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Objective: The study aimed to assess serum levels of syndecan-1 (SDC-1) and hyaluronan (HA) in patients with early- and late-onset preeclampsia (PE).Study Design: Blood samples were collected in the third trimester of pregnancy from 20 women with early-onset PE, 20 with late-onset PE, and 20 with normal pregnancy for the assessment of serum levels of SDC-1 and HA as markers of endothelial injury. PE was categorized as early-onset when diagnosed at <34 weeks of gestation and as late-onset when diagnosed at ≥34 weeks of gestation.Main Outcome Measures: The degree of endothelial injury in different forms of preeclampsia expressed by serum concentrations of SDC-1 and HA.Results: Concentration of HA was significantly higher and the level of SDC-1 was significantly lower in patients with PE than in the control group. However, the concentrations of both HA and SDC-1 did not differ significantly between the two groups of PE.Conclusions: Degree of endothelium injury is comparable in patients with early- and late-onset PE. [ABSTRACT FROM AUTHOR]

Introduction. The prevalence of endometriosis among reproductive age women is 7–17%; however, these figures reach 20–50% in patients suffering from infertility. Matrix metalloproteinases (MMPs) activity is thought to be particularly essential in the early phases of endometriosis development. Any changes in the equilibrium between MMPs activity and their tissue inhibitors (TIMPs) could be potentially harmful, promoting endometriosis development. The aim of this study was to investigate whether the MMP-2, MMP-9, TIMP-1 or TGF-B2 expression in eutopic endometrium from women with early endometriosis differ when compared with healthy subjects. The results were referred to the serum progesterone levels. Materials and method. Endometrial biopsy was taken from 42 patients (18 in the study group, 22 in thecontrol group) at the time of hysteroscopy for routine histology and for RT-PCR procedures. Comparison of the quantity of gene products was performed with a programme for densitometry and compared to GADPH product, which was a reference value. Results. The obtained results did not reveal any statistical difference in endometrial expression of MMP-2, MMp-9, TIMP-1, and TGF-β2 or serum progesterone level between women with endometriosis and without visible signs of this illness. Conclusion. Despite the lack of statistical differences, it was observed that both examined metalloproteinases expressed a tendency to higher gene expression in the eutopic endometrium of women with endometriosis. However, both TIMP-1 and TGF-β2 expressions had the same tendency – higher values in endometriosis patients. [ABSTRACT FROM AUTHOR]

Abstract Background A decrease in HOXA11 expression in eutopic mid-secretory endometrium has been found in women with endometriosis-associated infertility. Methods Using Real-time quantitative PCR (RQ-PCR) and western blotting analysis we studied the HOXA11 transcript and protein levels in mid-luteal eutopic endometrium from eighteen infertile women with minimal endometriosis, sixteen healthy fertile women and sixteen infertile women with fallopian tubal occlusion from the Polish population. We also evaluated transcript levels of DNA methyltransferases DNMT1, DNMT3A and DNMT3B in these groups of women. Results There were significantly lower levels of HOXA11 transcripts (p = 0.003, p = 0.041) and protein (p = 0.004, p = 0.001) in women with endometriosis as compared to fertile women and infertile women with tubal occlusion. Moreover, we found significantly higher methylation levels of the CpG region in the first exon of HOXA11 in infertile women with endometriosis compared with fertile women (p < 0.001) and infertile women with tubal occlusion (p < 0.001). We also observed significantly increased levels of DNMT3A transcript in women with endometriosis than fertile women (p = 0.044) and infertile women with tubal occlusion (p = 0.047). However, we did not observe significant differences in DNMT1 and DNMT3B transcript levels between these investigated groups of women. Conclusions We confirmed that reduced HOXA11 expression may contribute to endometriosis-associated infertility. Moreover, we found that DNA hypermethylation can be one of the possible molecular mechanisms causing a decrease in HOXA11 expression in the eutopic mid-secretory endometrium in infertile women with endometriosis.

Gold nanoparticles (GNPs) are widely used in the technological and biomedical industries, which is a major driver of research on these nanoparticles. The main goal of this study was to determine the influence of GNPs (at 20, 100, and 200 μg/mL concentrations) on the reactivity of human peripheral blood leukocytes. Flow cytometry was used to evaluate the respiratory burst activity and pyroptosis in monocytes and granulocytes following incubation with GNPs for 30 and 60 min. Furthermore, the concentration of interleukin-1β (IL-1β) in human blood samples was assessed using enzyme-linked immunosorbent assay (ELISA) after their incubation with GNPs for 24 h. Under the conditions tested in the study, the GNPs did not significantly affect the production of reactive oxygen species in the granulocytes and monocytes that were not stimulated using phorbol 12-myristate 13-acetate (PMA) in comparison to the samples exposed to PMA ( p < 0.05). Compared to the control sample, the greatest significant increase in the mean fluorescence intensity of the granulocytes occurred in the samples incubated with CGNPs = 100 and 200 µg/mL for tinc = 30 and 60 min ( p < 0.05). From our results, we conclude that the physicochemical properties of the nanoparticles, chemical composition, and the type of nanoparticles used in the unit, along with the unit and incubation time, influence the induced toxicity.

Preeclampsia (PE) is one of the leading causes of mortality and morbidity in pregnant women. Pregestational diabetes (PGDM) patients are prone to vascular complications and preeclampsia, whereas vascular exposure to hyperglycemia induces inflammation, vascular remodeling, and arterial stiffness. Corin is a serine protease, converting inactive pro-atrial natriuretic peptide (pro-ANP) into an active form. It also promotes salt and water excretion by activating atrial natriuretic peptide (ANP), and significantly increases trophoblast invasion. The study aimed to determine whether corin may be a predictor of PE in a high-risk group-women with long-term PGDM. The nested case-control prospective study involved 63 patients with long-term pregestational type 1 diabetes (PGDM). In total, 17 patients developed preeclampsia (the study group), whereas 43 patients without PE constituted the control group. To assess corin concentration, blood samples were collected at two time points: between 18th-22nd week of gestation and 28th-32nd week of gestation. PE patients presented significantly higher mid-gestation corin levels, urine protein loss in each trimester, serum creatinine in the third trimester, and lower creatinine clearance in the third trimester. The results of our study indicate that serum corin assessment may play a role in predicting preeclampsia. Thus, it may be included in the PE risk calculator, initially in high-risk groups, such as patients with PGDM., Competing Interests: The authors declare no conflict of interest.

Introduction: The most popular model of preeclampsia (PE) is a two-stage one in which the first stage involves a decreased perfusion of the placenta and the second stage is characterized by maternal endothelial injury and dysfunction. This model seems to be more appropriate for early-onset PE, than for the late-onset disease, as in the case of the latter the event of reduced placental perfusion seems is less obvious.The aim of the study was to assess the possible correlations between the serum levels of soluble FMS-like tyrosine kinase 1 (sFlt-1) and the components of endothelial glycocalyx (EG), namely syndecan -1 (SDC-1) and hyaluronan (HA), as the markers of endothelial damage, in patients with early- and late-onset PE., Materials and Methods: The study was conducted among 60 women in their late second and third trimester of the singleton pregnancy, including 20 patients with early-onset PE, 20 with late-onset PE, and 20 women with normal pregnancy, who served as the control group. All patients were hospitalized between 2015 and 2018 at the Division of Reproduction of Poznan University of Medical Sciences. The women in the control group were matched by gestational age with the patients in the study groups., Results: The median serum level of sFlt-1 was the highest in the patients with early-onset PE (3.53 (2.73-4.5) pg/ml) but it was not statistically different from the level in the patients with late-onset PE (3.14 (2.2-3.4) pg/ml). The mean serum level of SDC-1 also did not differ significantly between the two groups of patients with PE (6.17 ± 2.2 ng/ml in early-onset PE; 6.42 ± 2.2 ng/ml in late-onset PE). Both values of SDC-1 were significantly lower than that in the healthy pregnant women (11 ± 2.62 ng/ml, p  < .001). The median concentrations of HA did not differ between patients with early- (236.6 (101.1-351.9) ng/ml) and late-onset PE (234.7 (46.8-324.2) ng/ml). However, the levels in these study groups were significantly higher than in the control group (113.9 (30.9-379.8) ng/ml, p  < .001). There was no significant correlation found between the serum concentrations of sFlt-1 and both HA and SDC-1; however, such trend was noticed between the serum concentrations of sFlt-1 and HA in patients with early-onset PE, but not in those with the late-onset disease., Conclusions: Evaluation of serum concentrations of HA in patients with PE was found to be more useful in the assessment of endothelial injury, compared to the assessment of SDC-1.The degree of EG damage was comparable in patients with early- and late-onset PE. The pathomechanism of the damage seems to be more sFlt-1 dependent in patients withearly- onset PE than in the case of late-onset disease. The two-stage model of PE is more appropriate for early - onset PE, whereas the pathophysiology of the late-onset disease is rather more complex and heterogenous.

Preterm prelabor rupture of membranes (pPROM) accounts for nearly half of premature births. Although several risk factors have been identified, no markers allowing for effective prevention have been discovered. In this study, we investigated how the maternal serum levels of galectin-1 and galectin-9 change in patients with pPROM in comparison to uncomplicated pregnancies. A total of 75 patients were enrolled to both study and control group (37 vs. 38, respectively). The serum concentration of galectin-1 and galectin-9 were assayed in duplicate using an enzyme-linked immunoassay. All analyses were performed using PQ Stat v. 1.8.4 software. Galectin-1 levels were significantly higher in the controls (13.32 vs. 14.71 ng/mL, p = 0.02). Galectin-9 levels were similar in both groups (13.31 vs. 14.76 ng/mL, p = 0.30). Lower galectin levels were detected for early pPROM (before 32nd GW) in comparison to late pPROM and the controls (8.85 vs. 14.45 vs. 14.71 ng/mL, p = 0.0004). Similar trend was observed in galectin-9 levels, although no statistical significance was found (11.57 vs. 14.25 vs. 14.76 ng/mL, p = 0.26). Low galectin-1 maternal serum level is associated with the incidence of preterm prelabor rupture of membranes. Galectin-9 maternal serum levels were not significantly correlated with pPROM. However, in order to investigate gal-1 and gal-9 levels as potential, promising markers of pPROM, further clinical studies on larger groups are required.

Endometriosis is the cause of infertility. The eutopic endometrium of women with endometriosis showed an aberrant expression pattern of multitude genes. The role of TET1 protein in the pathogenesis of endometriosis and related infertility is not sufficiently known. Further, knowledge on TET1 transcriptional control still remains incomplete. The aim of the study was assessment of TET1 gene expression, DNA methylation and H3K27me3 level of its promoter region in eutopic endometrium of women with endometriosis and infertility. The study included 44 infertile patients with endometriosis (IWE) and 77 infertile (IW) and fertile (FW) patients without endometriosis. The research material was eutopic endometrium. The TET1 mRNA level was analyzed by qPCR. Western blot was used to evaluate the level of TET1 protein. The level of DNA methylation and H3K27me3 level of TET1 gene's promoter region were assessed using HRM and ChIP qPCR, respectively. The level of TET1 expression (TET1 mRNA; TET1 protein level) was lower in IWE during the implantation window (p < 0.001; p = 0.0329). The level of TET1 DNA methylation was higher in the secretory endometrium in mild and advanced IWE (p < 0.004; p < 0.008). H3K27me3 level did not differ between the study groups. The diminished expression of TET1 gene during the secretory phase, may account for the aberrant process of embryonic implantation in infertile endometriosis patients. DNA hypermethylation of TET1 gene is a potential relevant regulator of its expression. H3K27me3 occupancy does not affect the expression of TET1 gene in our study group., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

The paper presents results of preparation and modification of Ti20Nb5Zr foams by a thermal dealloying method followed by electrochemical modification. The first step of this study was the preparation of Ti20Nb5Zr30Mg nanopowder using mechanical alloying (MA). The second was forming green compacts by cold pressing and then sintering with magnesium dealloyed from the structure, which resulted in pores formation. The next step was surface modification by electrochemical etching and silver nanoparticle deposition. Porosity, morphology, mechanical properties as well as biocompatibility and antibacterial behavior were investigated. Titanium foam porosity up to approximately 60% and wide pore size distribution were successfully prepared. The new materials have shown positive behavior in the MTT assay as well as antibacterial properties. These results confirmed great potential for thermal dealloying in preparation of porous structures.

Objective: assessing the incidence of preeclampisa (PE) in women with diabetic kidney disease (DKD) and analyzing the significance of clinical characteristics and changes in laboratory findings throughout the pregnancy on the onset of PE. Methods: the study included 79 patients with DKD. All patients had elevated urinary protein loss (30-299 mg/24 h) or proteinuria (≥300 mg/24 h) in the first trimester of pregnancy. PE was diagnosed in 22,8% patients with DKD. Results: women with proteinuria and/or proliferative retinopathy at the admission developed preeclampsia significantly more frequently than those without these findings. The degree of proteinuria was significantly associated with the risk of PE development in each trimester of pregnancy. Patients with chronic hypertension developed PE significantly more frequently than those who had no chronic hypertension. Conclusion: chronic hypertension and the degree of primary kidney injury and dysfunction are crucial determinants of PE development in women with DKD. Proteinuria seems to be the best renal predictive factors of PE.

With regard to differences in the clinical symptoms of preeclampsia (PE), the degree of endothelial dysfunction may differ between early and late-onset preeclampsia (EOP and LOP). The authors of this study examined it by assessing the endothelial injury level in women with EOP (20 patients) and LOP (20 patients) and in normotensive pregnant women (20 patients) in their late second and third trimesters of pregnancy, using the two markers-the serum concentration of hyaluronan (HA) and the serum level of soluble vascular cell adhesion molecule-1 (sVCAM-1). The serum concentrations of HA and sVCAM-1 did not differ significantly between the EOP and LOP patients. However, these were statistically higher than that of the control group participants ( p < 0.05; p < 0.001). A significant correlation between the levels of HA and sVCAM-1 was found both in the entire group of patients with preeclampsia ( p = 0.0277) and in women with late-onset disease ( p = 0.0364), but not in the patients with early-onset preeclampsia ( p = 0.331). The obtained results indicated a comparable level of endothelial injury in the two types of PE. The presence of a similar degree of endothelial injury in patients with EOP and LOP should create awareness among all clinicians about the possible fatal complications in both groups of patients with PE.

Introduction: Endocan plays a role in the development of vascular tissue in health and disease and is an indicator of endothelial cells activation and angiogenesis. Objective: The aim of this study was to investigate the relationship between endocan serum levels and various types of hypertensive disorders in pregnant women. Patients and methods: We created three study groups (preeclampsia [ n  = 60], chronic hypertension [ n  = 39], gestational hypertension [ n  = 58]) and the control group consisting of 59 healthy pregnant women. The endocan serum concentration was assessed using commercially available ELISA kit. Results: There were no statistically significant differences in endocan serum levels (pg/mL) in each study group compared to controls. The multiple regression did not reveal significant differences between endocan levels in each study group after adjustment for prepregnancy BMI. We did not find any significant correlations between the endocan serum level and patients' age, gestational age (GA) at sample collection, prepregnancy BMI, systolic blood pressure, diastolic blood pressure, and 24-hour urinary protein excretion in each analyzed group. Moreover, in the preeclamptic participants, we did not observe a significant relationship between the endocan concentration and the features indicating the severity of the disease other than elevated blood pressure. There were no differences in endocan serum level in preeclampsia subgroups: early-onset versus late-onset and mild versus severe preeclampsia. Conclusions: Endocan is not involved in the pathogenesis of hypertensive disorders in pregnant women and could not be regarded as a marker of endothelial dysfunction in these cases.

Background Preterm premature rupture of membranes (pPROM) is associated with a high risk of prematurity and complications of fetal inflammatory response syndrome (FIRS). The aim of the study is to determine any correlations between the concentration of selected cytokines contained in the cervicovaginal secretion eluates and in the umbilical cord plasma in patients with pPROM and to find the noninvasive markers of FIRS in order to pinpoint the optimal time of the delivery. Methods The study included 80 patients with pPROM between the 24th and 34th week of gestation. The cervicovaginal fluid and umbilical cord blood were collected. Interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 19 (IL-19) and tumor necrosis factor-α (TNF-α) concentrations were measured in both materials. For the statistical analysis, SigmaStat3.5 software was used. Results There was no direct association in levels of IL-6, TNF-α, IL-10 and IL-19 between the cord blood and cervicovaginal secretions within the studied group. The cut-off point of IL-6 of 26.8 pg/mL in the vaginal fluid had high sensitivity and specificity in order to discriminate between newborns with and without FIRS (81.08%; 76.74%). Conclusion Further studies are needed on a larger group of participants to demonstrate that an elevated concentration of IL-6 above 26.8 pg/mL in the cervicovaginal secretion eluate is an indirect noninvasive marker of FIRS.

The paper presents a promising method of preparation of titanium-based foams by the thermal dealloying method. The first step of this study was the Ti-Ta-Mg based nanopowder preparation using the mechanical alloying (MA) process performed at room temperature. The next step was forming the green compacts by cold pressing and then sintering with magnesium dealloying from the titanium-based alloy structure. The mechanism of the porous structure formation was based on the removal of magnesium from the titanium alloy at a temperature higher than the boiling point of magnesium (1090 °C). The influence of the Mg content on the formation of the porous Ti-30Ta foam has been investigated. The sintering stage was performed in vacuum. During the dealloying process, the magnesium atoms diffuse from the middle to the surface of the sample and combine to form vapors and then evaporate leaving pores surrounded by the metallic scaffold. The porosity, the mechanical properties as well as biocompatibility have been investigated. The titanium-based foam of high porosity (up to 76%) and the pore size distribution from nano- to micro-scale have been successfully prepared. For the medical applications, the Ti-Ta metallic foams have shown a positive behavior in the MTT test. The as-shown results clearly exhibit a great potential for thermal dealloying in the preparation of porous structures.

Introduction: The etiology and pathogenesis of pregnancy-related hypertensive disorders is complex and multifactorial. The aim of our study is the investigation of the differences in the autoantibodies against angiotensin II type 1 receptor (AT1-AA) titers among pregnant patients with chronic hypertension, gestational hypertension, and preeclampsia compared to the healthy pregnant women. Patients and methods: We created three study groups (preeclampsia [ n  = 16], chronic hypertension [ n  = 13], gestational hypertension [ n  = 17]) and the control group consisting of 17 healthy pregnant women. Every compared group was matched for mother's age, parity, prepregnancy BMI, and gestational age at time of recruitment into study. The autoantibodies titer were assessed using commercially available ELISA kit. Results: We found a statistically higher AT1-AA titer in the group of patients with gestational hypertension (GH) and preeclampsia (PE) compared to healthy normotensive pregnant women (median 9.6 versus 7.8 ng/ml, p  = .01 and 10.9 ng/ml versus 7.8 ng/ml, p  = .02, respectively). There was no correlation between blood pressure values and AT1-AA titer in any group. We found no correlation in group with preeclampsia between urinary protein excretion and AT1-AA titer ( p  = .23, R = 0.32). Conclusions: We assume that pregnancy-related hypertensive disorders might be autoimmune diseases and AT1-AA contribute to the pathophysiology of the disease. Our study may have some therapeutic implications and shows the necessity of new research into the mechanisms involved in the production of AT1-AA. Such investigations might enable to inhibit the formation of these autoantibodies or elaborate another method for AT1-AA removal.

Objectives: Intrahepatic cholestasis of pregnancy (ICP) is the most common liver disorder during pregnancy. Cholestasisis associated with increased risk of fetal complications: prematurity, perinatal hypoxia and meconium stained amnioticfluid, and sudden intrauterine fetal death. The exact mechanisms associated with cholestasis fetal sequelae are not fullyunderstood. The aim of the study was the histopathological evaluation of placentas from patients with cholestasis andhealthy pregnant women to establish whether cholestasis is accompanied by changes in placental microstructure., Material and Methods: The effect of cholestasis on placental microstructure was investigated using placental tissue frompatients with cholestatsis treated with ursodeoxycholic acid (UDCA) and from uncomplicated pregnancies. Five placentalhistopathological features were analyzed: number of syncytial knots, number of capillaries per villous, structure of stroma,presence of Hofbauer cells, and villitis of unknown etiology., Results: There were no statistically significant differences in any of the studied parameters between cholestasis-affectedand healthy control groups., Conclusions: There are no diffrences in placental microstructure in cholestasis patients treated with UDCA and in patientswith uncomplicated pregnancy.

Objectives: The aim of our work was to assess the development of children with antenatally diagnosed idiopathic poly- hydramnios, over 12 months from the end of pregnancy., Material and Methods: The study included 91 healthy pregnant patients with idiopathic polyhydramnios. Diagnostic tests results and perinatal medical history were obtained retrospectively. Parents of children were contacted by phone and by mail. The answers were obtained from 64 (70%) parents. For statistical analysis SigmaStat3.5 software was used., Results: Ninety six percent of parents declared that in their opinion the development of children was normal. Abnormali- ties were found in 44% of the children. Thirty percent of neonates demonstrated mild abnormalities which may be due to organic or functional neuromuscular disorders: abnormal muscle tone, speech apparatus and development disorders, swallowing and breathing problems (manifested as vomiting, excessive regurgitation, idiopathic apnoeas). Isolated small malformations were diagnosed in 12 (19%) children. Two children (3%) with SGA were diagnosed with genetic syndromes. More than one of the abnormalities described above were diagnosed in 14% of children. Gestational age at the time of polyhydramnios diagnosis and its severity were not prognostic factors for abnormalities. Seventy percent of newborns were male., Conclusions: Despite the subjectively positive assessment of the development of children by the majority of parents, groups of common disorders requiring long-term follow-up have been identified. Functional disorders of the gastrointestinal tract, CNS and the group of neuromuscular disorders may be responsible for idiopathic polyhydramnios. SGA with co-existing idiopathic polyhydramnios is associated with the risk of genetic diseases. The more frequent incidence of idiopathic poly- hydramnios in male fetuses requires further research.

Little is known about the differences in ten-eleven translocation 1, 2, and 3 (TET1-3) expression in the endometrial phases in eutopic endometrium from infertile women with endometriosis (IWE) and fertile women without endometriosis (FW). Using RT-qPCR and western blot analysis, we assessed the TET expression in the mid-follicular and mid-luteal phases in eutopic endometrium from IWE (n = 38) and FW (n = 18). Both IWE and FW underwent laparoscopic and histological examinations for endometriosis. In the mid-luteal eutopic endometrium in IWE, compared to that of FW, we found significantly reduced levels of TET1 transcripts and proteins (p = .001 and p = .003, respectively) at the severity stage of I/II (p = .029 and p = .003, respectively) and transcripts only at the severity stage of III/IV (p = .003). In the mid-follicular eutopic endometrium of IWE, compared to that of FW, there was a statistically significant reduction in TET2 transcript levels at the severity stage of III/IV (p = .037). Compared to the mid-follicular endometrium, we found a statistically significant increase in TET3 transcript levels during the mid-luteal phase in the eutopic endometrium of all IWE (p = .034) and in the severity stage of III/IV (p = .025). We observed a change in the expression levels of TET1-3 in the eutopic endometrium of IWE.

Introduction: The surfaces of endoosseous dental implants have been subjected to numerous modifications in order to create a surface which can provide rapid bone healing and fast implant loading. Each modification has involved changes to the chemical composition and topography of the surfaces which have resulted in various biological reactions to the implanted material., Aim: The aim of this study was to evaluate the surface topography and chemistry of various modified titanium surfaces: (1) machined surface (MA), (2) alumina-blasted (Al2O3), (3) alumina-blasted and acid-etched (Al2O3 DE), (4) hydroxyapatite/tricalcium phosphate grit-blasted (HA/TCP) and (5) hydroxyapatite/tricalcium phosphate grit-blasted and acid-etched (HA/TCP DE) and to analyse the effects of surface roughness, and chemical composition on human osteoblast vitality, differentiation, morphology and orientation., Materials and Methods: The modified surfaces were subjected to topographic analysis using Scanning Electron Microscopy (SEM), optical profilometry, roughness analysis and chemical composition evaluation using Energy Dispersion Spectroscopy (EDS) analysis. The biological effects of the titanium modifications was analysed using human osteoblasts cell culture where the cell morphology, vitality (MTS assay) and differentiation (ALP activity) was analysed., Results: The machined surfaces were classified as anisotropic, smooth and composed of titanium and oxygen. The blasted surface samples along with the blasted and etched samples were found to be isotropic and rough. The grit-blasting procedure resulted in the incorporation of components from the blasting material. In the case of the blasted and etched samples, etching decreased the surface development as indicated by the Sdr and also reduced the amount of chemical compounds incorporated into the surfaces during the blasting procedure. The attached NHOst cells, proliferated the surfaces. With regard to the MA samples, the cells spread close to the titanium surface, with expanded cytoplasmic extensions and lamelipodia and were oriented in line with the groves left after machining. On the rough substrates, cells were less dispersed and exhibited numerous cytoplasmic extensions, filopodia and interconnections, they were not oriented with respect to the surfaces features. The cell viability of all samples except for Al2O3 decreased after the first day of culture. For all Al2O3, Al2O3 DE and HA samples the viability increased with culture time after an initial reduction. At the end of the culture period the ALP activity was slightly greater on Al2O3 and HA samples compared to the control with the HA DE sample having the same activity as the control. The Al2O3, HA and HA DE ALP samples showed comparable activity and were statistically different from MA and Al2O3 DE samples., Conclusions: In this study, variously treated titanium surfaces were correlated with osteoblastic cell viability, morphology and differentiation in comparison with the plastic and smooth titanium. All examined surfaces were found to be biocompatible. Favourable cell reactions were observed for Al2O3 and HA blasted surfaces. The surface roughness patterns influenced the growth orientation while the surface topography influenced osteoblast morphology. Further animal studies are necessary to compare the in-vivo effect on osseointegration of these modified titanium surfaces., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objectives: Genome-wide association studies in patients with endometriosis revealed ten significant single nucleo-tide polymorphisms (SNPs) in the Caucasian population, which include rs12700667 near NFE2L3, rs12037376 in WNT4, rs7521902 near WNT4, rs13394619 in GREB1, rs10859871 near VEZT, rs1537377 near CDKN2B-AS1, rs4141819 near ETAA1, rs7739264 near ID4, rs1519761 near RND3 and rs6542095 near IL1A., Material and Methods: We replicated ten polymorphisms among infertile women with endometriosis (n = 315) and healthy fertile women (n = 406) in the Polish Caucasian population. Genotyping was conducted either by high-resolution melting curve analysis or by a pre-designed TaqMan probe., Results: For all infertile women with endometriosis, the p values of the Cochran-Armitage trend test for the rs12700667 SNP was ptrend = 0.038 and the odds ratio (OR) for the risk allele frequency (RAF) of rs12700667 was 1.304 (95% CI = 1.009-1.685; p = 0.042). In patients with endometriosis with severity stages III/IV, ptrend for rs12700667 SNP was 0.036 and OR for the RAF was 1.394 (95% CI = 1.010-1.923; p = 0.043). In infertile women with endometriosis with severity stages III/IV for rs4141819 SNP, we observed ptrend = 0.026 and for RAF the OR = 1.350 (95% CI = 1.032-1.766; p = 0.029)., Conclusions: Our results demonstrate association of RAF of rs12700667 and rs4141819 SNPs with infertility in Polish women with advanced endometriosis.

Objectives: The development of endometriosis is associated with changes in the expression of genes encoding the 3β-hydroxysteroid dehydrogenase type II (HSD3B2) and 17β-hydroxysteroid dehydrogenase type II (HSD17B2), estrogen receptors 1 (ESR1) and 2 (ESR2) and the androgen receptor (AR). However, little is known about the expression of HSD3B2, HSD17B1, HSD17B2, ESR1 ESR2 and AR during the endometrial phases in eutopic endometrium from infertile women with endometriosis., Material and Methods: Using RT-qPCR analysis, we assessed the expression of the studied genes in the follicular and luteal phases in eutopic endometrium from fertile women (n = 17) and infertile women (n = 35) with endometriosis., Results: In the mid-follicular eutopic endometrium, we observed a significant increase in HSD3B2 transcript levels in all infertile women with endometriosis (p = 0.003), in infertile women with stage I/II endometriosis (p = 0.008) and in infertile women with stage III/IV endometriosis (p = 0.009) compared to all fertile women. There was a significant increase in ESR1 tran-scripts in all infertile women with endometriosis (p = 0.008) and in infertile women with stage I/II endometriosis (p = 0.019) and in infertile women with stage III/IV endometriosis (p = 0.023) compared to all fertile women. In the mid-luteal eutopic endometrium, we did not observe significant differences in HSD3B2, HSD17B1, HSD17B2, ESR1, ESR2 and AR transcripts between infertile women with endometriosis and fertile women., Conclusions: Observed significant increase in HSD3B2 and ESR1 transcripts in follicular eutopic endometrium from infer-tile women with endometriosis may be related to abnormal biological effect of E2 in endometrium, further affecting the development of human embryos.

Objectives: The aims of our study were to assess the correlation between the amniotic fluid index (AFI) value and the frequency and type of fetal anomalies., Material and Methods: The material included 94 patients at the third trimester of pregnancy, 60 with mild polyhydramnios, 19 with moderate one, and 15 with severe one. Polyhydramnios was diagnosed if AFI was > 24 cm. All patients were divided into three groups based on the value of AFI: 1) mild polyhydramnios with AFI between 24.1 and 29.9 cm, 2) moderate polyhydramnios with AFI between 30-34.9 cm, and 3) severe polyhydramnios with AFI ≥ 35 cm., Results: The incidence of fetal malformations correlated significantly with the degree of polyhydramnios and was the highest in patients with severe polyhydramnios (53.3%, p = 0.002). Congenital malformations of the gastrointestinal tract were the most frequent fetal anomalies in the whole group of patients (5.3%). Trisomy 18 was the most frequent aneuploidy found in women with polyhydramnios (2.1%)., Conclusions: The incidence of fetal congenital anomalies significantly increases with the degree of polyhydramnios, being most frequent in severe one and rather rare in a mild one. Congenital malformations of the gastrointestinal tract were the most frequent anomalies in patients with polyhy-dramnios, especially in women with severe polyhydramnios.

Endometriosis is considered to be an epigenetic disease. It has previously been reported that the DNA methyltransferase 3-like (DNMT3L) rs8129776 single nucleotide polymorphism (SNP) contributes to endometrioma. In the present study, high‑resolution melting curve analysis was used to investigate the risks associated with the DNMT3L c.910‑635A/G (rs8129776), c.832C/T (rs7354779), c.812C/T (rs113593938) and c.344+62C/T (rs2276248) SNPs on stage I‑II endometriosis‑associated infertility. Included in the present study were patients presenting with stage I‑II endometriosis‑associated infertility (n=154) and a control cohort of healthy patients with confirmed fertility (n=383). No significant association between the above‑listed DNMT3L SNPs and the development of endometriosis‑associated infertility was identified. The lowest P‑values generated from trend analysis were observed in the DNMT3L c.832C/T (rs7354779) SNP (Ptrend=0.114). Furthermore, haplotype analyses of the DNMT3L SNPs failed to reveal any risk association between the development of endometriosis‑associated infertility and the above‑listed polymorphisms, even when the SNPs were present in combinations. Finally, a meta‑analysis was performed to examine the association between the DNMT3L rs8129776 SNP and the development of endometrioma, from which no association between the two was identified. On the basis of these results, the present study has demonstrated that variations in the DNMT3L gene do not contribute to stage I-II endometriosis-associated infertility.

It has recently been reported that vitamin D blood plasma levels are associated with reduced risk of endometriosis. The present study aimed to investigate whether the vitamin D binding protein (GC), vitamin D receptor (VDR), and retinoid X receptor (RXR) gene variants may be genetic risk factors for endometriosis‑associated infertility. The subjects consisted of 154 women with endometriosis‑associated infertility and 347 controls. Using polymerase chain reaction restriction fragment length polymorphism and high resolution melt techniques, the GC rs1155563, rs2298849 and rs7041; RXRA rs10881578, rs10776909 and rs749759; VDR BsmI rs1544410; and FokI rs2228570 single nucleotide polymorphisms (SNPs) were investigated in the patients with endometriosis and the healthy controls. The results indicated that no significant differences were observed between the genotype and allele frequencies of all experimental SNPs in the vitamin D signaling pathway genes in women with endometriosis-associated infertility and controls. However, a significant association was present between the A‑T haplotype, consisting of VDR rs1544410 and rs222857 minor alleles, and endometriosis-associated infertility [OR=1.659 (1.122‑2.453), P=0.011]. The results of the present study suggested that VDR gene variants act as genetic risk factors for endometriosis‑associated infertility.

Data suggests that dopamine receptor DRD2 gene variants may contribute to hyperprolactinemia and that they may be risk factors for endometriosis-related infertility. The purpose of the present study was to determine whether nucleotide variants of the DRD2 gene may be associated with infertility related to endometriosis. Five DRD2 SNPs, rs1800497, rs6277, rs2283265, rs4245146 and rs4648317, which are located in different blocks of linkage disequilibrium, were studied in 151 cases and 381 controls. No significant differences between DRD2 rs1800497, rs6277, rs2283265, rs4245146 and rs4648317 genotype, allele nor haplotype frequencies were observed in women with endometriosis-related infertility compared with the control group. The present results did not confirm DRD2 gene variants to be genetic risk factors for endometriosis-related infertility.

There are inconsistent data on the contribution of vascular endothelial growth factor ( VEGF ) -460 C/T (rs833061), +405 G/C (rs2010963) and +936 C/T (rs3025039) single-nucleotide polymorphisms (SNPs) to endometriosis in different ethnicities. Therefore, using high-resolution melting curve analysis, the present study examined the distribution of these SNPs in females with endometriosis-related infertility and a control group. None of the three VEGF SNPs were associated with endometriosis-related infertility in the dominant and recessive models. The lowest P-values of the trend were observed for the VEGF +936 C/T (rs3025039) SNP in endometriosis-related infertility (P trend =0.149). Similarly, haplotype analyses of VEGF SNPs did not demonstrate any SNP combination as a risk for endometriosis-related infertility, and the lowest overall P-values, P=0.141 and P corr =0.395, were observed for a haplotype (TGT) of the above SNPs. Taken together, these results did not demonstrate the contribution of VEGF C/T, +405 G/C and +936 C/T SNPs to endometriosis-related infertility.

Unlabelled: Heparin has a beneficial effect in the treatment of recurrent miscarriages and positively affects implantation rates in the IVF procedure in women with reproductive disorders not associated with thrombophilia. Several studies have indicated that heparin, by blocking the enzymatic activity of heparanase, may affect the structure and function of the extracellular matrix (ECM) and related growth factors. Disturbances in the remodeling (ECM) are believed to be the potential cause of implantation disorders and recurrent miscarriages., Objectives: The aim of the study was the evaluation, on an in vitro model, of the effect of low molecular weight heparin (LMWH) on the expression of heparanase (HPSE) and, important for successful implantation and invasion of trophoblast, heparan sulfate (HS)--binding growth factors, i.e., heparin-binding epidermal growth factor-like (HB-EGF), vascular endothelial growth factor (VEGF-A), fibroblast growth factor (FGF2) in the endometrium, during the implantation window in women with recurrent miscarriage., Method: Biopsy samples, obtained from 10 patients with two or more unexplained miscarriages, were used to construct a co-culture of glandular epithelial cells and stroma. Endometrium in vitro model was supplemented with steroid hormones and enoxaparin 5 ug/ml. Using the qPCR, we assessed, relative levels of the HPSE, HB-EGF, VEGF-A and FGF2 transcripts in glandular epithelium and stroma in cell culture. Using ELISA, we measured con- centrations of the mentioned above factors in culture medium., Results: A statistically significant increase in the relative level of HPSE, HB-EGF VEGF-A, FGF2 transcripts in the cells of the glandular epithelium and stroma (p<0.001), as well as their increased concentration in the medium of cultures treated with steroid hormones (p<0.001) were observed. However we found no effect of LMWH supple- mentation on the level of the investigated factors., Conclusions: Our results show that the importance of the HPSE hydrolytic activity in the endometrium, during the implantation window, may have a secondary function, and/or that beneficial effects of LMWH in women with impaired reproduction have no significant, direct connection with the, catalyzed by HPSE, reconstruction of the ECM and with release of heparin-binding growth factors.

Objective: Endometriosis is recognized as an estrogen-dependent disease. There are conflicting data demonstrating single nuclear polymorphisms (SNPs) of CYP17 and CYP19 steroidogenic genes as related to endometriosis risk. We assessed the CYP17 5'-untranslated region -34 A/G (rs743572) and CYP19 Ex10 + C1558T (rs10046) SNPs in stage I-II endometriosis., Design: Case-control study., Setting: Division of reproduction at a university department in Poland., Population: A total of 115 women with diagnosed stage I-II endometriosis according to the revised American Society for Reproductive Medicine (rASRM) classification and 197 fertile women as controls., Methods: The SNPs CYP17 -34 A/G and CYP19 Ex10 + C1558T were identified by high-resolution melting curve analysis., Main Outcome Measures: Genotype prevalence and odds ratio for recessive and dominant genetic model for CYP17 and CYP19 SNPs., Results: We observed a significantly increased CYP17 GG and GA genotype frequency in women diagnosed with rASRM stage I-II endometriosis compared with fertile women (OR = 2.4; 95% CI 1.4-4.2, p = 0.002). We also found a significantly increased CYP17 G allele frequency in cases compared with controls (OR = 1.6; 95% CI 1.2-2.2, p = 0.004). There were no significant differences in the distribution of the CYP17 GG genotype and CYP19 Ex10 + C1558T polymorphism between women diagnosed with rASRM stage I-II endometriosis and controls., Conclusion: The CYP17 -34 G variant, previously associated with increased 17β-estradiol production, displayed a contribution to stage I-II endometriosis in women from a Polish population. Increased 17β-estradiol concentration in carriers of the CYP17 -34 G variant might contribute to endometriosis and associated pathological processes., (© 2013 Nordic Federation of Societies of Obstetrics and Gynecology.)

Objectives: Assessment of serum concentrations of antiangiogenic factors, triglycerides, glucose, insulin and SHBG in women with two forms of preeclampsia (placental and maternal)., Material and Methods: The study was conducted among 30 patients with placental preeclampsia and 20 women with maternal form of the disease. All patients were hospitalized at the Division of Reproduction, Poznan University of Medical Sciences, between 2010-2012. The placental form of preeclampsia was diagnosed in cases when Doppler signs of placental insufficiency were present. The maternal type of the disease was diagnosed in the absence of coexisting markers of placental insufficiency in the Doppler study ELISA was used to determine the concentrations of antiangiogenic factors (sFlt-1 and sEng)., Results: The placental form of preeclampsia was diagnosed significantly earlier than maternal type of the disease. In women with placental preeclampsia the gestational age at delivery and newborn birth weight were significantly lower than in patients with maternal preeclampsia. IUGR incidence (expressed as a percentage) was significantly higher in patients with placental preeclampsia as compared to the women with maternal form of the disease. Serum concentrations of sFlt-1 and sEng were significantly higher in women with placental preeclampsia. No differences in concentrations of glucose, insulin, triglycerides and SHBG were found between groups., Conclusions: 1. The Two Stage Model of preeclampsia, characterized by increased concentrations of antiangiogenic factors in maternal blood secondary to decreased placental blood flow seems to better explain the pathophysiology of the placental form of preeclampsia than the maternal one. 2. Late onset of clinical symptoms in maternal preeclampsia, lower incidence of IUGR, as well as lower concentrations of antiangiogenic factors in maternal blood, do not indicate the primary role of placental pathology in the pathogenesis of the disease. 3. In spite of no difference in metabolic abnormalities in third trimester of pregnancy between two types of preeclampsia, the patophysiology of the two forms of the disease seems to be different. 4. The obtained results of metabolic markers in women with two types of preeclampsia justify the need of further studies in this field in first trimester of pregnancy.

Introduction: Claudin-4 (CLDN4) is a transmembrane protein, responsible for cellular contact and organization. A different expression of claudin 4 in the endometrium, depending on the menstrual cycle and with peak at the aim of the 'implantation window', has been observed. CLDN4 is believed to play an important role in embryo implantation., The Aim: The aim of the study was to compare the mRNA CLDN4 expression levels in two subgroups of infertile women (idiopathic infertility or minimal endometriosis) and compare them to fertile controls., Method: The study included 36 women with idiopathic infertility and 24 with minimal endometriosis. The control group comprised 26 women. Eutopic endometrium samples were collected with a Pipelle device during the implantation window. Firstly mRNA was extracted from the endometrium and reverse transcribed into cDNA. Real time PCR was used for the assessment of relative expression levels., Results: The observed transcription level of CLDN4 did not differ statistically between the studied groups, but was significantly higher when compared to controls., Conclusions: Exceedingly high levels of CLDN4 might negatively influence fertility rates.

Objective: Overexpression of DNA methyltransferase 3A (DNMT3A) and aberrant methylation of various genes in eutopic endometrium have been demonstrated in women with endometriosis. We aimed to study whether DNMT3A polymorphisms could be a genetic risk factor for endometriosis and endometriosis-related infertility., Study Design: We studied 5 SNPs (rs2289195, rs7590760, rs13401241, rs749131 and rs1550117) located in the DNMT3A gene in 357 women with endometriosis and 640 controls., Results: We did not observe significant differences between genotype and allele frequencies of rs2289195, rs7590760, rs13401241, rs749131 and rs1550117 SNPs in women with endometriosis, endometriosis-related infertility, and controls. The lowest p values of the trend test were observed for DNMT3A rs1550117 in endometriosis and endometriosis-related infertility (p(trend)=0.049 and p(trend)=0.055, respectively)., Conclusions: Our results did not supply evidence for the contribution of SNPs located in DNMT3A to either endometriosis or endometriosis-related infertility., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Objectives: Assessment of uterine artery blood flow in women with congenital thrombophilia and antiphospholipid syndrome (APS) in the first half of pregnancy, Material and Methods: Uterine arteries blood flow was assessed in a Doppler examination in 20 women with thrombophilia (15 with congenital thrombophilia, 5 with APS) at 12 and 20 weeks gestation at the Division of Reproduction, Poznan University of Medical Sciences, between 2000 and 2012 The control group consisted of 20 multiparous pregnant women with no history of pregnancy complications. All patients with thrombophilia received enoxaparin or enoxaparin and aspirin before enrollment into the study Patients from the control group did not receive any antithrombotic prophylaxis. The mean Pulsatility Index (PI) of both uterine arteries and the presence or the absence of the "notch" was assessed, both at 12 and 20 weeks gestation in each patient from the study and from the control groups., Results: Mean PI values in the uterine arteries at 12 weeks in patients with thrombophilia and in controls were 1.82 (1.00-3.13) and 1.52 (1.30-1.88), respectively (p = 0.08). Mean PI value in the uterine arteries was 7.27 (0.61-2.48) in women with thrombophilia at 20 weeks, which turned out to be significantly higher (p = 0.026) than in the control group 1.07 (0.8-1.24). The bilateral "notch" was found at 12 weeks gestation in 40% of patients with thrombophilia vs. 0% in the control group (p = 0.03). There was no significant difference between the groups in this parameter at 20 weeks., Conclusions: 1. An increased impedance of flow was found in the uterine arteries in patients with thrombophilia at 12 and 20 weeks gestation in spite of antithrombotic prophylaxis. 2. Thrombotic episodes in patients with thrombophilia cannot be explained solely by the presence of placental thrombosis.

Aim: The aim of this study was to estimate the prevalence of factor V Leiden and prothrombin gene G20210A mutation among women with pregnancy loss in Poland., Material and Methods: we analyzed a group of 396 women (mean age of 30.4 (+/- 4.6) years), who experienced at least one pregnancy loss. Patients were recruited from 6 academic centers (Poznań, Białystok, Lublin, Wrocław Bydgoszcz, Gdańsk), and were divided into the following groups: 122 patients with 3 episodes of early recurrent pregnancy loss (group 1), 87 patients with late pregnancy loss (group 2) and 46 patients with intrauterine pregnancy loss (group 3). Patients who did not fulfill the above inclusion criteria were divided into additional groups. 50 healthy women (mean age of 29.2 (+/- 4.5) years), having at least one child, constituted the control group. Factor V Leiden mutation and prothrombin G20210A gene mutation were examined in all 396 women with pregnancy loss and 50 controls. For molecular analysis peripheral blood was tested. Genome DNA isolation from lymphocyte was performed with commercial assay QIAampDNA Blood Mini Kit., Results: Among 396 women with unexplained loss of at least one pregnancy 36 (9.1%) were carriers of inherited thrombophilia. Factor V Leiden mutation was present in 29 women (73%), prothrombin gene mutation G20210A in 6 (1.5%) and in 1 (0.3%) patient both mutations were detected. No coagulation defects were found in the control group. Factor V Leiden mutations was the most common disorder (21.7%) in patients with intrauterine demise and was significantly higher than in the group of women with early recurrent and late losses, p<0.011 and p<0,006 respectively The frequency of G20210 A prothrombin gene mutation did not differ substantially between the examined groups; the highest number (2.6%) was found in women with early and late pregnancy losses, and the lowest number (0.8%) was seen in women with early recurrent miscarriages., Conclusion: Factor V Leiden screening should be performed, regardless of negative history of thrombosis, in patients who experienced intrauterine fetal demise or recurrent early miscarriages.

The Aim: The aim of the study was the analysis of antiphospholipid syndrome frequency in Polish women with pregnancy loss., Material and Methods: We analyzed 352 women with average age of 31.4 (+/- 4.3) who experienced one or more pregnancy losses. Patients from 5 University centers: Poznań, Białystok, Lublin, Wrocław and Bydgoszcz were divided into 3 groups. In the 1st group there were 150 women with recurrent early pregnancy losses before 10 weeks gestation, in the 2nd group we included 160 women with late pregnancy losses, in the 3rd group we analyzed 42 women with labor complicated by preeclampsia or placental insufficiency within or before 34 gestational week. All 352 women were screened for the presence of anticardiolipin antibodies (aCL), anti beta2glikoprotein I (abeta2GPI) and lupus anticoagulant (LA). The first two antibodies were investigated with ELISA test and the last one with APPT based test. Only the second positive result qualified patients as antiphospholipid antibody (aPL) positive. Antibodies against cardiolipin and beta2glicoprotein I were analyzed in three classes--IgA, IgG and IgM--where the laboratory criteria of positivity were titers above 40 unitsU/ml or above 99 centile., Results: 13 patients (3.69%) in the screened population of 352 women met the criteria of the antiphospholipid syndrome. The frequency of APS in women with early and late pregnancy losses were 1.33% and 6.25%, respectively. The most common antibody found was lupus anticoagulant (LA). In 3 women with late pregnancy loss all three antibodies were found. In women with premature deliveries complicated with preeclampsia and/or placental insufficiency the frequency of APS was 2.58%., Conclusion: 1. Screening of antiphospholipid syndrome should be routinely performed in women with late pregnancy loss 2. We can conclude that APS is rarely found in women with pregnancy loss before 10 gestational week.

Background: The aim of the study was a histologic evaluation of placentas and chorions from pregnancies complicated by antiphospholipid syndrome (APS) and inherited thrombophilia in women treated with anticoagulants and in a group of women without the anticoagulant treatment., Material: 24 placentas from 18 patients with acquired and inherited thrombophilia and 23 chorions from 19 miscarriages in women with the same diagnosis were included in the study There were 33 chorions from miscarriages from healthy women and 25 placentas from uneventful pregnancies in the control group. Biopsies from placentas and chorions were stained with eosin and hematoxylin and evaluated for the presence of villous thrombosis, fibrin deposits, intraplacental hematomas, thrombosis in fetal circulation and other histological findings., Results: In the placentas from pregnancies complicated with APS and inherited thrombophilia, the presence of fibrin deposits in the basal and villous plate, stasis in the fetal circulation and thrombosis in the villous plate and villous vessels, were statistically more frequent (p < 0,001) comparing with physiological pregnancies. In the material from miscarriages the presence of fibrin deposits was statistically more common (p < 0,001) in women with thrombophilia. The number of physiological findings in placentas of women treated with anticoagulants in pregnancy and in the non-treated ones, whose pregnancies resulted in fetal demise, were not statistically different., Conclusions: 1. Fibrin deposits between villi and villous plate are the most recognizable features in placentas and chorions from APS pregnancies 2. In women with APS impaired trophoblast invasion seems to be the most likely cause of recurrent miscarriages 3. Anticoagulant therapy does not prevent either fibrin deposits collection or other placental changes

Objective: Endometriosis has been considered an epigenetic disease. Single nucleotide polymorphisms (SNPs) located in genes encoding enzymes of the folate and choline metabolism may affect DNA methyltransferase activity., Study Design: We studied 16 SNPs in 12 folate and choline metabolism genes, including BHMT (rs7356530 and rs3733890), BHMT2 (rs625879), CBS (844ins68), CHDH (rs893363 and rs2289205), CHKA (rs7928739), MTHFD1 (rs2236225), MTHFR (rs1801133), MTR (rs1805087), MTRR (rs1801394), PCYT1A (rs712012 and rs7639752), PEMT (rs4244593 and rs4646406) and TCN (rs1801198) in one hundred and sixty-three infertile women with minimal endometriosis and one hundred and fifty fertile women., Results: There were no significant differences between genotype and allele frequencies of these gene variants in infertile women with endometriosis (n=163) and controls (n=150). The lowest, but not statistically significant, p values of the trend test were observed for the CBS 844ins68 and MTR rs1805087 (ptrend=0.0527 and ptrend=0.0771, respectively) polymorphisms. However, the exhaustive multifactor dimensionality reduction analysis revealed an epistatic interaction between rs1801133 of MTHFR and rs4244593 of PEMT in endometriosis-associated infertility (p=0.0240)., Conclusions: Our results showed moderate evidence for the contribution of SNPs located in genes encoding folate and choline metabolism enzymes to infertility in women with endometriosis., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

In fertile women, HOXA-10 and HOXA-11 expression rises during the luteal phase, with the peak occurring during the implantation window, and stays at a high level until the end of the cycle. We evaluated the transcript and protein levels of HOXA-10 and HOXA-11 in the endometria of patients with idiopathic infertility (n = 15) and control patients (n = 10). The amounts of mRNA were determined by reverse transcription and real-time quantitative PCR. The protein levels were evaluated by Western blotting analysis. Using immunohistochemical techniques, we compared the localization of HOXA-10 and HOXA-11 proteins in the implantation window between the study and control groups. We observed statistically significantly decreased HOXA-10 and HOXA-11 transcript levels (p = 0.003, p = 0.012 respectively) in infertile patients compared to controls. There was no significant decrease in HOXA-10 protein levels between these groups (p = 0.074). However, we observed a significantly higher level of HOXA-11 protein in the endometria of infertile patients compared to controls (p = 0.015). HOXA-10 and HOXA-11 proteins were localized in the nuclei of the endometrial stromal cells. Immunohistochemical analyses did not reveal differences between amounts of HOXA-10 and HOXA-11 protein levels in infertility and control groups. Our results suggest that HOXA-10 and HOXA-11 gene expression in the endometrium during the implantation window may not be altered in patients with idiopathic infertility.

Implantation depends on two factors - embryo and endometrium. The period of maximal endometrial receptivity is a poorly understood phenomenon. We decided to look at three possible markers of implantation: pinopodes, leukemia inhibitory factor, and matrix metalloproteinase 2 and their correlations. We included in the study 23 idiopathic infertility patients and 21 patients with recurrent spontaneous abortions of unknown etiology. Twenty one fertile patients were also recruited. A biopsy was used for endometrial dating according to the Noyes and Hertig criteria, and assessed for the presence of pinopodes via a scanning electron microscope. Endometria were examined in Real Time-Polymerase Chain Reaction cycles for the mRNA expression of leukemia inhibitory factor (LIF) and matrix metalloproteinase 2 (MMP2). No difference was found in the stage of pinopodes development, nor in the coverage of endometrial surface between the studied groups. The expression level for LIF mRNA was lower in control patients compared to idiopathic infertility and recurrent miscarriage patients. No difference was detected in the expression of MMP2 between all studied groups. No correlation was found between pinopodes development stage and LIF and MMP2 expressions in endometrium. Of the studied factors, LIF and pinopodes show the most promise as potential markers of endometrial receptivity. However, the results achieved suggest that these markers are independent of each other.

Recent human and animal studies have suggested that reduced HOXA10 expression in the implantation window of eutopic endometrium may contribute to infertility in women with endometriosis. Therefore, we examined the HOXA10 transcript, protein and HOXA10 promoter methylation levels in midluteal eutopic endometrium from 17 infertile women with minimal endometriosis and 15 healthy fertile women from a Polish cohort. Real-time quantitative PCR (RQ-PCR) and western blotting analysis revealed significantly lower levels of HOXA10 transcript (P=0.019) and protein (P=0.048) levels in eutopic endometrium from infertile women with endometriosis as compared to healthy fertile women. Moreover, sodium bisulfite sequencing of three HOXA10 CpG islands showed significantly higher methylation levels of genomic DNA from midluteal eutopic endometrium from infertile women with endometriosis as compared to healthy fertile women (P=0.006). We confirmed that DNA hypermethylation can be one of the potential molecular mechanisms silencing HOXA10 expression in the midluteal endometrium associated with infertility in women with endometriosis., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

Human endometrium, deciuda and placenta have been shown to express factors that inhibit the complement activation cascade - decay-accelerating factor (DAF), membrane cofactor protein (MCP) and the C3 complement component. In the following study we have analyzed the transcripts levels for DAF, MCP and heparin-binding epidermal growth factor-like growth factor (HB-EGF), the C3 complement component and receptor for vascular endothelial growth factor (VEGFR1) as markers of endometrial unbalance between factors activating the complement system in women with consecutive miscarriages. Study enrolled 30 women with at least two consecutive miscarriages, and 19 healthly women, that comprised the control group. RNA was isolated from endometrial samples. Transcripts levels of DAF and MCP was higher in women with consecutive miscarriages compared to controls, 0.78 vs 5.08 (p<0.001) and 0.25 vs 0.17 (p=0.001) respectively. In consecutive miscarriages group, DAF and MCP expression was correlated with the C3 expression, with r=0.60; p<0.001 and r= 0.40; p=0.03 respectively. Correlation between DAF and C3 was also noted in controls, 0.70; p=0.001. In women with two or more consecutive miscarriages the analysis proved higher expression of genes that encode proteins that inhibit the complement cascade. Further studies are needed to confirm that this might be a reaction to increased presence of the complement factors, which like C3 that are synthesized in the endometrium.