gözel, Hilal; Keskin, İlknurPurpose: 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-beta, Notch1), DNA status and chromatin integrity of mural granulosa cells were analyzed.Results: TAS levels were positively correlated with the Hsp70 and Tgf-beta 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.Öğe Effects of density gradient sperm preparation on semen parameters and acrosomal status(Haydarpaşa Sample Training and Research Hospital, 2017) Keskin, İlknur; Karabulut, SedaAbstractIntroduction: The aim of this study was to observe changes in sperm parameters after density gradient sperm preparation. These data will help to decide upon preparation technique and to predict the outcome of semen parameters after using this technique.Methods: A total of 1402 men who were screened for infertility at the Florence Nightingale Hospital in vitro fertilization clinic between April 2011 and October 2016 were analyzed. Semen analysis was performed according to World Health Organization 2010 guidelines before and after semen preparation. Samples with <1 mil/mL sperm concentration were prepared using the density gradient centrifugation technique and the semen parameters (sperm concentration, total motility, progressive motility, normal morphology rate, and normal acrosomal status rate) were analyzed.Results: Although sperm concentration was reduced, the total motility, progressive motility, and normal morphology rate were higher after density gradient semen preparation. Sperm concentration decreased by 37.49% after semen preparation (p=0.01). Total motility rate increased significantly, by 21.47% (66.1% before and 87.57% after semen preparation), progressive motility rate increased by 28.12% (11.45% before and 39.57% after semen preparation), normal morphology rate increased by 5% (3% before and 8% after semen preparation), and normal acrosomal status rate increased by 14% (62% before and 76%) after semen preparation (p<0.01). Discussion and Conclusion: According to the results of this study, the sperm parameters of total motility, progressive sperm motility, normal morphology, and normal acrosomal status rate increased, but sperm concentration decreased after density gradient sperm preparation. These data will help the specialist to decide upon the sperm preparation technique to be used in assisted reproduction and to predict the outcome of semen parameters after using this technique.Öğe Effects of gallic acid on endometrial cancer cells in two and three dimensional cell culture models(Asian Pacific Organization for Cancer Prevention, 2021) Bülbül, Muhammet Volkan; Karabulut, Seda; Kalender, Mervenur; Keskin, İlknurBackground and Aim: Cell culture studies are an indispensable tools used to understand basic physiological, biophysical and biomolecular mechanisms. Although traditional two-dimensional (2D) cell culture models are more preferred in experimental studies, three-dimensional (3D) cell culture models, attract more attention due to several advantages including mimicking tumor physiology, biochemistry and biomechanics. We aimed to investigate the effects of Gallic Acid, an antimutagenic, antioxidant and anticarcinogenic agent, on both 2D and 3D cultured endometrial cancer cells for the first time. Methods: IC50 values were determined in 2D and 3D cultured endometrial cancer cells exposed to different doses of GA. In the 2D culture model exposed to GA, Caspase 3 expression levels were analyzed. In addition, the effect of GA on the migration of 2D cultured endometrium cancer cells was investigated. Results: IC50 value in the 3D model was found significantly higher than the 2D model. In 2D culture model, Caspase 3 expression and apoptosis was increased significantly in cells of GA exposed group compared to the control group. GA did not have a significant effect on the migration profile of cells. Conclusion: Gallic Acid is shown to induce apoptosis in Ishikawa cells via Caspase 3 activation. We demonstrated a significantly higher IC50 level in 3D model which provide evidence to prefer 3D models in drug-test trials. The data obtained in the current study will provide a basis for further experiments to analyze the effects of GA on endometrial cancer and to develop strategies for clinical treatment.Öğe Effects of human sperm cryopreservation on apoptotic markers in normozoospermic and non-normozoospermic patients(Cambridge University Press, 2018) Karabulut, Seda; Demiroğlu Zergeroğlu, Asuman; Yılmaz, Elif; Kutlu, Pelin; Keskin, IlknurThe 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.Öğe Erkek infertilitesinde oksidatif stresin CAMP aracılı PKA yolağı proteinlerine ve ıcsı parametrelerine etkisinin araştırılması(İstanbul Medipol Üniversitesi Sağlık Bilimleri Enstitüsü, 2020) Erdem Altun, Ceren; Karabulut, Sedaİnfertilite, çiftlerin en az 1 yıl süre ile korunmasız ve düzenli cinsel ilişkiye rağmen gebelik elde edilememesi olarak tanımlanmaktadır (DSÖ, 2010). Toplumun ~%15'inde görülen infertilite sorunu ile karşılaşan çiftler yardımla üreme tekniklerine başvurmakta ve ancak bu yolla gebelik sağlanabilmektedir. Oksidatif stresin, birçok hastalığın etiyolojisinde rol oynadığı gibi infertilitede de etkisi olduğu bilinmektedir. Sperm hücrelerinin fertilizasyon sürecinde (hiperaktivasyon, kapasitasyon) rol oynayan yolaklardan biri de cAMP bağımlı PKA yolağıdır. Çalışmamızda, seminal plazmada belirlenen oksidatif stres parametrelerinin sperm hücreleri üzerine hücresel, genetik ve fonksiyonel etkilerini ve bu spermler kullanılarak yapılan bir tüp bebek (ICSI) tedavisinde, tedavinin başarı ölçütlerine etkileri araştırılmıştır. Çalışmamıza, tüp bebek merkezine başvuran 40 çiftin erkek partnerlerinin semen parametreleri, oksidatif stres değerleri (TOS, TAS, MDA), DNA fragmentasyonu ve kromatin bütünlüğü değerleri, cAMP düzeyleri, PKA ve p-PKA proteinlerinin ekspresyon seviyeleri ve hücresel lokasyonları ve bu spermler kullanılarak yapılan bir ICSI tedavisi sonrası ICSI parametreleri araştırılmıştır. Oksidatif stresin ölçütlerinden biri olan MDA'nın, cAMP bağımlı PKA yolağı ile (p= 0,008) ve kromatin yapısı (p=0,607) ve DNA Fragmentasyonu (p=0,037) ile ilişkili olduğu belirlenmiştir. TAS düzeyi ile sperm motilitesi (p=0,047) ve blastosist gelişim oranı (p=0,023) arasında, TOS düzeyi ile de spermin progresif motilite oranı (p=0,053) arasında anlamlı ilişki bulunmuştur. PKA seviyesi ile progresif sperm motilitesi arasında anlamlı bir ilişki belirlenmiş (p=0,056), bu proteinin aktive formu olan p-PKA ile de fertilizasyon oranı arasında ilişkili belirlenmiştir (p=0,023). Bulgularımız doğrultusunda semendeki oksidatif stresin, cAMP aracılı PKA yolağını aktive ettiği, bu yolağın sperm DNA'sı ve kromatin yapısını bozduğu, ve sperm motilitesi ve ICSI sonrası fertilizasyon oranlarını düşürdüğü sonucuna ulaşılmıştır. Konu ile ilgili yapılacak kapsamlı çalışmalara ihtiyaç vardır.Öğe Erkek yaşı sperm parametrelerini etkiliyor mu? Türk popülasyonuna ait ilk seri(2020) Yıldırım, Berna; Keskin, İlknur; Sağıroğlu, Yusuf; Karabulut, SedaAMAÇ: Çalışmanın amacı, infertilite araştırma amacıyla elde edilmiş 1402 semen örneğinde, baba yaşı ile semen parametreleri (konsantrasyon, motilite, progresif motilite, morfoloji, akrozomal indeks, semen hacmi) arasındaki ilişkiyi araştırmak ve baba yaşının infertilite üzerine olası etkisinin araştırılmasına katkı sağlamaktır. GEREÇ ve YÖNTEMLER: Çalışma, Ocak 2017-Mayıs 2019 tarihleri arasında, İstanbul Florence Nightingale Hastanesi, Tüp Bebek Merkezi’ne başvuran ve semen analizi yapılan 1402 hastayı kapsamaktadır. Hastaların semen örnekleri WHO kriterlerine göre değerlendirilmiştir. Örnekler 40 yaş altı ve 40 yaş üstü olacak şekilde iki gruba ayrılmıştır. İki grupta sperm konsantrasyonu (mil/mL), toplam motilite (%), progresif motilite (%), normal morfoloji (%) ve akrozomal indeks (%) parametreleri karşılaştırılmıştır. BULGULAR: İki grup birbiriyle karşılaştırıldığında toplam motilite, progresif motilite, morfoloji ve akrozom indeks parametreleri arasında istatistiksel olarak anlamlı bir fark bulunmamasına rağmen sperm konsantrasyonunun 40 yaş üstü grupta istatistiksel olarak anlamlı derecede yüksek olduğu gözlemlenmiştir (p<0,05). SONUÇ: İleri anne yaşının infertiliteyi etkilediği daha açık olmakla birlikte, ileri baba yaşının infertiliteyle olan ilişkisi günümüzde araştırılmaya devam edilmektedir. Bu çalışma, Türkiye’de bu konuda yapılmış en büyük örneklem kümesine sahip ilk çalışmadır. Bulgular, ileri yaşla birlikte, semen parametrelerinden sperm konsantrasyonunun erkek yaşı ile birlikte arttığını, diğer parametrelerin ise değişmediğini göstermektedir. İleri baba yaşının infertiliteyle ilişkisini anlamak için genetik ve biyokimyasal seviyede daha ileri araştırmalara ihtiyaç vardır.Öğe Factors related to follicular oxidative stress in intracytoplasmic sperm injection cycles and its effects on granulosa cells(Cambridge University Press, 2021) Karabulut, Seda; Korkmaz, Oya; Kutlu, Pelin; Keskin, İlknurThe 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.Öğe Fresh versus frozen blastocyst transfer outcomes deriven from the same ICSI cycle in male factor infertility(IMR Press Limited, 2021) Karabulut, Seda; Kutlu, Pelin; Korkmaz, Oya; Bülbül, Muhammet Volkan; Keskin, İlknurThe aim of this study was to investigate the clinical outcomes of fresh and frozen-thawed blastocyst transfer deriven from the same ICSI cycle because of male infertility. 2372 couples undergoing a total of 5075 ICSI cycles (2372 fresh + 2703 frozen embryo transfer) were enrolled in the study. Female factor infertility cases were excluded to prevent from the contribution of maternal factors that would influence the outcomes dramatically. Only the cycles for which day 5 blastocyst transfer was performed, had at least 1 surplus blastocyst available for freezing, and had at least one frozen embryo transfer (FET) cycle were included in the study. Clinical pregnancy rates and live birth rates were compared between fresh and FET cycles. No statistically significant difference was obtained in clinical pregnancy rates and live birth rates of fresh versus FET cycles of day 5 transfers, regardless of the number of embryos transferred. According to the results of the recent study, frozen embryo cycles appear to be as effective as fresh ICSI cycles provided that the patients' embryos can develop into blastocysts, which is proven by using the same patient as the control group regardless of the result of previous ICSI attempt but further research is required to test the efficacy in terms of cost-effectiveness and the duration for conception.Öğe İleri baba yaşının sperm parametreleri, oksidatif stres, DNA fragmantasyonu, kromatin yapı ve mTOR ekspresyonuna etkilerinin incelenmesi(İstanbul Medipol Üniversitesi, Sağlık Bilimleri Enstitüsü, 2022) Atambay, Can; Karabulut, SedaÇocuk sahibi olmanın getireceği maddi yükümlülükler nedeniyle günümüzde çoğu birey aile kurma ve çocuk sahibi olma planlarını daha ileri yaşa ertelemektedir. Bu çalışmada ileri baba yaşının sperm kalitesine ve fertiliteye olası etkilerinin ortaya konması amaçlanmıştır. Çalışmada 40 yaş ve üzeri (n=20), 40 yaş altı (n=20) olmak üzere sağlıklı bireylerden alınan toplam 40 adet semen örneği incelenmiştir. Alınan örneklerde gruplar arasında semen parametreleri, kromatin yapı bütünlüğü, DNA fragmantasyonu, oksidatif stres indeksi ve mTOR protein ekspresyonu seviyeleri karşılaştırılmıştır. İleri baba yaşındaki bireylerde toplam motilite ve ileri hareketli sperm oranları istatistiksel olarak anlamlı derecede daha düşük, kromatin hasarı ve DNA fragmantasyonunun ise istatistiksel olarak anlamlı derecede yüksek olduğu bulunmuştur. Sperm konsantrasyonu, toplam sperm sayısı ve hareketsiz sperm oranları, oksidatif stres indexi ve mTOR proteini ekspresyonu değerleri ise ileri baba yasındaki bireylerde daha yüksek olmasına rağmen, bu farklılıklar istatistiksel olarak anlamlılık düzeyinde bulunmamıştır. Araştırmanın sonucunda ileri baba yaşının DNA hasarına neden olduğu ve kromatin bütünlüğüne zarar verdiği ancak oksidatif stres ve mTOR sinyal yolağında anlamlı bir etki göstermediği tespit edilmiştir.Öğe Intracytoplasmic morphologically selected sperm injection, but for whom?(Cambridge University Press, 2019) Karabulut, Seda; Aksünger, Özlem; Korkmaz, Oya; Eren Gözel, Hilal; Keskin, İlknurIntracytoplasmic sperm injection (ICSI) is performed in cases of infertility by injecting a motile and morphologically normal sperm cell under a routine ×400 magnification at which is hard to distinguish morphologically healthy sperm. Recently, the use of high-powered differential interference contrast optics gave the opportunity to select a sperm under ultra-high magnification of ×10,160. The aim of the present study was to evaluate the efficacy of the intracytoplasmic morphologically selected sperm injection (IMSI) technique in different infertility populations undergoing ICSI. Main outcome measures of routine ICSI were compared with IMSI in three different groups of patients (1, non-selected; 2, male infertility; and 3, repeated implantation failure group). Results were analysed to evaluate the effects of the IMSI procedure and to find the most suitable group of patients who may benefit from the procedure. IMSI caused a significant increase in the fertilization and top quality embryo rates in the male infertility group and a significant increase in fertilization and pregnancy rates in the repeated implantation failure group, whereas no effect was observed in the non-selected group with patients of various indications. A positive effect of IMSI on the outcome of male factor infertility and repeated implantation failure patients was observed. Data observed confirmed that the application of IMSI was beneficial for a selected group of patients with male factor infertility and repeated implantation failure.
