The Effects of Glutathione on Clinically Essential Fertility Parameters in a Bleomycin Etoposide Cisplatin Chemotherapy Model.

Chemotherapeutic agents used in the treatment of testicular cancer cause damage to healthy tissues, including the testis. We investigated the effects of glutathione on sperm DNA integrity and testicular histomorphology in bleomycin etoposide cisplatin (BEP) treated rats. Twelve-week-old male rats of reproductive age (n = 24) were randomly divided into three groups, the (i) control group, (ii) BEP group, and (iii) BEP+ glutathione group. Weight gain increase and testes indices of the control group were found to be higher than that of the BEP group and BEP+ glutathione group. While the BEP treatment increased sperm DNA fragmentation and morphological abnormalities when compared to the control group, GSH treatment resulted in a marked decrease for both parameters. Moreover, BEP treatment significantly decreased serum testosterone levels and sperm counts in comparison to the control group, yet this reduction was recovered in the BEP+ glutathione treated group. Similarly, seminiferous tubule epithelial thicknesses and Johnsen scores in testicles were higher in the control and BEP+ glutathione groups than in the BEP-treated group. In conclusion, exogenous glutathione might prevent the deterioration of male reproductive functions by alleviating the detrimental effects of BEP treatment on sperm quality and testicular histomorphology.

Rescue IVM of Denuded GV- and MI-Stage Oocytes of Premenopausal Rats with Oncostatin M, Insulin-like Growth Factor I, and Growth Hormone.

Immature oocytes are retrieved and matured through in vitro maturation (IVM). Maturation, fertilization rates, and embryo development via IVM are all lower than those found in vitro fertilization (IVF) cycles. We investigated the effects of oncostatin M (OSM), insulin-like growth factor-1 (IGF-I), and growth hormone (GH) in rescue IVM. A total of 111 germinal vesicle (GV) and 17 metaphase I (MI) oocytes were obtained after conventional IVF from 28 female Wistar albino rats. Denuded immature oocytes were cultured in maturation medium supplemented with OSM, IGF-1, or GH. The quantities of metaphase II (MII) oocytes matured from the GV stage were 17 of 30 (56.6%), 15 of 28 (53.5%), 10 of 30 (33.3%), and 7 of 23 (30.3%), in control, OSM, IGF-I, and GH groups, respectively. Maturation rates in control and OSM groups were higher than those in IGF-I and GH groups (p = 0.001). The quantities of MII oocytes matured from MI stage were 7 of 7 (100%), 4 of 4 (100%), 1 of 1 (100%), and 1 of 5 (20%) in control, OSM, IGF-I, and GH groups, respectively. Maturation rates from MI to MII stages in control, OSM, and IGF-I groups were higher than those in the GH group (p = 0.004). Acceptable maturation rates are observed with OSM in rat oocytes in rescue IVM.

Early Compaction Might Be a Parameter to Determine Good Quality Embryos and Day of Embryo Transfer in Patients Undergoing Intracytoplasmic Sperm Injection.

INTRODUCTION: Compaction is the first event in embryo morphogenesis. Blastocyst transfer on day five or six has been widely performed in the last decade. We investigated the clinical value of early compaction on day three for evaluation of the transferred embryo quality and pregnancy. METHODS: Four hundred patients with female factor infertility and 776 fresh embryo transfers were included. Two groups were formed: Early compaction group had embryo transfer with at least one day-three embryo exhibiting early compaction. Transferred embryos without early compaction comprised the control group. Embryo transfer was performed on day three or five after the assessment of embryo compaction by a time-lapse technology system. Each patient underwent only a single cycle of embryo transfer. We analyzed fertilization, pregnancy, and live birth rates. RESULTS: We detected significantly higher numbers of the retrieved oocytes, metaphase II (MII) oocytes, and fertilized oocytes in the early compaction group. Moreover, the transfer of the early compacting embryos on day three resulted in higher pregnancy and live birth rates. CONCLUSION: Our data suggest that early compaction might be a factor to determine good quality embryos and embryo transfer day.

Anti-Müllerian hormone as a predictor of pregnancy in women under 35 years with unexplained infertility undergoing ICSI: a retrospective study.

BACKGROUND: Anti-Müllerian hormone (AMH) is a well-established marker for the determination of ovarian reserve. However, its role in the prediction of pregnancy is still under debate. In this retrospective study, we aimed to evaluate the relationship of serum AMH levels with pregnancy rates in patients with unexplained infertility undergoing ICSI. Moreover, we compared the predictive value of AMH with that of antral follicle count (AFC). METHODS: Records of 76 patients under 35 years of age with AMH levels between 1 and 3.5 ng/mL were examined retrospectively. Participants were divided into groups based on their AMH level and age. RESULTS: AMH levels in women under 30 years were found significantly higher than those in women over 30 years (P=0.033). Fifty-seven of 76 patients (75%) were pregnant. Age did not have a significant effect on the pregnancy rates in the selected study group (P=0.252). On the other hand, despite the poor predictive accuracy, serum AMH was shown to have a predictive value with a cut-off point of 1.95 ng/mL. Logistic regression tests demonstrated a higher pregnancy rate (3.396 fold) with an AMH level 1.95 or above. There was no significant relationship between AFC and pregnancy. CONCLUSIONS: AMH might have a role in the prediction of pregnancy after ICSI in patients under 35 years with unexplained infertility.

Sperm Selection and Embryo Development: A Comparison of the Density Gradient Centrifugation and Microfluidic Chip Sperm Preparation Methods in Patients with Astheno-Teratozoospermia.

In recent years, microfluidic chip-based sperm sorting has emerged as an alternative tool to centrifugation-based conventional techniques for in vitro fertilization. This prospective study aims to compare the effects of density gradient centrifugation and microfluidic chip sperm preparation methods on embryo development in patient populations with astheno-teratozoospermia. In the study, the semen samples of the patients were divided into two groups for preparation with either the microfluidic or density gradient methods. Selected spermatozoa were then used to fertilize mature sibling oocytes and the semen parameters and embryo development on days 3 and 5 were assessed. While the density gradient group was associated with a higher sperm concentration, motility (progressive and total) was significantly higher in the microfluidic chip group. No significant differences were observed in the fertilization rates or grade 1 (G1) and grade 2 (G2) proportions of the third-day embryos. Furthermore, while the proportions of the poor, fair and good blastocysts on day 5 did not differ significantly, excellent blastocysts (indicating high-quality embryos) were observed in a significantly higher proportion of the microfluidic chip group. When compared to the classical density gradient method, the microfluidic chip sperm preparation yielded sperm with higher motility and higher quality blastocysts at day 5; in patients with astheno-teratozoospermia.

Comparison of the rates for reaching the blastocyst stage between normal and abnormal pronucleus embryos monitored by a time-lapse system in IVF patients

Objective: To compare the rates of blastocyst stage development between embryos fertilized after one (MPN) or more than two pronucleus (PN) (3PN, 4PN-multiPN) with those after 2PN in the same patients. Material and Methods: The embryos of patients who had both abnormal PN (MPN, 3PN or 4PN) and normal fertilized (2PN) embryos after intracytoplasmic sperm injection (ICSI) fertilization, were followed with a time-lapse system following the ICSI procedure. The rates of reaching the blastocyst stage were compared between normal and abnormally fertilized embryos. Results: One thousand eight hundred and twenty oocytes were collected from 140 patients and 1280 (70.3%) of them were fertilized. MPN, 2PN and 3PN, 4PN (multiPN) ratios of the embryos in the pronuclear stage were 11.4%, 83.13% and 5.47%, respectively. The rates of reaching the blastocyst stage among these embryos were 17.1%, 60.8% and 42.8% for MPN, 2PN and multiPN, respectively. The proportion reaching blastocyst development was significantly higher following 2PN compared to those after MPN and multiPN (p<0.05). Embryos developing after multiPN had significantly higher rates of reaching the blastocyst stage compared to those after MPN (p<0.01). Conclusion: The majority of abnormally pronucleated embryos arrest without reaching the blastocyst stage. MultiPN embryos have a higher rate of blastocyst development than MPN embryos.

IVF outcomes after hysteroscopic metroplasty in patients with T- shaped uterus.

BACKGROUND: T- shaped uterus may be associated with infertility and adverse pregnancy outcomes. Hysteroscopic metroplasty may improve the reproductivity for these cases. To our knowledge, there is no data in literature about the clinical consequences of in vitro fertilization (IVF) in patients undergoing hysteroscopic metroplasty for T-shaped uterus. The principal objective of the current study is to assess the impact of hysteroscopic metroplasty for T-shaped uterus on the reproductive outcomes of IVF. METHODS: IVF outcomes of 74 patients who underwent hysteroscopic metroplasty for T- shaped uterus and 148 patients without any uterine abnormalities and with diagnosis of unexplained infertility (control group) were retrospectively analyzed. RESULTS: Patients in metroplasty and control groups were comparable with respect to age, BMI, partner's age and duration of infertility. Number of patients with a history of pregnancy beyond 20 weeks of gestation was significantly lower in the metroplasty group (4.1% vs 18.2%; p < 0.05). Number of previous unsuccessful cycles and percentage of patients with ≥3 unsuccessful IVF cycles (35.1% vs 17.6%; p < 0.05) were significantly higher in the metroplasty group. There were no significant differences in the reproductive outcomes such as the pregnancy rate, clinical pregnancy or live birth rate between the metroplasty and control groups. There were non-significant trends for higher rates of miscarriage (18.8% vs 8%, p > 0.05) and biochemical pregnancy (20.0% vs 10.7%, p > 0.05) in the metroplasty group compared to the control group. CONCLUSIONS: Reproductive results of the IVF cycles after hysteroscopic correction of T-shaped uterus were comparable to those of the patients without any uterine abnormalities and with diagnosis of unexplained infertility. Hysteroscopic metroplasty may contribute to improved IVF outcomes in patients with T-shaped uterus.

Evaluation of Sperm DNA Fragmentation via Halosperm Technique and TUNEL Assay Before and After Cryopreservation.

AIM: Human sperm DNA fragmentation is one of the factors suggested for male infertility. The ratio of sperm DNA damage in semen may adversely affect both the fertilization rate and the embryo development of in vitro fertilization/ intracytoplasmic sperm injection cycles. Sperm cryopreservation both increases the success rates in assisted reproductive techniques (ARTs) and contributes to the preservation of fertility before testis surgery, chemotherapy, and radiotherapy. The aim of the current study is to determine sperm DNA fragmentation, following cryopreservation. METHODS: A cross-sectional, observational study was conducted at a university hospital infertility clinic. One hundred (n = 100) volunteer fertile men (ages between 21 and 39 years) with normozoospermic sperm parameters were involved in the current study. Sperm DNA damage was evaluated with the Halosperm technique and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Fresh samples were studied in liquid form. The remaining samples were kept frozen and then thawed after 1 month and reevaluated with the Halosperm technique and TUNEL assay. Results were then compared between the fresh and frozen samples. RESULTS: Sperm DNA fragmentation results with the Halosperm technique both before and after cryopreservation were 25% (5%-65%) and 40% (6%-89%), respectively, with a statistically significant increase (15%; P < .001). Sperm DNA fragmentation results by TUNEL assay before and after cryopreservation were 17% (3%-43%) and 36% (7%-94%), respectively, with a statistically significant increase (19%; P <.001). CONCLUSION: The current data demonstrate increased sperm DNA damage after cryopreservation. Further studies may contribute to development of less harmful techniques and cryoprotectants in order to improve the results of ART.

Clinical pregnancy after uterus transplantation.

OBJECTIVE: To present the first clinical pregnancy after uterus transplantation. DESIGN: Case study. SETTING: Tertiary center. PATIENT(S): A 23-year-old Mayer-Rokitansky-Kuster-Hauser syndrome patient with previous vaginal reconstruction and uterus transplantation. INTERVENTION(S): Eighteen months after the transplant, the endometrium was prepared for transfer of the thawed embryos. MAIN OUTCOME MEASURE(S): Implantation of embryo in an allografted human uterus. RESULT(S): The first ET cycle with one day 3 thawed embryo resulted in a biochemical pregnancy. The second ET cycle resulted in a clinical pregnancy confirmed with transvaginal ultrasound visualization of an intrauterine gestational sac with decidualization. CONCLUSION(S): We have presented the first clinical pregnancy in a patient with absolute uterine infertility after uterus allotransplantation. Although the real success is the delivery of a healthy near-term baby, this clinical pregnancy is a great step forward and a proof of concept that the implantation phase works.

Preliminary results of the first human uterus transplantation from a multiorgan donor.

OBJECTIVE: To describe the first-year results of the first human uterus transplantation case from a multiorgan donor. DESIGN: Case study. SETTING: University hospital. PATIENT(S): A 21-year-old woman with complete müllerian agenesis who had been previously operated on for vaginal reconstruction. INTERVENTION(S): Uterus transplantation procedure consisting of orthotopic replacement and fixation of the retrieved uterus, revascularization, end to site anastomoses of bilateral hypogastric arteries and veins to bilateral external iliac arteries and veins was performed. MAIN OUTCOME MEASURE(S): Resumption of menstrual cycles. RESULT(S): The patient had menarche 20 days after transplant surgery. She has had 12 menstrual cycles since the operation. CONCLUSION(S): We have described the longest-lived transplanted human uterus to date with acquirement of menstrual cycles.

Immunolocalization of fibronectin during reparative dentinogenesis in rat molor teeth after pulp capping with mineral trioxide aggregate or calcium hydroxide.

The aim of this study was to evaluate the immunolocalization of fibronectin during reparative dentinogenesis in rat teeth after pulp capping with mineral trioxide aggregate (MTA) or calcium hydroxide (Ca(OH)2). The pulps of 72 upper and lower first molar teeth from 18 male Wistar rats were experimentally exposed. The pulps were capped with MTA or (Ca(OH)2); final restoration followed with zinc oxide and eugenol cement. The animals were euthanized at, respectively, one, three, seven and fourteen days postoperatively. At day one, all groups showed varying degrees of inflammation, from mild to severe. There was no positive reaction for fibronectin at day one. After three days, a partial acute pulpitis was observed in the Ca(OH)2 group. There was less inflammation in the MTA group (p<0.05), and a layer of fibrin barrier was observed along the pulp walls of the MTA material. The layer of fibrodentin formation showed positive reaction for fibronectin. At seven days, the Ca(OH)2 group showed mild inflammation and demonstrated more immunostaining for fibronectin than the Ca(OH)2 group (p<0.05) at three days. Pulps capped with MTA at seven days showed thicker fibrin barrier formation than the MTA group at three days and more immunostaining for fibronectin in whole groups (p<0.05). At fourteen days, there was no positive reaction for fibronectin in either the MTA or Ca(OH)2 group. It seems MTA showed better biocompability properties with the dental pulp tissue, inducing the expression of reparative molecule fibronectin compared with Ca(OH)2. Therefore, MTA may be a better choice for pulp capping procedures.

Perindopril, atenolol, and amlodipine prevent aortic ultrastructural changes in rats exposed to ethanol.

BACKGROUND: The effects of perindopril, an angiotensin-converting enzyme inhibitor, atenolol, a beta adrenergic receptor blocker, and amlodipine, a calcium channel blocker, were investigated in chronic alcohol administered rats. MATERIAL/METHODS: Adult male Wistar rats (240-320 g) were used in the present study. Alcohol was given to rats on a modified liquid diet for 21 days. Perindopril (2.5 and 5 mg/kg), atenolol (5 and 10 mg/kg), and amlodipine (5 and 10 mg/kg) were injected into rats in different groups intraperitoneally for 21 days. Control rats were pair fed an isocaloric liquid diet containing sucrose as a caloric substitute for alcohol. Saline was injected into the control rats for 21 days. The hearts were removed after the rats were anesthetized by ether, and 1-mm3 samples from the ascending aortas were fixed. Five fields per aorta were examined and photographed with a transmission electron microscope. Blood alcohol levels were also measured spectrophotometrically. RESULTS: Daily alcohol consumption of the rats was in the range of 12.09-15.50 g/kg. Blood alcohol concentrations were 145.63 mg/dl on the 21st day of alcohol consumption. Chronic alcohol consumption caused some marked aortic wall injuries. Perindopril, atenolol, and amlodipine at high doses, but not low doses, produced some significant beneficial effects on alcohol-induced aortic wall damage. CONCLUSIONS: These results imply that perindopril, atenolol, and amlodipine may have protective effects on heavy chronic alcohol consumption-induced aortic wall injury in rats only in high doses.

Retrieval of zona-free immature oocytes in a woman with recurrent empty follicle syndrome: a case report.

BACKGROUND: Different ovulation trigger methods such as gonadotropin releasing hormone-agonist (GnRH-a) and recombinant human chorionic gonadotropin (r-hCG) plus rescue oocyte retrieval might reveal oocytes in patients with recurrent empty follicle syndrome. CASE: Endogenous luteinizing hormone was triggered with a GnRH-a (Buserelin [Suprefact pro-injection, Aventis-Pharma, Turkey], 250 microg subcutaneously) in a GnRH antagonist (Cetrorelix [Cetrotide 0.25, SeronoTurkey], 0.25 mg/d, starting on day 6), down-regulated cycle. At the first scheduled retrieval, 3 cumulus-oocytecorona complexes were recovered from the left ovary. During chemical denudation with hyaluronidase, 2 of them underwent lysis. The third was a zona-free, germinalvesicle-stage oocyte after mechanical denudation. Oocyte pickup was stopped, and recombinant human chorionic gonadotropin (hCG) (250 microg subcutaneously) was injected. Five cumulus-oocyte-corona complexes were retrieved from the right ovary 24 hours after rescue with recombinant hCG. Only mechanical denudation was done, and 4 zona-free oocytes with germinal vesicle breakdown were seen. All oocytes underwent intracytoplasmic sperm injection, and none of them were fertilized. CONCLUSION: Oocyte maturation defects should be included in etiologic mechanisms for counseling patients with empty follicle syndrome.

The prevention of myocardial ultrastructural changes by perindopril, atenolol and amlodipine in chronic alcohol administered rats.

The effects of perindopril, an angiotensin converting enzyme inhibitor, atenolol, a beta adrenergic receptor blocker and amlodipine, a calcium channel blocker were investigated in chronic alcohol administered rats. Adult male Wistar rats (240-320 g) were used in the present study. Alcohol was given to rats by a modified liquid diet for 21 days. Perindopril (2.5 and 5 mgkg(-1)), atenolol (5 and 10 mg kg(-1)) and amlodipine (5 and 10 mg kg(-1)) were injected to rats in different groups intraperitoneally for 21 days. Control rats were pair fed by an isocaloric liquid diet containing sucrose as a caloric substitute for alcohol. Saline was injected to control rats for 21 days. Rats were anesthetized with ether. Their hearts were removed and 1 mm3 samples from left ventricles were fixed. Five fields per heart were examined and photographed with transmission electron microscope. Blood alcohol levels were also measured spectrophotometrically. Daily alcohol consumption of the rats was in a range of 12.09-15.5 g kg(-1). Blood alcohol concentrations were found as 145.63 mg dl(-1) at 21st day of alcohol consumption. Chronic alcohol consumption caused some marked myocardial injuries. Perindopril and atenolol but not amlodipine produced some significant beneficial effects on alcohol-induced myocardial damages. Our results imply that perindopril and atenolol but not amlodipine have protective effects on heavy chronic alcohol consumption-induced myocardial injury in rats.

Serum and follicular fluid levels of soluble Fas and soluble Fas ligand in IVF cycles.

OBJECTIVE: To determine follicular fluid (FF) and serum levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients undergoing IVF cycles. STUDY DESIGN: A prospective comparative study among patients with endometriosis (n=12), infertility due to male factor (n=12) and poor responders (n=32) undergoing IVF cycles in Centrum IVF Clinic. Individual FF and serum samples were collected from patients during transvaginal ultrasonography-guided follicle aspiration. Patients were classified as poor responder patients undergoing IVF cycles with GnRHa, triptorelin and GnRH antagonist, cetrotide, patients with endometriosis and patients with infertility due to male factor. sFas, sFasL levels in both FF and serum samples and their correlations with clinical outcomes of IVF were measured in each study group. RESULTS: Serum and FF levels of sFas, sFasL were similar in the poor responder and male factor groups. There were no differences between the serum and FF levels of both sFas and sFasL among poor responder patients receiving either GnRH agonist or antagonist therapies. Serum levels of sFas were significantly lower in the endometriosis group compared to the male factor group. Serum and FF levels of sFas, sFasL were similar among patients with or without clinical pregnancy. CONCLUSION: sFas and sFasL are detected in both serum and follicular fluid samples from IVF cycles, their levels are similar between poor responder and male factor groups as well as between GnRH agonist and antagonist treatment groups. These soluble apoptotic factors may not be predictive for the outcomes of IVF. Decreased serum levels of sFas, suggests increased apoptosis in endometriosis.

Metformin treatment in patients with polycystic ovary syndrome undergoing in vitro fertilization: a prospective randomized trial.

In the present study, we investigated the impact of metformin therapy on in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcomes in patients with polycystic ovary syndrome (PCOS). Metformin does not lead to any improvement in IVF/ICSI outcomes among patients with PCOS.

Serum and follicular fluid levels of soluble Fas, soluble Fas ligand and apoptosis of luteinized granulosa cells in PCOS patients undergoing IVF.

BACKGROUND: There are limited data about the levels of soluble apoptotic factors and their modulation with therapeutic regimens in IVF cycles. The aim of the current study was to determine follicular fluid, and serum levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in PCOS patients undergoing IVF/ICSI cycles; also to investigate the effects of metformin on these factors and on apoptosis of luteinized granulosa cells. METHODS: We investigated the serum and follicular fluid levels of sFas and sFasL in patients with PCOS (n = 28) and compared them with those of the patients with infertility due to male factor (n = 12) undergoing IVF cycles. Effects of metformin therapy on these parameters and apoptosis of luteinized granulosa cells were also investigated among the patients with PCOS. RESULTS: Serum levels of sFas were significantly lower in the PCOS group compared to those in women with infertility due to male factor. Metformin therapy in PCOS patients preceding IVF cycles increased serum levels of sFas and decreased follicular fluid levels of sFasL compared to those on placebo. Follicular fluid from PCOS patients demonstrated luteinized granulosa cell DNA fragmentation in agarose gel, whereas a similar pattern was not observed among PCOS patients undergoing metformin therapy. CONCLUSION: Decreased serum levels of sFas and luteinized granulosa cell DNA fragmentation is observed in patients with PCOS undergoing IVF cycles. Metformin therapy preceding IVF demonstrates an antiapoptotic effect with increased serum levels of sFas, decreased follicular fluid levels of sFasL and prevention of luteinized granulosa cell DNA fragmentation.

Relationship between extracorporeal shock wave lithotripsy and semen parameters in patients with lower ureteral stones.

The objective of this study was to investigate whether extracorporeal shock wave lithotripsy can affect semen parameters in patients with distal ureteral stones. We concluded that extracorporeal shock wave lithotripsy could reduce sperm concentration and motility permanently in men with lower ureteral stones.