Levels of serum ß-human chorionic gonadotropin after embryo transfer and subsequent miscarriage, pre-eclampsia, and intrauterine growth restriction.

Background: This study aimed to examine maternal serum concentration of ß-human chorionic gonadotropin (ß-hCG) on Day 16 after embryo transfer and risk of miscarriage, pre-eclampsia, and intrauterine growth restriction (IUGR). Methods: In this study, we evaluated 125 pregnancies following in vitro fertilization (IVF). ß-hCG concentrations were measured on the morning of Day 16 after embryo transfer. Baseline characteristics of the study participants were also recorded. Results: Concentrations of ß-hCG on Day 16 after embryo transfer were inversely associated with the higher risk of miscarriage (p < 0.001), but did not with pre-eclampsia and IUGR (p > 0.05). Spearman's correlation coefficient showed a reverse and significant association between ß-hCG and higher risk of miscarriage (σ = 0.531 and p < 0.001). There was a significant association between frozen embryo transfer and the risk of IUGR and pre-eclampsia (p = 0.005 and p = 0.023, respectively). Conclusions: Maternal serum concentrations of ß-hCG on Day 16 after IVF/embryo transfer were associated with the higher risk of miscarriage, but not pre-eclampsia and IUGR.

Association between follicular fluid bacteria with inflammatory markers of the complete blood count and the outcomes of assisted reproductive technology in women with endometriosis: A case-control study.

Background and Aims: Endometriosis is a common reason for infertility and poor outcomes of assisted reproductive technology (ART). Inflammation is involved in the pathogenesis of this disease. The presence of microorganisms in women with endometriosis may increase levels of inflammatory markers. The purpose of this study is to determine the relationship between the presence of bacteria in the follicular fluid with the inflammatory markers of the complete blood count (CBC) and the outcomes of in vitro fertilization (IVF) in women with endometriosis. Methods: This case-control study was conducted on 74 patients undergoing IVF, referred to Al-Zahra Hospital in Rasht (Iran) in 2021. The patients were divided into two case groups including 37 women with endometrioma and the control group, including 37 infertile women with a male factor and normal ultrasound. In total, 74 follicular fluids were collected from the case and control groups and were cultured in the laboratory. The relationship between culture results with IVF outcomes and the levels of CBC inflammatory markers including the number of white blood cells (WBCs), lymphocytes, neutrophils, neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR), erythrocyte sedimentation rate (ESR), and c-reactive protein (CRP) was analyzed. Results: There was no significant statistical difference between the frequency of bacteria present in the follicular fluid (p = 0.861), the mean rate of fertilization (p = 0.363), the frequency of CRP (p = 0.999), and the mean WBCs, lymphocytes, neutrophils, NLR, LMR, and PLR in the two groups. There was a significant statistical difference between the mean number of oocytes of metaphase II (p = 0.034) and the mean ESR (p = 0.018) in the two groups. Conclusions: It seems necessary to evaluate follicular fluid as a biological substance that is considered an optimal factor for predicting oocyte quality, fertilization rate, embryo quality, and the success rate of ART.

Pregnancy outcome in long- versus short-acting gonadotropin-releasing hormone agonist cycles in participants with normal ovarian reserve: An RCT.

Background: There is no agreement on which of the 2 gonadotropin-releasing hormone (GnRH) agonist protocols are the most efficient, neither there is any consensus on which one yields a better clinical pregnancy percentage. Objective: The present study aims to compare the effectiveness of reduced dosages of long- and short-acting GnRH agonists on pregnancy outcomes. Materials and Methods: In this randomized controlled clinical trial, 400 women were randomly assigned to 2 groups (n = 200/group): the reduced dosage of long-acting GnRH agonist group (group 1, 1.25 mg Decapeptyl) and the short-acting GnRH agonist group (group 2, 0.5 mg/day Buserelin Acetate). The study was conducted at Mehr Medical Institute, Rasht, Iran between July 2019 and July 2020. Biochemical and clinical pregnancy were compared between groups. Results: No significant differences were observed in the endometrial lining, the total number of retrieved and metaphase-II oocytes, progesterone, and serum estradiol levels on human chorionic gonadotropin day, fertilization rate, and top-quality embryos between the groups. The duration of induction (10.8 ± 1.7 vs. 10 ± 2.1, p < 0.001) and the total dosage of gonadotropins (2939.4 ± 945.9 vs. 2441 ± 1247.1, p < 0.001) were significantly greater in group 2 than in group 1. No significant differences were observed between the 2 groups in terms of implantation rate, chemical pregnancy rate, and clinical pregnancy rate. A higher percentage of ovarian hyperstimulation syndrome was observed in group 2 (p = 0.005). Conclusion: Due to a lower percentage of ovarian hyperstimulation syndrome in group 1 and similar assisted reproductive technology outcomes in both groups, the long protocol was found to be superior to the short protocol.

Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system.

PURPOSE: Development of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cytoarchitecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model. METHODS: The testicular cells were harvested from the three brain-dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT-PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post-meiotic marker and apoptotic genes of Bax (BCL2-Associated X Protein) and Bcl-2 (B-cell lymphoma 2), respectively by using RT-qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC). RESULTS: Relative expression of SCP3, PRM2 and Bcl-2 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast-like cells. CONCLUSION: Our findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence of SCF. Developed organoids are capable of recapitulating many important properties of a stem cell niche.

A report of two cases with successful oocyte retrieval after wrongly earlier injection of ovulation induction trigger: Case report.

Oocyte maturation is induced by trigger administration and is acritical step in the success of assisted reproductive technology (ART)treatment. The ideal time interval between trigger administration and oocyte retrieval varies in the literature. Extremely short or long time intervals are both knownto cause unfavorable outcomes in oocyte collection. Accurate control over theinterval between trigger injection and oocyte retrieval is very important forwomen undergoing in vitro fertilization (IVF), to avoid unexpected premature ovulation. In this report, we present two infertile women who mistakenly injected the triggering dose of gonadotropin releasing hormone agonist (GnRHa) 12 h earlier. Case 1 and case 2 were 23 and 30 years old, respectively. Therewas no intervention to prevent pre-operative ovulation, and oocyte retrievalhas been done 48-50 h after trigger injection. oocytes and embryos quality wereacceptable. In conclusion, in patients who have the wrong trigger injection, oocyte retrieval is recommended after consulting the patient about theadvantages and disadvantages of the oocyte retrieval operation.

Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet-rich plasma.

BACKGROUND: Effective culture systems for attachment, migration, proliferation, and differentiation of spermatogonial stem cells (SSCs) can be a promising therapeutic modality for preserving male fertility. Decellularized extracellular matrix (ECM) from native testis tissue creates a local microenvironment for testicular cell culture. Furthermore, platelet-rich plasma (PRP) contains various growth factors for the proliferation and differentiation of SSCs. METHODS: In this study, human testicular cells were isolated and cultured for 4 weeks, and SSCs were characterized using immunocytochemistry (ICC) and flow cytometry. Human testicular tissue was decellularized (0.3% SDS, 1% Triton), and the efficiency of the decellularization process was confirmed by histological staining and DNA content analysis. SSCs were cultured on the human decellularized testicular matrix (DTM) for 4 weeks. The viability and the expression of differentiation genes were evaluated by MTT and real-time polymerase chain reaction (PCR), respectively. RESULTS: Histological evaluation and DNA content analysis showed that the components of ECM were preserved during decellularization. Our results showed that after 4 weeks of culture, the expression levels of BAX, BCL-2, PLZF, and SCP3 were unchanged, while the expression of PRM2 significantly increased in the cells cultured on DTM supplemented with PRP (ECM-PRP). In addition, the expression of GFRA1 was significantly decreased in the ECM group compared to the control and PRP groups. Furthermore, the MTT test indicated that viability was significantly enhanced in cells plated on DTM supplemented with PRP. CONCLUSION: Our study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs.

Evaluation of PGK2 and ACR proteins in seminal plasma: suggestion of potential new biomarkers for prediction of sperm retrieval in non-obstructive azoospermia patients.

This study aimed to assess the role of testis-specific proteins, PGK2 and ACR, in the prediction of sperm retrieval results by microdissection testicular sperm extraction (micro-TESE) in men with non-obstructive azoospermia (NOA). This was a case-control study including 48 semen samples of NOA patients undergoing the micro-TESE procedure, 15 semen samples from normozoospermic men as the positive control, and 12 semen samples from obstructive azoospermia/post-vasectomy (OA/PV) as negative controls. We investigated the levels of PGK2 and ACR proteins by ELISA tests in seminal plasma samples. The ELISA results revealed a significantly higher concentration of PGK2 and ACR in the NOA patients with successful sperm retrieval (NOA+) in comparison to NOA patients with failed sperm retrieval (NOA-) group (p = 0.0001 in both cases). For the first time, the data from this study suggests that a seminal PGK2 concentration of 136.3 pg/ml and ACR concentration of 21.75 mIU/ml can be used as cut-off values for the prediction of micro-TESE outcomes in NOA patients. These findings may be useful to avoid unnecessary micro-TESE operations. Overall, the seminal levels of the PGK2 and ACR proteins may be useful in predicting sperm retrieval success by micro-TESE in NOA patients.

The association between reproductive history and menopausal symptoms: an evidence from the cross-sectional survey.

BACKGROUND: During menopause, women experience annoying symptoms which may affect their daily activities and quality of life. This study aimed to determine whether reproductive history, an important indicator of estrogen exposure across the lifetime, is associated with the severity of menopausal symptoms in women. METHODS: This study was a cross-sectional study conducted on 214 women aged 35-65 who were randomly selected, and data was collected by a predesigned structured questionnaire. Each item was graded by subjects and a total score was obtained by summing all subscale scores. RESULTS: There was a significant association between the somatic, psychological, and urogenital menopausal symptoms and reproductive characteristics. Women with a history of abortion had greater total (ß = 0.194, p = 0.009), and psychological (ß = 0.230, p = 0.002) symptoms score. Women with higher number of children were more likely to have higher somatic (ß = 0.212, p = 0.005) symptoms than others. CONCLUSIONS: Our findings showed reproductive factors may have an influence on the severity of menopausal symptoms. After confirmation by further studies, these findings may help target women at risk of more severe menopausal symptoms at later ages.

Cell-free mtDNA level and its biomarker potency for ART outcome are different in follicular fluid of PCOS and non-PCOS women.

INTRODUCTION: Lack of reliable biomarkers for estimating the outcome is one of the current gaps in ART. In this study, we assessed whether cell-free mitochondrial DNA within the follicular fluid (FF cf-mtDNA) of PCOS patients has biomarker applicability or not. Furthermore, probable involved mechanisms in the FF cf-mtDNA pathway were evaluated. METHODS: The level of FF cf-mtDNA was compared between 50 PCOS patients and 50 women without any certain reproductive disorder, and analyzed for correlations with ART outcome. The associations between levels of FF cf-mtDNA and TFAM, POLG, and RNase H1 genes expression in mural granulosa cells (MGCs), as well as IL-6, and TNFα in follicular fluid (FF) were assessed. RESULTS: We identified that FF cf-mtDNA level was significantly lower in PCOS women and was accompanied by a reduction in the expression of mtDNA biogenesis genes in MGCs of the patients. Although a significant association between FF cf-mtDNA level and ART outcome was observed in the control group, no correlation could be proved in the PCOS group. Moreover, the expression level of TFAM was negatively associated, while amounts of IL-6 and TNFα were positively correlated with FF cf-mtDNA level in both groups. CONCLUSION: PCOS patients present a lower FF cf-mtDNA level in comparison with non-PCOS women. FF cf-mtDNA has biomarker applicability for ART outcome in women without any certain reproductive disorder, but not for those with PCOS. It seems that mtDNA packaging dysfunction results in elevated FF cf-mtDNA, and subsequent effects are triggered by increasing the inflammatory cytokines.

Sertoli cell-only syndrome: etiology and clinical management.

Almost 50% of infertility cases are due to male factors, and spermatogenesis failure is one of the most severe forms of male infertility. Sertoli cell-only syndrome (SCOS) also known as germ cell aplasia is characterized by azoospermia in which the seminiferous tubules of testicular biopsy are lined only with Sertoli cells. The definitive diagnosis of SCOS is by diagnostic testicular biopsy. Although SCOS may be a result of Klinefelter syndrome, most of the SCOS men have a normal karyotype. Along with genetic aberrations, signaling pathways and endocrine processes might be major factors in the development of SCOS. Sperm retrieval and intracytoplasmic sperm injection (ICSI) are available treatments for SCOS. However, some SCOS patients do not have therapeutic options to help them having a biological child. This review aims to summarize our present knowledge about SCOS and to highlight the importance of future researches in the diagnosis and treatment of this disorder.

The Germ Cell-Specific Markers ZPBP2 and PGK2 in Testicular Biopsies Can Predict the Presence as well as the Quality of Sperm in Non-obstructive Azoospermia Patients.

To assess the role of three testis-specific genes including ZPBP2, PGK2, and ACRV1 in the prediction of sperm retrieval result and quality of retrieved sperm by microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients. This was a case-control study including 57 testicular samples of NOA patients including 32 patients with successful sperm retrieval (NOA+) and 25 patients with failed sperm retrieval (NOA-), and 9 samples of men with normal spermatogenesis in the testes as the positive control (OA). We investigated the expression of candidate genes by RT-qPCR and germ cell population patterns by DNA flow cytometry in testicular biopsy samples. The association between PGK2 expressions with the quality of retrieved spermatozoa was also evaluated. The RT-qPCR data revealed a significantly higher expression of ZPBP2 and PGK2 in the NOA+ in comparison to NOA- group (P = 0.002, and P = 0.002, respectively). Flow cytometry results revealed that the haploid cell percentage was significantly higher in NOA+ vs. NOA- group (P = 0.0001). In samples with a higher percentage of haploid cells, expression levels of ZPBP2 and PGK2 were higher (P = 0.001). The PGK2 expression was significantly associated with retrieved sperm quality (P = 0.01). Our results contribute to the search for the biomarkers for predicting the presence of testicular sperm and would be useful to avoid unnecessary multiple micro-TESE. Overall, the expression pattern of the ZPBP2 and PGK2 may be useful in predicting sperm recovery success and quality of retrieved sperm in NOA patients.

Three-dimensional co-culture of human spermatogonial stem cells with Sertoli cells in soft agar culture system supplemented by growth factors and Laminin.

Application of a three-dimensional (3D) culture system for in vitro proliferation and differentiation of human spermatogonial stem cells (SSCs) is a useful tool for the investigation of the spermatogenesis process and the management of male infertility particularly in prepubertal cancer patients. The main purpose of this study was to investigate the proliferation of human SSCs co-cultured with Sertoli cells in soft agar culture system (SACS) supplemented by Laminin and growth factors. Testicular cells were isolated from testes of brain-dead patients and cultured in two-dimensional (2D) culture system for 3 weeks. After 3 weeks, functional SSCs were evaluated by xenotransplantation and also identification of cells was assessed by immunocytochemistry, flow cytometry, and RT-PCR. Then, SSCs and Sertoli cells were transferred to the upper layer of SACS for 3 weeks. After 3 weeks, the number of colonies and the expression of specific SSCs and Sertoli cell markers, as well as apoptotic genes were evaluated. Our results showed that transplanted SSCs, migrated into the basement membrane of seminiferous tubules of recipient mice. The expression of PLZF, α6-Integrin, and Vimentin proteins in SSCs and Sertoli cells were observed in 2D and 3D culture systems. The expression rate of PLZF, α6-Integrin, Bcl2, and colony number in SACS supplemented by Laminin and growth factors group were significantly higher than non-supplemented groups (P ≤ 0.01), but the expression rate of c-kit and Bax in supplemented group were significantly lower than non-supplemented groups (P ≤ 0.05). This 3D co-culture system decreased apoptosis and increased propagation of human SSCs. Therefore, this designed system can be utilized to increase the proliferation of human SSCs in prepubertal male cancer and azoospermic men to obtain an adequate SSCs number to outotransplant success and in vitro spermatogenesis.