Fetal cells and cell-free fetal DNA in maternal blood: new insights into pre-eclampsia

The examination of fetal cells, specifically erythroblasts, and cell-free fetal DNA from the blood of pregnant women is currently the subject of intense research with the aim of developing new risk-free methods for prenatal diagnosis. An unexpected finding made during these studies was that the traffic of fetal erythroblasts into the maternal peripheral circulation was enhanced in pre-eclampsia. Independent prospective studies examining samples collected in the second trimester indicated that this perturbation in fetal cell trafficking occurs early in pregnancy, well before the onset of pre-eclampsia symptoms. The quantitative analysis of cell-free fetal and maternal DNA levels indicated that these concentrations were elevated in a co-ordinate manner in manifest pre-eclampsia, and that these elevations corresponded to disease severity. On the other hand, analysis of prospectively collected samples indicated that only cell-free fetal but not maternal DNA levels were elevated before onset of symptoms in pregnancies which subsequently developed pre-eclampsia. These data support hypotheses suggesting that pre-eclampsia is a multi-step disorder, initiated by a placental lesion that occurs early in pregnancy and which subsequently leads to a systemic maternal inflammatory response and associated endothelial cell damag

Decrease in lipid levels of syncytiotrophoblast micro-particles reduced their potential to inhibit endothelial cell proliferation

Background: Preeclampsia is characterized by damage to the maternal endothelium that has been suggested to be mediated in part by elevated shedding of inflammatory placental syncytiotrophoblast micro-particles (STBM) into the maternal circulation. Previously, we have shown that STBM, prepared by three different methods: mechanical dissection, in vitro placental explants culture and perfusion of placenta, can inhibit endothelial cell proliferation. Only mechanically prepared STBM induced apoptosis in the endothelial cells. Now, we have examined lipid levels in the three STBM preparations and their differential responses on endothelial cells. Methods: We examined the lipid levels in the three STBM preparations using thin layer chromatography. Furthermore, the effects of reduced lipid levels in the three STBM preparations using the pharmacological agent methyl-β-cyclodextrin were examined on endothelial cell proliferation and apoptosis. Results: Among the three STBM preparations, mechanical STBM contained highest levels of lipids. The reduction in lipid levels in mechanical STBM reduced their potential to inhibit human umbilical vein endothelial cells (HUVEC) proliferation and blocked their potential to induce apoptosis. No similar effect was observed following lipid reduction in the two other STBM preparations. Conclusions: As it has been suggested that mechanically derived STBM may more closely resemble placental micro-particles generated in preeclampsia, our data suggest that lipid content may play a role in the anti-endothelial defects present in this diseas

Role of placentally produced inflammatory and regulatory cytokines in pregnancy and the etiology of preeclampsia

Human pregnancy is a metabolic and immune challenge for the mother who has to accommodate in her womb a semi-allogeneic fetus whose energy needs increase tremendously with gestation. Recent compelling research has suggested that proper inflammatory changes and oxidative balance are a requisite for successful pregnancy. The placenta is an integral component of this inflammatory response as it actively produces a variety of cytokines and immunomodulatory hormones. In preeclampsia, a life-threatening disorder of pregnancy that is characterized by widespread damage and dysfunction of the maternal endothelium, placental oxidative stress and aberrant cytokine expression induces an exaggerated maternal systemic inflammatory response to pregnanc

Diagnosis and treatment of iron-deficiency anaemia during pregnancy and postpartum

Introduction: Iron-deficiency anaemia during pregnancy and postpartum occurs frequently and may lead to severe maternal and foetal complications. New treatment regimens include intravenous iron administration in particular clinical situations. The aim of the study was to determine optimal diagnostic and therapeutic approaches to iron-deficiency anaemia during pregnancy and postpartum. Methods: The evidence from data available from published studies and recommendations regarding diagnosis and treatment were reviewed. As conclusions, recommendations are given by an expert panel. Results: During pregnancy, oral iron therapy is given as first-line treatment. In cases with lack of efficacy, unwarranted side effects or very low haemoglobin values, intravenous iron treatment with iron carboxymaltose is a preferable alternative, although data regarding safety are limited. In the postpartum period, haemoglobin values less than 95g/L are treated ideally by intravenous carboxymaltose, leading to more rapid haemoglobin recovery. Conclusion: New intravenous iron preparations such as iron carboxymaltose have an excellent efficacy, side effect profile and advantages as compared to oral iron preparations for particular clinical indication

Correlation of perineal ultrasound and lateral chain urethrocystography in the anatomical evaluation of the bladder neck

Although perineal ultrasound has been widely used, no standard values have been published. In 52 women with urinary stress incontinence the following parameters were measured during resting and straining: a) with ultrasound: distance (Dy) between bladder neck and central line of the symphysis, distance (Dx) between bladder neck and lower border of the symphysis, and retrovesical angle β; b) with urethrocystography (UCG): distance H between the bladder neck and the SCIPP line, inclination angle α and retrovesical angle β. Dy and H correlated during resting (r=0.608; p<0.001) and straining (r=0.575; p<0.001). The distance Dy of 11mm corresponded to a bladder neck position on the SQIPP line. A rotational descent was detectable when Dx increased from 13mm (± 6.5) during resting to 16mm (±7.8) during straining (p=0.009). It was concluded that perineal ultrasound is valuable for the anatomical evaluation of the bladder neck. This is the first report to better define the position of the bladder based on ultrasonographic measurement

Is the quantity of circulatory cell-free DNA in human plasma and serum samples associated with gender, age and frequency of blood donations?

Circulatory cell-free DNA (cf-DNA) is increased in a variety of clinical pathologic conditions; therefore, these markers could be widely used as markers for detecting and monitoring several disorders. To better understand the biology of this molecule, we analysed the relationship between the level of circulatory cf-DNA and physiological parameters such as gender, age and frequency of blood donations. Paired plasma and serum samples were obtained from 87 blood donors and 50 healthy adults who had never donated blood. Cf-DNA was extracted from plasma and serum samples using the MagNA Pure LC Instrument. Quantity determination of circulatory cf-DNA was performed by TaqMan real-time PCR for the ubiquitous GAPDH gene. Our data showed that the concentration of cf-DNA in serum was about eightfold higher than that in plasma. Regarding the level of these circulatory species, no significant differences were observed between the age-matched men and women and gender-matched, different-age cohorts, except in women who were older than 60years of age. Frequent blood donations did not increase the circulatory species. Circulatory cf-DNA in plasma and serum samples is not correlated with human gender and human age except in women who are older than 60years of age. Frequent blood donation did not affect the quantity of circulatory cf-DNA. The explanation for the latter most likely is the short half-life time of free fetal DNA in maternal circulatio

Risk factors in prolonged postpartum urinary retention: an analysis of six cases

Purpose: Prolonged first and second stage of labor, isolated prolongation of the second stage, forceps delivery or vacuum extraction, perineal laceration, nulliparity and epidural anesthesia are known risk factors for developing prolonged postpartum urinary retention (PUR). The aim of our study was to analyze number and constellations of these risk factors, in prolonged postpartum urinary retention (PPUR) in our own unit to facilitate the identification of patients at high risk and thus to prevent bladder overdistension by early intervention. Methods: We performed a retrospective analysis of all our cases with PPUR between 2003 and 2008 including variables like age weight, height, body mass index, fetal birth weight and head circumference. Results: The incidence of PPUR at our institution is low being 0.06%. No woman combined all six risk factors. The majority had five risk factors, all had at least four. An isolated prolonged second stage of labor was common to all patients with PPUR. Five women had an epidural anesthesia, three were nulliparous and only two women delivered spontaneously. All but one woman suffered from perineal tears. Interestingly, fetal head circumference was larger than 36cm in four of six cases. Conclusion: In contrast to simple PUR, the prolonged form of PUR could be the result of a cumulative effect of different single risk factor

Macrophage migration inhibition factor is elevated in pregnancy, but not to a greater extent in preeclampsia

Background: Maternal serum concentrations of macrophage migration inhibitory factor (MIF) have recently been reported to be elevated in cases with preeclampsia. These findings may be important in increasing our understanding of the underlying events leading to the development of preeclampsia, as this cytokine is also expressed in the placenta, where it has been shown to possess immunemodulatory activities. For this reason we attempted to independently verify this report. Methods: Plasma levels of MIF were assessed by ELISA in plasma samples collected from normal healthy male and female blood donors (n=20 per group), as well as healthy normal pregnant women in all three trimesters of pregnancy (n=60). In addition, MIF levels were examined from cases with mild and severe preeclampsia (n=20 per study cohort) and matched normotensive pregnancies (n=20). Results: MIF levels were found to be elevated in pregnancy (median=10.1ng/ml) when compared to non-pregnant controls (median=1.7ng/ml). A moderate, but not significant, elevation was found to occur from the first to the third trimester of pregnancy. No significant difference was found to occur between the two preeclampsia study groups when compared to the normotensive control group. Conclusions: Our data suggest that circulatory MIF concentrations are elevated throughout pregnancy, but are not further increased in preeclampsi

Neutrophil NETs: a novel contributor to preeclampsia-associated placental hypoxia?

Recent studies have suggested that the innate immune system is involved in the pathogenesis of preeclampsia. Its pathogenesis involves neutrophil activation and increased levels of cell-free DNA in the maternal plasma. Activation of neutrophils has recently been shown to induce DNA containing neutrophil extracellular traps (NETs) which trap and kill bacteria. Massive NETs induction by the placentally derived factors (IL-8 and placental micro-debris) and their increased presence in preeclamptic placenta suggest that NETs might be involved in the pathogenesis of preeclampsia. Therefore, increased presence of NETs in preeclampsia may play a role in the deficient placental perfusion associated with this disorde