Investigation of the Effect of Seminal Plasma Exosomes from the Normal and Oligoasthenoteratospermic Males in the Implantation Process.

Background: Seminal plasma exosomes are now recognized to play a complex role in the regulation of the female reproductive system infertility. The objective of this study was to assess the effect of exosomes derived from the sperm of men with oligoasthenoteratozoospermia on endometrial implantation-related genes. Methods: To isolate the exosomes, we employed an ultracentrifugation method on samples derived from 10 fertile men with normal sperm parameters and 10 men with oligoasthenoteratozoospermia. The size distribution and ultrastructure of the exosomes were then characterized using transmission electron microscopy and dynamic light scattering. We detected an exosome marker using western blot analysis and confirmed the cytoplasmic localization of the exosomes by incubating them with DiI dye and visualizing them using fluorescence microscopy. After 6 hours of in vitro treatment of endometrial epithelial cells with 100 µg/ml seminal exosome, the endometrial receptivity genes were examined using qRT-PCR. To perform data analysis and quantification, we utilized Image J and Prism software. P< 0.05 were considered statistically significant. Results: After 6 hours of treatment, the mRNA levels of MUC1, LIF, G-CSF, CX3CL1, and VEGF were significantly downregulated in the endometrial epithelial cells treated with oligoasthenoteratozoospermia exosomes compared to the normal group. Although changes were observed in the mean mRNA levels of IL8 and TGF-ß genes in the oligoasthenoteratozoospermia group compared to the normal group, these differences did not reach statistical significance (p > 0.05). Conclusions: Oligoasthenoteratozoospermia exosomes have a distinct effect on endometrial receptivity compared to normal exosomes, leading to reduced expression of implantation-related genes.

Diagnostic value and agreement of transrectal in comparison with transvaginal sonography among women with abnormal uterine bleeding.

AIMS: Transvaginal sonography (TVS) is not applicable for evaluating the endometrium in some cases with abnormal uterine bleeding (AUB) such as virgin women. The aim of the present study was to investigate the diagnostic value of transrectal sonography (TRS), as a non-invasive alternative for TVS, among women with AUB. METHODS: In this cross-sectional study, 40 women over 40 years of age with AUB first underwent TRS and then TVS. Endometrial biopsy was also performed as a gold standard method to evaluate endometrial pathologies. The findings of the two endometrial ultrasound methods were compared with the biopsy results. Also, the findings of the two methods were compared in terms of the agreement on other uterine markers and adnexal findings. RESULTS: There was a high agreement between the findings of TVS and TRS for uterine size, endometrial thickness and homogeneity, endomyometrial junction, myometrial homogeneity, uterine lesions, uterine lesions size, ovarian size and ovarian echogenicity. When comparing the results of these two methods with biopsy findings, sensitivity, specificity as well as positive and negative predictive values of both methods were similar (82%, 100%, 100%, and 0%, respectively). CONCLUSION: When it is not possible to use TVS in women with AUB, TRS is a suitable, non-invasive and inexpensive alternative method for evaluating the endometrium and other genital organs.

Anti-Müllerian Hormone Predictive Levels to Determine The Likelihood of Ovarian Hyper-Response in Infertile Women with Polycystic Ovarian Morphology.

BACKGROUND: The objective of this study was to investigate serum levels of anti-Müllerian hormone (AMH) in normal-ovulatory infertile women with polycystic ovarian morphology (PCOM) and their association with ovarian hyper-response. MATERIALS AND METHODS: This prospective cohort study was carried out on 100 infertile women with PCOM who were treated with an antagonist/agonist triggered stimulation protocol at Shahid Akbar-Abadi Hospital IVF Centre, Tehran, Iran. Serum AMH levels were measured before starting the assisted reproductive technology (ART) cycle and the ovarian hyper-response was evaluated by retrieved oocyte numbers, ooestradiol levels on the triggering day, and the incidence of ovarian hyper-stimulation syndrome (OHSS) clinical signs and symptoms. Logistic regression and the area under the curve (AUC) were used to estimate the effects of AMH and the accuracy of the test. RESULTS: Receiver operating characteristic (ROC) curve analysis showed that AMH could significantly predict ovarian hyper-response in PCOM patients (AUC=0.73). The estimated threshold value was 4.95 ng/ml, with a specificity of 74.58% (95% confidence interval [CI]: 50.85, 93.22) and sensitivity of 73.17% (95% CI: 48.78, 92.68). Logistic regression results showed a significant interaction between AMH and body mass index (BMI, P=0.008), which indicated that BMI had a moderation effect. CONCLUSION: Individualized stimulation protocols for patients with isolated PCOM and AMH greater than 4.95 ng/ml may significantly reduce the chances of developing OHSS. However, the AMH cut-off values to predict ovarian hyperresponse differ for different BMI categories among PCOM patients; thus, it becomes a more precise predictive marker with increasing BMI.

Differences in and Correlates of Sexual Function in Infertile Women with and without Polycystic Ovary Syndrome.

BACKGROUND: The aim of this study was to examine sexual function and its correlates among infertile women with polycystic ovary syndrome (PCOS) in comparison with their non-PCOS counterparts. MATERIALS AND METHODS: In this case-control study, 209 infertile women (116 PCOS and 93 non-PCOS) from Tehran, Iran, were evaluated in February and March 2018. Female sexual function index (FSFI), hormonal status, and documented reports of hyperandrogenic manifestations of the patients were investigated. RESULTS: The mean age of the patients was 32.00 ± 5.00 years old. Eighty-four (40.2%) patients including 42.2% of the PCOS patients and 37.6% of the non-PCOS cases (P>0.05), were suspected of female sexual dysfunction (FSD). The most impaired functions in both groups were desire and arousal. Sexual function was not significantly different between the groups. However, PCOS women had more orgasm problems and acne worsened their sexual function. Total FSFI was positively associated with prolactin level but negatively associated with central obesity in the non-PCOS group; it was negatively correlated with marital duration in the PCOS group. Luteinizing hormone (LH) and pain, prolactin level and lubrication, and central obesity and arousal were correlated in the non-PCOS women. Prolactin level and orgasm, marital duration and arousal, and marital duration and the total FSFI were correlated in the PCOS women. CONCLUSION: Sexual function was similarly low in infertile PCOS and non-PCOS women. However, orgasm problems and the negative effect of acne varied between the two groups. Further investigations may target how hormonal profile may affect sexual function. Practitioners should scrutinize the specific impaired sexual domains and their correlated conditions in PCOS women, notably orgasm, acne, and prolactin level. Interventions should be well tailored based on particular needs of infertile PCOS women.

Effect of Vitamin D3 on Mitochondrial Biogenesis in Granulosa Cells Derived from Polycystic Ovary Syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrine disorder diagnosed by anovulation hyperandrogenism. Hyperandrogenism increases apoptosis, which will eventually disturb follicular growth in PCOS patients. Since mitochondria regulate apoptosis, they might be affected by high incidence of follicular atresia. This may cause infertility. Since vitamin D3 has been shown to improve the PCOS symptoms, the aim of study was to investigate the effects vitamin D3 on mtDNA copy number, mitochondrial biogenesis, and membrane integrity of granulosa cells in a PCOS-induced mouse model. MATERIALS AND METHODS: In this experimental study, the PCOS mouse model was induced by dehydroepiandrosterone (DHEA). Granulosa cells after identification by follicle-stimulating hormone receptor (FSHR) were cultured in three groups: 1. granulosa cells treated with vitamin D3 (100 nM for 24 hours), 2. granulosa cells without any treatments, 3. Non-PCOS granulosa cells (control group). Mitochondrial biogenesis gene (TFAM) expression was compared between different groups using real-time PCR. mtDNA copy number was also investigated by qPCR. The mitochondrial structure was evaluated by transmission electron microscopy (TEM). Hormonal levels were measured by an enzymelinked immunosorbent assay (ELISA) kit. RESULTS: The numbers of pre-antral and antral follicles increased in PCOS group in comparison with the non-PCOS group. Mitochondrial biogenesis genes were downregulated in granulosa cells of PCOS mice when compared to the non-PCOS granulosa cells. However, treatment with vitamin D3 increased mtDNA expression levels of these genes compared to PCOS granulosa cells with no treatments. Most of the mitochondria in the PCOS group were spherical with almost no cristae. Our results showed that in the PCOS group treated with vitamin D3, the mtDNA copy number increased significantly in comparison to PCOS granulosa cells with no treatments. CONCLUSION: According to this study, we can conclude, vitamin D3 improves mitochondrial biogenesis and membrane integrity, mtDNA copy number in granulosa cells of PCOS mice which might improve follicular development and subsequently oocyte quality.

Vitamin D3 affects mitochondrial biogenesis through mitogen-activated protein kinase in polycystic ovary syndrome mouse model.

Polycystic ovarian syndrome (PCOS) is a disorder characterized by oligomenorrhea, anovulation, and hyperandrogenism. Altered mitochondrial biogenesis can result in hyperandrogenism. The goal of this study was to examine the effect of vitamin D3 on mitochondrial biogenesis of the granulosa cells in the PCOS-induced mouse model. Vitamin D3 applies its effect via the mitogen-activated pathway kinase-extracellular signal-regulated kinases (MAPK-ERK1/2) pathway. The PCOS mouse model was induced by the injection of dehydroepiandrosterone (DHEA). Isolated granulosa cells were subsequently treated with vitamin D3, MAPK activator, and MAPK inhibitor. Gene expression levels were measured using real-time polymerase chain reaction. MAPK proteins were investigated by western blot analysis. We also determined reactive oxygen species (ROS) levels with 2', 7'-dichlorofluorescein diacetate. Mitochondrial membrane potential (mtMP) was also measured by TMJC1. Mitochondrial biogenesis (peroxisome proliferator-activated receptor gamma coactivator 1-α and nuclear respiratory factor), antioxidant (superoxide dismutase, glutathione peroxidase, and catalase), and antiapoptotic (B-cell lymphoma-2) genes were upregulated in the PCOS mice that treated with vitamin D3 compared with the PCOS mice without any treatment. Vitamin D3 and MAPK activator-treated groups also reduced ROS levels compared with the nontreated PCOS group. In summary, vitamin D3 and MAPK activator increased the levels of mitochondrial biogenesis, MAPK pathway, and mtMP markers, while concomitantly decreased ROS levels in granulosa cells of the PCOS-induced mice. This study suggests that vitamin D3 may improve mitochondrial biogenesis through stimulation of the MAPK pathway in cultured granulosa cells of DHEA-induced PCOS mice which yet to be investigated.

Causal Effects of Body Mass Index and Maternal Age on Oocyte Maturation in Assisted Reproductive Technology: Model-Average Causal Effect and Bayesian LASSO Method.

BACKGROUND: Body Mass Index (BMI) and maternal age are related to various disorders of the female reproductive system. This study aimed to estimate the causal effects of BMI and maternal age on the rate of meta-phase II oocytes (MII) using a new statistical method based on Bayesian LASSO and model averaging. METHODS: This investigation was a historical cohort study and data were collected from women who underwent assisted reproductive treatments in Tehran, Iran during 2015 to 2018. Exclusion criteria were gestational surrogacy and donor oocyte. We used a new method based on Bayesian LASSO and model average to capture important confounders. RESULTS: Overall, 536 cycles of 398 women were evaluated. BMI and Age had inverse relationships with the number of MII based on univariate analysis, but after adjusting the effects of other variables, there was just a significant association between age and the number of MII (adjusted incidence rate ratio (aIRR) of age =0.989, 95% CI: [0.979, 0.998], P=0.02). The results of causal inference based on the new presented method showed that the overall effects of age and BMI of all patients were significantly and inversely associated with the number of MII (both P<0.001). Therefore the expected number of MII decreased by 0.99 for an increase of 1 year (95% CI: [-1.00, -0.97]) and decreased by 0.99 for each 1-unit increase in BMI (95% CI: [-1.01, -0.98]). CONCLUSION: Maternal age and BMI have significant adverse casual effects on the rate of MII in patients undergoing ART when the effects of important confounders were adjusted.

The myo-inositol effect on the oocyte quality and fertilization rate among women with polycystic ovary syndrome undergoing assisted reproductive technology cycles: a randomized clinical trial.

PURPOSE: The aim of the present study was to evaluate the effect of myo-Inositol administration on oocyte quality, fertilization rate and embryo quality in patients with PCOS during assisted reproductive technology (ART) cycles. METHODS: Fifty infertile PCOS patients were randomly designated in two groups. In the study group, patients received daily doses of 4 g myo-Inositol combined with 400 mg folic acid and in the control group patients received only 400 mg folic acid from 1 month before starting the antagonist cycle until the day of ovum pick up. Oocyte and embryo qualities were assessed according to European Society of Human Reproduction and Embryology (ESHRE) guidelines. The gene expression of PGK1, RGS2 and CDC42 as a factor of oocyte quality in granulosa cells was analyzed using real-time RT-PCR. Levels of total antioxidant capacity (TAC) and reactive oxygen species (ROS) were evaluated by chemiluminescence assay in follicular fluid. RESULTS: The percentage of metaphase II oocyte, fertilization rate and embryo quality significantly improved in the study group (p < 0.05), but the number of retrieved oocytes and follicle count were not statistically different between groups. Furthermore, the gene expression of PGK1, RGS2 and CDC42 was significantly higher in the study group (p < 0.05) but no differences were found between two groups in terms of TAC and ROS levels. CONCLUSIONS: The present study findings suggest that myo-Inositol alters the gene expression in granulosa cells and improves oocyte and embryo quality among PCOS patients undergoing ART.

Comparison of uterine and ovarian stromal blood flow in patients with polycystic ovarian syndrome.

Polycystic ovarian syndrome (PCOS) is one of the most common endocrine disorders. The aim of this study was to find the correlation between color Doppler ultrasound and serum tests as auxiliary diagnostic criteria in areas where there is no possibility of some tests. A total of 108 patients were enrolled. They were divided into three groups including patients with PCOS, patients with PCOA ultrasound, patients with ovaries and normal hormone tests. Transvaginal sonography was performed from three groups and the results were evaluated in gray scale. The volume of the ovary, the number of follicles and the placement of follicles were recorded using using Doppler spectrum of uterine artery and ovarian stroma. Their arterial resistance index was also calculated. In the next step, serum samples were evaluated to determine the level of LH, FSH, free testosterone, DHEAS and 17-OHP hormones in the early follicular phase. Gray scale ultrasonographic findings (volume and number of ovarian follicles) as well as LH values were higher in patients with PCOS than those in the other two groups. These results proved the reliability of using these factors in the prediction of PCOS. In this study, Doppler indexes did not correlate with the size of the ovaries, the number of ovarian follicles and the measured hormone levels. The findings of transvaginal ultrasound and investigating the relationship with clinical and laboratory outcomes, a more suitable pattern could be chosen for more accurate patient selection and, leading to timely treatment and reducing the complications of the disease.

Comparison of the pregnancy outcomes and the incidence of fetal congenital abnormalities in infertile women treated with letrozole and clomiphene citrate.

OBJECTIVES: The aim of the study was to evaluate the incidence of congenital fetal anomalies reported among infertile Iranian women treated with letrozole compared to those treated with Clomiphene Citrate (CC). METHODS: This retrospective case-control study was performed based on information available from infertile patients referred to the Akbar-Abadi Hospital IVF Center, Tehran, Iran, undergoing ovulation induction (OI) or intrauterine insemination (IUI) cycles from 2007 to 2014.We compared sonographic findings, fertility results, pregnancy complications and congenital anomalies in two groups of women treated with letrozole and CC. RESULTS: Overall, the results of the 2009 cycles including 1237 CC cycles and 772 letrozole cycles were evaluated. In spite of a higher chance of ovulation following CC treatment, overall fertility rates were similar in the two treatment groups. The frequency of adverse outcomes of pregnancy was similar in the two groups, except for the incidence of first trimester abortion, which was significantly higher in the CC-treated group. The frequency of fetal anomalies and major chromosomal abnormalities in the Letrozole group was 4.76% (five cases), and in the CC group, it was 2.12% (three cases). This difference was not statistically significant. CONCLUSION: Based on the findings of this study, it seems that the incidence of congenital anomalies in offspring after OI with letrozole is not increased compared to the CC group.

Breast Cancer and Ovulation Induction Treatments.

BACKGROUND: This study was performed to determine whether the use of ovulation induction drugs in treatment of infertility have a significant effect on the risk of breast cancer. PATIENTS AND METHODS: This case control study (928 cases, 928 controls), was performed in the gynecology and oncology clinics of Shahid Beheshti University of Medical Sciences between 2011 and 2013. Data were collected via in-person interviews using a questionnaire, which included demographic and gynecologic information. Statistical analysis was performed using SPSS statistics software version 20 (IBM Corp). RESULTS: The use of ovulation induction drugs was not significantly associated with an increased risk of breast cancer (odds ratio [OR], 1.13; 95% confidence interval [CI], 0.7-1.855) among women with infertility (OR, 1.28; 95% CI, 0.8-1.95). CONCLUSION: We observed no statistically significant relationship between infertility and ovulation induction drugs with the risk of breast cancer, except for significant increases in the risk of breast cancer among patients who had used fertility drugs for >6 months.

The relationship between serum vitamin D level and premenstrual syndrome in Iranian women.

BACKGROUND: Premenstrual syndrome (PMS) is among the most unfavorable problems in women in reproductive age; however its pathophysiology is still not fully confirmed. Vitamin D as an immunomodulator could prevent inflammatory state before and during menstruation. OBJECTIVE: The aim was to investigate whether there is any relationship between serum vitamin D levels and PMS. MATERIALS AND METHODS: In total, 82 women participate in this case-control study which was conducted in Shahid Akbar-abadi hospital from November 2013 to March 2015. Categorization was based on an Iranian version of the premenstrual symptoms screening tool (PSST). Levels of 25 hydroxy-vitamin D3 (25OHD) were determined by using 25-OH Vitamin D ELISA kit in luteal phase. Characteristics of participants and vitamin D levels were compared between two groups by using independent sample t-test. RESULTS: Menarche age of women with PMS was significantly lower than normal women (p=0.04). Body mass index was not statistically different between groups. We observed a high rate of vitamin D deficiency and also its severe deficiency in both PMS and non-PMS groups. However, our study demonstrated no significant difference in the levels of serum 25OHD between the two groups. CONCLUSION: It seems there is no association between PMS and serum levels of vitamin D3; however, the high rate of vitamin D deficiency among young Iranian women emerges special health care considerations in this group.

Treatment results of high dose cabergoline as an adjuvant therapy in six patients with established severe ovarian hyper stimulation syndrome.

BACKGROUND: The beneficial role of cabergoline as a prophylactic agent to prevent ovarian hyper stimulation syndrome (OHSS) among high-risk patients has been demonstrated in previous studies. But data for its role as a treatment for established severe OHSS is still limited. We represent the treatment results of high dose oral cabergoline in management of six patients after the syndrome is established. CASE: High-dose oral cabergoline (1 mg daily for eight days) was prescribed as an adjuvant to symptomatic treatment for six hospitalized patients with established severe OHSS following infertility treatment cycles. In two cases OHSS resolved rapidly despite the occurrence of ongoing pregnancy. CONCLUSION: Considering the treatment outcomes of our patients, high dose cabergoline did not eliminate the need for traditional treatments, but it was a relatively effective and safe therapy in management of established severe OHSS, and prevented the increase in its severity following the occurrence of pregnancy.

Women's knowledge and attitude towards mode of delivery and frequency of cesarean section on mother's request in six public and private hospitals in Tehran, Iran, 2012.

AIM: The rate of cesarean section (CS) has been reported to be as high as 40% among Iranian women in the year 2009. The aim of this study was to determine the rate of cesarean delivery on mother's request (CDMR) and to determine maternal attitude and knowledge about various modes of delivery in private and public (university) hospitals in Tehran. MATERIAL AND METHODS: All primiparous mothers delivering in six selected hospitals between April 2010 and March 2011 were included. Trained investigators handed a predesigned questionnaire to mothers 1 day after delivery to be filled out in the presence of the investigator. RESULTS: From 600 deliveries, 501 (83.5%) were CS and 99 (16.5%) were normal vaginal delivery. The CS rates in university hospitals versus private hospitals were 78.5% and 91.9%, respectively. In total, mothers' knowledge scores were poor, intermediate, and good in 55.6%, 37.9%, and 6.5% of cases, respectively, and no significant difference in knowledge was observed between mothers attending private or public hospitals. The overall rate of CDMR was 20.8%; and the most frequent reason was fear of pain. Women with CDMR were at higher marital age, education, insurance coverage, and socioeconomic status compared with the women with vaginal delivery. CONCLUSION: Prompt action is needed to reduce the unacceptably high rate of unwarranted cesarean deliveries. Improving women's knowledge about the risks and benefits of different modes of delivery can lead to a positive maternal attitude towards vaginal delivery.

Super infection of an ovarian dermoid cyst with actinomyces in an infertile woman.

We present super infection of an ovarian dermoid cyst with actinomyces in an infertile patient. This is a case-report study for evaluation a couple with male factor infertility, who was a good candidate for intracytoplasmic sperm injection (ICSI), while a 10 cm dermoid cyst was found in the woman's right ovary. Patient complained of pelvic pain, intermittent fever, dysmenorrhea, and dyspareunia. The cyst was extracted using laparoscopy, whilst in histopathological examination, an actinomycosis super infection was reported. Actinomyc super infection of an ovarian dermoid cyst is a very rare incident which can also occur in women with no history of intrauterine device (IUD) usage or previous fertility.