Human uterine fluid lavage-derived extracellular vesicle isolation: a comparative study for minimally invasive endometrial receptivity assessment.

RESEARCH QUESTION: Does pre-implantation uterine fluid lavage (UFL) of patients undergoing IVF and frozen embryo transfer (FET) affect implantation and clinical pregnancy rates? Which methods among ultracentrifugation, sucrose cushion and qEV column are suitable for isolating UFL extracellular vesicles? DESIGN: First, UFL was collected from 20 patients undergoing IVF and FET 2 days before embryo transfer as the case group. The control group consisted of 20 patients undergoing IVF and FET patients without lavage. All patients were monitored for 6 weeks. In the next step, the UFLs (n = 30) were collected and pooled. The UFL-derived extracellular vesicles were extracted by ultracentrifugation, sucrose cushion and qEV column methods and characterized. RESULTS: Preimplantation uterine lavage sampling did not affect implantation and clinical pregnancy rates. Extracellular vesicles were successfully isolated from UFL by all three methods. Scanning electron microscopy and dynamic light scattering analysis showed that the isolated vesicles were morphologically spherical. The qEV technique showed that they were smaller and homogenized in size. SDS-PAGE of extracellular vesicles showed a weaker albumin band in the qEV column. Western blot analysis indicated that the isolated extracellular vesicles by the qEV column were more immunoreactive for all the common extracellular vesicle markers (CD81, CD9, CD63, and TSG101). Six reference genes were compared by real-time polymerase chain reaction in the isolated extracellular vesicle subpopulations, and lowest cycle threshold value was observed for the 18SrRNA gene. CONCLUSIONS: The isolation of endometrial secretome extracellular vesicles is a minimally invasive procedure for individual assessment of endometrial receptivity and can be carried out during conception cycles along with transvaginal ultrasonography. Molecular analysis of UFL-derived extracellular vesicle components could suggest biomarkers to determine precise extracellular vesicle timing.

Prevalence of generalized anxiety disorder and its related factors among infertile patients in Iran: a cross-sectional study.

BACKGROUND: Generalized anxiety disorder (GAD) is one of the most prevalent anxiety disorders among infertile patients. This study aimed to determine the prevalence of GAD and its associated factors among infertile patients in Tehran, Iran. METHODS: This cross-sectional study included 1146 infertile patients in a referral fertility center in Tehran, Iran between May and October 2017. GAD was measured using the Generalized Anxiety Disorder-7 (GAD-7) scale. The associations between GAD and demographic/fertility characteristics were estimated using simple and multiple logistic regression with odds ratio (OR) and 95% confidence interval (CI). RESULTS: The mean total GAD-7 score was 6.61 (SD = 5.32). Using a cut-off value of 10, the prevalence of GAD was 28.3%. In adjusted analysis, female sex (OR = 2.54, 95% CI = 1.88-3.42, P < 0.001), low educational level (OR = 1.45, 95% CI = 1.08-1.94, P = 0.012), high infertility duration (OR = 1.05, 95% CI = 1.01-1.09, P = 0.013), and treatment failure (OR = 1.52, 95% CI = 1.13-2.04, P = 0.006) were associated with GAD. CONCLUSIONS: The prevalence of GAD is relatively high in infertile patients. We conclude that all infertile patients should be screened for symptoms of GAD and treated for this disorder as need arises.

Factors associated with poor quality of life among Iranian infertile women undergoing IVF.

Infertility is a medical and social condition and has a considerable impact on a person's quality of life (QoL). The aim of the study was to determine the QoL of women with fertility problem, and identify factors associated with poor QoL. This cross-sectional study included 155 women with fertility problems undergoing IVF in a referral fertility center in Tehran, Iran. The Fertility Quality of Life, the Hospital Anxiety and Depression Scale and demographic and fertility information questionnaire were administered to all women. The mean total QoL score was 62.57 ± 16.89. Multivariate analysis showed that the anxiety (ß = -1.59, p < 0.001) and depression (ß = -2.09, p < 0.001) had a negative impact on QoL. Cause of infertility and failure in previous treatment were also significant factors of poorer QoL. The findings indicate that the QoL was worse in women with high depression and anxiety level, failure in previous treatment and unknown cause of infertility. Thus, a comprehensive approach, including psychosocial interventions and support, is absolutely essential to improve the QoL in these women.

Joint Modeling of In Vitro Fertilization Outcomes among A Population of Iranian Infertile Couples: A Historical Cohort.

BACKGROUND: Women who undergo in vitro fertilization (IVF) cycles should successfully go via multiple stages (i.e., clinical pregnancy, no abortion under 12 weeks, no abortion under 20 weeks, and delivery) to achieve a live birth. In this study, data from multiple IVF cycles and its multiple stages were reanalyzed to illustrate the success factors associated with various stages of IVF cycles in a population of Iranian infertile women. MATERIALS AND METHODS: This historical cohort study includes 3676 assisted reproductive technology (ART) cycles. Covariates take into account in this study were women's age, type of infertility (primary, secondary), body mass index (BMI), cause of infertility, history of abortion, duration of infertility, number of oocytes, number of embryos, fertilization rate, semen factors (Spermogram) and having polycystic ovarian syndrome (PCOS) during IVF cycles. Joint modeling was fitted to apply informative cluster size. RESULTS: Increasing age un women was associated with an increase in the BMI and a positive history of abortion and PCOS, and also, an increase in the number of treatment cycles, while in men was associated with the negative spermogram. With the increase in the number of treatment cycles, the result of the IVF success decreased, but with the increase in the number of embryos, fertilization rate and also, quality and / or quantity parameters of spermogram, we encountered with an increase in the IVF success rate. CONCLUSION: It seems that a joint model of the number of treatment cycles and the result of IVF is a valuable statistical model that does not ignore the significant effect of cycle numbers, while this issue is ignored usually in the univariate models.

TIMPs Expression as A Maternal Cell Free Plasma Biomarker of Severe Preeclampsia: A Case-Control Study.

OBJECTIVE: Preeclampsia (PE) is a pregnancy related disorder with prevalence of 6-7%. Insufficient trophoblastic invasion leads to incomplete remodeling of spiral arteries and consequent decrease in feto-placental perfusion. Altered placental expression of tissue inhibitors of matrix metalloproteinase (TIMPs) is considered to be involved in this process while the balance between matrix metalloproteinases (MMPs) and TIMPs contributes to remodeling of the placenta and uterine arteries by degradation and refurbishing of extracellular matrix (ECM). Therefore, TIMPs, fetal expression pattern was evaluated with the aim of its potential to be used as a determinant for the (early) detection of PE. MATERIALS AND METHODS: In this case-control study, cell free fetal RNA (cffRNA) released by placenta into the maternal blood was used to determine expression patterns of TIMP1, 2, 3 and 4 in the severe preeclamptic women in comparison with the normal pregnant women. Whole blood from 20 preeclamptic and 20 normal pregnant women in their 28-32 weeks of gestational age was collected. The second control group consisted of 20 normal pregnant women in either 14 or 28 weeks of gestation (each 10). cffRNA was extracted from plasma and real-time polymerase chain reaction (PCR) was done to determine the expression levels of TIMP1, 2, 3 and 4 genes. RESULTS: Statistical analysis of the results showed significant higher expression of TIMP1-4 in the preeclamptic women in comparison with the control group (P=0.029, 0.037, 0.037 and 0.049, respectively). Also, an increased level of TIMPs expression was observed by comparing 14 to 28 weeks of gestational age in the normal pregnant women in the second control group. CONCLUSION: An increased cffRNA expression level of TIMPs may be correlated with the intensity of placental vascular defect and may be used as a determinant of complicated pregnancies with severe preeclampsia.

Role of gender in explaining metabolic syndrome risk factors in an Iranian rural population using structural equation modelling.

Many factors can lead to an increase in the prevalence of metabolic syndrome (MetS) in different populations. Using an advanced structural equation model (SEM), this study is aimed to determine the most important risk factors of MetS, as a continuous latent variable, using a large number of males and females. We also aimed to evaluate the interrelations among the associated factors involved in the development of MetS. This study used data derived from the Fasa PERSIAN cohort study, a branch of the PERSIAN cohort study, for participants aged 35 to 70 years with 10,138 males and females. SEM was used to evaluate the direct and indirect effects, as well as gender effects of influencing factors. Results from the SEM showed that in females most changes in MetS are described by waist circumference (WC), followed by hypertension (HP) and triglyceride (TG), while in males most changes in MetS are described by WC, followed by TG then fasting blood glucose (FBG). Results from the SEM confirmed the gender effects of social status on MetS, mediated by sleep and controlled by age, BMI, ethnicity and physical activity. This study also shows that the integration of TG and WC within genders could be useful as a screening criterion for MetS in our study population.

Risk Factors Associated with Recurrent Pregnancy Loss and Outcome of Pre-Implantation Genetic Screening of Affected Couples.

BACKGROUND: Recurrent pregnancy loss (RPL) is a multifactorial disorder which affects up to 5% of couples around the world. Several factors are considered to be involved in RPL; but, the etiology remains unexplained in 35-60% of cases. The aim of this study was to assess the frequency of risk factors associated with RPL in a group of our clinic clients, and their pre-implantation genetic screening (PGS) outcome. MATERIALS AND METHODS: We designed a retrospective descriptive study among, 602 Iranian couples referred to the Royan Reproductive Clinic (Tehran-Iran) from 2006 to 2018. Their karyotyping test and PGS outcomes were analyzed. PGS had been applied by array comparative genomic hybridization (array-CGH) on embryos from these patients. Also, karyotyping test had been performed using standard cytogenetic techniques. RESULTS: G-banding analysis revealed a frequency of 15.61% chromosomal abnormalities in RPL couples. Also, the reciprocal translocations were more frequent (33/1204 cases) compared to the other structural abnormalities. Pregnancy rate per embryo transferred were 50% with array-CGH approach. CONCLUSION: Our findings could confirm a positive correlation between chromosomal abnormalities and RPL rate. Applying PGS for the RPL couples, leads to improvement of pregnancy success rate.

Factors Associated with In Vitro Fertilization Live Birth Outcome: A Comparison of Different Classification Methods.

BACKGROUND: In vitro fertilization (IVF) is a useful assisted reproductive technology to achieve pregnancy in infertile couples. However, it is very important to optimize the success rate after IVF by controlling for its influencing factors. This study aims to classify successful deliveries after IVF according to couples' characteristics and available data on oocytes, sperm, and embryos using several classification methods. MATERIALS AND METHODS: This historical cohort study was conducted in a referral infertility centre located in Tehran, Iran. The patients' demographic and clinical variables for 6071 cycles during March 21, 2011 to March 20, 2014 were collected. We used six different machine learning approaches including support vector machine (SVM), extreme gradient boosting (XGBoost), logistic regression (LR), random forest (RF), naïve Bayes (NB), and linear discriminant analysis (LDA) to predict successful delivery. The results of the performed methods were compared using accuracy tools. RESULTS: The rate of successful delivery was 81.2% among 4930 cycles. The total accuracy of the results exposed RF had the best performance among the six approaches (ACC=0.81). Regarding the importance of variables, total number of embryos, number of injected oocytes, cause of infertility, female age, and polycystic ovary syndrome (PCOS) were the most important factors predicting successful delivery. CONCLUSION: A successful delivery following IVF in infertile individuals is considerably affected by the number of embryos, number of injected oocytes, cause of infertility, female age, and PCOS.

The Birth Satisfaction Scale-Revised Indicator (BSS-RI): a validation study in Iranian mothers.

OBJECTIVE: Birth Satisfaction Scale-Revised Indicator (BSS-RI) is a short selfreport instrument designed to measure satisfaction of the childbearing women's experiences of labour and its outcomes. The aim of this study was to examine the reliability and validity of the Persian version of BSS-RI in Iranian mothers. METHODS: This cross-sectional study was conducted on 396 mothers in Tehran, Iran, between July and September 2017. The mothers were administered the BSS-RI, and a demographic questionnaire. Internal consistency of the BSS-RI was examined with Cronbach's alpha, construct validity was evaluated via exploratory factor analysis (EFA), and divergent validity was examined by correlating the BSS-RI with gestational age. RESULTS: The EFA results demonstrated a two-factor structure corresponding to the Stress of Childbearing and Quality of Care domains of the structure proposed by provider. The Cronbach's alpha for Stress of Childbearing and Quality of Care subscales and total BSS-RI were 0.665, 0.847, and 0.563, respectively. The mean of BSS-RI total score was 6.16 (SD = 2.60), and the Stress of Childbearing and Quality of Care subscales were 2.71 (SD = 2.39), and 3.45 (SD = 1.11), respectively. The BSS-RI showed no significant correlation with the gestational age, confirming divergent validity. CONCLUSION: Like the original English version, the Persian version of the BSS-RI is a reliable and valid instrument for measuring birth satisfaction in Iranian mothers. It can also be used as short and easy to administer tool for assessment of birth satisfaction in large sample survey research.

Safety and Efficacy of Allogeneic Adipose Tissue Mesenchymal Stromal Cells in Amyotrophic Lateral Sclerosis Patients, Single-Center, Prospective, Open-Label, Single-Arm Clinical Trial, Long-Term Follow-up.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder with very limited treatment options. Stem cells have been raised as a new treatment modality for these patients. We have designed a single-center, prospective, open-label, and single arm clinical trial to assess the safety, feasibility, and rather efficacy of administrating allogeneic adipose-derived mesenchymal stromal cells (Ad-MSCs) in ALS patients. We enrolled 17 patients with confirmed ALS diagnosis with ALS Functional Rating Scale-Revised (ALSFRS-R) ≥24 and predicted forced vital capacity (FVC) ≥40%. Allogeneic Ad-MSCs were transplanted intravenously for all patients. Follow-ups were done at 24 hours, 2, 4, 6, and 12 months after cell infusion by checking adverse events, laboratory tests, and clinically by ALSFRS-R and FVC. Patients were also followed five years later and ALSFRS-R score was recorded in the survived individuals. There was no report of severe adverse events related to cell infusion. Two patients experienced dyspnea and chest pain 36 and 65 days after cell infusion due to pulmonary emboli. The progressive decrease in ALSFRS-R and FVC levels was recorded and three patients died in the first year. During five years follow up, despite a notable decrease in functional scores, 5 patients survived. Intravenous (IV) infusion of allogeneic Ad-MSCs in ALS patients is safe and feasible. The survival rate of the patients is more than IV autologous MSCs (Registration number: IRCT20080728001031N26).

The Four-item Patient Health Questionnaire for Anxiety and Depression: A Validation Study in Infertile Patients.

BACKGROUND: The most common mental disorders in infertile patients are depression and anxiety. The four-item Patient Health Questionnaire-4 (PHQ-4) is a widely used tool that consists of the PHQ-2 depression and Generalized Anxiety Disorder-2 (GAD-2) scales. Given that PHQ-4 has not been validated in infertile patients, this study aimed to examine its reliability and validity in this population. MATERIALS AND METHODS: Participants in this cross-sectional study consisted of 539 infertile patients from a referral fertility centre in Tehran, Iran. The PHQ-4, Hospital Anxiety and Depression Scale (HADS), World Health Organisation- Five Well-Being Index (WHO-5), Penn State Worry Questionnaire (PSWQ) and demographic/infertility questionnaires were administered to all participants. Factor structure and internal consistency of PHQ-4 were evaluated via confirmatory factor analysis (CFA) and Cronbach's alpha, respectively. The convergent validity of this scale was examined by its relationship with HADS, WHO-5 and PSWQ. RESULTS: CFA results provided support for a two-factor model of PHQ-4. Internal consistency of the PHQ-4 and its subscales both were elevated with Cronbach's alpha coefficients of 0.767 (PHQ-4), 0.780 (PHQ-2) and 0.814 (GAD-2). Inter-item correlations were between 0.386 and 0.639, and corrected item-total correlations were between 0.576 and 0.687. PHQ-4, PHQ-2 and GAD-2 showed positive correlations with measures of HADS-anxiety, HADS-depression, and PSWQ and negative correlations with WHO-5, which confirmed convergent validity. Among demographic/fertility variables, we observed that gender, infertility duration, and failure in previous treatment were correlated with PHQ-4 and its subscales scores. CONCLUSION: The PHQ-4 is a reliable and valid ultra-brief screening instrument for measuring both anxiety and depressive symptoms in infertile patients.

The First Report of Differences in Gut Microbiota Composition between Obese and Normal Weight Iranian Subjects

Background: OObesity is a complex disorder influenced by various genetic and environmental factors. It has been shown that gut microbiota, which colonizes gastrointestinal tract, has a substantial role as an environmental factor in the pathophysiology of obesity. Since the composition of gut microbiota alters with regard to different criteria, such as ethnicity, geographical location, diet, lifestyle, age, and gender, we aimed to determine firmicutes/bacteroidetes (F/B) ratio and the abundance of important gut microbiota members, Akkermansia muciniphila, Faecalibacterium prausnitzii, Roseburia, Bifidobacterium, and Prevotella in Iranian obese and normal weight individuals, for the first time. Methods: In this study, 50 normal and 50 obese subjects were recruited and classified based on their BMI into normal weight and obese groups. Stool samples were collected. Following DNA extraction from the samples, quantitative PCR was conducted based on 16s rDNA universal primers. Finally, the correlation between the bacterial abundance and obesity was analyzed by statistical analyses. Results: We observed a significant increase of F/B ratio in the obese group, compared to the normal weight group (p = 0.002). Although A. muciniphila (p = 0.039) and Bifidobacterium (p = 0.049) abundance significantly decreased, the abundance of F. prausnitzii (p = 0.046) significantly elevated with BMI increase in the studied groups. Conclusion: Owing to the importance of the gut microbiota composition in obesity development, determination and targeted restoration of gut microbiota pattern could be valuable in the control and treatment of obesity in certain populations.

Differential expression of progesterone receptor isoforms related to PGR +331g/a polymorphism in endometriosis: A case-control study.

BACKGROUND: Endometriosis are defined as a progesterone-resistance disease. Two progesterone receptor (PR) isoforms, namely PR-A and PR-B, mediate the special effects of progesterone. One of the most effective polymorphism in the promoter region of PGR is the +331G/A. OBJECTIVE: The differential expression level of PR isoforms due to +331G/A polymorphism may be able to influence the function of progesterone and reduce the susceptibility of endometriosis. MATERIALS AND METHODS: This analytic, case-control study was carried out at Royan Institute, Tehran, Iran. Whole-blood samples were collected from 98 infertile women undergoing laparoscopy for endometriosis and 102 healthy fertile women. After DNA extraction, genotype frequencies were determined by polymerase chain reaction-restriction fragment length polymorphism. Then, RNA was extracted from the selected eutopic tissue samples of endometriosis patients. Analysis of PR-A and PR-B mRNA expressions were performed using Real-time polymerase chain reaction. RESULTS: The frequency distribution of GG, GA genotypes in +331G/A polymorphism was 98.04%, 1.96% in the patients and 97.96%, 2.04% in the control groups, respectively (p = 0.968). Although our data did not show any significant association with +331G/A in the patient and control groups, we were able to demonstrate significantly higher expression level of PR-B and no significant lower expression level of PR-A isoforms in patients by favoring GA to GG genotypes (p = 0.017, p = 0.731, respectively). CONCLUSION: Our findings show that patients with GA genotypes had a higher expression level of PR-B compared to patients with GG genotypes.

The Effect of Imbalanced Progesterone Receptor-A/-B Ratio on Gelatinase Expressions in Endometriosis.

BACKGROUND: Gelatinases degrade extracellular matrix (ECM) components to allow for physiological remodeling and contribute to pathological tissue destruction in endometriosis. It is known that the function of gelatinases is resistant to suppression by progesterone in endometriosis. The ability of progesterone to impact gene expression depends on the progesterone receptor-A/-B(PR-A/PR-B) ratio. An imbalanced PR-A/PR-B ratio in endometriotic tissue may be the result of the differential expression of MMP-2 and MMP-9, which could be important in the etiology and pathogenesis of the disease. Hence, we decided to study the association of PR-A/PR-B ratio and gelatinases expression in endometriosis. MATERIALS AND METHODS: In this prospective case-control study, we enrolled 40 women, 20 in the case group who were diagnosed with stage III/IV endometriosis and 20 normal subjects without endometriosis (controls) who referred to Royan Institute, Tehran, Iran during 2013-2014. We obtained 60 tissue samples [ectopic (n=20), eutopic (n=20), and normal endometrium (n=20)]. RNA was extracted from the tissue samples in order to analyze PR-A, PR-B, MMP-2, and MMP-9 mRNA levels through real-time polymerase chain reaction (PCR). RESULTS: There was significantly lower expression of the PR-B isoform in ectopic tissues compared to the control (P=0.002) and eutopic endometrium (P=0.006) tissues. PR-A expression was higher, but not significantly so, in the same ectopic and eutopic endometrium tissues compared to the control tissues (P=0.643). There was significant overexpression of MMP-9 in ectopic samples compared to control (P=0.014) and eutopic endometrium (P=0.012) samples. The PR-A/PR-B ratio was not significantly higher in either eutopic or ectopic samples compared to the control samples (P=0.305). CONCLUSION: Our findings support an altered PR-B expression in endometriosis, which may be associated with MMP-9 overexpression. This finding can be important for disease pathogenesis.

Gene expression analysis of MMPs in women with preeclampsia using cell-free fetal RNA in maternal plasma.

OBJECTIVE: Nucleic acids released from the placenta into the mother's blood circulation system provide a valuable source of potential biomarkers for early detection of pregnancy complications such as preeclampsia (PE). PE affects nearly 5-10% of pregnancies worldwide and is a major contributor to the maternal and neonatal mortality and morbidity. It is known that altered placental expression of matrix metalloproteinases (MMPs) may cause shallow cytotrophoblastic invasion and ultimately lead to preeclampsia. The present study aimed to evaluate pattern of placental/fetal expression of the MMP family (MMP-2, MMP-9, MMP-14, MMP-15 and MMP-26) in preeclamptic women and compare it to normal pregnancies, using cell free fetal RNA (cff-RNA). METHODS: Blood samples were obtained from 20 pregnant women diagnosed with severe PE (28-32 weeks) and 40 control healthy pregnant women in two groups of either matched gestational age (N = 20) or 14 and 28 weeks pregnancies (each 10). cff-RNA was extracted from plasma, followed by reverse transcription of cff-RNA. Expression of MMP genes was measured using quantitative reverse transcription PCR (qRT-PCR). RESULTS: The expression levels of MMP-2, MMP-9 and MMP-15 were significantly increased, while MMP-14 expression level was significantly reduced and the expression of MMP-26 showed a relative increase in PE pregnancies compared to the control group. Additionally, increased level of MMPs expression was observed by comparing 14 and 28 weeks gestation age in normal pregnancy. CONCLUSION: Using cff-RNA, circulatory expression level of MMP-2, MMP-9, MMP-14 and MMP-15 were significantly altered in preeclampsia compared to normal pregnancies.

Detection of Partial AZFc Microdeletions in Azoospermic Infertile Men Is Not Informative of MicroTESE Outcome.

BACKGROUND: Microdeletions of the Yq chromosome are among the most frequent genetic etiological factor of male infertility which spans the azoospermia factor regions (AZFa, AZFb and AZFc). Microdeletions are mostly seen in the AZFc region and usually cover genes actively involved in spermatogenesis. Partial AZFc microdeletions may also occur with various spans, namely gr/gr, b2/b3 and b1/b3. It is known that the outcome of microtesticular sperm extraction (TESE), the surgical process for sperm retrieval from the testis in infertile azoospermic men, may be predicted based on the type of AZF microdeletion. We therefore aimed to evaluate the correlation between partial AZFc microdeletions and microTESE results. MATERIALS AND METHODS: In this cross-sectional study, 200 infertile azoospermic men referred to the Royan Institute were examined for the presence of partial AZFc microdeletions before undergoing microTESE. Partial AZFc microdeletions were detected by multiplex polymerase chain reaction (PCR) of seven different sequence-tagged site (STS) markers. The data were analyzed with the Chi-square test. RESULTS: Among the 90 patients (45%) with a positive microTESE outcome, 9 (10%) showed a partial microdeletion in AZFc region. Of the 110 (55%) patients with a negative microTESE outcome, 7 (6.3%) had an AZFc partial microdeletion. With respect to the span of the microdeletions, among the 200 patients, 11 (5.5%) were gr/gr and 5 (2.5%) were b2/b3. Statistical analysis showed no significant difference between the patients with and without partial AZFc microdeletions with respect to microTESE outcome. CONCLUSION: Partial AZFc microdeletions is not a predictor of microTESE outcome in azoospermic men.

Association between The Number of Retrieved Mature Oocytes and Insulin Resistance or Sensitivity in Infertile Women with Polycystic Ovary Syndrome.

BACKGROUND: The objective of this study was to describe the association between luteinizing hormone (LH)/ follicle-stimulating hormone (FSH) ratio and demographic variables and maturation stage of oocytes in insulinresistant and insulin-sensitive patients with polycystic ovary syndrome (PCOS) in comparison with control group. MATERIALS AND METHODS: In this case-control study, 60 patients with in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) indication were subdivided into 3 groups as follow: 20 subjects were assigned to control (fertile women with male infertility history) group, 20 subjects with PCOS were insulin resistant (IR) and 20 subjects with PCOS were insulin sensitive (IS). After puncture, retrieved oocytes were classified into metaphase II (MII) as mature and in metaphase I (MI) or germinal vesicle stage (GV) as immature. Regression analyses were used to explore the association between MII oocyte number and demographic and clinical variables. RESULTS: LH/FSH ratio was significantly higher in PCOS-IR women compared to controls but not significantly different from that of PCOS-IS group. PCOS-IR women had lower MII oocyte number compared with that of controls. According to multiple regression analysis, the number of previous assisted reproductive technology (ART) cycles was negatively associated with the number of MII oocytes. CONCLUSION: Insulin resistance can be associated with reductions in MII oocyte number in patients with PCOS.

Modeling In Vitro Fertilization Data Considering Multiple Outcomes Observed among Iranian Infertile Women.

BACKGROUND: Women undergoing in vitro fertilization (IVF) cycles should successfully go through multiple points during the procedure (i.e., implantation, clinical pregnancy, no spontaneous abortion and delivery) to achieve live births. In this study, data from multiple cycles and multiple points during the IVF cycle are collected for each individual to model the effects of factors associated with success at different stages of IVF cycles in Iranian infertile women. MATERIALS AND METHODS: This historical cohort study includes 996 assisted reproductive technology (ART) cycles of 511 infertile women. Covariates considered in this study were women's age, type of cycle (fresh or frozen embryo transfer), number of embryos transferred and having polycystic ovarian syndrome during IVF cycles. Generalized estimating equations were used for calculation of odds ratio (OR) and 95% confidence intervals (95% CI) of success at different stages during IVF cycles. Cluster-weighted generalized estimating equations (CWGEE) was also fitted to handle informative cluster size. RESULTS: After adjusting for potential confounders, it was seen that receiving frozen embryo transfer was associated with higher odds of success compared to receiving fresh embryo transfer (adj OR: 2.26, 95% CI: 1.66-3.07); however, cycles with fresh embryo transfer exhibited better results in clinical pregnancy compared to those receiving frozen embryo. Being in the age category of 38 to 40 was associated with lower odds of success compared to the reference category (p<35) in CWGEE model (adj OR: 0.67, 95% CI: 0.45-1.00). The number of embryos transferred was positively associated with the odds of success in CWGEE (adj OR: 1.21, 95% CI: 1.03-1.42) as well as the GEE model. CONCLUSION: Receiving frozen embryo was positively associated with odds of success compared to cycles with fresh embryo. The number of embryos transferred and women's age were significantly associated with odds of success.

Comparison of In Vitro Fertilization/Intracytoplasmic Sperm Injection Cycle Outcome in Patients with and without Polycystic Ovary Syndrome: A Modified Poisson Regression Model.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a frequent condition in reproductive age women with a prevalence rate of 5-10%. This study intends to determine the relationship between PCOS and the outcome of assisted reproductive treatment (ART) in Tehran, Iran. MATERIALS AND METHODS: In this historical cohort study, we included 996 infertile women who referred to Royan Institute (Tehran, Iran) between January 2012 and December 2013. PCOS, as the main variable, and other potential confounder variables were gathered. Modified Poisson Regression was used for data analysis. Stata software, version 13 was used for all statistical analyses. RESULTS: Unadjusted analysis showed a significantly lower risk for failure in PCOS cases compared to cases without PCOS [risk ratio (RR): 0.79, 95% confidence intervals (CI): 0.66-0.95, P=0.014]. After adjusting for the confounder variables, there was no difference between risk of non-pregnancy in women with and without PCOS (RR: 0.87, 95% CI: 0.72-1.05, P=0.15). Significant predictors of the ART outcome included the treatment protocol type, numbers of embryos transferred (grades A and AB), numbers of injected ampules, and age. CONCLUSION: The results obtained from this model showed no difference between patients with and without PCOS according to the risk for non-pregnancy. Therefore, other factors might affect conception in PCOS patients.

Determinants of The Grade A Embryos in Infertile Women; Zero-Inflated Regression Model.

OBJECTIVES: In assisted reproductive technology, it is important to choose high quality embryos for embryo transfer. The aim of the present study was to determine the grade A embryo count and factors related to it in infertile women. MATERIALS AND METHODS: This historical cohort study included 996 infertile women. The main outcome was the number of grade A embryos. Zero-Inflated Poisson (ZIP) regression and Zero-Inflated Negative Binomial (ZINB) regression were used to model the count data as it contained excessive zeros. Stata software, version 13 (Stata Corp, College Station, TX, USA) was used for all statistical analyses. RESULTS: After adjusting for potential confounders, results from the ZINB model show that for each unit increase in the number 2 pronuclear (2PN) zygotes, we get an increase of 1.45 times as incidence rate ratio (95% confidence interval (CI): 1.23-1.69, P=0.001) in the expected grade A embryo count number, and for each increase in the cleavage day we get a decrease 0.35 times (95% CI: 0.20-0.61, P=0.001) in expected grade A embryo count. CONCLUSIONS: There is a significant association between both the number of 2PN zygotes and cleavage day with the number of grade A embryos in both ZINB and ZIP regression models. The estimated coefficients are more plausible than values found in earlier studies using less relevant models.