Fertile Control Group Research Articles

Serum pentraxin-3 expression varies according to polycystic ovary syndrome phenotypes.

Pentraxin-3 (PTX-3) is a multibiological protein involved in cumulus cell expansion, fertilization, and implantation. This study was designed to analyze how circulating PTX-3 levels change in women with polycystic ovary syndrome (PCOS). A total of 50 Turkish participants, 35 of whom had PCOS and 15 of whom were fertile, were included in the study. Patients in the PCOS group were divided into 4 different phenotypes according to the NIH criteria (phenotypes A-D). The number of patients in phenotype A was the highest 13 (37.1%). In the calculations made without phenotyping, the serum ptx3 levels of the PCOS group were found to be significantly lower compared to the fertile control group (3.32 ± 0.73 ng/mL vs 4.97 ± 1.29 ng/mL; P < .001). The ptx3 value of phenotypes A and B was significantly lower than phenotype D (P = .008 and P = .009, respectively). When the phenotypes were compared with the fertile control group, the PTX-3 levels of phenotypes A and B were significantly lower than the fertile group. Although the ptx3 levels of phenotypes C and D were lower than the fertile group, the difference did not reach statistical significance. This is the first study to investigate serum ptx3 levels by phenotype in PCOS. While serum PTX-3 levels decreased in phenotypes A and B, ptx3 levels in phenotypes C and D were similar in fertile patients.

High levels of bisphenol A among infertile men can impair spermatogenesis by oxidative stress and elevated levels of microRNA-337

BackgroundStudies have shown that Bisphenol A may interfere with the process of spermatogenesis and result in a decrease in the quality of semen. Nevertheless, the fundamental processes remain unclear. This study was done to investigate the connections between exposure to Bisphenol A, spermatogenesis with microRNA-337, and malondialdehyde in infertile men.MethodsThis study was a case–control study on 73 participants. Infertile group (1a): azoospermia (n = 16), infertile group (1b): oligozoospermia (n = 22), and control group (2): normospermic (n = 35) were enrolled in this study. Full history, local examination, semen analysis, and urine and blood samples were taken from all participants. Urinary Bisphenol A, malondialdehyde, and serum microRNA-337 were measured.ResultsThe mean Bisphenol A level in azoospermia group shows statistically significant increase comparing to fertile control group. The mean microRNA-337 level in oligozoospermia and azoospermia group shows statistically significant increase comparing to fertile controls. The mean malondialdehyde level in infertility groups shows statistically significant increase comparing to fertile control group. No linear correlations were recorded between Bisphenol A levels with semen quality parameters, hormonal profile, and microRNA-337.ConclusionWhile there is no significant change in the levels of Bisphenol A between normal fertile males and infertile males with oligozospermia, a significant elevation in the BPA level was observed in infertile males with azoospermia. A significant upregulation of the miRNA-337 gene expression in infertile males either oligozospermia or azoospermia was also observed. In addition, lipid peroxidation as evident by the significant elevation of MDA levels was marked among infertile patients.

Altered N6-methyladenosine methylation level in spermatozoa messenger RNA of the male partners is related to unexplained recurrent pregnancy loss.

Understanding the pathogenesis of unexplained recurrent pregnancy loss is paramount for advancing effective treatments. Various biological processes, including spermatogenesis and embryo development, are tightly regulated by N6-methyladenosine modifications. However, few studies have focused on the impact of sperm N6-methyladenosine modifications on embryonic development. Therefore, we aimed to study altered N6-methyladenosine-mediated messenger RNA methylation modifications in the spermatozoa of male partners from couples experiencing unexplained recurrent pregnancy loss, to identify potential diagnostic markers and explore their potential molecular mechanisms in pregnancy loss and embryogenesis. Methylated RNA immunoprecipitation (MeRIP) sequencing and RNA sequencing were conducted on the spermatozoa of men from couples in the 'unexplained recurrent pregnancy loss' group (n=6), and the fertility control group (n=6). To identify the role of the detected key genes, zebrafish model embryos were studied, and multi-omics (transcriptomics, proteomics, and metabolomics) analyses helped to explore the molecular mechanism of abnormal embryogenesis. Comparing unexplained recurrent pregnancy loss with the fertility control group, 217 N6-methyladenosine peaks were significantly upregulated, and 40 were downregulated in the spermatozoa. The combined analyses of spermatozoa-methylated RNA immunoprecipitation sequencing and RNA sequencing indicated that N6-methyladenosine methylation and the expression of SEMA5A, MT-ATP6, ZNF662, and KDM4C were significantly different. In zebrafish embryos, the altered expression of the four genes increased embryonic mortality and malformations by disturbing several key signaling pathways and zygotic genome activation. This study highlights the paternal epigenome, which could be one of the reasons for faulty embryogenesis leading to pregnancy loss. The N6-methyladenosine modification, the most prevalent RNA modification, contributes to the exploration and understanding of the paternal epigenome in the maintenance of pregnancy and fetal growth and development. The four genes identified in this study may serve as potential diagnostic markers and elucidate novel molecular mechanisms of embryogenesis.

Association between infertility and risk of cardiovascular diseases in young women

Scientists and medical experts are beginning to understand the significant role that a woman’s past reproductive experiences play in her potential risk of developing heart disease. Reproductive history is seldom considered when assessing the cardiovascular risk. Infertility, high blood pressure, and hypertension are risk factors for heart disease. Additional analyses were conducted to determine whether the apparent increase in risk could be influenced by other risk factors associated with infertility such as irregular menstruation, thyroid conditions, and waist circumference. This study aimed to investigate the association between cardiovascular risk and infertility in women and those who are fertile. The objectives of this study were to estimate the levels of serum creatinine kinase (MB) in both infertile and fertile control groups, as well as to estimate the LDL and HDL levels in the same groups. The study group comprised 78 participants, of which 39 were fertile and 39 were infertile. CK (MB), HDL, LDL, Uric acid, and high sensitivity C reactive Protein (hs-CRP) levels were analyzed. In this study, we discuss how infertility may share common pathways with cardiovascular diseases. Numerous mechanisms may be involved in mediating infertility, including ovulatory abnormalities, endometriosis, and uterine fibroids. For example, in addition to having lower HDL levels, women with polycystic ovary syndrome (PCOS) are more likely to have higher levels of total cholesterol, triglycerides, and LDL. PCOS patients typically exhibit elevated UA levels and hyperuricemia, which are commonly associated with increased androgen levels.

Upregulated RPA2 in endometrial tissues of repeated implantation failure patients impairs the endometrial decidualization.

To investigate the expression and underlying mechanism of RPA2 in endometrium of patients with repeated implantation failure (RIF). In this study, we retrieved the expression profiles from GEO databases and filtered the differentially expressed genes between RIF and the fertile control group. Ultimately, RPA2 was confirmed as a target gene. RPA2 expression in endometrial tissues of RIF patients, the control group, and different phases was detected by RT-qPCR, immunohistochemistry, and Western blotting. The role of RPA2 in endometrial decidualization was performed by in vitro decidualization inducing by 8-Br-cAMP and MPA. Furthermore, RT-qPCR was used to detect changes in the decidual biomarkers after transfection of RPA2 overexpression vector in human endometrium stromal cell (HESC). RPA2 was significantly upregulated in the mid-secretory endometrium of patients with RIF. As a proliferation-related gene, RPA2 was obviously higher expressed at proliferative phase during the normal menstrual cycles. Moreover, the downregulation of RPA2 was discovered during decidualization of HESC. Furthermore, RPA2 overexpression impaired decidualization by inhibiting the expression of prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Our finding indicated that aberrant upregulation of RPA2 attenuated decidualization of HESC in RIF women and provided new potential therapeutic targets.

Preliminary analysis of hormonal and seminal sperm parameters of subfertile and fertile stallions treated with the GnRH agonist lecirelin

Subfertility is relatively common in stallions and few treatments are available to improve the semen characteristics of affected animals. The GnRH therapy, however, has been reported to improve sperm quality in individual clinical cases. The aim of this study was to evaluate therapeutical effects of daily GnRH analogue lecirelin treatment on sperm characteristics of fertile and subfertile stallions and to assess differences in the endocrine response among groups. Stallions were divided into untreated (n=3), fertile control (n=4) and subfertile (n=4) groups. Fertile and sub-fertile stallions received saline or 0.05 mg lecirelin, respectively, daily for 60 days and were evaluated for >90 days post-treatment. Ejaculates were collected biweekly, but semen and serum samples were evaluated weekly. Fertile stallions were characterized as having normal seminal parameters and a 75% conception rate. Subfertile stallions had low total and progressive motility (< 50% and 20%, respectively), sperm count, abnormal morphology (>30% of major defects), and ≈20% conception rate in the previous breeding season. Testicular response was assessed by determination of the concentrations of testosterone and estrone sulphate (interstitial compartment), anti-Müllerian hormone (AMH) and inhibin-B (tubular compartment) and correlated with sperm kinetics and production. Sperm kinetics were evaluated by CASA (IVOS Version 12 Hamilton Thorne Research, MA, USA), sperm concentration by Neubauer hemocytometer (Optik Labor®, Lancing, England), serum concentrations of inhibin- B, AMH and estrone sulfate weredetermined by ELISA and testosterone by RIA. Data were analyzed by ANOVA for repeated measures. In the subfertile group, testosterone concentrations decreased from week 0 to 6 (667 vs 302 pg/ml, respectively P<0.05), but tended to recover 1 month after treatment ceased (469 to 977pg/ml, respectively, P<0.08). Concentrations of all hormones were positively correlated with one another and with total sperm number (P<0.01), the highest between inhibin-B and estrone sulphate (R2=0.72) (P<0.01). No statistically significant effects on other hormones or semen characteristics were detected irrespective of group. Despite non statistical difference between groups, it is important to note that total sperm number increased by 60% 30 days after the beginning of the treatment and remained elevated by the end of the evaluation period in the fertile group. These data suggest that the treatment increased sperm production, but further investigations are underway. In conclusion, lecirelin treatment did not affect hormonal and seminal parameters in treated stallions, and did not affect testicular function. The correlation between hormones and sperm output indicates that these hormones are potentially valuable biomarkers for testicular function. Acknowledgements: São Paulo Research Foundation (FAPESP, grant #2019/01119-8 and #2022/03405-0), John P. Hughes Endowment and Clinical Endocrinology Laboratory, UC-Davis.

Mst1-mediated phosphorylation of Nur77 improves the endometrial receptivity in human and mice.

Successful embryo implantation requires the attachment of a blastocyst to the receptive endometrial epithelium, which was disturbed in the women with recurrent implantation failure (RIF). Endometrial β3-integrin was the most important adhesion molecule contributing to endometrial receptivity in both humans and mice. Nur77 has been proven indispensable for fertility in mice, here we explore the role of Nur77 on embryo-epithelial adhesion and potential treatment to embryo implantation failure. The expression and location of Mst1 and Nur77 in endometrium from fertile women and RIF patients were examined by IHC, qRT-PCR and Western blotting. Invitro kinase assay following with LC-MS/MS were used to identify the phosphorylation site of Nur77 activated by Mst1. The phosphorylated Nur77 was detected by phos-tag SDS-PAGE assay and specific antibody against phospho-Nur77-Thr366. The effect of embryo-epithelium interaction was determined in the BeWo spheroid or mouse embryo adhesion assay, and delayed implantation mouse model. RNA-seq was used to explore the mechanism by which Nur77 derived peptide promotes endometrial receptivity. Endometrial Mammalian sterile 20 (STE20)-like kinase 1 (Mst1) expression level was decreased in the women with RIF than that in the fertile control group, while Mst1 activation in the epithelial cells promoted trophoblast-uterine epithelium adhesion. The effect of Nur77 mediated trophoblast-uterine epithelium adhesion was facilitated by active Mst1. Mechanistically, mst1 promotes the transcription activity of Nur77 by phosphorylating Nur77 at threonine 366 (T366), and consequently increased downstream target β3-integrin expression. Furthermore, a Nur77-derived peptide containing phosphorylated T366 markedly promoted mouse embryo attachment to Ishikawa cells ([4 (2-4)] vs [3 (2-4)]) and increased the embryo implantation rate (4 vs 1.4) in a delayed implantation mouse model by regulating integrin signalling. Finally, it is observed that the endometrial phospho-Nur77 (T366) level is decreased by 80% in the women with RIF. In addition to uncovering a potential regulatory mechanism of Mst1/Nur77/β3-integrin signal axis involved in the regulation of embryo-epithelium interaction, our finding provides a novel marker of endometrial receptivity and a potential therapeutic agent for embryo implantation failure. National Key Research and Development Program of China (2018YFC1004400), the National Natural Science Foundation of China (82171653, 82271698, 82030040, 81971387 and 30900727), and National Institutes of Health grants (R01HL103869).

TOP2A deficit-induced abnormal decidualization leads to recurrent implantation failure via the NF-κB signaling pathway

BackgroundSuccessful implantation is a complex process that is influenced by embryo quality, endometrial receptivity, immune factors, and the specific type of in vitro fertilization protocol used. DNA topoisomerase IIα (TOP2A) is a well-known protein involved in cell proliferation; however, its expression and effect on the endometrium in recurrent implantation failure (RIF) have not been fully elucidated.MethodsThe human endometrial tissues of healthy controls and patients with RIF were collected. A proteomic analysis was performed to evaluate the differentially expressed proteins between the RIF group and the fertile control group. The expression patterns of TOP2A in the human preimplantation endometrium of the patients with RIF were determined by immunohistochemical staining, Western blotting and qRT-PCR. TOP2A knockdown (sh-TOP2A) T-HESCs were generated using lentiviruses. The expression of TOP2A in T-HESCs was manipulated to investigate its role in decidualization. The TOP2A-related changes in decidualization were screened by mRNA sequencing in decidualized TOP2A knockdown and control T-HESCs and then confirmed by Western blotting and immunofluorescence staining. TOP2A-deficient mice were generated by injection of TOP2A-interfering adenovirus on GD2.5 and GD3.5.ResultsWe performed a proteomic analysis of endometrial tissues to investigate the potential pathogenesis of RIF by comparing the patients with RIF and the matched controls and found that TOP2A might be a key protein in RIF. TOP2A is ubiquitously expressed in both stromal and glandular epithelial cells of the endometrium. The data indicate that TOP2A expression is significantly lower in the mid-secretory endometrium of women with RIF. TOP2A expression was downregulated under stimulation by 8-bromo-cAMP and MPA. Ablation of TOP2A resulted in upregulated expression of decidual biomarkers and morphological changes in the cells. Mechanistic analysis revealed that TOP2A regulates the NF-κB signaling pathway in decidualized T-HESCs. The TOP2A-deficient mice exhibited lower fetal weights.ConclusionsOur findings revealed that abnormal expression of TOP2A affects decidualization and changes the “window of implantation”, leading to RIF. TOP2A participates in the processes of decidualization and embryo implantation, functioning at least in part through the NF-κB pathway. Regulating the expression of TOP2A in the endometrium may become a new strategy for the prevention and treatment of RIF.

P-394 Peripheral and uterine natural killer cell cut-off levels and their correlation in fertile controls and women with unexplained recurrent implantation failure (RIF)

Abstract Study question What is cut-off value for peripheral and uterine natural killer (pNK, uNK) cells and any correlation between them in fertile and RIF women? Summary answer Cut-off values were 7% for uNK and 11.6% for pNK in fertile controls. No significant correlation observed among fertile and infertile women with RIF. What is known already Association of uNK and pNK cells with infertility is debatable. It is not clear whether measuring NK cell levels in RIF has a role. pNk cell levels however has been used in various studies to guide the need of immunotherapy in RIF women. Definition of normal range of uNK cell numbers has also not been stated clearly because of lack of standardised protocol. Literature reveals the cut off values for pNk cells to be 12% or 18% and for uNK cells between 5-12.9%. Correlation between uNK and pNK cells in fertile or RIF women has not been studied earlier. Study design, size, duration A prospective study was conducted at ART Centre, Department of Obstetrics and Gynecology at tertiary care institute during a period from January 2019 to January 2021. Thirty women with RIF and fifty fertile controls with age &amp;lt;35 years having regular menstrual cycles and no hormonal treatment in last 3 months were enrolled in the study for uNK and pNK cell testing. The permission was obtained from the Institute Ethics Committee. Participants/materials, setting, methods Subjects included women with RIF with age 20-35 years, tubal and unexplained factors, normal ovarian reserve, normal karyotype and normal uterine cavity. In both subjects and controls, midluteal endometrial biopsy sample was taken for Immunohistochemistry staining of CD 56+ cells to determine uNK cells. Peripheral venous blood was also obtained at the time of biopsy for flow cytometry in peripheral blood mononuclear cells to determine pNK cells. Cut off values were derived from fertile controls. Main results and the role of chance The mean age and BMI were comparable between fertile control and study group(29.45±3.3 vs 31.17±3.3 years, 22.97±1.89 vs 23.21±2.2 kg/m2; p-value &amp;gt;0.05). The control group had median value of 7% for uNK cell levels and 11.6% for pNK cell levels. In RIF subjects, the median value for uNK cells was 13.5% and even though higher compared to controls it was not statistically significant(p-value &amp;gt;0.05) . The median pNK values were comparable among controls and RIF group (11.6% vs. 12%). All RIF subjects did not have elevated NK cell levels. 18 (60%) subjects had elevated uNK cell levels (more than 7%) whereas 40% had below the reference value. pNK cells were also elevated in only 53.3% of subject population. There was positive correlation found between uNK and pNK cell levels in RIF subjects, however it was not statistically significant (r 0.312, p-value 0.09). In fertile controls, there was insignificant negative correlation between uNK and pNK cell levels (r -0.231, p-value 0.33). Limitations, reasons for caution The small sample size is the foremost limitation of study. Wider implications of the findings uNK cell testing may be worthwhile in women with unexplained RIF so as to guide immunotherapy to improve pregnancy outcomes. As no significant correlation has been found between pNK and uNK cells in fertile and RIF women, pNK cell testing in RIF women may be done with caution. Trial registration number Not Applicable

AURKC Gene Polymorphism (rs58264281) Associated with Idiopathic Male Infertility Risk in Northeast of Iran: A Case-Control Study

Background: Infertility is a major public health and social problem in human reproduction that is known as a multifactorial complex disorder. Genetic background and mutations and single nucleotide polymorphisms (SNPs) on the genes involved in sperm development are the important causes of male infertility. Objectives: In this study, we evaluated the association of AURKC gene polymorphism (rs58264281) and idiopathic male infertility in the Iranian Azeri population. Methods: This study was performed among 100 men with idiopathic infertility (case group) and 100 healthy men with successful fertility (control group) from East Azerbaijan, Iran. Genomic DNA extraction was carried out from peripheral blood samples by the proteinase K method. Genotype analysis was conducted by the tetra-primer amplification refractory mutation system-polymerase chain reaction (Tetra-ARMS PCR). SPSS version 21 was used for the analysis of the obtained data. Results: We observed that the CA and AA genotypes were significantly increased in patients with infertility as compared to healthy controls. Our results demonstrated that the mutant allele of AURKC gene polymorphism (rs58264281) was a significant risk factor in male infertility. Conclusions: We suggested a significant correlation between the AURKC gene rs58264281 polymorphism and male infertility in the Iranian Azeri population. However, further studies are required among other ethnicities, races, and geographic areas with larger sample sizes.

Follicle-stimulating hormone (FSH) receptor gene polymorphisms in Iraqi patients with non-obstructive azoospermia

Background: Follicle-stimulating hormone (FSH) is a pivotal hormone for male fertility, and its action on gonads is exerted by FSH receptors (FSHRs). Objectives: To examine whether the presence of FSHR gene single nucleotide polymorphisms (SNPs), G919A and A2039G, involved in non-obstructive azoospermia (NOA) in Iraqi infertile men. Methods: Two common SNPs, A919G and A2039G, in the FSHR gene were analyzed in 104 subjects (70 infertile patients with NOO: 33 NOA patients were not receiving treatment and 37 were on infertility treatment, and 34 normozoospermic fertile men as controls). Results: The results revealed that the homozygous wild genotype (AA) of rs6165 FSHR gene SNP was more abundant than (AG) and (GG) genotypes in both groups of infertile NOA patients with a frequency of 49% in those who untreated, 81% in patients undergoing treatment and in the control group 41%. Whereas, the highest percentage of heterozygous genotype (AG) in the fertile control group was 41% when compared to NOApatients with a genotype frequency of 24% (for those who untreated) and 11% (for patients on treatment), respectively; with (A) allele frequency of 86% and the observed frequency of (G) allele was only 14% in the patients’ group as compared to that of controls that were (65 %) and (35 %), respectively. The rs6166 genotyping revealed that the homozygous wild genotype (GG) of FSHR gene was more abundant than (AG) and (AA) genotypes in NOA patients receiving infertility treatment with a frequency of (68%), in NOA patients who didn’t receive treatment 49%, while the lowest frequency was detected in the healthy fertile control group (47%). Conclusions: These results support the evidence that rs6165 and rs6166, FSHR SNPs, might be involved in the pathogenesis and protection against NOA, respectively.

Association of the single nucleotide polymorphism C1858T of the PTPN22 gene with unexplained recurrent pregnancy loss: A case-control study

Background Lymphoid-tyrosine-phosphatase which is encoded by the protein tyrosine phosphatase non-receptor 22 (PTPN22) gene plays a pivotal role in the regulation of immune responses by dephosphorylating several signaling intermediates of immune cells.ObjectiveSince a balanced immune response has been shown to be important during pregnancy, the purpose of this research was to compare the frequency of the PTPN22 C1858T polymorphism in women with unexplained recurrent pregnancy loss (URPL) vs. in a control group for the first time.Materials and MethodsGenomic DNA from 200 individuals with URPL and 200 individuals without URPL (the control group) at the infertility center in Yazd, Iran was isolated using the salting-out method. The PTPN22 C1858T polymorphism of the two groups was analyzed using polymerase chain reaction-restriction fragment length polymorphism. Genotype frequencies in the women with URPL and the fertile control group were compared using the Chi-square test.ResultsThere were significant differences in the frequency of the PTPN22 1858T polymorphism in the URPL individuals vs. the healthy controls, i.e. 32.0% and 21.5%, respectively (p = 0.01).ConclusionOur findings suggest that the PTPN22 1858T polymorphism could play a role in recurrent pregnancy loss. Therefore, genotyping of the mentioned polymorphism can help clinicians to predict the probable risk of URPL.

P–375 Intralipid supplementation in women with unexplained recurrent implantation failure and elevated uterine natural killer cell levels - A randomized placebo controlled trial

Abstract Study question Does intralipid supplementation in women with unexplained recurrent implantation failure (RIF) with elevated uterine natural killer cell (uNK) levels improve pregnancy outcomes during IVF? Summary answer Intralipid supplementation appears to improve clinical pregnancy rate in women with unexplained RIF with elevated uNK cell levels. What is known already The increased numbers of uNK cells in peri-implantation endometrium have been reported in women with recurrent miscarriage (RM) and RIF after IVF. However, reports are contradictory when it comes to correlation of increased numbers of uNK cells with pregnancy outcome. Current opinion suggests there is a potential for intralipid therapy in improving reproductive outcome, although data on live birth rate is very limited. No studies have assessed the effect of intralipid on IVF outcomes in RIF women based on elevated uNK cells. Identified studies have all used pNK cell testing as preferred diagnostic tool for analysis of NK cell levels. Study design, size, duration A randomized placebo controlled trial was conducted at Division of Reproductive Medicine at tertiary care institute. Thirty women with RIF and fifty fertile controls with age &amp;lt;35 years having regular menstrual cycles and no hormonal treatment in last 3 months were enrolled in the study from January 2019 to December 2020 for uNK cell testing. Randomization was done using random numbers and sealed envelopes. Only the subjects were masked and allocation concealment was done. Participants/materials, setting, methods Subjects included RIF 20–35 years, normal ovarian reserve, unexplained and tubal factors, normal karyotype and normal uterine cavity. Cut off for uNK cells was derived from fertile controls by immunohistochemical staining of CD56+ cells from midluteal endometrial biopsy sample. Subjects with elevated uNK cell levels were randomized during IVF to group A (Intralipid) or group B ( saline ). The infusion was repeated within one week of positive pregnancy test and then every 2 weeks. Main results and the role of chance The mean age and BMI were comparable between fertile control and study group(29.45±3.3 vs 31.17±3.3 years, 22.97±1.89 vs 23.21±2.2 kg/m2 ;p&amp;gt;0.05). The median uNK cell levels was 7%(used as cut off) in fertile controls and 13.5% in RIF. 18 women (60%, 18/30) with RIF who had elevated uNK cell level (&amp;gt;7%) were randomized. Four women were lost to follow up. The median age, BMI, number of previous failed cycles and duration of infertility were comparable between Group A(n = 7) and Group B(n = 7){30(IQR:27–31) vs 33(IQR:30–34)years, 22.7(IQR:21.08–24.4) vs 22.6(IQR:21.37–24.2)kg/m2, 2(IQR:2–3) vs 2(IQR:2–3), 8(IQR:7–8) vs 8(IQR:7–10)years}. The median FSH, AMH and AFC were 5.86(IQR:5.13–7.67)mIU/l, 2.4(IQR:2.16–6.12)ng/ml, 10(IQR:8–12) in Group A which were comparable with Group B {6.2(IQR:4.78–6.5)mIU/l, 4.8(IQR:2.67–6.25)ng/ml, 12(IQR:12–16) }. All patients underwent antagonist protocol. The clinical pregnancy rate was 57.14%(4/7) in group A which was significantly higher as compared to 28.6%(2/7) in group B(p &amp;lt; 0.05). None of the patients reported any side effects due to intralipid. Limitations, reasons for caution The limitation of present study is its small sample size. However, the study is currently recruiting more RIF patients, and these are the interim results of the same. More RCTs with larger sample size are required to assess the efficacy of intralipid in this specific subset of population. Wider implications of the findings: The present study suggests the beneficial effect of intralipid in women with unexplained RIF with elevated uNK cell levels in increasing the chemical and clinical pregnancy rate. However, ongoing pregnancy rate and live birth rate should be investigated further in this subset of population. Trial registration number CTRI/2019/01/017213

Calpain7 Negatively Regulates Human Endometrial Stromal Cell Decidualization in EMS by Promoting Foxo1 Nuclear Exclusion by Increasing the Phosphorylation of AKT1

Background: Endometrial receptivity damage caused by impaired decidualization may be one of the mechanisms of infertility in endometriosis (EMs). Our previous study demonstrated that CAPN7 is abnormally overexpressed in EMs. Whether CAPN7 affects the regulation of decidualization remains to be determined. Methods: The expression of CAPN7 during decidualization in endometria was examined by IHC and western blotting. The transduction of a CAPN7 recombinant adenovirus into human endometrial stromal cells was performed to investigate relevant decidualization marker genes. An artificial decidualization experiment was performed to study the role of CAPN7 in decidualization in vivo. The potential molecular mechanisms were further confirmed through immunofluorescence and coimmunoprecipitation analyses. IHC analysis was performed to evaluate differentially expressed proteins, including CAPN7, p-AKT1 and p-FoxO1, between the fertile control and EMs groups. Findings: CAPN7 expression decreased during human endometrial stromal cell (HESC) decidualization in vitro. CAPN7 negatively regulated decidualization in vitro and in vivo. In addition, CAPN7 notably promoted the phosphorylation of FoxO1 at Ser319, Ser256 and Thr24, leading to increased FoxO1 exclusion from nuclei in decidualized HESCs. Furthermore, the overexpression of CAPN7 obviously promoted the phosphorylation of AKT1, which is upstream of FoxO1, at Ser473. We also identified one conserved potential PEST sequence in the AKT1 protein and found that CAPN7 was able to hydrolyse AKT1. Interpretation: CAPN7 might serve as a promising therapeutic target for improving endometrial receptivity in women with EMs. Funding: This work was supported by the National Key Research and Development Program of China (2018YFC1004400), the National Natural Science Foundation of China (No. 81070492, No. 81871165 and No. 31872846), a grant from Jiangsu Provincial Key Research and Development (BE2018602), and a special grant for principal investigators from the Health Department of Jiangsu Province (ZDRCA2016070). Declaration of Interest: We declare no potential conflicts of interest concerning the research, authorship, and/or publication of this article. Ethical Approval: All samples were collected with the informed consent of the patients, and approval from the ethics committee was obtained for this study.

Effect of trace elements on the seminal oxidative status and correlation to sperm motility in infertile Saudi males.

The impact of trace elements, especially zinc, selenium, copper, and magnesium, on male fertility has gained great interest and significance. Increased oxidative stress and altered trace element levels are probable etiological factors underlying male reproductive dysfunction and infertility. The present study focused on the evaluation of seminal oxidative markers, such as reactive oxygen species (ROS), malondialdehyde (MDA), and total antioxidant capacity (TAC), and trace element levels in the normozoospermic fertile control group (n = 40) and asthenozoospermic infertile group (n = 30). Semen from infertile men exhibited significantly higher ROS and MDA levels accompanied with significant decline in TAC and trace element (zinc and magnesium) levels. Furthermore, a significant correlation was observed between trace elements and oxidative markers with sperm motility. The current study revealed increased lipid peroxidation and oxidant-reductant imbalance that leads to deterioration of semen quality and male infertility. Thus, oxidative stress and trace elements can be considered important biomarkers of male infertility. Measurement of seminal oxidative stress with conventional seminological parameters must be integrated in fertility assessment from early stages to ensure healthy semen characteristics and fertility in men.

Talin1 regulates endometrial adhesive capacity through the Ras signaling pathway

AimsBlastocyst implantation is mainly depended on the adhesion between cells and cell matrix. Endometrial adhesion plays an important role in establishing embryo implantation, but the underlying mechanisms are remains unclear. Talin1 is a local adhesion complex protein that is necessary for cell adhesion and movement. However, the role and mechanisms of Talin1 in embryo implantation are still unclear. Main methodsThe expression of Talin1 and Integrin αvβ3 was measured in the receptive endometrium from the RIF (Recurrent implantation failure) cohort and NC (normal fertile control group) cohort. A JEG-3 trophoblast and endometrial epithelial cell adhesion model and pregnant mouse model were established. The molecular mechanism of Talin1-mediated cell adhesion was explored by RNA sequencing, RT-qPCR, as well as western blotting assays. Key findingsTalin1 enhances endometrial cell adhesion by regulating the Ras signaling pathway, and ultimately facilitates embryo implantation. SignificanceThis study revealed the molecular mechanisms of regarding the pathogenesis of RIF caused by endometrial receptivity insufficiency. Further pharmacological research on the Ras signaling pathway would be valuable and might provide new therapeutic targets for RIF patients.

Role of uterine artery Doppler in assessment of unexplained infertility

BackgroundThe study aimed to compare the values of uterine artery Doppler indices including resistance index (RI), pulsatility index (PI), and systolic/diastolic (S/D) ratios in fertile female and female with unexplained infertility and investigate their association with unexplained infertility. This study included 70 women divided into two groups. Study group included 40 women with unexplained infertility and control group included 30 fertile women.ResultsThe mean ages of study and control groups were 26.9 and 28.5 years respectively. In infertile group, the mean values of uterine arteries resistance index (RI), pulsatility index (PI), and systolic/diastolic (S/D) ratios were 0.9, 2.9, and 8.0 respectively. In the fertile control group, mean RI and PI values and S/D ratio were 0.6, 1.5, and 2.7 respectively. There was statistically significant difference as regards RI, PI, and S/D ratios between study and control groups. The best cut off values of RI, PI, and S/D ratios for predicting increased uterine blood flow impedance were more than 0.67, 1.95, and 3 respectively with sensitivity of 100%, 95%, and 100% respectively, specificity of 96.7%, 86.7%, and 96.7% respectively, and diagnostic accuracy of 98.6%, 91.42%, and 98.57% respectively.ConclusionUterine artery Doppler indices mean values were higher in unexplained infertility women than fertile women. Uterine artery Doppler indices have high sensitivity and specificity for diagnosis of high uterine blood flow impedance. High uterine blood flow impedance diagnosed by uterine artery Doppler may contribute in the etiology of unexplained infertility. Uterine artery Doppler should be included in investigation of unexplained infertility.

FEMALE SEXUAL DYSFUNCTION IN SUBFERTILITY PATIENTS VERSUS THOSE SEEKING FERTILITY CONTROL

Background: Female sexual dysfunction (FSD) and subfertility are common problems affecting approximately 43 and 20% of women respectively. Studies on association of female sexual dysfunction and infertility is not much .the presented study compare the prevalence of female sexual dysfunction in patients on assessment for sub-fertility and those either seeking or already on fertility control services at a private sonolive clinic in Benha. Methods:This was an analytical cross sectional study. Eligible women of reproductive age (20–43 years), attending the private sonolive infertility clinic in Benha Egypt in the period from January 2019 to July 2020 with complaints of subfertility and those seeking fertility control services (as controls)were requested to fill a general demographic tool containing personal data and the Female Sexual Function Index (FSFI) questionnaire after informed consent. Sexual dysfunction was calculated as a percentage of patients not achieving an overall FSFI score of 26.55. Results:The prevalence of female sexual dysfunction was 45% in the subfertile group and 35% in fertility control group. The difference was statistically significant (p = 0.006).the important affected items were desire and lubrication also infrequent coitus in the fertility window Conclusion:The present study demonstrated a significant association between the fertility status and the prevalence female sexual dysfunction. Subfertility type was associated with sexual dysfunction especially the desire and lubrication which affect the whole score of sexual dysfunction.

The Role of Arachidonic and Linoleic Acid Derivatives in Pathological Pregnancies and the Human Reproduction Process.

The aim of the available literature review was to focus on the role of the proinflammatory mediators of AA and LA derivatives in pathological conditions related to reproduction and pregnancy. Arachidonic (AA) and linoleic acid (LA) derivatives play important roles in human fertility and the course of pathological pregnancies. Recent studies have demonstrated that uncontrolled inflammation has a significant impact on reproduction, spermatogenesis, endometriosis, polycystic ovary syndrome (PCOS) genesis, implantation, pregnancy and labor. In addition, cyclooxygenase-mediated prostaglandins and AA metabolite levels are higher in women’s ovarian tissue when suffering from PCOS. It has been demonstrated that abnormal cyclooxygenase-2 (COX-2) levels are associated with ovulation failure, infertility, and implantation disorders and the increase in 9-HODE/13-HODE was a feature recognized in PCOS patients. Maintaining inflammation without neutrophil participation allows pregnant women to tolerate the fetus, while excessive inflammatory activation may lead to miscarriages and other pathological complications in pregnancies. Additionally AA and LA derivatives play an important role in pregnancy pathologies, e.g., gestational diabetes mellitus, preeclampsia (PE), and fetal growth, among others. The pathogenesis of PE and other pathological states in pregnancy involving eicosanoids have not been fully identified. A significant expression of 15-LOX-1,2 was found in women with PE, leading to an increase in the synthesis of AA and LA derivatives, such as hydroxyeicozatetraenoic acids (HETE) and hydroxyoctadecadiene acids (HODE). Synthesis of the metabolites 5-, 8-, 12-, and 15-HETE increased in the placenta, while 20-HETE increased only in umbilical cord blood in women with preeclampsia compared to normal pregnancies. In obese women with gestational diabetes mellitus (GDM) an increase in epoxygenase products in the cytochrome P450 (CYP) and the level of 20-HETE associated with the occurrence of insulin resistance (IR) were found. In addition, 12- and 20-HETE levels were associated with arterial vasoconstriction and epoxyeicosatrienoic acids (EETs) with arterial vasodilatation and uterine relaxation. Furthermore, higher levels of 5- and 15-HETE were associated with premature labor. By analyzing the influence of free fatty acids (FFA) and their derivatives on male reproduction, it was found that an increase in the AA in semen reduces its amount and the ratio of omega-6 to omega-3 fatty acids showed higher values in infertile men compared to the fertile control group. There are several studies on the role of HETE/HODE in relation to male fertility. 15-Hydroperoxyeicosatetraenoic acid may affect the integrity of the membrane and sperm function. Moreover, the incubation of sperm with physiologically low levels of prostaglandins (PGE2/PGF2α) improves the functionality of human sperm. Undoubtedly, these problems are still insufficiently understood and require further research. However, HETE and HODE could serve as predictive and diagnostic biomarkers for pregnancy pathologies (especially in women with risk factors for overweight and obesity). Such knowledge may be helpful in finding new treatment strategies for infertility and the course of high-risk pregnancies.

Talin1 Regulates Endometrial Adhesive Capicity Through the Ras Signaling Pathway

Background: Blastocyst implantation is mainly depended on the adhesion between cells and cell matrix. Endometrial adhesion plays an important role in establishing embryo implantation, but the underlying mechanisms are remains unclear. Talin1 is a local adhesion complex protein that is necessary for cell adhesion and movement. However, the role and mechanisms of Talin1 in embryo implantation are still unclear. Methods: The expression of Talin1 and Integrin αvβ3 was measured in the receptive endometrium from the RIF (Recurrent implantation failure) cohort and NC (normal fertile control group) cohort. A JEG-3 trophoblast and endometrial epithelial cell adhesion model and pregnant mouse model were established. The molecular mechanism of Talin1-mediated cell adhesion was explored by RNA sequencing, RT-qPCR, as well as western blotting assays. Findings: Talin1 enhances endometrial cell adhesion by regulating the Ras signaling pathway, and ultimately facilitates embryo implantation. Interpretation: This study revealed the molecular mechanisms of regarding the pathogenesis of RIF caused by endometrial receptivity insufficiency. Further pharmacological research on the Ras signaling pathway would be valuable and might provide new therapeutic targets for RIF patients. Funding: This study was supported by grants from the National Natural Science Foundation of China and the Natural Science Foundation of Guangxi Zhuang Autonomous Region and the Guangxi Medical University Training Program for Distinguished Young Scholars Declaration of Interests: The authors declare that they have no conflicts of interest. Ethics Approval Statement: All participants provided written informed consent, and the study was approved by the Ethics Committee of the First Affiliated Hospital of Guangxi Medical University. All animal experimental procedures were approved by the animal ethics committee of Guangxi Medical University in accordance with animal research guidelines.