Heterogeneous Immunolocalisation of Zinc Transporters ZIP6, ZIP10 and ZIP14 in Human Normo- and Asthenozoospermic Spermatozoa.

Zinc (in the form of Zn2+) is necessary for male fertility. Both Zn2+ quantity and its localisation have been detected in seminal plasma and ejaculated spermatozoa, suggesting its active uptake via zinc import transporters (ZIPs). Immunofluorescence was used to characterise the expression and localisation of three distinct types of ZIP transporters in ejaculated spermatozoa of normo- and asthenozoospermic sperm samples. ZIP6, ZIP10 and ZIP14 showed heterogeneous sperm cell expression and different compartmental distribution. In both types of sperm samples, ZIP6 and ZIP14 were predominantly localised in the sperm head, while ZIP10 was found along the sperm tail. Compartmental localisation of ZIPs in asthenozoospermia was not changed. However, regarding sub-compartmental localisation in sperm head regions, for ZIP6 asthenozoospermia only decreased its acorn/crescent-like pattern. In contrast, ZIP14 immunostaining was altered in favour of crescent-like, as opposed to acorn-like and acorn/crescent-like patterns. The specific ZIPs localisation may reflect their different roles in sperm cell integrity and motility and may change over time. This is the first report of their specific compartmental and sub-compartmental localisation in ejaculated human sperm cells. Further research will lead to a greater understanding of the roles of ZIPs in sperm cell biology, which could positively influence procedures for human infertility therapy.

Predictors of IVF/ICSI success following treatment of endometriosis as the cause of primary infertility.

OBJECTIVES: Treatment of endometriosis prior to IVF/ICSI could be followed by the significant reduction of ovarian reserve. The aim is to identify potential markers of the IVF/ICSI outcome in patients with endometriosis associated infertility and to evaluate their clinical significance. MATERIAL AND METHODS: The prospective cohort study included 73 patients with primary infertility caused by endometriosis that were subjected to 77 IVF/ICSI cycles. Patients were classified into two groups. In the first group some type of treatment had previously been applied, and in the second group patients were immediately subjected to the IVF/ICSI procedures. RESULTS: When pregnancy was achieved, there were significantly more patients under 35 years of age, more patients with primary infertility duration up to 3 years, and more patients with endometriosis that was previously treated (77.4%) (p < 0.039). In the cases of the successful outcome Endometriosis Fertility Index > 7, lower basal FSH and FSH/LH ratio were found, as well as significantly higher basal E2, basal P4 and AMH. Significantly lower doses of gonadotropins were needed in cases of the successful outcome, and long protocol with agonists was more frequently used. Multivariate logistic regres-sion analysis showed that previous therapy of endometriosis, P4 ≥ 0.7 ng/mL, AMH ≥ 0.9 ng/mL, A class of embryos, and the use of long protocol with agonists were predictors of the successful IVF/ICSI outcome. CONCLUSIONS: Therapy for endometriosis, AMH and P4 levels appeared to be predictors for the successful IVF/ICSI outcome and the use of long protocol with agonists could be advised in these cycles.

Reproductive outcomes of IVF after comprehensive endometriosis treatment: a prospective cohort study.

OBJECTIVES: To evaluate the impact of pharmacological and surgical endometriosis treatment on IVF reproductive outcomes in patients with primary infertility. MATERIAL AND METHODS: The study, conducted over a five year period, included 73 patients with endometriosis associated primary infertility subjected to 77 cycles. Group I included patients treated for endometriosis before the IVF (subgroups A: surgical and pharmacological treatment and B: only surgical treatment). Group II included patients immediately subjected to IVF. Assessed outcomes were pregnancy rate (PR) per started cycle, fertilization rate (FR), implantation rate (IR) and live birth rate (LBR). RESULTS: Group IA included 25 patients, Group IB 21 and Group II 27 patients. FR and IR showed no significant differences between groups. PR was significantly higher in the Group I than Group II (49% vs 25%, p = 0.030). PR per started cycle was the highest in the Group IA and the lowest in the Group II (p = 0.040). LBR was significantly higher in whole Group I (p = 0.043) and subgroup IA (p = 0.020) than Group II. Group IA and IB did not differ regarding examined outcomes. Regression analysis showed that endometriosis pretreatment method can impact both achieving pregnancy (p = 0.036) and having a live born child (p = 0.008) after IVF. The combined surgical and pharmacological endometriosis treatment, shorter infertility duration, lower EFI score, using long protocol with FSH+HMG gonadotropins increase the probability of successful IVF. CONCLUSIONS: A combined surgical and pharmacological endometriosis treatment had a positive impact on IVF reproductive outcomes, both on pregnancy and on live birth rates.

Predictive Value of Basal Serum Progesterone for Successful IVF in Endometriosis Patients: The Need for a Personalized Approach.

The data regarding the role of progesterone (P4) in reproductive events of endometriosis patients are limited. This prospective study aimed to examine the predictive value of basal P4 serum levels for successful in vitro fertilization (IVF) in patients with primary infertility and endometriosis. The study included 73 patients divided according to endometriosis treatment (surgery vs. control-no treatment). The general data, basal hormonal status, and pregnancy rates were determined for every patient. Clinical pregnancy was achieved in 40.3% of patients, and more often in patients treated for endometriosis before IVF. The regression analysis showed that higher basal P4 serum levels were associated with achieving pregnancy through IVF. When regression was adjusted for the patient and IVF characteristics, higher basal P4 serum levels were associated with pregnancy achievement in both groups of women, along with the basal serum levels of FSH, LH, and AMH; EFI score; and stimulation protocol. The ROC analysis showed that the basal P4 serum level for successful IVF should be ≥0.7ng/mL. The basal P4 serum level cut-off for IVF success in endometriosis patients was determined for the first time. Constructed models for IVF success prediction emphasize the importance of determining the basal P4 serum levels for the personalized treatment of endometriosis-related infertility.

Clinical and ultrasonographic parameters in assessment of labor induction success in nulliparous women.

Purpose: Evaluation of simplified Bishop score and ultrasound cervicometry in the assessment of labor induction success in nulliparous women.Methods: Prospective cohort study included 146 nulliparous women with singleton pregnancy and indications for labor induction. Prior to labor induction, cervicometry and Bishop score were determined. Upon delivery, patients were classified as those delivered vaginally and by cesarean section (CS) after unsuccessful labor induction.Results: Bishop score >5 was found in 47.95% of vaginally delivered women and 12.33% of patients delivered by CS (p < .01). Cervicometry had appropriate findings in 34.2% of vaginally delivered women and 75.3% of those delivered by CS (p < .01). Bishop score (>5 versus ≤5) had lower sensitivity (52.05%) and specificity (12.33%) than cervicometry (good versus unfavorable findings) (sensitivity 65.75%, specificity 75.34%) for prediction of labor induction success. If Bishop score was ≤5, cervicometry had 50.0% sensitivity and 78.13% specificity, while if Bishop score was >5, 82.86% sensitivity and 55.56% specificity. Obtained model for predicting labor induction outcome in nulliparous women based on their clinical and ultrasonographical characteristics identified the Bishop score as the most important predictor.Conclusions: Study confirmed the usefulness of simplified Bishop score and ultrasound cervicometry in the assessment of labor induction success in nulliparous women.

Toxic Metal and Trace Element Concentrations in Blood and Outcome of In Vitro Fertilization in Women.

The aim of this study was to investigate the association of trace element and toxic metal concentrations in blood and the outcome of in vitro fertilization (IVF). The study included 104 consecutive patients that underwent assisted reproductive technology (ART) procedures. The following parameters were determined: cadmium (Cd), mercury (Hg), arsenic (As), and lead (Pb); and copper (Cu), zinc (Zn), selenium (Se), and magnesium (Mg). Serum samples were obtained before commencing stimulation. Patients with smoking habit had significantly higher Pb concentrations (P = 0.022), as well as higher concentrations of As and Hg but not significantly. All subjects were divided into groups of pregnant and nonpregnant patients. Pregnant patients had lower mean values of Mg (P = 0.009), As (P < 0.05), and Pb (P = 0.034), compared to nonpregnant, and a significant correlation between pregnancy outcome and concentrations of Mg, Cd, and Pb was found. Women who had had delivered had lower Mg (P = 0.009) and Cd (P = 0.014) concentrations. There was a significant correlation of the negative outcome of IVF procedure with higher concentrations of Pb (P = 0.046) and Cd (P = 0.012). In conclusion, our results suggest that there is a difference in Mg, Pb, and Cd concentrations between pregnant and nonpregnant women. There was no association between toxic metals and number and quality of oocytes and embryos, while there was with fertilization rate. Concerning trace elements, we did not find the correlation of trace elements with oocyte number and quality, nor with a number of fertilized oocytes, except for Cu. Patients who were pregnant had lower concentrations of Mg.

Oxidative Stress Markers in GnRH Agonist and Antagonist Protocols in IVF.

BACKGROUND: Our aim was to study the effect of GnRH agonist and antagonist protocols of ovarian stimulation on oxidative stress parameters in serum and the influence of oxidative stress parameters change on the outcome of IVF cycles. METHODS: This prospective study included 82 patients who underwent IVF procedures. We determined SOD, MDA and SH groups in serum. Serum samples were obtained between the second and fourth day of the cycle and on the day of HCG administration during ovarian stimulation. RESULTS: Patients were divided into two groups depending on the protocol of stimulation. The mean total and mature oocytes number and number of fertilized oocytes were higher in GnRH agonist group. There was no significant difference in biochemical pregnancy, miscarriage and live-birth rate in both groups. Mean serum SOD was significantly lower, while mean serum MDA and SH groups were significantly higher after ovarian stimulation. Delivery rate was higher in patients without OS while miscarriage rate was higher in patients with OS. CONCLUSIONS: Our study confirmed that there is a difference in the concentration of oxidative stress parameters before and after ovarian stimulation. IVF outcome is better in patients without OS after ovarian stimulation. However, the protocol of ovarian stimulation is neither associated with a change in oxidative stress parameters nor with the outcome of ART procedures.

Survivin and VEGF as Novel Biomarkers in Diagnosis of Endometriosis.

BACKGROUND: The aim of this study was to investigate the role of peripheral blood markers as additional diagnostic tools to transvaginal ultrasound (TVU) findings in the diagnosis of endometriosis. METHODS: This study included 40 patients undergoing laparoscopy for suspected endometriosis from January to December 2012. Preoperative levels of serum CA125, CA19-9, CEA and mRNA expression levels for survivin and VEGF were obtained. Real-time PCR was used to determine relative gene expression. A new diagnostic score was obtained by deploying the peripheral blood markers to the TVU findings. Statistical methods used were Chi-square, Fisher's, Student's t-test or the Mann - Whitney test. RESULTS: There was a statistically significant difference in serum CA125, survivin and VEGF levels in patients with endometriosis and those without endometriosis (p<0.001, p=0.025 and p=0.009, respectively). False negative TVU findings were noted in 3/13 patients (23.1%) with peritoneal endometriosis without ovaries involvement. High sensitivity (93.3%), specificity (90.0%), PPV (96.6%), NPV (81.8%) and accuracy (92.5%) were obtained for a diagnostic score based on TVU and significant peripheral blood markers (CA125, survivin and VEGF). CONCLUSIONS: Determination of serum CA125, mRNA expression levels for survivin and VEGF along with TVU can contribute to higher accuracy of the noninvasive diagnostic tools for endometriosis.

[Poor Ovarian Response to Stimulation for In Vitro Fertilization].

The term "poor respond (POR) patients" is used for the group of women who respond badly to usual doses of gonadotropins in in vitro fertilization (IVF) treatments; the consequence is low pregnancy rate. A consensus was reached on the minimal criteria needed to define POR. At least two of the following three features must be present: 1. advanced maternal age (40 years or more) 2. previous POR (3 or less oocytes with a conventional stimulation protocol) 3. abnormal ovarian reserve (AMH 0.5-1.1 ng/ml or AFC 5-7).The aim is to find better therapeutic options for these patients. Increased levels of day 3 follicle stimulating hormone (FSH) and estradiol (E2), as well as decreased levels of anti-Müllerian hormone (AMH) and antral follicle count (AFC), can be used to assess ovarian reserve, as indirect predictive tests. A larger number of well designed, large scale, randomized, controlled trials are needed to assess the efficacy of different management strategies for poor responders: flare up gonadotropin releasing hormone (GnRH) agonist protocols, modified long GnRH agonist mini-dose protocols, luteal initiation GnRH agonist stop protocol, pretreatment with estradiol--GnRH antagonist in luteal phase, natural cycle aspiration or natural cycle aspiration GnRH antagonist controlled, adjuvant therapy with growth hormone or dehydroepiandrosterone (DHEA). The results of up to now used protocols are unsatisfactory and stimulation of the ovulation in poor responders remains a challenge, especially when bearing in mind that in the majority of cases the patients will be menopausal in relatively short period of time.

Ruptured corpus luteum cyst in early pregnancy: a case report.

INTRODUCTION: Transvaginal sonography and human chorionic gonadotropin (hCG) testing are cornerstones of modern clinical practice in cases with the suspected ectopic pregnancy. In unclear cases, if the level of hCG is above the discriminatory zones, the use of uterine curettage is recommended.There is an increasing concern that strict observation of the guidelines would potentially harm otherwise normal early intrauterine pregnancies in certain cases. CASE OUTLINE: A 35-year-old woman was admitted to hospital due to a severe lower abdominal pain. Based on the positive pregnancy test and sonographic exams which failed to demonstrate intrauterine pregnancy, the diagnosis of ectopic pregnancy was presumed. Laparoscopy revealed ruptured corpus luteum cyst and the diagnosis was confirmed on histopathological finding. Postoperatively, normal intrauterine gestation was visualised. CONCLUSION: Since the diagnosis of early pregnancy and its complications can be misleading, in unclear cases, we support the expectative "wait and see" management consisting of serial hCG testing and repeated ultrasound examinations. Avoidance of uterine curettage in such unclear cases would further reduce the possibility of normal early pregnancy interruption.