Effects of different drug treatments on the proliferation of human ovarian carcinoma cell line MDAH-2774

Background/aim: In this study, the effects of resveratrol as a natural polyphenol compound, gemcitabine as an antimetabolite that has nucleoside structure analogous to deoxycytidine, and para-aminophenol-derived paracetamol were investigated with single and combined applications in monolayers of the MDAH-2774 human ovarian cancer cell line. Materials and methods: Drugs were evaluated in cell culture with respect to cell proliferation, cell cytotoxicity (trypan blue dye exclusion test), synthesis phase of cell cycle, and cell structure in 24, 48, 72, and 96 h. Result: Resveratrol and gemcitabine diminished both cell proliferation and cell cycle synthesis phase indication in monolayer cell cultures (P < 0.05). All combination groups showed similar effects that were mainly more effective in respect to single usage of resveratrol and gemcitabine in monolayer cell cultures. Conclusion: The effects of gemcitabine, resveratrol, and paracetamol were investigated in monolayers of the MDAH-2774 human ovarian cancer cell line and a decrease in cell number in cell cycle synthesis phase, prevention of cell proliferation, and destruction of cell structure were observed.

Social stigma and familial attitudes related to infertility.

OBJECTIVE: To determine the perceived social stigma and familial attitides and perception of sexuality in infertile couples attending infertility clinics. MATERIALS AND METHODS: Infertile couples attending infertility clinics between the years of 2014 and 2015 were requested to complete detailed evaluation forms including questions related to the social stigma on their infertility, their familial attitudes, and perception of sexuality. Any partner of the infertile couple accepting to enroll in the study was given the evaluation forms. Their scores related to answers and demographics, and parameters related to infertility were analyzed. RESULTS: A total 598 partners of infertile couples enrolled in the study, 58% represented 177 couples. Their infertility was primary in 98.3% and the duration of marriage and infertility was 9.81±5.58 and 9.76±5.53 years, respectively. The perception of social exclusion was present in 38% (p<0.001) of infertile couple, which was more significantly pronounced in female partners (p=0.013). Fifteen percent of the infertile couples thought themselves as isolated in public and losing value in public (p<0.001). However, sixty percent of infertile couples thought that they would achieve a notable place in community after having a baby (p<0.001). Infertility was accepted as a reason of divorce in only 13% of infertile couples (p<0.001). The majority of perticipnats, irrespective of sex, rejected that infertile women or men lost sexual appeal (86%; p<0.001). CONCLUSIONS: There is significant effect of infertility on familial attitudes and perception of sexuality of infertile couples. Unfortunately, there is significant negative social stigma on infertile couples.

Antioxidant activity of CAPE (caffeic acid phenethyl ester) in vitro can protect human sperm deoxyribonucleic acid from oxidative damage.

PURPOSE: Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage. METHODS: Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 µmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy. RESULTS: Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ±â€¯0.15). Incubation of samples with 100 µmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ±â€¯0.12) (P < 0.0033). Incubation of all samples with CAPE (10 µmol/L, 50 µmol/L, 100 µmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels. CONCLUSIONS: The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.

Subclinical hypothyroidism: Is it important in intracytoplasmic sperm injection cycles?

OBJECTIVE: To compare intracytoplasmic sperm injection (ICSI) outcomes of women with subclinical hypothyroidism with those of euthyroid women. MATERIALS AND METHODS: A retrospective case-control study was conducted. Out of 2529 ICSI cycles evaluated, 41 women with hypothyroidism, 28 women with hyperthyroidism, and 128 women with subclinical hyperthyroidism were excluded, and 2336 cycles were analyzed. Women were identified as having subclinical hypothyroidism (case group, n=105) in the presence of a thyroid-stimulating hormone level >4.5 mU/L and normal free T4 and compared with euthyroid controls (n=2231). RESULTS: The mean age, body mass index, day 3 follicle-stimulating hormone level, and antral follicle count of the study patients were similar to the control group (p>0.5). The cycle cancellation rate of the study group was similar to the control group (13.3% vs. 7.6%, p=0.1). The clinical pregnancy rate was 21.2% in the study group, which was significantly lower than the 35.8% in the control group (p=0.04). The take-home baby rate was also significantly lower in the study group compared with the control groups (13.5% vs. 31.4% respectively, p=0.01). CONCLUSION: Both the clinical pregnancy rate and the take-home baby rate is lower in women with subclinical hypothyroidism at the time of ICSI cycle.

Microdose Flare-up Gonadotropin-releasing Hormone (GnRH) Agonist Versus GnRH Antagonist Protocols in Poor Ovarian Responders Undergoing Intracytoplasmic Sperm Injection.

BACKGROUND: Microdose flare-up GnRH agonist and GnRH antagonist have become more popular in the management of poor ovarian responders (POR) in recent years; however, the optimal protocol for POR patients undergoing in vitro fertilization has still been a challenge. METHODS: In this observational study design, two hundred forty four poor ovarian responders were retrospectively evaluated for their response to GnRH agonist protocol (group-1, n=135) or GnRH antagonist protocol (group-2, n=109). Clinical pregnancy rate was the primary end point and was compared between the groups. Student t-test, Mann Whitney U test and χ (2)-test were used to compare the groups. The p<0.05 was considered to show a statistically significant result. RESULTS: The mean total gonadotropin doses were 3814±891 IU in group 1 and 3539±877 IU in group 2 (p=0.02). The number of metaphase-II oocytes (3.6±2.4 vs. 2.8±1.9, p=0.005) and implantation rates (27.8% vs. 18.8%, p=0.04) in group 1 and group 2, respectively were significantly different. The fertilization rate in group 1 and group 2 was 73% vs. 68%, respectively (p=0.5) and clinical pregnancy rate was 19.8% vs. 14.4%, respectively (p=0.13). CONCLUSION: The GnRH agonist microdose flare-up protocol has favorable outcomes with respect to the number of oocytes retrieved and implantation rate; nevertheless, the clinical pregnancy rate was found to be similar in comparison to GnRH antagonist protocol in poor ovarian responders. GnRH antagonist protocol appears to be promising with significantly lower gonadotropin requirement and lower treatment cost in poor ovarian responders.

Progesterone vaginal capsule versus vaginal gel for luteal support in normoresponder women undergoing long agonist IVF/ICSI cycles.

OBJECTIVES: The aim of the study was to investigate the effects of two different vaginal progesterone forms, administered for luteal phase support, on pregnancy outcomes in normoresponder women aged < 35, who underwent long agonist IVF/ICSI-ET cycles. MATERIAL AND METHODS: A retrospective cohort analysis was designed. Normoresponders with primary infertility, who un-derwent IVF/ICSI-ET cycles employing GnRH analogue and who received progesterone as either capsule or gel form for LPS following a single embryo transfer, were analyzed. The cycles were categorized into two groups: micronized progesterone vaginal capsule 600 mg/day (Group 1, n = 78) and progesterone vaginal gel 180 mg/day (Group 2, n = 99). Positive ß-hCG, clinical pregnancy and ongoing pregnancy rates were analyzed. RESULTS: Both, demographic and stimulation characteristics were comparable between the groups. No difference was observed between the capsule and the gel groups regarding positive ß-hCG (33.3% and 28.3%, respectively; p = 0.580), clinical pregnancy (26.9% and 22.2%, respectively; p = 0.584), and ongoing pregnancy rates (21.8% and 20.2%, respectively; p = 0.942) after treatment completion. CONCLUSIONS: In long agonist IVF/ICSI-ET cycles, positive ß-hCG, clinical pregnancy and ongoing pregnancy rates do not significantly differ between normoresponder patients receiving micronized progesterone vaginal capsule and those receiv-ing progesterone vaginal gel for LPS.

Characteristics and outcomes of in vitro fertilization in different phenotypes of polycystic ovary syndrome.

OBJECTIVE: The aim of this study was to investigate whether polycystic ovary syndrome (PCOS) phenotype without polycystic ovaries (PCO) differs in terms of in vitro fertilization (IVF) outcomes compared with classic phenotypes. MATERIALS AND METHODS: This retrospective controlled study included 262 patients who underwent IVF treatment with an indication of unexplained or tubal factor infertility (control group), ovulatory patients with PCO morphology (group 1), PCOS phenotype with oligoanovulation and hyperandrogenemia (group 2), PCOS phenotype with PCO morphology and oligoanovulation (group 3). Outcomes and baseline characteristics of IVF-embryo transfer treatments were compared among all groups. RESULTS: PCOS phenotype without PCO morphology had similar IVF stimulation characteristics compared with classic phenotypes; however, a higher total gonadotropin dose was needed to achieve similar results compared with patients with PCO morphology with or without PCOS. Basal follicle-stimulating hormone level (beta coefficient=0.207, p=0.003), group (beta coefficient=-0.305, p<0.001) and age (beta coefficient=0.311, p<0.001) were significantly associated with the total gonadotropin dose. The number of good quality embryo on transfer day was significantly lower in patients with isolated PCO morphology and PCO morphology with oligoanovulation than in those with PCOS phenotype without PCO morphology. CONCLUSION: PCO morphology provides easier stimulation, whereas hyperandrogenemia provides better results as good quality embryos. However, the end point is similar in terms of biochemical, clinical, and ongoing pregnancy rates.

Can blood or follicular fluid levels of presepsin predict reproductive outcomes in ART; a preliminary study.

Many stages of COH protocols are considered to potentiate a state of systemic inflammation. The limit beyond which inflammation has negative impacts on the formation of conception and the reproductive outcomes are compromised still remains unclear. Presepsin is a novel biomarker for diagnosing systemic inflammation and sepsis. We aimed to investigate whether plasma and follicular fluid presepsin values on oocyte pick-up (OPU) day, embryo transfer (ET) day and pregnancy test (PT) days could predict reproductive outcomes during IVF treatment in women with UEI. Patients were assigned to two groups according to pregnancy test results; pregnant (Group 1) and non-pregnant (Group 2). From all patients included in the study, 2 cc of venous blood was sampled on the three days and follicular fluid (FF) was collected during oocyte retrieval. Plasma presepsin, CRP and WBC values and FF presepsin values were measured and compared between the 2 groups. There was no significant difference between FF and plasma presepsin levels on the OPU day (298±797.4 ve 352.9±657.1; P=0.701, respectively). Plasma WBC, CRP and presepsin levels on the OPU, ET and PT days and FF presepsin levels on OPU day were not different between the 2 groups. Plasma presepsin course on the separate 3 days were different between the groups.

Vaginal micronized progesterone capsule versus vaginal progesterone gel for lutheal support in normoresponder IVF/ICSI-ET cycles.

OBJECTIVE: To compare the outcomes of luteal phase support by micronized progesteron vaginal capsule 600mg/day and progesterone vaginal gel 180mg/day in the normoresponder IVF/ICSI-ET cycles of the patients down-regulated via GnRH agonist long protocol or fixed antagonist protocol below 40 years of age. METHODS: A total of 463 normoresponder cycles between January 2013 and December 2013 were retrospectively analyzed. Those with a BMI>28 kg/m(2), any kind of uterine, ovarian or adnexial pathology, any significant systemic, endocrine or metabolic disease or who were reported as azoospermia, were excluded from the study. The patients were grouped according to the usage of micronized progesterone vaginal capsule 600mg/day (Group 1) or progesterone vaginal gel 180mg/day (Group 2) as luteal phase support. Treatment cycle characteristics and pregnancy outcomes were compared between groups. RESULTS: Group-I included 220 cycles and group 2 included 243 cycles. Although the MII oocyte percentage among the total number of MII oocytes was significantly higher in Group-II (77.5% and 80.2%; p=0.034), positive ß-hCG (32.3% and 21.8%; p=0.015) and clinical pregnancy (27.3% and 17.7%; p=0.018) rates were significantly higher in Group-I. No difference was observed between groups regarding the ongoing pregnancy rates (23.2% and 17.3%; p=0.143). CONCLUSION: Micronized progesterone vaginal capsule 600mg daily used for luteal support in the IVF/ICSI-ET cycles was observed to significantly increase the biochemical and clinical pregnancy rates compared to progesterone vaginal gel 180mg daily. However, no difference was observed between two groups regarding ongoing pregnancy rates.

Effects of salpingectomy and antituberculosis treatments on fertility results in patients with genital tuberculosis.

AIM: The aim of this study was to demonstrate the effects of tubal surgery and antituberculosis treatments in patients with genital tuberculosis. MATERIAL AND METHODS: A total of 38 infertile women who had been diagnosed with pelvic tuberculosis and who had had laparoscopy and hysteroscopy were recruited into the study. All women with female genital tuberculosis were divided into two groups: group 1 (salpingectomized, n=21) and group 2 (not salpingectomized, n=15). Both of the groups were treated with antitubercular therapy for 6-12 months. RESULTS: There was no significant difference in level of gonadotrophins used, estradiol levels on human chorionic gonadotrophin day, mean and mature oocytes retrieved, mean embryos transferred, or cancellation and fertilization rates. Only the number of days of stimulation was statistically significantly higher in group 1 compared to group 2 (10.4±2.3 vs 9.2±1.8; P=0.048). Although it did not reach the statistically significant level, clinical pregnancy rate was higher in group 1 (37.5%, 12/32 vs 23.8%, 5/21; P=0.306). Although not statistically significant, number of ongoing pregnancies per embryo transfer, spontaneous abortion rates before 20 weeks of gestation and take-home baby rates were higher in group 1 compared to group 2 (15.5%, 12/77 vs 6.6%, 3/45; P=0.150; 28.1%, 9 vs 23.8%, 5; P=0.600; 9%, 3 vs 0; P=0.160, respectively). CONCLUSION: Salpingectomy is an option for treatment in patients diagnosed with pelvic tuberculosis and infertility to improve both clinical pregnancy rates and take-home baby rates in patients treated with antituberculosis therapy for 12 months.

Effect of lower than expected number of oocyte on the IVF results after oocyte-pickup.

OBJECTIVES: To investigate whether a lower than expected number of oocyte after ≥14 mm follicle aspiration during OPU has any effect on pregnancy outcomes Methods: This is a retrospective study done between 2010 and 2013 at the IVF Unit of the Zeynep Kamil Women and Children Diseases Education and Research Hospital, dealing with the medical records of infertile patients who underwent IVF cycle and controlled ovarian stimulation with long agonist or fix antogonist protocol. The patients included into the study were those diagnosed with a primary infertility, aged between 23 and 39, at a BMI of 22-28 kg/m(2) and having received the first or second IVF treatment. Male factor, presence of uterine anomaly, patients with serious endometriosis and patients with low ovarian reserve were all excluded from the study. Typically, oocyte pick-up was performed in all the patients 35.5 hours after the hCG implementation. Single or double embryo transfer was performed, where available. Patients were classified into two groups. Group 1 consisted of those with no difference between ≥14 mm aspirated follicle number and expected number of oocyte or with 1 missing number of oocyte at the most. Group 2 consisted of those with at least ≥2 missing number of oocyte between aspirated follicle number and expected number of oocyte. Statistical analysis was performed using Student's t test for continuous variables and chi-square test for categorical variables. Additionally, a Linear regression analysis was conducted between the total number of oocyte and pregnancy. RESULTS: In total, 387 treatment cycles were included into the study. Group 1 consisted of 134 patients and Group 2 consisted of 252 patients. Antral follicle number (12.8 ± 4.3 and 14.5 ± 4.1, P = 0.0007), hCG day E2 value (1990.7 ± 1056.4 and 2515.2 ± 1332.7, P < 0.0001) and the the number of aspirated follicle during OPU (9.1 ± 4.4 and 13.7 ± 5.5, P < 0.0001) were significantly higher in Group 2; whereas on the other hand, daily gonadotropin dose (290.9 ± 79.9 and 273.4 ± 74.4, P = 0.034) and total gonadotropin doses (2545 ± 1031.8 and 2247.7 ± 901.9, P = 0.004) were significantly higher in Group 1. The pregnancy rate was significantly higher in Group 1 (29.1% and 19.4%, P = 0.041). No correlation was observed between the number of oocyte and pregnancy (r = 0.082, P = 0.107). CONCLUSIONS: The number of aspirated follicles during IVF treatment being higher than the collected number of oocyte leads to a statistically significant fall in the pregnancy rates. There is no correlation between the number of oocyte and pregnancy.

Blood serum and seminal plasma selenium, total antioxidant capacity and coenzyme q10 levels in relation to semen parameters in men with idiopathic infertility.

In this case-control study, we aimed to evaluate the serum and seminal plasma levels of Selenium (Se), total antioxidant capacity (TAC), and Coenzyme Q10 (CoQ-10) and determine their relationship with sperm concentration, motility, and morphology in men with idiopathic infertility. A total of 59 subjects were enrolled in the study. Forty four patients were diagnosed with idiopathic male infertility and had abnormal sperm parameters, and 15 subjects had normal sperm parameters with proven fertility. Serum Se, semen Se, and semen TAC levels were significantly different in the fertile and infertile groups (p<0.01, p<0.001, and p<0.001, respectively). However, serum TAC, serum, and seminal plasma CoQ-10 levels did not differ between fertile and infertile groups. When the levels of the measured parameters were compared in serum and seminal plasma, serum levels of Se were found to be correlated positively with the semen levels in all subjects included into the study (N=59) (r=0.46, p<0.01). A relationship was found between neither serum and semen levels of TAC nor between serum and semen levels of CoQ-10. Correlations among measured serum and semen parameters with sperm parameters demonstrated that both the serum and semen levels of Se were correlated positively with spermatozoa concentration, motility, and morphology. Additionally, seminal plasma levels of TAC correlated positively with all these sperm parameters. On the other hand, seminal plasma levels of CoQ-10 correlated only with sperm morphology but not with concentration or motility. No relationship was observed between serum levels of TAC or serum levels of CoQ-10 and sperm parameters. In conclusion, serum and seminal plasma Se deficiency may be a prominent determinant of abnormal sperm parameters and idiopathic male infertility. Measurement of serum Se levels may help determine nutritional status and antioxidant capacity in infertile patients, which may help distinguish those patients who will benefit from supplementation therapy.

The incidence of preeclampsia in ICSI pregnancies.

OBJECTIVE: We aimed to evaluate the association between infertility etiology in ICSI pregnancies and preeclampsia; besides, we aimed to discuss the effect of the paternal factor in the pathogenesis of preeclampsia. HYPOTHESIS: We hypothesized that preeclampsia is more common in ICSI pregnancies with male factor. It is known that maternal exposure to paternal sperm cells over a time period has a protective effect against preeclampsia. Male partners with azospermia have no sperm cells in their seminal fluid, whose female partners will not be able to develop some protective immunity against preeclampsia. We hypothesized that the infertile couples with male factor (partner with azoospermia and also oligospermia) would be an ideal model to test the partner-specific protective immunity against preeclampsia, as the women had no chance to develop adequate protective immunity via the partner's sperm exposure. METHODS: This Single-center, retrospective study included 508 infertile couples admitted to our IVF center between January 2001 and March 2008. The data regarding the maternal age, etiology of the infertility, the pregnancy rates, abortus ratio and viable pregnancy rates was collected from the case files. Antenatal complications such as preeclampsia, placenta previa, abruptio placenta, premature rupture of membranes, premature labor, oligohydramnios, gestational diabetes, postmaturity, postpartum complications and neonatal outcomes were evaluated via the file records and phone interviewing. The study population was divided into two main groups according to the etiology of infertility. 301 of the study population (group 1) was infertile due to male factor and 207 of the study population (group 2) was female factor and unexplained infertility cases.Group 1 patients were divided further into two subgroups: group 1a included 56 cases in which TESE (testicular sperm extraction) was used to obtain the sperm cells as the male factor was severe and as there was no sperm cells in seminal fluid. Group 1 b consists of 245oligospermic cases who obtained sperm cells via conventional methods. RESULTS: The mean ages of women in Group one and two were 30.22±5.06 and 31.58±4.36 years respectively (p=0.001). 129 cases (42,8%) from group one and 106 cases (51,2%) from Group two ended in first trimester and early second trimester (<24 gestational weeks) pregnancy loss. In group one, only 172 cases of 301 pregnancies passed over 24 weeks of gestational age, whereas in group two, 101 cases of 207 patients passed over 24 gestational weeks. There was no significant difference between two groups regarding chemical pregnancies and early pregnancy loss (p=0.314). There was no significant difference between the groups regarding placenta previa, gestational diabetes, oligo hydramnios and intrauterine growth retardation. One one pregnancy was 1.5 times more vulnerable for preeclampsia. CONCLUSION: Pregnancies with azoospermic and oligospermic partners had an increased risk for developing preeclampsia.

Association between serum estradiol level on the hCG administration day and IVF-ICSI outcome.

BACKGROUND: Estradiol (E2) is required for follicular development and play an important role in embryo implantation. OBJECTIVE: The aim of this study was to assess the impact of serum E2 levels on the day of hCG administration in IVF-ICSI patients who are performed controlled ovarian hyperstimulation (COH). MATERIALS AND METHODS: A total of 203 women who were undergone one time IVF cyclus were evaluated in this cross sectional study. All the patients were treated either with long protocol or with microdose flare protocol. The patients were categorized into five groups according to the serum E2 levels on the day of hCG administration. RESULTS: The mean number of the retrieved oocytes was (NRO) 10.6±6.7, mean fertilization rate was 55.7±24.8, and implantation rate was 9.0±19.2. Of 203 patients, 43 (21%) patients were pregnant. When the overall results are examined, the number of the retrieved oocytes and the number of transferred embryos were better in patients with serum E2 levels >4000 pg/ml and these values were statistically significant. There were no statistical difference in patients 37 years or older. In women ≤36 years old, the IVF-ICSI outcomes were better in patients with serum E2 levels >4000 pg/ml. CONCLUSION: In spite of the lack of high quality evidence to support a positive association between serum E2 levels and IVF-ICSI outcomes, this study shows that high E2 levels during COH might be associated with an increased potential of pregnancy depending on better ovarian response. When the overall results are examined, the best scores were in patients with serum E2 levels >4000 pg/ml.

Comparison of ovulation induction and pregnacy outcomes in IVF patients with normal ovarian reserve who underwent long protocol with recombinant-FSH and highly purified-hMG.

OBJECTIVE: Gonadotropins used in controlled ovarian stimulation have been increasing in number. Beside the recombinant preparations such as rec-FSH, rec-LH and h-hMG human-derived preparations have entered the market. We decided to compare the effects of rec-FSH and HP-hMG with GnRHa on embryo quality and pregnancy outcome in women undergoing an IVF cycle. MATERIAL AND METHODS: In this study, data of 87 patients who had applied to our center from 2007 to 2008 and who had met all inclusion criteria, were analyzed. The patients underwent controlled ovarian hyperstimulation with HP-hMG, rec-FSH following down-regulation with a GnRHa in a long protocol, selected according to determined criteria and acquired embryo via IVF transfer. RESULTS: Of the 87 patients, 44 were stimulated with rec-FSH and 43 with HP-hMG. Distribution of infertility causes was similar between the groups. Duration of gonadotropin administration (p=0.677, Student's t-test) and the total dose of gonadotropin received (p=0.392, Student's t-test) were similar between the two groups. The fertilization rate of the rec-FSH group was significantly higher than the HP-hMG group (p=0.001, Mann-Whitney U test). No significant differences were observed between the study groups in biochemical, clinical and ongoing pregnancy parameters. CONCLUSION: The higher oocyte yield with rec-FSH does not result in higher quality embryos. LH activity in combination with FSH activity positively affected the oocyte and embryo maturation. Therefore, when we consider the clinical and ongoing pregnancy rates there is no inferiority of HP-hMG in controlled ovarian stimulation.