Oxidative stress and female infertility: the role of follicular fluid soluble receptor of advanced glycation end-products (sRAGE) in women with endometriosis.

Oxidative Stress (OS) relates to the pathophysiology of endometriosis by activation of the inflammation process in the ovary, abdomen, peritoneum and endometrium. Advanced Glycation end-products (AGEs) cause oxidative damage to the follicles of the ovary. This study aims to investigate the correlation of follicular fluid soluble receptor of AGEs (FF sRAGE) with fertility-related parameters in infertile women with endometriosis. From January 2012 to July 2015 twenty-four women diagnosed with mild to moderate endometriosis aged 28-38 years underwent assisted reproduction. sRAGE levels measured in FF were related to lifestyle factors, sociodemographic characteristics, gynaecological and obstetric parameters, hormonal status and fertilization. sRAGE was inversely associated with BMI (r = -0.503, p = 0.012). No significant association of sRAGE with age (p = 0.714) or alcohol consumption (p = 0.882) was found. Pearson's r correlation coefficient revealed that sRAGE was positively associated with serum AMH (r = 0.518, p = 0.009), FF AMH (r = 0.630, p = 0.001), number of follicles >15mm (r = 0.601, p = 0.002), total number of follicles aspirated (r = 0.698, p < 0.001), total number of MII oocytes obtained, (r = 0.757, p < 0.001) and the number of embryos with good embryo scoring (suitable for ET) (r = 0.522, p = 0.009). It seems that measurement of FF RAGE might be a useful predictive marker for IVF success in infertile women with endometriosis undergoing assisted reproduction.

Imbalance of growth factors mRNA expression associated with oxidative stress in the early pregnancy loss.

OBJECTIVE: The aim of this study was to examine the role of growth factors associated with angiogenesis and oxidative stress in the pathogenesis of spontaneous miscarriage. METHODS: We performed a comparative mRNA expression analysis of VEGF, PlGF, Flt-1, Angiogenin and Endoglin using Real-Time PCR, in the placenta and decidua collected from 12 patients presenting with spontaneous abortion and from 14 women undergoing induced abortion, during the first and second trimester of pregnancy. RESULTS: The mRNA expression of Flt-1 was significantly upregulated in the placenta of spontaneous abortions (5.17-fold, IQR: 2.72-9.11, p < 0.01). The placental expression of the soluble isoforms of Flt-1, sFlt-1 e15a and sFlt-1 i13, was also significantly upregulated in spontaneous abortions (sFlt-1 e15a: 2.35-fold, IQR: 0.98-2.83, p < 0.01; sFlt-1 i13: 3.47-fold, IQR: 2.37-5.08, p < 0,05). Placental tmFlt-1, PlGF and Endoglin showed a tendency of higher expression levels in spontaneous abortions, although they did not reach statistical significance (tmFlt-1: 7.42-fold, IQR: 3.58-14.32; PlGF: 2.36-fold, IQR: 0.90-4.12; Endoglin: 1.97-fold, IQR: 1.18-2.43). VEGF and Angiogenin mRNA expression in induced, as well as in spontaneous abortions, did not convey any statistically significant difference. In the decidua, the expression levels of Flt-1 and its splice variants sFlt-1 e15a, sFlt-1 i13 and tmFlt-1 did not show any statistically significant differences, as was the case for the rest of the herein examined growth factors. CONCLUSIONS: In this study, we observed higher levels of sFlt-1 mRNA expression in the placenta of spontaneous abortions, while expression of other growth factors in placenta and decidua remained constant. This suggests that an imbalance of sFlt-1 expression in the placenta might contribute to the pathogenesis of spontaneous abortion, probably via oxidative stress, providing a possible biomarker for prompt identification of this condition.

Expression of Delta Like Ligand 4 (DLL4) in endometrial carcinomas and tumor vasculature.

PURPOSE: Delta like ligand 4 (DLL4) is a transmembrane ligand of the Notch Signalling pathway, that regulates blood vessel sprouting and maturation. We investigated the expression of DLL4 in endometrial cancer. METHODS: DLL4 was assessed in the plasma (with ELISA) and tissues (with immunohistochemistry) 33 patients with endometrial cancer, treated with radical hysterectomy for stage I endometroid carcinoma. The angiogenic activity (AA) of endometrial cancer was quantified by assessing the CD31+ microvessel density (MVD) in the invading tumor front. Vascular maturation index (VMI), defined as the percentage of CD31+ microvessels expressing DLL4, was calculated as the ratio of the CD31+ MVD to the DLL4+ MVD. RESULTS: The angiogenic activity was directly related with the histological grade (p=0.01). The VMI ranged from 0.1 to 0.7 (median 0.34). The concentration of DLL4 in the plasma ranged from 55-81pg/ml (mean 62.8) before, and dropped to 55-62 (mean 58.2) after hysterectomy (p<0.05). DLL4 was also expressed by cancer cells in 17/33 cases. No correlation between DLL4-related parameters with histopathological variables was noted. CONCLUSION: This pilot study shows that DLL4 is overexpressed in endometrial cancer cells, vasculature and is also elevated in the plasma of a fraction of patients before surgery. The percentage of DLL4+ vessels in the penetrating sample ranged from 10-70%, indicating a large difference in the quality of angiogenesis produced between the endometrial tumors of the same histological type and differentiation.

Investigating the impact of different strategies for endometrial preparation in frozen cycles considering normal responders undergoing IVF/ICSI cycles: a multicenter retrospective cohort study.

Uncertainty exists concerning the type, adjunct, or dose of regimen to offer in frozen cycles in infertile women undergoing IVF/ICSI. Current systematic reviews have failed to identify one method of endometrial preparation as being more effective than another, whereas many IVF Units use variable and mixed protocols mainly based on their experience and convenience of use. Thus, we performed a four-center two-arm retrospective cohort study, encompassing 439 cycles in 311 women. The modalities analyzed were: Modified natural cycle without and with luteal support (Groups 1,2) and Hormone Replacement cycle (HRC) with and without GnRHa suppression (Groups 3,4). Various schemes of progesterone and estradiol were used and compared. χ2 tests for categorical data and t-tests for continuous data were employed, stratifying by exposure, along with univariate and multivariable Logistic Regression models and subgroup analyses, according to the number of embryos transferred (1 vs. ≥2) and day of transfer (d2 vs. d5). Group 3 presented with statistically significant higher live birth and miscarriage rates in comparison to Group 4 (RR = 5.87, 95%CI: 2.44-14.14 and RR = 0.19, 95%CI: 0.06-0.60, respectively), findings that persisted in subgroup analyses according to the day of transfer and the number of embryos transferred. Progesterone administration through the combination of vaginal tabs and gel was associated with lower clinical pregnancy rates when compared to tabs (RR = 0.19, 95%CI: 0.05-0.71). The stable estrogen protocol compared to increasing estrogen at day 5 was associated with a higher positive hCG test and clinical pregnancy rate, while the progesterone through vaginal tabs was linked with lower miscarriages compared either with gel or combinations. In conclusion, HRC with GnRHa appears to be superior to HRC without GnRHa, concerning live birth and miscarriage, especially when the number of embryos transferred are ≥2 and irrespective of day of transfer. The use of progesterone vaginal tabs compared to gel or combinations is associated with better outcomes. Age is a significant predictor of a negative hCG test and clinical pregnancy rates. A properly conducted RCT is needed to evaluate the optimal frozen embryo transfer preparation strategy.Abbreviations: SD: standard deviation; BMI: body mass index; PCOS: polycystic ovarian syndrome; IQR: interquartile range; FSH: follicle-stimulating hormone; LH: luteinizing hormone; TSH: thyroid-stimulating hormone.

Administration of low-molecular-weight heparin in patients with two or more unsuccessful IVF/ICSI cycles: a multicenter cohort study.

To compare the effects of the administration of low-molecular-weight heparin (LMWH) in subfertile patients with two or more unsuccessful IVF/ICSI cycles. In this six-center two-arm retrospective cohort study, the study population (230 women) underwent a GnRH-antagonist protocol and was classified into two groups, according to the couse of LMWH or not. Groups were compared regarding the clinical and IVF/ICSI cycle characteristics and reproductive outcomes, whereas clinical pregnancy and miscarriage constituted the primary endpoints. Logistic regression analysis was performed to determine the potential predictors of clinical pregnancy, miscarriage and live birth rates using the Enter method. Baseline characteristics were comparable in the two groups. There was no statistically significant difference between the two study groups with regard neither to clinical pregnancy and miscarriage rates (33/133 vs. 20/97, p = .456 and 15/133 vs. 9/97, p = .624, respectively), nor to the secondary outcomes preset for this study (all p values >.05). Logistic regression revealed that age of the woman and ICSI and dose of gonadotrophins used were predictors of clinical pregnancy and live birth, respectively. In conclusion, there is no evidence to support the standard addition of LMWH in patients with two or more unsuccessful IVF/ICSI cycles.

Endometrial injury for RIF patients undergoing IVF/ICSI: a prospective nonrandomized controlled trial.

To evaluate the effect of endometrial injury on clinical outcomes in subfertile women with repeated implantation failures (RIF) undergoing assisted reproduction. In this prospective nonrandomized controlled trial, 103 subfertile women with RIF were included. Fifty-one underwent endometrial injury through hysteroscopy in the early follicular phase of the previous cycle and 52 underwent the standard protocol without any intervention. Live birth and miscarriage were the primary outcomes. Clinical and in vitro fertilization (IVF) cycle characteristics, were also compared between groups. Both groups were comparable in terms of baseline and cycle characteristics. Live birth rates were significantly higher in the study, compared with the control group (18/51 vs. 8/52, odds ratio (OR) = 0.25; 95% confidence interval (CI) = 0.10-0.64; p = 0.020), although miscarriage rates were similar (7/51 vs. 10/52, OR= 0.25; 95%CI= 0.12-0.66; p = 0.452). The rest of the outcomes parameters were comparable between groups. Logistic regression analysis revealed that endometrial injury and duration of subfertility were independent predictors of live birth after control of other variables (OR = 2.818; 95%CI = 1.044-7.605; p = 0.041 and OR = 0.674; 95%CI = 0.461-0.985, p = 0.042, respectively). Endometrial injury induced through office hysteroscopy in the preceding cycle in subfertile women with RIF improves live birth rates.

Fetal ovarian cysts. Our clinical experience over 16 cases and review of the literature.

OBJECTIVE: Fetal ovarian cysts are intra-abdominal structures frequently diagnosed prenatally, tending to present as isolated unilateral lesions in normal fetuses in the third trimester. These cysts may present with complications and their diameter and echogenicity are the main criteria for establishing their prognosis. Spontaneous regression of fetal ovarian cysts is very usual. In the present study, we present our clinical experience on fetal ovarian cyst surveillance and treatment, as well as a review of the literature in the same field. MATERIAL AND METHOD: In this study, we reviewed pre- and postnatal medical records and ultrasonography of 16 fetuses that were diagnosed with ovarian cysts, in Obstetrics Department of University Hospital of Alexandroupolis, between January 2000 and April 2010. We have also reviewed the available literature about fetal ovarian cysts. RESULTS: In a total of 16 cases, postnatal surgery was performed in one infant due to ovarian cyst torsion. In the remaining 15 cases, cysts regressed completely in two fetuses during pregnancy and all the rest of the cysts, including four complex ones, resolved spontaneously after birth. CONCLUSIONS: When fetal ovarian cysts are detected, they should be followed up by serial ultrasonographic examinations. The majority of them will regress spontaneously in a period of 12 months after birth, independent of their sonographic findings. Only symptomatic cysts or cysts with a diameter >5 cm, which do not regress or enlarge, should be treated.

The contribution of rapid intraoperative cytology in the evaluation of endometrial cancer spread.

INTRODUCTION: Peritoneal washing cytology and imprint cytology of pelvic lymph nodes samples were used to evaluate the rapid cytologic detection of peritoneal and retroperitoneal spread of endometrial cancer. MATERIALS AND METHODS: We undertook a study on 194 endometrial cancer patients who underwent primary treatment in the Gynecologic Clinic, Democritus University of Thrace. All patients were subjected to peritoneal washing (PW) cytology and imprint cytology performed on lymph node sampling. The cytologic specimens were stained by May-Grünwald Giemsa (MGG) and Haematoxylin eosin (HE) techniques. Cell-blocks prepared from peritoneal washings (PWs) and the lymph node samples were sent for histologic examination. The cytologic fi ndings were correlated to histologic results. RESULTS: Rapid intraoperative cytology provides a useful diagnostic technique for the assessment of endometrial cancer spread. HE and MGG stain presented different values of sensitivity and specifi city in the detection of peritoneal and retroperitoneal spread of endometrial cancer. CONCLUSION: Cytologic assessment of intraperitoneal and retroperitoneal spread of endometrial cancer is a rapid, intraoperative procedure, which provides the surgeon with useful information regarding the stage of the disease and the subsequent therapeutic approach.

Transvaginal aspiration of ovarian cysts: our experience over 121 cases.

We performed an evaluation of ultrasound-guided transvaginal aspiration of ovarian cysts as a viable alternative to surgery in 104 reproductive and 17 postmenopausal women. One-hundred and twenty-one patients with a simple >4 cm diameter ovarian cyst, with a benign appearance on ultrasound as well as on clinical and blood examination, underwent transvaginal fine needle aspiration of the cyst under ultrasonographic control. One-hundred and four patients were of reproductive age and 12 were postmenopausal. Sixty women who were of reproductive age and in which OCP treatment was not contraindicated followed a six-month therapy with oral contraceptives after the intervention. In the group of patients of reproductive age under OCP treatment the cyst persisted in nine of the 60 women (recurrence rate 15%). In the other group of patients of reproductive age,under no OCP treatment, the recurrence rate was 47% (21 of the 44 women). In the group of postmenopausal patients, the cyst persisted in ten out of 17 cases (recurrence rate 58,9%). Transvaginal aspiration of ovarian cysts is a reliable alternative to surgery with many advantages such as excellent tolerance, low risk and cost of complications and recurrence. The OCP treatment after aspiration seems to increase the success rate of expectant management.

Comparison of the ultrasonic scalpel to CO(2) laser in cervical conization.

The aim of the present study was to compare the efficacy of CO(2) laser with that of ultrasonic scalpel in cervical conization in terms of intraoperative and postoperative outcome. One-hundred and two patients were submitted to cervical conization by ultrasonically activated scalpel, while 97 patients were submitted to cervical conization by use of CO(2) laser. Comparison of mean estimated blood loss, mean operative time and mean cone volume between the two groups was performed with Student t test. Postoperative complications were compared by x(2) test. There was no statistical significance regarding the mean operating time, mean blood loss, mean cone volume and postoperative complications in the two methods. However, thermal artifacts at the cone margins were minimal in the harmonic group (2/102 cones, 1.96%), while in the laser group they were considerably more (18/97 cones, 18.5%) (p < 0.05). Conization using the harmonic scalpel is as safe and effective as the CO(2) laser procedure. It is cheaper, produces less smoke, better visual field and less thermal artifacts in the cone margins. It is a reliable method that overcomes most problems associated with the CO(2) laser, as well as the other conventional conization procedures.

The contribution of hysteroscopy to the detection malignancy in symptomatic postmenopausal women.

Hysteroscopic evaluation of endometrial cavity is widely used in cases of abnormal uterine bleeding. The aim of the present study is to compare the hysteroscopic and histological findings in women suffering from postmenopausal bleeding. Between 1990 and 2009, 425 women aged 47-83 years were included in the study. None of the women had received hormonal therapy or had any malignancy in the past. All women underwent diagnostic hysteroscopy and histologic sampling of the endometrial cavity. Hysteroscopy was successfully completed in 423 women and was suggestive of malignant lesions in 23 cases, which were confirmed histologically. The hysteroscopic findings in three cases were suggestive of atrophic endometrium, atypical hyperplasia and endometrial carcinoma, and the histologic diagnosis confirmed endometrial carcinoma (two cases) and one uterine sarcoma. Hysteroscopy proves to be a safe and effective technique in the diagnosis and management of abnormal uterine bleeding.

Fetal distress evaluation using and analyzing the variables of antepartum computerized cardiotocography.

OBJECTIVE: In this study, we tried to establish cut-off values for more than one parameters of computerized cardiotocography (c CTG) in the prediction of fetal distress during labor, using a group of pregnant women with low-risk pregnancies. METHOD: A retrospective study was performed. Data were collected from 167 patients for measurements of fetal heart rate (FHR) variables and perinatal outcome. Computerized CTG was performed with an Oxford Sonicaid monitor with connection to a 8000 system for CTG spontaneous analysis. The following c CTG variables were considered: FHR, number of accelerations, the presence and the number of episodes of high and low variation, the number of decelerations, short-term variation (STV), peaks of contractions (per hour) and fetal movements assessed by maternal perception (per hour). Computerized CTG recordings started not earlier than the beginning of week 38 of gestation. Immediately after delivery, blood sample was collected from umbilical artery for umbilical artery blood gas analysis (UBGA). The main UBGA parameter in cord umbilical artery that was considered for analysis was pH. pH values<7.25 were considered as suspicious for acidemia and pH values>or=7.25 as normal. RESULTS: Women suspicious for fetal distress during labor presented significantly lower fetal movements (P=0.026), accelerations (P=0.018), variability (P<0.001), number of high episodes (P<0.001), higher values of FHR baseline (P<0.001) and low episodes (P<0.001). Only the number of decelerations did not differ significantly between the two groups (P=0.545). The cut-off points of 5.00 for STV and 3.00 for high episodes were determined to classify women with fetal distress, which yielded high sensitivities (34 and 52%) and specificities (96.6 and 94.9%), with positive predictive values of 81.0 and 81.3% and negative predictive values of 77.4 and 82.2%, respectively. CONCLUSIONS: In conclusion, we believe that not only STV but also other components of the cCTG, mainly the presence and the number of episodes of high variation, are related to pregnancy's outcome as measured by an umbilical artery pH.

Acute abdomen as initial presentation of gestational choriocarcinoma.

Choriocarcinoma is the most malignant tumor of gestational trophoblastic neoplasia. Choriocarcinoma presenting as postpartum hemorrhage and spontaneous uterine perforation with intra-abdominal hemorrhage is very rare. We present a 29-year-old woman with spontaneous uterine rupture due to choriocarcinoma following a live birth pregnancy. The long time interval (2 years) between the previous live birth pregnancy and the diagnosis of the disease, the acute onset of the disease by uterine rupture as the first symptom and the negative urine hCG test are presented and discussed in this case report.

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

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

Differentiation of myomas by means of biomagnetic and doppler findings.

AIM: To elucidate the hemodynamics of the uterine artery myomas by use of Doppler ultrasound and biomagnetic measurements. METHOD: Twenty-four women were included in the study. Sixteen of them were characterised with large myomas whereas 8 of them with small ones. Biomagnetic signals of uterine arteries myomas were recorded and analyzed with Fourier analysis. The biomagnetic signals were distributed according to spectral amplitudes as high (140-300 ft/ radicalHz), low (50-110 ft/ radicalHz) and borderline (111-139 ft/ radicalHz). Uterine artery waveform measurements were evaluated by use of Pulsatility Index (PI) (normal value PI < 1.45). RESULTS: There was a statistically significant difference between large and small myomas concerning the waveform amplitudes (P < 0.0005) and the PI index (P < 0.0005). Specifically, we noticed high biomagnetic amplitudes in most large myomas (93.75 %) and low biomagnetic amplitudes in most small ones (87.5 %). CONCLUSION: It is suggested that the biomagnetic recordings of uterine artery myomas could be a valuable modality in the estimation of the circulation of blood cells justifying the findings of Doppler velocimetry examination.

Nonlinear analysis of biomagnetic signals recorded from uterine myomas.

OBJECTIVE: To determine if there is any non-linearity in the biomagnetic recordings of uterine myomas and to find any differences that may be present in the mechanisms underlying their signal dynamics. METHODS: Twenty-four women were included in the study. Sixteen of them were characterised with large myomas and 8 with small ones. Uterine artery waveform measurements were evaluated by use of Pulsatility Index (PI) (normal value PI<1.45). RESULTS: Applying nonlinear analysis to the biomagnetic signals of the uterine myomas, we observed a clear saturation value for the group of large ones (mean = 11.35 +/- 1.49) and no saturation for the small ones. CONCLUSION: The comparison of the saturation values in the biomagnetic recordings of large and small myomas may be a valuable tool in the evaluation of functional changes in their dynamic behavior.

Cytologic differential diagnostic problems in ulcerative cervicitis.

OBJECTIVE: To improve the detection rates in the determination of signs consistent with ulcerative cervicitis against signs of intraepithelial neoplasia, in PAP smears. STUDY DESIGN: We investigated the frequency of establishing a diagnosis of a simple ulcerative cervicitis by histology in a cohort of 58 females with cervical cytology suggestive of intraepithelial neoplasia. RESULTS: There was found a detection rate of 81.58% in the determination of ulcerative cervicitis by cytology and a rate of 100% in the determination of HPV-infection, CIN1, CIN2 and CIN3 respectively. There was no statistically significant difference between cytology and histology in the diagnosis of ultracerative cervicitis (p>0.5, chi-square). CONCLUSIONS: There is an overestimation of the cytological signs in the diagnosis of ulcerative cervicitis 18.42%). There is also an excellent agreement in the diagnosis of HPV-infection and CIN grades (100%) between cytology and histology, discordant with the literature (2.4%-71%).

Fetal magnetoencephalography in intrauterine growth retarded pregnancies.

OBJECTIVE: To investigate the fetal brain activity in pre-eclamptic and normal pregnancies. Biomagnetic measurements were performed by means of a superconducting quantum interference device (SQUID) in an electrically shielded room of low magnetic noise. METHODS: The study population included 28 pre-eclamptic (34-37 weeks' gestation) and 19 normal pregnancies (34-37 weeks' gestation). The pre-eclamptic group consisted of 11 cases with birth weight below the 10th percentile (1969 +/- 129 g) and 17 cases above the 10th percentile (2496 +/- 257 g). Biomagnetic signals (waveforms) recorded from fetal brains were expressed in terms of magnetic power spectral amplitudes. RESULTS: The total corresponding spectral amplitudes in the frequency range 2-7 Hz were high (719 +/- 69 Ft/ radical Hz in neonates <10th percentile and 537 +/- 105 Ft/ radical Hz in neonates >10th percentile) in most (82%) pre-eclamptic pregnancies and low (382 +/- 35 Ft/ radical Hz) in most normal pregnancies (95%). The difference between normal and all pre-eclamptic pregnancies was highly significant (Student's t-test: p < 0.0001). Comparing the two groups of pre-eclampsia we found also a highly significant difference (Student's t-test: p < 0.0001). CONCLUSION: Biomagnetic measurements of fetal brain activity could be useful in the assessment of fetal central nervous system development.

Interpregnancy interval and the risk of preterm birth in Thrace, Greece.

OBJECTIVE: To examine the influence of short interpregnancy interval on the prevalence of preterm birth in two, ethically different, Greek populations. STUDY DESIGN: We studied 652 urban Christian women and 578 rural, Romany, Muslim women who had had two consecutive, singleton pregnancies. We related the prevalence of preterm birth to the interpregnancy intervals (cut-off point, 6 months). Student's t-test, x(2)-test and relative risk estimation were used. RESULTS: Preterm birth and interpregnancy intervals less than 6 months occurred more often among Muslims than Christians. Among Muslims, an interval of <6 months was associated with greater prevalence of preterm birth (16% versus 7.3%, P=0.013, RR=2.4 and 95% C.I. 1.3-4.7). Christians did not demonstrate a similar relationship. CONCLUSIONS: A short interpregnancy interval seems to be a risk factor for preterm birth in the population of rural, Romany, Muslim women.