International Journal of Fertility and Sterility

100-107

Authors: Mohammad Reza Mirinezhad; Maliheh Aghsizadeh; Mohammadreza Fazl Mashhadi; Sara Moazedi; Maryam Mohammadi Bajgiran; Hamideh Ghazizadeh; Shayan Yaghouti; Mahdi Mohammadian Ghosooni; Mohammad Amin Mohammadi; Elahe Hasanzadeh; Ali Ebrahimi Dabagh; Arezoo Rastegarmoghadam Ebrahimian; Ensieh Akbarpour; Habibollah Esmaily; Gordon A Ferns; Tayebeh Hamzehloei; Alireza Pasdar; Majid Ghayour‐Mobarhan

Background: Premature menopause (PM) is the cessation of ovarian function before age 40. PM women are more likely to have cardiovascular diseases (CVDs), diabetes, and mental disorders. This is the first study that assessed the association of single nucleotide polymorphisms (SNPs) with anti-heat shock protein 27 (Hsp27), High-sensitivity C-reactive protein (hs- CRP), and PM and serum pro-oxidant-antioxidant balance (PAB), as putative risk factors for CVDs. We aimed to explore the association of oxidative stress markers with eight different SNPs shown to be related to premature menopause. Materials and Methods: In this cross-sectional research, we included 183 healthy women and 117 premature menopausal women. We determined baseline characteristics for all participants and measured serum hs-CRP, anti-HSP-27 antibody titer, and PAB levels using the established methods. Genotyping for eight SNPs was done using the tetra amplification refractory mutation system polymerase chain reaction (Tetra-ARMS PCR) and allele-specific oligonucleotide PCR (ASO-PCR) methods. Results: We found a significant difference between mean serum PAB levels and the genetic variant of rs16991615 (P=0.03). ANCOVA showed a significant effect of the genotypes rs4806660 and rs10183486 on hs-CRP serum levels in the case and control groups, respectively (P=0.04 and P=0.007). ANCOVA also showed an association between rs244715 genotypes and anti-hsp27 serum levels in the case group (P=0.02). There was a significant effect of the genotypes of rs451417 on the serum hs-CRP level in the control group (P=0.03). Conclusion: There was a significant association of the genetic variants related to PM with oxidative stress and inflammatory markers (serum PAB, anti-hsp27 antibody, and hs-CRP). Accordingly, this seems to be an effective approach to predicting susceptible subjects for cardiovascular and mental disorders as well as various cancers.

115-122

Authors: Mahsa Afrough; Roshan Nikbakht; Mahmoud Hashemitabar; Elham Ghalambaz; Sareh Amirzadeh; Adel Zardkaf; Sahar Adham; Maryam Mehdipour; Parvin Dorfeshan

Background: This research was aimed at assessing the relationship between the follicular fluid (FF) antioxidants activity, aging and in vitro fertilization (IVF) outcome. Materials and Methods: The present cross-sectional study was carried out on 65 women undergoing IVF/intracytoplasmic sperm injection (IVF/ICSI) cycles due to unexplained infertility. Ovarian stimulation was performed using the long gonadotropin-releasing hormone (GnRH) agonist protocol. After ovum pickup, FF was collected and processed to measure the level of superoxide dismutase (SOD), catalase (CAT) activity, total antioxidant capacity (TAC) and glutathione (GSH). Day 3 after ICSI, fresh embryos were transferred and later, possible pregnancy was assessed. Patients participating in this study were divided into four groups on the basis of age and pregnancy outcome. Results: SOD activity was not significantly different between the groups (P=0.218). GSH in the group whose participants were aged ≤35 years and were pregnant was higher than that in other groups. CAT activity in groups with younger participants was higher compared to the other groups. The mean TAC was higher in groups with pregnant participants compared to the non-pregnant women. Correlation analysis showed that: GSH level had a significant negative correlation with age (P<0.001, R -0.55) and a significant positive correlation with pregnancy (P=0.015, R=0.30). CAT level also had a significant negative correlation with age (P<0.001, R=-0.42) and the level of TAC had a significant positive correlation with pregnancy (P<0.001, R=0.59). Conclusion: According to our results, the levels of TAC, GSH and CAT in younger and pregnant women were higher compared with those undergoing ICSI cycles. Given the correlation of FF antioxidant activity with age and pregnancy, it is necessary to carry out more research on these compounds and the maintenance of pregnancy.

128-134

Authors: Mahtab Moti; Leila Amini; Hamid Haghani; Mohammad Reza Nateghi

Background: The aim of this study was to evaluate the effects of thiamine (vitamin B1) on general health and infertility treatment outcomes in women with polycystic ovary syndrome (PCOS). Materials and Methods: The study is a triple-blinded, randomized, placebo-controlled clinical trial performed on 64 infertile women with PCOS referred to Sarem Hospital in Tehran, Iran. The primary outcomes of the study were general health and infertility treatment outcomes. Eligible women were randomly assigned to the vitamin B1 group (n=32, vitamin B1 tablet at a dose of 300 mg/day for 4 weeks) or the placebo group (n=32, placebo tablet daily for 4 weeks). A general health questionnaire was completed before and after the intervention by both groups, and treatment success was evaluated at the end of the study. Data were analyzed using SPSS software ver.16 P<0.05 was considered statistically significant. Results: The mean age of participants in the vitamin B1 (VB1) group was 30.4 ± 3.27 years and in the placebo (Pl) group was 29.1 ± 2.66 years with the mean duration of marriage 12.7 ± 3.01 and 13.2 ± 2.97 years respectively. Our results showed that there were significant differences between the two groups in overall score (P<0.001) and scores for all domains of the general health questionnaire including somatic symptoms (P<0.001), anxiety and insomnia (P<0.001), social dysfunction (P=0.028), and severe depression (P<0.001) after the intervention. Four weeks consumption of vitamin B1 also resulted in higher numbers of positive pregnancy tests (P=0.006), although the number of fetuses was not significantly different between the two groups after the intervention. Conclusion: The results of the current study support a possible favourable effect of vitamin B1 on improving general health, infertility treatment outcome, and retrieved follicle count without changing the number of fetuses in women with polycystic ovary syndrome (registration number: IRCT201510266917N3).

140-145

Authors: Fatemeh Ghahghayi; Abolfazl Payandeh; Aida Najafian; Marzieh Ghasemi; Ayob Jabari Jabari

Background: It is difficult to obtain healthy oocytes in poor ovarian responders with conventional treatment methods. Thus, the need to investigate new methods is essential. This study aims to investigate ovulation induction outcomes in patients with decreased ovarian reserve (DOR) in two groups treated with double stimulation (DuoStim) during the follicular and luteal phases in comparison with the antagonist cycle. Materials and Methods: This was a randomised clinical trial that enrolled the patients with reduced ovarian reserve. The patients referred for in vitro fertilization (IVF) at Molud Infertility Clinic, Ali Ebn Abitalib (AS) Hospital, Zahedan, Iran from 2020 to 2021. Participants were randomly divided into two groups, those who underwent treatment with DuoStim during the follicular and luteal phase (case group) and those who received the conventional antagonist cycle (control group). Results: The mean number of metaphase II (MII) eggs was 7.7 ± 3.1 in the case group and 6.1 ± 3.9 in the control group (P=0.063). The mean total number of retrieved eggs in the case group was 9.2 ± 3.7 and in the control group, it was 6.9 ± 4.4 (P=0.023). The mean number of embryos obtained in the case group was 6.5 ± 3.9; in the control group, it was 4.7 ± 2.8 (P=0.016). Conclusion: The DuoStim method can effectively play a role in increasing the total number of retrieved eggs and embryos (registration number: IRCT20120817010617N8).

153-161

Authors: Javad Najafi Salehi; Hussein Eimani; Abdolhossein Shahverdi; Mehdi Totonchi; Rouhollah Fathi; Seyed Akbar Moosavi; Seyed Mohamad Javad Taher Mofrad; Leila Sadat Tahaei

Background: The parallel and continued improvements in both infertility treatment and the management of malignancy cases have brought to the forefront the potential for fertility preservation. Using ovarian follicular resources can effectively improve reproductive capacity and prevent infertility. The primary aim of this research was to try to generate an appropriate in vivo environment for the growth of the mouse follicles. Hence, the possible effects of the ovarian parenchyma cell suspension were explored on the growth and maturation of preantral follicles in vitro. Materials and Methods: In this experimental study, ovarian parenchymal cells were mechanically dissociated from preantral follicles of 12-14 days-old NMRI mice and then divided into 5 experimental groups (G1: Control, G2: Fresh follicle with fresh parenchyma cell suspension, G3: Vitrified-warmed follicle with fresh parenchyma cell suspension, G4: Fresh follicle with frozen-thawed parenchyma cell suspension, and G5: Vitrified-warmed follicle with frozenthawed parenchyma cell suspension). The diameter of the follicles and immature oocytes, viability, antrum formation, resumption of meiosis, in vitro fertilization (IVF), and Gdf9, Bmp6, and Bmp15 gene expression were examined on different periods. Results: The diameter of the follicles and the oocytes on days 4 and 8, as well as the survival rate of the follicles up to day 12, were significantly higher in G2 and G4 compared to the Ctrl group (G1: 73.66%, G2:87.99%, G3: 82.70%, G4: 94.37%, and G5: 78.59%). Expression of growth marker genes for G3, and G5 groups was significantly higher than other groups, which indicated the protective effects of parenchyma cell suspension on follicles damaged by vitrification solutions. Conclusion: The growth, survival, and maturation of preantral follicles could be enhanced by co-culturing them with ovarian parenchyma cells. Further studies are needed to optimize the conditions for a successful parenchyma cell suspension-induced in vitro maturation (IVM) to occur in infertility clinics.

167-172

Authors: Ali Hamidi Madani; Mohammad Pakdaman; Gholamreza Mokhtari; Ehsan Kazemnezhad; Mohammad Hamidi Madani; Ali Fathollahi; Nadia Rastjou Herfeh; Roozbeh Roohinezhad

Background: Varicocele is one of the most common treatable causes of male infertility, and its treatment may be beneficial for fertility. This study aimed to evaluate fertility rate and DNA fragmentation index (DFI) following varicocelectomy in primary infertile men with clinical varicocele. Materials and Methods: This prospective longitudinal study was conducted on primary infertility men, in a tertiary center from December 2018 to December 2019 with one-year follow-up. Data of the semen parameters, DFI (%), and fertility rate were gathered before, as well as 4 and 12 months after undergoing varicocelectomy. For data analysis, SPSS software and analytical test were used. Results: Out of 76 patients who were analyzed, 22 (29%) became fertile and 54 (71%) remained infertile. Semen parameters and DFI (%) were improved significantly following varicocelectomy (P<0.001). Smoking history, occupational heated exposure, body mass index (BMI), and infertility duration were determined as predictors associated with fertility status (P<0.05). Conclusion: Although varicocele repair improved the DFI, the fertility rate was achieved in less than one-third of patients; it seems that the other parameters, such as the history of smoking, occupational heated exposure, overweight, and duration of infertility should be considered as predictors of fertility status, in primary infertile men who are a candidate for varicocelectomy.

180-184

Authors: Elham Poursafari Talemi; Seyedeh-Hanieh Hosseini; Hamid Gourabi; Marjan Sabbaghian; Anahita Mohseni Meybodi

Background: Infertile men with multiple morphological abnormalities of the sperm flagella (MMAF) phenotype exhibit mosaic sperm flagella abnormalities such as short, bent, coiled, and irregular flagella or absent flagella. Sperm flagellum has an ultrastructurally axonemal structure that contains a large number of proteins. A-Kinase Anchoring Protein 3 (AKAP3) is expressed in spermatozoa. It may function as a regulator of motility and the acrosome reaction. This study aimed to compare genetic changes in infertile men suffering MMAF phenotype with the control group. Materials and Methods: In this case-control study, genetic variants of the AKAP3 gene were evaluated in 60 infertile men with MMAF phenotype and 40 fertile men, as control. As exon five of the AKAP3 gene encodes the functional domain of this protein, its genetic variants were studied. Therefore, polymerase chain reaction (PCR)-sequencing was undertaken on the DNA extracted from control and patients’ blood samples. Results: Sixty infertile men with MMAF phenotype and 40 normozoospermic men, as control, were enrolled in this study. Four haplotype variants 1378T>C (rs10774251), 1391C>G (rs11063266), 1437T>C (rs11063265), and 1573G>A (rs1990312) were detected in all patients and controls. On the other hand, a missense mutation 1499T>C (rs12366671) was observed in four patients with the homozygous form while seven patients carried the heterozygous form. No mutation was identified in the controls (P=0.04). The difference between the variation allele frequencies was assessed in the patient and control groups by the Fisher Exact Test. Conclusion: In the homozygous form, this mutation changed Isoleucine to Threonine. This alternation occurred inside the AKAP4 binding domain of the AKAP3 protein. The observed variants caused no significant deviation in the secondary structure of AKAP3 protein and probably its function in spermatozoa flagella. So, these variants cannot be considered as the causes of MMAF phenotype in the studied patients.