Emiliana Konova

Association of inherited thrombophilia with embryonic and postembryonic recurrent pregnancy loss

To investigate the impact of maternal-inherited thrombophilia: effects of factor V Leiden (FVL) and prothrombin gene mutation (FII 20210G>A) on the development of recurrent pregnancy loss in embryonic and postembryonic periods. A total of 153 patients were analysed for FVL and FII 20210G>A according to placenta gestation: 94 women with embryonic loss prior 10 weeks of gestation and 59 women with postembryonic (early fetal) loss occurring between 10 and 14 weeks of gestation. The control group consisted of 100 healthy women, with at least one uncomplicated full-term pregnancy. FVL prevalence was not significantly associated with pregnancy loss prior to 10 weeks of gestation (9.6%) compared with controls (7%) [odds ratio (OR) 1.41; 95% confidence interval (CI) 0.454–4.416, P > 0.05], but it was much more pronounced in women with postembryonic loss (10–14 weeks of gestation) – 18.6% (OR 3.05; 95% CI 1.010–9.387, P = 0.047). FII 20210G>A was significantly higher in both groups with embryonic (17%) and early fetal losses (16.9%) as compared to controls (3%) (OR 6.63; 95% CI 1.731–29.752, P = 0.003; OR 6.60; 95% CI 1.572–31.856, P = 0.006). FII 20210G>A is significantly associated with an increased risk of early recurrent pregnancy loss throughout the entire first trimester. FVL was significantly higher only in early fetal period after starting of the placentation process, but not associated with embryonic recurrent pregnancy loss. These results suggested that the first trimester should be viewed rather as a heterogeneous interval, with different relation to FVL in the embryonic and postembryonic fetal period. Genetic testing should be applied according to the diverse contribution of thrombophilic markers to embryonic and postembryonic period.

Inherited thrombophilia and IVF failure: the impact of coagulation disorders on implantation process.

In connection with the embryo acceptance process after IVF procedure, endometrial cells surface receptors, extracellular matrix (ECM) molecules, endothelium and blood circulation factors were involved in remodelling of endometrium. Plasminogen activator inhibitor type 1 plays a significant role during the early phases of placental vascular remodelling and regulates the trophoblast invasion through controlling plasmin activity. Endometrial cell surface protein integrin alphaV/beta3, responsible for the adhesion of the embryo, has had also the same subunit beta3, which is component of integrin alphaIIb/beta3 connected with platelet aggregability. Prothrombin, furthermore, has had a debatable effect upon endothelial and mesenchymal cells and possible contribution on embryo vascular development. Confoundable data have been present about the role of coagulation factor V and its role for implantation. These and other coagulation factors have relatively common gene polymorphisms that enhanced their activity. This review discusses the effect of increased coagulation activity on implantation process, which is not yet fully determined. The establishment of the positive or negative impact of mother hypercoagulability on the success of embryo implantation after assisted reproduction technology could determine the timing of preventing anticoagulant therapy in women with history of early embryo loss.

Age-Related Changes of Anti-Elastin Antibodies in Senescence-Accelerated Mice

Background: Antibodies recognizing the elastin precursor tropoelastin (ATEAb) or degradation products α-elastin (AEAb) are found in the serum of healthy human subjects, as a part of a homeostatic mechanism which assembles new or clears altered elastin structures. Serum ATEAb (reflecting elastin synthesis) and AEAb (reflecting elastin destruction) appear to correlate with the production and breakdown of the elastic tissue, respectively. Objective: The aim of this study was to investigate plasma levels of AEAb and ATEAb in senescence-accelerated prone (SAMP8) and senescence-accelerated resistant (SAMR1) mice, compared with imprinting control region (ICR) mice in order to evaluate their age-related changes. Methods: The levels of AEAb and ATEAb were measured by home-made ELISA in plasma of SAMP8, SAMR1, and ICR mice, grouped according to their age (3 and 9 months) and sex. The specificity of AEAb and ATEAb activity in mouse plasma, and elastin-derived peptides (EDP) in sera of ICR mice at 3 and 9 months of age were tested by competitive ELISA. Results: The specificity of AEAb and ATEAb in mouse plasma was confirmed by the competitive investigations. The levels of AEAb in the plasma of SAMR1 and SAMP8 were increased compared to the levels measured in ICR on the matched ages (p < 0.001). Age-related increase of the levels of AEAb and ATEAb was established in the 3 strains (p < 0.001). Significantly higher levels of AEAb were established in female 9-month-old mice compared to males in all strains. The ATEAb:AEAb ratio was significantly lower in the SAM compared to the ICR strain. Positive correlation was established between the levels of serum AEAb and EDP in mouse sera of ICR mice. Conclusion: Variations with age in the plasma levels of AEAb and ATEAb were established in SAM compared with ICR, and in SAMP8 compared with SAMR1. Our findings suggest that increased anti-elastin IgG autoantibodies could be used as a marker of aging in SAM and possibly contribute to the processes of aging. The absence of a difference between SAMP8 and SAMR1 regarding the ATEAb:AEAb ratio raises the question if SAMR1 are an appropriate control of SAMP8 in terms of the senescence of the elastic tissues.

Anti-elastin antibodies and elastin turnover in normal pregnancy and recurrent pregnancy loss.

Problem  The aim of this study was to investigate elastin turnover and autoimmunity in patients with a history of recurrent pregnancy loss (RPL) and during normal pregnancy.

Upregulation of the CD40―CD40 ligand system in pre-eclampsia

To investigate the CD40–CD40 ligand (CD40L) system in women with pre‐eclampsia.

Polymorphism A1/A2 in the cell surface integrin subunit β3 and disturbance of implantation and placentation in women with recurrent pregnancy loss.

Polymorphism A1/A2 in the β3 subunit of integrins αIIb/β3 and αV/β3 is implicated in the risk of development of embryonic and fetal recurrent pregnancy loss (RPL). In 191 women with RPL, polymorphism A1/A2 was statistically significantly associated with RPL at <10 weeks of gestation (29.3% versus 16.4% in controls), but it was much more pronounced in 67 women with RPL between 10 and 20 weeks of gestation (41.8%), illustrating its role in recurrent fetal loss.

Flow Cytometric Investigation of CD40-CD40 Ligand System in Preeclampsia and Normal Pregnancy

Our aim was to investigate the CD40-CD40 ligand system in preeclamptic women. We also studied CD62P and platelet—monocyte aggregates, which have been closely linked to the CD40—CD40L system. Platelet expression of CD40L and CD62P and expression of CD40 on monocytes and platelet—monocyte aggregates were determined by flow cytometry in whole blood from 23 preeclamptic women, 23 normotensive pregnant women, and 23 nonpregnant women. The preeclamptic women showed a significant increase in CD40L and CD62P on platelets and in CD40 on monocytes when compared with normotensive pregnant women and nonpregnant women (all P < .001). There was a significant increase in platelet—monocyte aggregates in preeclamptic women (P < .001) and normotensive pregnant women (P = .003) compared with nonpregnant women. Preeclampsia is associated with activation of the CD40—CD40L system. The activation of this system may contribute to the development or maintenance of the proinflammatory and prothrombotic milieu found in preeclampsia.

Non-Enzymatic Glycation of Human Fibrillin-1

Non-enzymatic glycation of proteins is one of the key mechanisms in the pathogenesis of diabetic complications and may be significant in the age-related changes of tissues. We isolated and investigated the in vitro glycation of human aortic fibrillin-1. Fibrillin-1 was prepared from thoracic aortas of 9 accident victims distributed in three age groups. The purity of isolated fibrillin-1 was proved. It was glycated by incubating with different glucose concentrations in 0.2 M phosphate buffer, pH 7.4, for 30 days, at 37°C. The degree of early products of glycation was measured by two colorimetric methods, i.e. nitroblue tetrazolium and 2-thiobarbituric acid. Advanced glycation end products (AGEs) were determined by fluorescence measurement. The highest level of early products of glycation was found on day 2 after the beginning of incubation for the fibrillin-1 isolated from the youngest group. Fluorescence in the age groups, as an index of advanced glycation, consistently increased between days 6 and 24. The fibrillin-1 isolated from the youngest group had the highest capacity to form fructosamine and AGEs under glycation in vitro. The capacity of glycation of the ‘oldest’ fibrillin did not increase significantly during the incubation. Investigation of the properties of glycated fibrillin-1 will help to understand the importance of this long-lived protein to age-related changes in tissues and for diabetic complications.

Levels of platelet-leukocyte aggregates in women with both thrombophilia and recurrent pregnancy loss.

The aim of our study was to investigate the significance of platelet-leukocyte aggregates (PLA) in women with recurrent pregnancy loss (RPL) as well as to identify association between common thrombophilic factors and whole blood levels of PLA in these patients. We measured PLA by whole blood flow cytometry in 66 nonpregnant women with hereditary and/or acquired thrombophilia and RPL, classified to 3 study groups, according to the type of losses (first, second, and third trimester) and 35 age-matched healthy controls. Platelet-leukocyte aggregates levels in all study groups were significantly increased compared to the control group (median values 2.13%, 2.32%, and 2.41%, vs median value in the control group 1.39%, P < .05 for all comparisons). Women with a single thrombophilic factor and women with combination of thrombophilic factors did not differ significantly as regards the PLA levels (2.13% vs 2.27%, P = .4). This study suggests that PLA may have a role in the pathogenesis of RPL in women affected by hereditary or acquired thrombophilia.

ORIGINAL ARTICLE: Anti-Elastin Antibodies and Elastin Turnover in Normal Pregnancy and Recurrent Pregnancy Loss

Problem  The aim of this study was to investigate elastin turnover and autoimmunity in patients with a history of recurrent pregnancy loss (RPL) and during normal pregnancy.