Knowledge and beliefs about oocyte cryopreservation for medical and social reasons in female students: a cross-sectional survey.

BACKGROUND: With the increasing number of young women surviving cancer and a growing trend among highly educated women to postpone childbearing for educational or professional pursuits, there is a rising demand for egg freezing services to ensure a successful pregnancy. This study aims to assess the knowledge and beliefs surrounding oocyte cryopreservation, both for medical and social reasons, among female students in Tehran, Iran. METHODS: An online cross-sectional survey was carried out from March to August of 2022, involving a total of 1279 childless students pursuing master's and doctoral degrees at universities in Tehran. The participants were between the ages of 18 and 38. Knowledge and beliefs about medical and social oocyte cryopreservation were assessed through Fertility Preservation Survey (FPS) instrument. RESULTS: The mean age of the participants was 26.38 ± 4.9. The majority of students expected to be "30-34 years" when they become pregnant with their first child (41.1%, M: 30.3 ± 4.13 years) and "35-39 years" when they give birth to their last child (46.7%, M: 35.28 ± 4.18 years). The students agreed with preserving fertility with medical (93.3%) and social (86.9%) indications and believed the medical (95.1%) and social (87.4%) costs of cryopreservation should be covered by the healthcare system. Among the participants, 75.6% considered cost to be a definite or probable factor in their decision to pursue fertility preservation. The oncology team's recommendation was identified as the most important factor in deciding on medical egg freezing (92.6%, M: 3.46 ± 0.71). The overall correct response rate for the knowledge questions was 57.7%. The majority of participants (95.5%) agreed that physicians should routinely provide information about egg freezing to women of childbearing age during their regular healthcare visits. CONCLUSIONS: The research results revealed that female students in Tehran universities have a positive attitude towards medical and social egg freezing, but lack sufficient knowledge about the ideal timing of childbearing. Health professionals could provide detailed information about fertility preservation and age-related infertility as part of routine healthcare visits or reproductive health planning. Additionally, expanding supportive policies and incentives for childbearing established by the government to cover the costs of fertility preservation would be beneficial.

Commitment Theory as the Theoretical Framework in Third-Party Reproduction.

Assisted reproductive technology (ART) and third-party reproduction provide the opportunity for infertile couples to have children through different genetic links. This type of treatment has created many challenges for infertile couples. With this treatment, the infertile couple will have a child who is biologically related to the gamete/embryo donor. Accordingly, the transformation that occurs in the structure of traditional families and the concept of parenthood is one of the main consequences and challenges which requires in-depth research. In spite of the successful expansion of infertility treatment and third-party reproduction, there is still no proper social context for implementing third-party infertility treatments in Iran. Therefore, despite the need to use the technology, some couples refuse the treatment unless their confidentiality is preserved. Many couples follow the practice surreptitiously by keeping the donation treatment confidential, to get rid of the existing social stigma and protect their identity, the child and the donor's identity. Commitment theory as a theoretical strategy is proposed to solve the problems of all parties involved in this type of "social and non-biological" parenting. Commitment theory in the context of third-party reproduction expresses the commitment to the contract accepted by the donor and the recipient of the gamete/embryo, based on which, the recipients consider the resulting child as their own, and are committed to all the related paternal-maternal rights and duties such as "alimony" and "inheritance". On the other hand, the donors undertake to waive all their paternal-maternal rights and duties by donating gamete/embryo.

Step-by-step decision-making process in third party assisted reproduction: a qualitative study.

This study explores the experience of decision-making about the third party assisted reproduction in Iranian infertile patients using a qualitative approach. Data were collected in 2017-2019 through in-depth interviews of 20 infertile participants (3 couples and 14 individuals) who became pregnant or had children through third-party reproduction and 12 specialists. The interviews were transcribed verbatim, the data were managed using the MAXQDA 10 and analysed using a conventional content analysis approach. The main category that emerged was 'step-by-step process of adjustment to third party assisted reproduction'. From this, five sub-categories were extracted: (i) 'parental preference for biological children over non-biological children'; (ii) 'parental preference for non-biological children over childlessness threats'; (iii) 'strong preference for having partial genetic link with children'; (iv) 'flexibility in removing or overcoming barriers to third party reproduction'; and (v) 'parental preference for adopted children over childlessness'. The findings in the present study suggest the importance of lineage and biological relatedness in Iranian culture. Patients adjusted to non-biological parenting as a treatment option only if they have to. Using a step-by-step decision-making process helped participants make more thoughtful decisions by organising relevant information and defining alternatives. The results of the present study can help healthcare providers with counselling regarding parenting with or without genetic ties.

The Impact of Third Party Reproduction on Family and Kinship.

The development of in vitro fertilization (IVF) in the UK, in 1978, proved a major breakthrough in the process of human reproduction, which had remained constant in human history. The impact of IVF and the ensuing assisted reproductive technologies (ARTs) has not been limited in revolutionizing the "natural" practice of biological reproduction, but has reached out to and affected almost every institution in society. Family and kinship, as the social expression of reproduction and the institutions which are the most transparently structured realm of human life are those most profoundly affected by ARTs. Although literature on the implications of ARTs is in general abundant, this article presents new insights on their impact on family and kinship in Iran, which remains a unique case in the Muslim world. It explores the particular way ARTs, especially third-party donation, have been endorsed and practiced in Iran, and their consequences for the family, the infertile individuals, and their position vis-à-vis their kin and social group. The conclusion points to the lack of clarity concerning the initial rulings by the Islamic jurists, who allowed the practice of ARTs, and which has led to a number of unintended consequences regarding the legal, religious, cultural, and ethical issues, affecting the family, its structure and the relationship between the kin group. These consequences range, inter alia, from the question of the anonymity of third-party donor, to the permissibility of gamete donation between blood relatives, and to the absence of enforceable legislation.

Testis biopsy in absolute head teratospermia: Is it a logical option?

INTRODUCTION: Absolute head teratospermia (100% abnormal head morphology) is associated with poor fertility and assisted reproductive techniques results. We aimed to find if it is possible to bypass this disorder using sperm retrieved by testis biopsy. METHODS: Multiple testis biopsies were performed in patients with infertility and absolute head teratospermia who were not able to provide semen on the injection day from 2006 to 2018. Then, the found sperms were evaluated based on being proper or not proper for intracytoplasmic sperm injection. RESULTS: Only 2 patients, of a total of 22 (9%), had relatively proper sperms for microinjection. DISCUSSION: There is no benefit to performing testis biopsy in non-azoospermic patients with absolute abnormal head morphology.

Assessing the potential of HSPA2 and ADAM2 as two biomarkers for human sperm selection.

Selection of the best sperm, with the least defects, is a critical factor in the success of ART especially in male factor infertility. This study assessed the potential Heat shock protein (HSPA2) and metallopeptidase domain2 (ADAM2) biomarkers for sperm selection. Sperm were obtained from 72 asthenoteratozoospermic and 42 normospermic ejaculates. The semen characteristic, DNA fragmentation (DFI), chromatin maturation index (CMI), ADAM2 and HSPA2 levels on sperm, and their correlation with embryo quality were assessed in both groups. Results showed the significant reduction in HSPA2 and ADAM2 in asthenoteratozoospermic compared to normazoospermic ejaculates regarding the cut-off value of 14 and 13% for these two biomarkers. The specificity of HSPA2 and ADAM2 separately, and the combination of these two biomarkers, were 95.2, 90.5 and 93.5%, respectively, for sperm from normozoospermic ejaculates. However, they were 48.6, 50.0 and 54.5% for asthenoteratozoospermic ones. A significant correlation was observed with HSPA2, ADAM2 and a combination of these two biomarkers with CMI, DFI and embryo quality. Although a combination of these two biomarkers have the potential to be a good choice for selecting sperm with the lowest level of chromatin damage, it seems that selection according to HSPA2 has priority over ADAM2 or a combination of the two.

Monoclonal and Polyclonal Antibodies Specific to Human Fibromodulin.

BACKGROUND: The unique expression of fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL) has been previously reported. Detecting FMOD in CLL patients using specific anti-FMOD mAbs might provide a promising method in detection, monitoring, and prognosis of CLL. OBJECTIVES: In this study, we aimed for producing specific antibodies against FMOD to facilitate further cohort study of CLL, thus addressing FMOD as a potential target of detection. MATERIALS AND METHODS: Human FMOD gene (1087 bp) was extracted from genome of the CLL patients, and was cloned into the expression vector of pET-22b (+). The recombinant FMOD protein (rFMOD) was expressed in Escherichia coli. The purified rFMOD protein was used as an immunogen in rabbit and mice. Hybridoma technology was used to develop the monoclonal antibodies (mAbs). Polyclonal antibody (pAb) was purified from the rabbit sera using affinity column. The reactivity of anti-FMOD antibodies was assessed in ELISA, immunocytochemistry (ICC) and Western blot. RESULTS: ICC results showed that the anti-FMOD antibodies specifically detected FMOD in CLL PBMCs and cell lines. The developed anti-FMOD pAb detected FMOD in CLL lysates, compared to healthy PBMCs, in Western blot and ELISA. CONCLUSIONS: The developed anti-FMOD mAbs, and pAb specifically detect FMOD in CLL samples and might be used as research tools for further investigations in CLL.

Anchored Fibromodulin as a Novel Target in Chronic Lymphocytic Leukemia: Diagnostic and Therapeutic Implications.

BACKGROUND: We have previously reported the aberrant expression of Fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL). Although FMOD has been considered as a cytoplasmic or secretory protein, we discovered the cell surface expression of FMOD in leukemic B cells via anchoring with glycosylphosphatidylinositol (GPI). OBJECTIVE: To evaluate FMOD as a new biomarker in CLL patients in comparison with healthy individuals. METHODS: A monoclonal antibody was generated against human FMOD. The cell surface expression of FMOD in 52 CLL patients and 45 healthy individuals were compared by flow cytometry. A bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) was used to determine the cell surface localization of FMOD using ELISA and flow cytometry techniques. Annexin V-FITC and propidium iodide (PI) was used to detect apoptosis induction in CLL PBMCs following in vitro incubation with anti-FMOD mAb. RESULTS: The results demonstrated the widespread cell surface expression of GPI-anchored FMOD in CLL patients (median: 79.9 %), although healthy individuals had low FMOD expression (median: 6.2 %) (p≤0.0001). The cut-off value of FMOD expression was estimated with high sensitivity and specificity at 17.9 %. Furthermore, in vitro apoptosis induction of leukemic cells following incubation with anti-FMOD mAb showed a direct apoptosis of CLL cells (27.9%) with very low effect on healthy PBMCs (6%). CONCLUSION: The membrane-anchoring of FMOD by means of a GPI moiety in leukemic cells supports FMOD as a highly potential diagnostic and therapeutic target in CLL patients.

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Practical Difficulties in Estimating The Prevalence of Primary Infertility in Iran.

BACKGROUND: According to the World Health Organization (WHO)'s clinical, epidemiological and demographic definitions, infertility is an inability to become pregnant within one, two or five years of exposure to pregnancy, respectively. Inconsistent infertility-related definitions and various methodological approaches make it difficult to compare quantitative data in this regard and consequently, have negatively influenced estimating the prevalence of infertility. The present study reviewed the results of a large population-based survey on how the clinical, epidemiological and demographic definitions of infertility produce different results in terms of infertility prevalence in Iran and subsequently, compared the findings in order to find the right time of treatment-seeking by couples. MATERIALS AND METHODS: This community-based, cross-sectional study was carried out by Avicenna Research Institute in the urban and rural parts of Iran between 2010 and 2011. Using cluster sampling, the reproductive history of 17,187 married women aged 20-40 years, was recorded. Totally, 1011 clusters were randomly selected according to post office codes, proportional to the population of the province. Descriptive and inferential statistical analysis of the data was carried out by SPSS statistical software. RESULTS: The prevalence of primary infertility based on the WHO's clinical, epidemiological and demographic definitions were 20.2, 12.8 and 9.2%, respectively. In addition, secondary infertility rate was 4.9%. CONCLUSION: Infertility estimates over a two-year exposure period made a 50% decrease in infertility rate; however, increasing exposure period to five years made no significant difference in infertility rate. The findings showed that most of the couples will get pregnant within two years of unprotected sexual intercourse and thus, need no treatment. Due to practical difficulties in estimating the prevalence of primary infertility, the reference limit for time to pregnancy, should be reconsidered and giving more time to younger women to become pregnant, seems reasonable.

Fibronectin as a new biomarker for human sperm selection in assisted reproductive technology.

OBJECTIVE: Fibronectin (FN) is a multifunctional diametric glycoprotein on the surface of the sperm that plays an important role in the sperm-oocyte interaction and fertilization process. The aim of the present study was to assess the FN levels as a sperm surface biomarker for sperm selection in assisted reproductive technology. MATERIAL AND METHODS: Polyclonal antibody against human FN was produced in rabbit. Its quality, purity, and immune reactivity were assessed by SDS-PAGE and western blot. In addition, the presence of FN on the sperm surface was assessed through immunocytochemistry and flow cytometry. The amount of FN and the sperm quality were assessed in normozoospermia (N) (42 men) and asthenoteratozoospermia (AT) (72 men) groups through sperm chromatin dispersion (SCD), sperm chromatin structure assay (SCSA), and chromatin maturation index (CMI). RESULTS: The results showed the distribution of FN protein on the equatorial region of the human sperm surface. In addition, the FN levels were found to have a significant difference between the two groups with 24.64±9.08% in N and 16.90±7.27% in AT (p≤0.0001). In addition, FN level negatively correlated with SCD (p≤0.0001), SCSA (p≤0.0001), and CMI (p≤0.001). Threshold values of FN level and DNA fragmentation index (DFI) percentage were 16 and 30 and were identified as cut-off values to determine the N group with a specificity of 83.3% and 81.0% and a sensitivity of 16.8% and 19.0%, respectively. The specificity and sensitivity of FN-DFI were 91.2% and 8.8%, respectively. CONCLUSION: It appears that FN can be used for the selection of sperm with suitable quality, although future studies are recommended.

The effect of paternal age on semen quality and fertilization outcome in men with normal sperm DNA compaction, reactive oxygen species, and total antioxidant capacity levels.

OBJECTIVE: Studies showed a decrease of the semen analysis parameters and an increase in the average age of first-time fathers over the past several decades. The aim of the present study was to assess the influence of paternal age on semen quality and fertilization outcomes in men with normal sperm DNA fragmentation and chromatin maturation index (DFI and CMI), reactive oxygen species (ROS), and total antioxidant capacity (TAC) levels. MATERIAL AND METHODS: The study was performed on 70 men with their wife's age ≤38 years and normal sperm DFI, CMI, ROS, and TAC levels. None of the couples had a history of genital inflammation, chronic diseases, endocrine abnormality, chromosomal aberrations, Y chromosome microdeletion, azoospermia, and leukocytospermia. These men were separated into 2 groups according to their age (group A: age <45 years and group B: age ≥45 years). Semen analysis and fertilization outcome after using the intracytoplasmic sperm injection were assessed in both groups. RESULT: Sperm concentration showed a significant reduction in group B (p=0.04). Although semen volume, sperm normal morphology, and progressive motility were decreased in group B, the reduction was not significant when compared with group A (p=0.09, p=0.47, and p=0.77, respectively). In addition, the differences of embryo quality with grades A, B, and C and 8-cell embryo formation were not statistically significant between the 2 groups. CONCLUSION: These results demonstrated that in men with normal sperm DFI, CMI, ROS, and TAC levels, there were no significant changes in semen parameters and fertilization outcomes with an increasing age.

Reactive oxygen species-induced alterations in H19-Igf2 methylation patterns, seminal plasma metabolites, and semen quality.

PURPOSE: This study was conducted in order to investigate the effects of reactive oxygen species (ROS) levels on the seminal plasma (SP) metabolite milieu and sperm dysfunction. METHODS: Semen specimens of 151 normozoospermic men were analyzed for ROS by chemiluminescence and classified according to seminal ROS levels [in relative light units (RLU)/s/106 sperm]: group 1 (n = 39): low (ROS < 20), group 2 (n = 38): mild (20 ≤ ROS < 40), group 3 (n = 31): moderate (40 ≤ ROS < 60), and group 4 (n = 43): high (ROS ≥ 60). A comprehensive analysis of SP and semen parameters, including conventional semen characteristics, measurement of total antioxidant capacity (TAC), sperm DNA fragmentation index (DFI), chromatin maturation index (CMI), H19-Igf2 methylation status, and untargeted seminal metabolic profiling using nuclear magnetic resonance spectroscopy (1H-NMR), was carried out. RESULT(S): The methylation status of H19 and Igf2 was significantly different in specimens with high ROS (P < 0.005). Metabolic fingerprinting of these SP samples showed upregulation of trimethylamine N-oxide (P < 0.001) and downregulations of tryptophan (P < 0.05) and tyrosine/tyrosol (P < 0.01). High ROS significantly reduced total sperm motility (P < 0.05), sperm concentration (P < 0.001), and seminal TAC (P < 0.001) but increased CMI and DFI (P < 0.005). ROS levels have a positive correlation with Igf2 methylation (r = 0.19, P < 0.05), DFI (r = 0.40, P < 0.001), CMI (r = 0.39, P < 0.001), and trimethylamine N-oxide (r = 0.45, P < 0.05) and a negative correlation with H19 methylation (r = - 0.20, P < 0.05), tryptophan (r = - 0.45, P < 0.05), sperm motility (r = - 0.20, P < 0.05), sperm viability (r = - 0.23, P < 0.01), and sperm concentration (r = - 0.30, P < 0.001). CONCLUSION(S): Results showed significant correlation between ROS levels and H19-Igf2 gene methylation as well as semen parameters. These findings are critical to identify idiopathic male infertility and its management through assisted reproduction technology (ART).

Sortilin as a Novel Diagnostic and Therapeutic Biomarker in Chronic Lymphocytic Leukemia.

BACKGROUND: The overexpression of sortilin/neurotensin receptor 3 has previously been reported in various human solid tumors but not in hematological malignancies. Here, we report the overexpression of sortilin in leukemic cells from patients with Chronic Lymphocytic Leukemia (CLL). METHODS: Flow cytometry was used to compare the expression of sortilin in CLL patients (n=52) and healthy individuals (n=26). Also, in vitro apoptosis induction was assessed in CLL Peripheral Blood Mononuclear Cell (PBMCs) following directly targeting of sortilin. RESULTS: The results showed a significant expression of sortilin on the surface of CLL PBMCs (range from 2.2 to 71.5%) in comparison to healthy individuals (range from 0.03 to 7.4%) (p≤0.0001). The optimal cut-off value of sortilin expression was determined at 7.2% with high sensitivity and specificity. Treatment of leukemic cells with anti-sortilin antibody could induce apoptosis without any effect on normal cells. CONCLUSION: Apoptosis induction in CLL cells together with a significant correlation between the expression of sortilin and CD23 represent a possible functional role of sortilin in leukemogenesis of CLL cells. Therefore, sortilin might be considered as a promising novel biomarker in diagnosis, monitoring, and therapy of patients with CLL.

Upstream or swim up processing technique: which one is more effective to select human sperm with high chromatin integrity.

BACKGROUND: Sperm processing methods separate motile sperms with good morphology from dead and abnormal forms of sperms, immature germ cells, and non-sperm cells. OBJECTIVE: The propose of this study was to compare the efficacy of upstream and swim-up processing techniques to separate sperms with the high quality especially in relation to sperm chromatin integrity. MATERIALS AND METHODS: This experimental study used semen samples from 60 normozoospermic men. Specimens were divided into equal aliquots for processing by swim up (group A), and upstream (group B) methods and compare with control by raw semen (group C). Sperm concentration, morphology, motility, DNA fragmentation and chromatin maturation were measured in these three groups. RESULTS: The results revealed that sperm concentration in the swim up samples was significantly greater than upstream samples (p≤0.04). as addition, motile sperm recovery including the percentage of progressive motility and a total number of motile sperm was better in the swim-up compared to an upstream method and raw semen (p≤0.001). The cell debris and seminal fluid were equally removed by both methods and the percentage of normal forms was also similar in both procedures (p≥0.4). In addition, sperm DNA fragmentation and chromatin maturation were not significantly different between the three groups (p≥0.1). CONCLUSION: According to results, apparently the upstream method had no significant efficiency to separate good quality sperms compare to swim up. Therefore, swim up seems to be a simple, inexpensive, reliable and widely available method with an efficient yield to separate motile sperm with good morphology and better chromatin integrity for insemination in the infertility clinics.

Evaluating the Potential of Three Sperm Surface Antigens as Egg-adhesion Biomarkers for Human Sperm Selection.

BACKGROUND: The selection of sperm with good genomic integrity and surface antigens is suggested for improving assisted reproductive technology (ART) outcome. The aim of this study was evaluating the heat shock protein (HSPA2), Dj-1 and serum amyloid P compound (SAP) three sperm surface proteomes as biomarkers for this purpose. METHODS: In this study, semen samples were obtained from 114 men who presented at Avicenna Fertility Clinic for their treatment. The semen characteristics, DNA fragmentation Index (DFI), chromatin maturation index (CMI), biomarker levels, and their embryo quality were considered. The paired-samples t-test and independent-samples t-test were used for analyzing the data and p-values<0.05 were considered significant. RESULTS: Outcomes exhibited the major reduction in HSPA2, DJ-1 and SAP following reduction in sperm quality and DNA integrity (p<0.001) with cut-off value of 14% (HSPA2), 12% (DJ-1) and 10% (SAP). The specificity of these three biomarkers was 95.2, 73.8 and 88.1%, respectively. Also, DFI (p<0.001), CMI (p<0.05), cleavage (p<0.05), and embryos quality (p<0.001) decreased significantly in abnormal spermiogram (ANS) group in compared with normal spermiogram (NS) group. It was shown that DFI was 97.1% in HSPA2, 76.5% in DJ-1 and 94.1% in SAP, and CMI was 95.0%, 75.50% and 87.5%, respectively. The significant correlation was found between of the three biomarkers and CMI (p<0.001), DFI (p<0.001) and embryos quality (p<0.001). CONCLUSION: By comparing the efficiency of these three biomarkers for selecting sperm with the lowest level of chromatin damages, it seems that selection based on HSPA2 has significance over others.

Transplantation of Spermatogonial Stem Cells Suspension into Rete Testis of Azoospermia Mouse Model.

PURPOSE: The loss of spermatogonia following chemo-or radiotherapy leading to temporary or permanent infertility of the patient is a well known and unwanted side effect of many oncological therapies. MATERIALS AND METHODS: In this study, germ cells were isolated from 4 days old mouse testis cells. Busulfan treatment was used to the eliminate proliferating cells in the testis of recipient mice. The donor cells suspended in DMEM, were introduced into the rete testis of recipient mice via microinjection method. To distinguish the progeny of the transplanted donor stem cells from endogenous germ cells, BrdU-labeled cells were used. In addition, real time PCR was performed to determine expression levels of ngn3 and LIN28 (spermatogonia stem cells markers)before and after transplantation. Western blot analysis was further performed to detect an increase in - ngn3 expression after transplantation. RESULTS: Transplantations of stem cells into rete testis of the recipients was done. Our results clearly showed a significant increase in spermatozoa number in epididymal luman Spermatogonial stem cells (SSCs) did not show alkaline phosphatase activities while ngn3 and LIN28 were clearly expressed. Ngn3 and LIN28 expression were reduced after busulfan treatment compared to untreatmented mice. However, the expression of ngn3 and LIN28 increased after transplantation . BrdU-labeled testis cells were successfully transplanted into rete testis of recipient mice. These cells remained in rete testis of all recipient mice up to two months after transplantation. CONCLUSION: The present study clearly confirme that a regeneration after cytotoxic treatment was based on morphological criteria. We demonstrated the increase in stem cell numbers during regeneration and after transplantation. Transplantation of spermatogonial stem cells suspension by the injection of cells via the rete testis of recipient azoospermia model considerably enhances the efficiency of this procedure.

Efficacy of Intrauterine Injection of Granulocyte Colony Stimulating Factor (G-CSF) on Treatment of Unexplained Recurrent Miscarriage: A Pilot RCT Study.

BACKGROUND: Endometrium undergoes several changes in structure and cellular composition during pregnancy. Granulocyte Colony-stimulating Factor (GCS-F) is an important cytokine with critical role in embryo implantation and pregnancy. The aim of the present study was to evaluate the impact of intrauterine injection of G-CSF in patients who suffer from unexplained recurrent miscarriage (RM). METHODS: In the present randomized clinical trial, a total of 68 patients were randomly allocated into two study groups including intrauterine G-CSF (n=23, 300 µg) injection and control group (n=27, no G-CSF injection). Eighteen out of 68 patients were excluded from the final analysis due to different reasons. All patients were in Ovulation Induction (I/O) cycle. In G-CSF group, intrauterine injection of G-CSF was done twice in the cycle. All enrolled patients were under 40 years old and had at least two unexplained pregnancy losses. Pregnancy was evaluated by titer of ßhCG, presence of gestational sac (implantation) and fetal heart rate (clinical pregnancy) was assessed by vaginal ultrasonography. Student's T test and Mann-Whitney U were used for analysis. The p≤0.05 was determined as statistically significant. RESULTS: No significant differences were observed between the two study groups when the rates of chemical pregnancy (26.1% vs. 29.6%, p=0.781), implantation (26.1% vs. 22.2%, p=0.750), clinical pregnancy (17.4% vs. 11.1%, p=0.689) and abortion (33% vs. 37.5%, p=0.296) were compared. CONCLUSION: In our study, no significant difference was observed between the two study groups when the rates of chemical pregnancy, implantation, clinical pregnancy and abortion were compared.

Role of vitamin E and D3 supplementation in Intra-Cytoplasmic Sperm Injection outcomes of women with polycystic ovarian syndrome: A double blinded randomized placebo-controlled trial.

BACKGROUND & AIMS: Ovulation induction has been proven to impose oxidative stress during ICSI treatment cycle. Also, data indicates that PCOS women show higher oxidative markers. Available data are not convincing about which antioxidant supplements have the potential to overcome oxidative stress in PCOS subjects. The aim of this trial was to investigate the possible role of combined vitamin E and D supplementation in the ICSI outcomes (oocyte number and quality, embryo number and quality, pregnancy rate) of PCOS subjects. METHODS: A total of 105 PCOS infertile women scheduled for ICSI were enrolled in a double-blinded RCT to treatment group (vitamin E, 400 mg/day - and vitamin D3, 50,000 IU/one in two weeks, n = 52) or placebo group (n = 53) for 8 weeks. The primary outcomes were implantation rate, pregnancy and clinical pregnancy rate. Secondary outcomes included oocyte quality, embryo quality, fertilization rate, alteration in serum MDA, TAC and vitamin D3 after treatment. Further, association between serum and follicular fluid Malondialdehyde (MDA), Total Antioxidant Capacity (TAC), and serum vitamin D3 level were assessed. RESULTS: Pregnancy, clinical pregnancy and implantation rate were significantly higher in treatment group (P < 0.001). Data analysis in both groups revealed a significant increase in serum MDA compared to baseline and a significant decrease in serum TAC compared to baseline after treatment. Further analysis showed that there is a positive weak association between vitamin D level, implantation rate (P = 0.015) and increased clinical pregnancy (P = 0.037). No significant association was detected between either follicular fluid or serum MDA and TAC and ICSI outcomes. CONCLUSIONS: In conclusion, the findings of this trial do not add clinical support to the evidence that vitamins E and D3 may play a role in the success rate of IVF via an antioxidant mechanism. REGISTRY CODE: IRCT2014081018662N1.

Ooplasmic transfer in human oocytes: efficacy and concerns in assisted reproduction.

BACKGROUND: Ooplasmic transfer (OT) technique or cytoplasmic transfer is an emerging technique with relative success, having a significant status in assisted reproduction. This technique had effectively paved the way to about 30 healthy births worldwide. Though OT has long been invented, proper evaluation of the efficacy and risks associated with this critical technique has not been explored properly until today. This review thereby put emphasis upon the applications, efficacy and adverse effects of OT techniques in human. MAIN BODY: Available reports published between January 1982 and August 2017 has been reviewed and the impact of OT on assisted reproduction was evaluated. The results consisted of an update on the efficacy and concerns of OT, the debate on mitochondrial heteroplasmy, apoptosis, and risk of genetic and epigenetic alteration. SHORT CONCLUSION: The application of OT technique in humans demands more clarity and further development of this technique may successfully prove its utility as an effective treatment for oocyte incompetence.