Implantation Failure In Women Research Articles

Interplay of “leukemia inhibitory factor receptor gene” (rs3099124) polymorphism, leukemia inhibitory factor and ovarian steroids with unexplained infertility

Objective of the study To explore the association of leukemia inhibitory factor receptor (LIFR) gene variant rs3099124, ovarian steroids, and leukemia inhibitory factor with unexplained infertility in Pakistani females. Methodology A case–control investigation in which eighty-one (81) females with unexplained infertility and one hundred and sixty-two (162) fertile counterparts (age and body mass index compared) were recruited between October 2016 and 2018. Ten milliliters of venous blood was collected from all participants. “Genomic DNA” was taken out from lymphocytes in peripheral blood samples. “Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS-PCR)” was constructed through software “Primer-I”. Amplification was carried out by “T-ARMS-PCR” followed by subsequent sequencing for confirmation and extensive consonance. Estradiol, Progesterone and Leukemia Inhibitory Factor (LIF) were measured in serum by ELISA. Results Statistically significant difference was noticed in genotype frequency in “LIFR-gene variant; rs3099124” (χ2 = 28.222, P value < 0.01) between research participants. Although, rs “3099124” “AA” (OR = 0.000; 95%CI = 0–0) and “GA” genotypes (OR = 0.525; 95%CI = 0.226–1.22) showed non-significant safety/protection against unexplained infertility yet minor/risk allele “A” frequency was greater in women with unexplained infertility suggesting a possible explanation of implantation failure. LIF concentration varied between fertile and infertile groups (χ2 = 9.857, P < 0.05) revealing significant threat of unexplained infertility in women with decreased LIF concentration (OR = 2.316, 95%CI = 1.214–4.416). Progesterone was significantly related to unexplained infertility in both study groups (χ2 = 20.347, P < 0.05). High progesterone reduced the possibility of unexplained infertility (OR = 0.306; 95% CI = 0.166–0.567). Conclusion LIFR gene variation (rs3099124) and reduced LIF secretion may cause implantation failure in women with unexplained infertility.

An In Vivo Screening Model for Investigation of Pathophysiology of Human Implantation Failure.

To improve current infertility treatments, it is important to understand the pathophysiology of implantation failure. However, many molecules are involved in the normal biological process of implantation and the roles of each molecule and the molecular mechanism are not fully understood. This review highlights the hemagglutinating virus of Japan (HVJ; Sendai virus) envelope (HVJ-E) vector, which uses inactivated viral particles as a local and transient gene transfer system to the murine uterus during the implantation period in order to investigate the molecular mechanism of implantation. In vivo screening in mice using the HVJ-E vector system suggests that signal transducer and activator of transcription-3 (Stat-3) could be a diagnostic and therapeutic target for women with a history of implantation failure. The HVJ-E vector system hardly induces complete defects in genes; however, it not only suppresses but also transiently overexpresses some genes in the murine uterus. These features may be useful in investigating the pathophysiology of implantation failure in women.

MiR-297 inhibits expression of progesterone receptor and decidualization in eutopic endometria of endometriosis.

Progesterone resistance is an epigenetic factor that reduces endometrial receptivity and causes implantation failure in women with endometriosis. In addition, dysregulated miRNAs contribute to the underlying pathogenic mechanisms of endometriosis. This study aimed to determine the effect of miR-297 on the progesterone receptor (PR) expression and on insufficient decidualization of endometrial stromal cells (ESCs) within the eutopic endometria of infertile women with minimal or mild endometriosis. ESCs were isolated from infertile endometriosis and normal patients and were transfected with miR-297 mimic or miR-297 inhibitor or respective control. qRT-PCR and western blot were conducted to quantify the expression of miR-297 and PR. The effect of miR-297 on ESCs decidualization was investigated by induced decidualization in vitro. We observed an increase in miR-297 expression and a decrease in the expression of PR in the ESCs from endometriosis patients. Moreover, the expression of PR, most notably PRB, was found to be downregulated following transfection with miR-297 mimic and upregulated following treatment with miR-297 inhibitor. In addition, overexpressed miR-297 inhibited the decidualization of ESCs in vitro. We further determined that miR-297 exerts direct regulatory effects on PR expression. We demonstrated that miR-297 interferes with fertility by repressing the expression of PR and preventing efficient decidualization in eutopic endometria. Further, miR-297 directly contributes to progesterone resistance in minimal or mild cases of endometriosis. Thus, regulation of miR-297 may prove to be a promising therapeutic strategy for endometriosis.

MISCLASSIFICATION OF PREIMPLANTATION GENETIC TESTING FOR ANEUPLOIDY: A SYSTEMATIC REVIEW AND META-ANALYSIS

Aneuploidy accounts for the majority of miscarriages, congenital anomalies and implantation failure in women. Preimplantation genetic testing for aneuploidy (PGTA) is a screening test applied to embryos created from an in vitro fertilization cycle to diminish the chance of an aneuploid conception. Embryos that screen negative (euploid) are transferred and those that screen positive (aneuploid) are discarded. There is a lack of consensus regarding risk of aneuploidy after euploid embryo transfer, as well as chance of euploidy after aneuploid classification.

Endocrine disrupting chemicals interfere with decidualization of human primary endometrial stromal cells in vitro

Multiple studies have shown associations between exposure to endocrine disrupting chemicals (EDCs) and reduced fertility in women. However, little is known about the target organs of chemical disruption of female fertility. Here, we focus on the hormone-sensitive uterine lining, the endometrium, as a potential target. Decidualization is the morphological and functional change that endometrial stromal cells undergo to support endometrial receptivity, which is crucial for successful implantation, placentation, and pregnancy. We investigated the effect of nine selected EDCs on primary human endometrial stromal cell decidualization in vitro. The cells were exposed to a decidualization-inducing mixture in the presence or absence of 1 μM of nine different EDCs for nine days. Extent of decidualization was assessed by measuring the activity of cAMP dependent protein kinase, Rho-associated coiled-coil containing protein kinase, and protein kinase B in lysates using photoluminescent probes, and secretion of prolactin into the media by using ELISA. Decidualization-inducing mixture upregulated activity of protein kinases and prolactin secretion in cells derived from all women. Of the tested chemicals, dichlorodiphenyldichloroethylene (p,p’-DDE), hexachlorobenzene (HCB) and perfluorooctanesulfonic acid (PFOS) significantly reduced decidualization as judged by the kinase markers and prolactin secretion. In addition, bisphenol A (BPA) reduced prolactin secretion but did not significantly affect activity of the kinases. None of the EDCs was cytotoxic, based on the assessment of total protein content or activity of the viability marker casein kinase 2 in lysates. These results indicate that EDCs commonly present in the blood circulation of reproductive-aged women can reduce decidualization of human endometrial stromal cells in vitro. Future studies should focus on detailed hazard assessment to define possible risks of EDC exposure to endometrial dysfunction and implantation failure in women.

Вплив антропометричних факторів та даних акушерсько-гінекологічного анамнезу на ризик невдалої імплантації після запліднення in vitro

Infertility is a significant demographic and socio-economic problem, which is solved with assisted reproductive technologies. Relevance of the infertility problem continues to grow, which leads to the search for new methods to improve the success of assisted reproductive technologies. Purpose — to analyze the effect of anthropometric factors, obstetrical and gynaecological history on the risk of implantation failure in women with infertility who have fertilization in vitro. Materials and methods. An analysis of 106 infertile women who were treated with assisted reproductive technologies in September 2019 — February 2021. The control group consisted of women who received infertility treatment according to the standard treatment regimens: controlled ovarian stimulation, gamete sampling, intracytoplasmic sperm injection and embryo transfer. The study group included patients who received antiplatelet agents, anticoagulants and intravenous immunoglobulins G in addition to the standard therapy. Results. In this study, no differences were found in the two study groups regarding age, height, weight, body mass index, menarche, menstrual duration, coitus (p&gt;0.05). During the analysis, it was found that with increasing age of a woman, the risk of unsuccessful implantation increases, OR=1.11 (95% CI 1.01–1.21) for each year, with increasing patient height, the risk decreases (p=0.006), OR=0.85 (95% CI 0.76–0.96) for each centimetre, with an increase in the number of sexual intercourses per week (p=0.012), OR=1.71 (95% CI 1.12–2.60) the risk increases by each unit. Implantation success with using modified treatment regimens with additional aspirin, low molecular weight heparins, intravenous immunoglobulins G, was higher (p=0.049), OR=0.37 (95% CI 0.14–0.99) compared to the standard infertility treatment scheme. Conclusions. This study found unfavourable effects of age, woman's height, and sexual intercourses on embryo implantation. Additional use of aspirin, low molecular weight heparin, intravenous immunoglobulin G as a supplemental therapy to standard treatment regimens of infertility with IVF had a successful impact on the effectiveness of embryo implantation. The research was carried out in accordance with the principles of the Helsinki declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interest was declared by the authors. Key words: implantation failure, infertility, assisted reproductive technologies, in vitro fertilization (IVF).

Circulating MMP-7 and VEGF as potential predictive biomarkers for recurrent implantation failures.

Recurrent implantation failure (RIF) is considered to be one of the major limiting factors of assisted reproductive technology (ART) programme success. The current study focused on the investigation of matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), cytokines and cell adhesion molecules in peripheral blood (PB) and follicular fluid (FF) obtained from 44 women aged between 25 and 39 years old and undergoing intracytoplasmic sperm injection (ICSI). These women were divided into two groups: 22 RIF women with embryo implantation failures after the transfer of at least four fresh or frozen-thawed good quality embryos in a minimum of three ICSI cycles, and 22 ICSI success women (controls) who achieved a clinical pregnancy at their first ICSI attempt. The PB and FF samples were obtained from each patient on the day of oocyte retrieval. MMP-1, -2, -3, -7, -9, TIMP-1, -2, vascular endothelial growth factor (VEGF), leukaemia inhibitory factor (LIF), vascular cell adhesion molecule 1 (VCAM1) and intercellular adhesion molecules 1 (ICAM1) were analyzed using enzyme-linked immunosorbent assay of PB and FF. Our results showed significant decreases in PB MMP-7 and PB VEGF in the RIF group compared with controls [281.11 (33-614) pg/ml vs 119.92 (27-441) pg/ml; P-value = 0.030] and [82.54 (25.94-210.20) pg/ml vs 30.93 (13.62-193.33) pg/ml; P-value = 0.022; respectively]. Receiver operating characteristic (ROC) curve analysis showed informative area under the curve values for PB MMP-7, as well as for PB VEGF, making them able to be proposed as biomarkers of the RIF. Therefore, circulating MMP-7 and VEGF seem to play an interesting role in embryo implantation in in vitro fertilization (IVF)/ICSI cycles and could be proposed as circulating biomarkers of the RIF. These results could be helpful for clinicians and patients to choose the best rescue strategy and treatment to minimize implantation failure in women undergoing IVF/ICSI procedures after the first attempt.

ENDOMETRIAL BCL6 EXPRESSION IS NOT ASSOCIATED WITH IMPLANTATION OUTCOMES

B-cell CLL/lymphoma 6 (BCL6) is a proto-oncogene that is overexpressed in the secretory endometrium of women with endometriosis. It has been postulated that BCL6 may serve as a surrogate marker for occult endometriosis, as well as a predictor of endometrial dysfunction and poor implantation. Therefore, BCL6 expression has been proposed as an adjunct test for women undergoing IVF. However, this test has not yet been validated in the general IVF population. The aim of this study was to determine whether increased endometrial BCL6 expression is associated with implantation failure in women undergoing IVF.

Chloride intracellular channel 4 is dysregulated in endometrium of women with infertility and alters receptivity

The endometrium remodels in each menstrual cycle to become receptive in preparation for embryo implantation which occurs in the mid-secretory phase of the cycle. Failure of blastocyst adhesion and implantation cause infertility. We compared chloride intracellular channel 4 (CLIC4) expression in human endometrium from women with normal fertility and primary unexplained infertility in the mid-secretory/receptive phase of the menstrual cycle. CLIC4 localised to both the epithelial and stromal regions of the endometrium of fertile tissues across the cycle. CLIC4 expression was significantly reduced in the luminal and glandular epithelium and remained unchanged in the stromal region of mid-secretory infertile endometrium compared to fertile endometrium. siRNA knockdown of CLIC4 significantly compromised adhesive capacity of Ishikawa cells (endometrial epithelial cell line). This reduced adhesion and CLIC4 expression was associated with elevated SGK1, p53, SIRT1, BCL2 and MCL1 gene expression in the Ishikawa cells. CLIC4 expression was increased in primary human endometrial stromal cells during decidualization, however, siRNA knockdown of CLIC4 did not affect decidualization. Our data provide evidence that CLIC4 may regulate receptivity and facilitate blastocyst attachment initiating implantation. Reduced CLIC4 levels may be causative of implantation failure in women.

Beyond Acephalic Spermatozoa: The Complexity of Intracytoplasmic Sperm Injection Outcomes.

This review analyses the genetic mechanisms of acephalic spermatozoa (AS) defects, which are associated with primary infertility in men. Several target genes of headless sperms have been identified but intracytoplasmic sperm injection (ICSI) outcomes are complex. Based on electron microscopic observations, broken points of the sperm neck are AS defects that are based on various genes that can be classified into three subtypes: HOOK1, SUN5, and PMFBP1 genes of subtype II; TSGA10 and BRDT genes of subgroup III, while the genetic mechanism(s) and aetiology of AS defects of subtype I have not been described and remain to be explored. Interestingly, all AS sperm of subtype II achieved better ICSI outcomes than other subtypes, resulting in clinical pregnancies and live births. For subtype III, the failure of clinical pregnancy can be explained by the defects of paternal centrioles that arrest embryonic development; for subtype I, this was due to a lack of a distal centriole. Consequently, the embryo quality and potential ICSI results of AS defects can be predicted by the subtypes of AS defects. However, this conclusion with regard to ICSI outcomes based on subtypes still needs further research, while the existence of quality of oocyte and implantation failure in women cannot be ignored.

Platelet Rich Plasma in Repeated Implantation Failure in Women with Thin Endometrium Thickness

Background: repeated implantation failure (RIF) is a major challenge in reproductive medicine. Platelet rich plasma (PRP) was able to promote the endometrial growth and improve pregnancy outcome of patients with thin endometrium. Aim of the study: it was to evaluate the effectiveness of intrauterine infusion of PRP in treatment of thin endometrium in frozen-thawed embryo (FTE) transfer. Patients and Methods: the study included 10 infertile patients with thin endometrium ( 7mm on the day of progesterone administration. Sixty percent of the patients became pregnant; 83.3% continued pregnancy and 16.7% experienced missed abortion. Conclusion: intrauterine PRP may be effective in promoting endometrial growth, endometrial thickness, and improving pregnancy out come in repeated implantation failure.

Platelet Rich Plasma in Repeated Implantation Failure in Women with Thin Endometrium Thickness

Background: repeated implantation failure (RIF) is a major challenge in reproductive medicine. Platelet rich plasma (PRP) was able to promote the endometrial growth and improve pregnancy outcome of patients with thin endometrium. Aim of the study: it was to evaluate the effectiveness of intrauterine infusion of PRP in treatment of thin endometrium in frozen-thawed embryo (FTE) transfer. Patients and Methods: the study included 10 infertile patients with thin endometrium ( 7mm on the day of progesterone administration. Sixty percent of the patients became pregnant; 83.3% continued pregnancy and 16.7% experienced missed abortion. Conclusion: intrauterine PRP may be effective in promoting endometrial growth, endometrial thickness, and improving pregnancy out come in repeated implantation failure.

FOXO1 regulates uterine epithelial integrity and progesterone receptor expression critical for embryo implantation.

Successful embryo implantation requires a receptive endometrium. Poor uterine receptivity can account for implantation failure in women who experience recurrent pregnancy loss or multiple rounds of unsuccessful in vitro fertilization cycles. Here, we demonstrate that the transcription factor Forkhead Box O1 (FOXO1) is a critical regulator of endometrial receptivity in vivo. Uterine ablation of Foxo1 using the progesterone receptor Cre (PgrCre) mouse model resulted in infertility due to altered epithelial cell polarity and apoptosis, preventing the embryo from penetrating the luminal epithelium. Analysis of the uterine transcriptome after Foxo1 ablation identified alterations in gene expression for transcripts involved in the activation of cell invasion, molecular transport, apoptosis, β-catenin (CTNNB1) signaling pathway, and an increase in PGR signaling. The increase of PGR signaling was due to PGR expression being retained in the uterine epithelium during the window of receptivity. Constitutive expression of epithelial PGR during this receptive period inhibited expression of FOXO1 in the nucleus of the uterine epithelium. The reciprocal expression of PGR and FOXO1 was conserved in human endometrial samples during the proliferative and secretory phase. This demonstrates that expression of FOXO1 and the loss of PGR during the window of receptivity are interrelated and critical for embryo implantation.

GM-CSF (sargramostim) treatment in women with recurrent implantation failure undergoing transfer of single healthy blastocyst after pgs: a randomized controlled trial

The GM-CSF is a cytokine promoting leukocyte growth as well as trophoblast development. We described that this cytokine may be used in the treatment of recurrent abortion. We tested in this randomized controlled trial the use of GM-CSF (sargramostim) in the treatment for recurrent implantation failure in women undergoing IVF. A controlled randomized study conducted on women with Recurrent Implantation Failure. The study was conducted to the CERM, Rome, Italy, from the January 2016 to December 2017 on 73 women with recurrent implantation failure after IVF cycles. This study was approved by IRB. Inclusion criteria were: at least 9 good embryos previously transferred, women less than 38 years old, absence of systemic diseases. These women underwent IVF cycle and PGS on developed blastocysts. Single healthy blastocyst transfer was performed in the next cycle using only chromosomally healthy blastocysts. Patients were randomly divided in two groups: one (36 women) treated with subcutaneous GM-CSF 1.5mg/kg/daily (60-100) from the day of embryo transfer to the day of β-hcg day and if it was positive the treatment was continued for other 40 days: the control group (37 women) was treated with subcutaneous saline solution infusion in the same way of the study group. Primary outcome was the pregnancy rate. Epidemiological data of the two groups did not show statistically significant differences. Pregnancy rate in the group treated with GM-CSF was 75.0% (27/36) whereas in the control group was 43.2% (16/37), P= 0.0087. No side effects were observed. The clinical use of GM-CSF in women experienced implantation failure may be useful, even though more studies are needed to confirm these findings.

The endometria of women with endometriosis exhibit dysfunctional expression of complement regulatory proteins during the mid secretory phase

The control of complement activation within embryo-endometrium environment is critical for embryo survival. Cell evasion from complement attack requires interaction of complement regulatory proteins (CRPs) with cell adhesion αvβ3 integrin. We aim to compare the expression of CRPs in endometria of women with and without endometriosis and to examine the molecular interaction of decay accelerating factor (DAF) with αvβ3 integrin. Endometrial expression of Membrane cofactor protein (CD46), Decay accelerating factor (DAF), Membrane attack complex inhibitory factor (CD59) and β3 integrin subunit were determined through menstrual cycle by immunohistochemistry. DAF protein quantity was determined by Western blot and mRNA levels measured in epithelial cells isolated by laser capture microdissection (LCM). Using in vitro assay, we examined DAF and β3 integrin expression through paracrine regulation between endometrial compartments. To determine whether β3 integrin and DAF interacts in vivo, endometrial samples were subjected to immunoprecipitation and colocalization using dual immunofluorescence technique. DAF and β3 integrin expression were significantly low in samples from women with endometriosis during mid secretory phase. This observation was supported by decreased DAF protein quantity; faint DAF and β3 integrin interaction and reduced mRNA levels in cells dissected by LCM. Moreover epithelial DAF and β3 integrin expression through paracrine regulation by progesterone from stromal compartment was disrupted in endometriosis. Endometria from women with endometriosis exhibits aberrant expression of complement proteins. The abnormal DAF expression potentially compromises embryo survival, contributing to understand the implantation failure in women with endometriosis.

Dysregulated genes involved in implantation failure in women with endometriosis

In most women, endometriosis leads to infertility. Infertility in women with endometriosis may be due to poor embryo implantation. A number of mechanisms are associated in preparing the endometrium for implantation such as cell proliferation, cellular differentiation, immunological response, apposition and production of growth factors. These processes along with the genes involved may be altered in women with endometriosis. A prospective case-control study was conducted to assess expression of dysregulated genes contributing to implantation failure, in eutopic endometrium of women with endometriosis compared to that of women without endometriosis. Eutopic endometrial biopsies in mid-luteal phase from 14 women with endometriosis and 13 women without endometriosis were obtained by Pipelle curette between 31 March 2016 and 31 October 2016 in Akanksha IVF centre. Paralleled gene expression profiling using high density oligonucleotide microarrays was applied to investigate dysregulated genes in eutopic endometrium of women with endometriosis compared to women without endometriosis. Data analysis was conducted by GeneSpring version 4.0.4 and ArrayMining. T-test using P value <0.05 and a fold-change expression >2 was applied. Select dysregulated genes were randomly validated by RT-PCR. This study has shown that in eutopic endometrium from women with endometriosis, genes involved in implantation failure were significantly dysregulated. Genes involved in production of oxidative stress, apoptosis and immune response were upregulated, whereas genes involved in promoting adhesion, cellular proliferation and growth were downregulated. Dysregulation of genes involved in embryo implantation likely contributes to decreased pregnancy rates in women with endometriosis. Increased immune response may contribute to increased inflammation leading to oxidative stress. Increased oxidative stress leads to DNA damage and apoptosis, thereby impairing embryo implantation. Moreover, upregulation of genes promoting apoptosis and downregulation of genes promoting adhesion, cellular proliferation and growth in the endometrium may reduce endometrial receptivity in women with endometriosis.Tabled 1Dysregulated genes in eutopic endometrium of women with endometriosisGene SymbolGene NameFold ChangeGene FunctionDAPK1Death-associated protein kinase 12.61ApoptosisC4BPAComplement component 4 binding protein,α5.18Immune responseERO1LERO1-like2.65Oxidative stressHELLSHelicase-2.48Cellular proliferationIGF1Insulin-like growth factor 1-3.9Cellular growthITGA6Integrin, alpha 6-2.6Cellular adhesion Open table in a new tab

Immunosuppression with tacrolimus improved implantation and rescued expression of uterine progesterone receptor and its co-regulators FKBP52 and PIASy at nidation in the obese and diabetic mice: Comparative studies with metformin

Diabesity is often associated with subfertility and recurrent miscarriages. Evidence links systemic and local uterine cytotoxicity to the pathogenesis of implantation failure (IF) in diabetes. Immunosuppression with tacrolimus improved pregnancy outcomes in obese and diabetic mice and repeated IF in women with elevated Th1/Th2 blood cell ratios. However the mode of action of tacrolimus in protecting against IF and the molecular mechanisms associated with recurrent miscarriages in the obese and diabetic subjects are yet to be elucidated. Here we administered tacrolimus (FK506) (0.1 mg/kg) for four consecutive weeks to the NONcNZO10/LtJ mice, a model of human PCOS, chronically fed with 60% kCal fat for 16 consecutive weeks to simulate human obesity-associated T2DM. Compared to those immunosuppressed with tacrolimus and their normative controls, high-fat fed (HFD) diabetic NONcNZO mice exhibited higher rates of peri- and post-implantation resorption and had aberrant expression of uterine IFNγ and progesterone receptor (PGR) and its immunophilin co-chaperone FKBP52 at nidation. Immature uterodomes and lack of activation of uterine STAT3 and NFκB at implantation were characteristics of IF in the HFD-dNONcNZO dams also low in the deciduogenic factors IL11 and GM-CSF. Therapeutic interventions with tacrolimus or metformin normalized the expression of decidual IFNγ, PGR and FKBP52, increased co-localization of protein inhibitor of activated STATy (PIASy) to PGR and resulted in the upregulation of uterine IL11and LIF. Rescued phosphorylation of STAT3 and NFκBp65 and uterodome maturation at nidation defined implantation success in treated dams. To our knowledge this is the first report to show that the impact of HFD on the hemochorial implantation is at least in part mediated through disruption of PGR signaling at nidation and that immunosuppression with tacrolimus or treatment with metformin restores PGR-mediated influences during implantation in the obese and diabetic subjects.

Effect of bisphenol A on reproductive processes: A review of in vitro, in vivo and epidemiological studies.

As bisphenol A (BPA) is characterized by a pronounced influence on human hormonal regulation, particular attention has been aimed at understanding its role in reproductive processes in males and females, as well as on fetal development. Owing to the increasing number of alarming reports on the negative consequences of the presence of BPA in human surroundings, more and more studies are being undertaken to clarify the negative effects of BPA on human reproductive processes. The aim of this work was to collect and summarize data on the influence of BPA exposure on reproductive health. Based on an analysis of selected publications it was stated that there is strong proof confirming that BPA is an ovarian, uterine and prostate toxicant at a level below the lowest observed adverse effect level (50 mg kg-1 bodyweight) as well as a level below the proposed safe level (4 μg kg-1 bodyweight). It seems there is also reliable evidence in relation to the negative effect of BPA on sperm quality and motility. Limited evidence also pertains to the case of the potential of BPA to affect polycystic ovary syndrome occurrence. Although in epidemiological studies this disease was common, in studies on animal models such results were still not confirmed. No unambiguous results of epidemiological studies and with animal models were obtained in relation to the evaluation of associations between BPA and implantation failure in women, evaluation of associations between BPA and sexual dysfunction in men, and impact of BPA on birth rate, birth weight and length of gestation. Copyright © 2017 John Wiley & Sons, Ltd.

Особливості експресії ядерних стероїдних рецепторів в ендометрії у жінок з невдалими спробами запліднення in vitro в анамнезі

Structural and functional maturity of the endometrium is formed in a condition of dynamic fluctuations of ovarian steroid hormones – estrogens and progesterone during the menstrual cycle. The objective: of the research was to determine the reactivity of estrogene core receptors-б and progesterone core receptors in endometrial stroma and glands. Patients and methods. Endometrial samples were obtained during hysteroscopy in 42 patients with infertility on day LH + 6 – LH + 11 of the menstrual cycle. The control group (CG) consisted of 18 healthy women who had at least one physiological pregnancy that resulted in the birth of a healthy child. Receptivity of the endometrium to the hormones was studied by immunoperoxidase method using mouse MAbs for estrogen receptors-a (ER-a) (clone ID5, «DAKO», Denmark) and progesteron receptor (RP) (clone A6, «DAKO», Denmark). The color intensity was assessed by tree-point system. The expression levelі of ER-a and RP were set by calculating semiquantitative index: IRS = SI ґ PP, where IRS – immunoreactivity index; SI – optical intensity staining; PP – the percentage of positively stained nuclei. Results. Normal ER-a and RP immunoreactivity was observed in 8 (19.05%) women with failed in vitro fertilization attempts in history, total reduction of ER-a and RP immunoreactivity (medium and severe) in glands and stroma was observed in 6 (14.29%) of infertile patients. Normal levels of ER-a and RP immunoreactivity were observed among KG women. Noteworthy is the presence of comorbidity. So in 4 of the six cases of endometrial polyp appeared on the background of chronic endometritis. In 4 women diagnosed with simple hyperplasia without atypia the defective rejection of the endometrium from the previous cycle and the incompatibility of endometrium structure to the day of cycle were found. In the group of infertile women high reactivity of receptors was observed only in 8 (19.05%) biopsies. Deviation of the morphological structure of the endometrium in this subgroup was observed significantly more often than in KG: incompatibility of morphological endometrial structure to the day of menstrual cycle was found in 12.50% of cases, simple hyperplasia without atypia – of 55.00%, endometrial polyp – 12.50%, chronic endometritis – 25.00%. Conclusions. The change of immunoreactivity of estrogen receptor-б and progesterone plays an important role in the formation of implantation failure in women with unsuccessful in vitro fertilization attempts. The obtained data confirm the hypothesis of multiple factors of development regulation of the endometrium in natural cycles and in IVF cycles. High frequency of structural and functional inferiority of the endometrium is the indication for in-depth study. Correction of the endometrial condition need to be done before pregnancy. Key words: infertility, failed in vitro fertilization attemp, immunoreactivity, receptors estrogen-a, progesterone receptors.

High Amounts of Total and Extracellular Vesicle-Derived Soluble HLA-G are Associated with HLA-G 14-bp Deletion Variant in Women with Embryo Implantation Failure.

Human leukocyte antigen-G (HLA-G) expression is related to 14-bp insertion/deletion polymorphism at the 3'UTR of the HLA-G gene. Soluble forms of HLA-G are released as free molecules or via extracellular vesicles (EVs). Due to the crucial role of HLA-G during pregnancy, we analyzed the 14-bp polymorphism and the two secreted forms in implantation failure women (IF) and in fertile women (FW). For the genetic analysis, 49 IF and 34 FW were genotyped. For sHLA-G quantification, serum samples from 35 IF and 23 FW were available. ExoQuick(™) kit was used for EVs precipitation. The total soluble HLA-G (sHLA-Gtot ) and vesicular sHLA-GEV were quantified by ELISA. The EVs size and concentration were determined by nanoparticle tracking analysis (NTA). An increased proportion of IF presented high levels of sHLA-Gtot (P = 0.02) and vesicular sHLA-GEV (P = 0.0003) compared to FW. The 14-bp deletion allele is more frequent in IF (P = 0.0002) and associated with high levels of sHLA-Gtot and vesicular sHLA-GEV . The high expression of sHLA-Gtot and sHLA-GEV , together with the presence of the 14-bp deletion allele, might be involved in implantation failure.