Ekkehard Schleussner

Trophoblast invasion: the role of intracellular cytokine signalling via signal transducer and activator of transcription 3 (STAT3)

Trophoblast cells display a very unique capability: they physiologically invade into the surrounding tissue. This capability is widely associated with tumours, and, indeed, the invasive behaviour of both is rather similar. The imposing difference is that trophoblast cell invasion is temporally and locally controlled in contrast to unlimited tumour invasion. It initiates immediately after embryo implantation into the endometrium. Parallel to tumours, trophoblasts secrete proteases, such as matrix metalloproteinases, which dissolve the extracellular matrix and the surrounding tissue. Thereby, these proteases prepare and allow true invasion of trophoblasts. The invasive capacities of trophoblasts are positively and negatively regulated by numerous cytokines including leukaemia inhibitory factor (LIF), interleukin-6, hepatocyte growth factor, granulocyte macrophage-colony stimulating factor and others. They interact via specific receptors with the trophoblast cells, in which they activate intracellular signalling cascades. These will then induce expression of invasion relevant genes. One of these signalling pathways is the Janus kinase/signal transducers and activators of transcription (STAT) pathway. Especially phosphorylated STAT3 enhances invasiveness of tumours and trophoblast cells, where it is mainly activated by LIF. One of its most efficient physiological antagonists is suppressor of cytokine signalling 3. The balance of these two intracellular molecules seems to be a key regulator of tumour and trophoblast invasion.

Multicentre study of fetal cardiac time intervals using magnetocardiography

Objective A database with reference values of the durations of the various waveforms in a magnetocardiogram of fetuses in uncomplicated pregnancies is assessed. This database will be of help to discriminate between pathologic and healthy fetuses. A fetal magnetocardiogram is a recording of the magnetic field in a location near the maternal abdomen and reflects the electric activity within the fetal heart. It is a non‐invasive method, which can be used with nearly 100% reliability from the 20th week of gestation onward.

mTOR mediates human trophoblast invasion through regulation of matrix-remodeling enzymes and is associated with serine phosphorylation of STAT3

The intracellular signaling molecule mammalian target of rapamycin (mTOR) is essential for cell growth and proliferation. It is involved in mouse embryogenesis, murine trophoblast outgrowth and linked to tumor cell invasiveness. In order to assess the role of mTOR in human trophoblast invasion we analyzed the in vitro invasiveness of HTR-8/SVneo immortalized first-trimester trophoblast cells in conjunction with enzyme secretion upon mTOR inhibition and knockdown of mTOR protein expression. Additionally, we also tested the capability of mTOR to trigger signal transducer and activator of transcription (STAT)-3 by its phosphorylation status. Rapamycin inhibited mTOR kinase activity as demonstrated with a lower phosphorylation level of the mTOR substrate p70 S6 kinase (S6K). With the use of rapamycin and siRNA-mediated mTOR knockdown we could show that cell proliferation, invasion and secretion of matrix-metalloproteinases (MMP)-2 and -9, urokinase-like plasminogen activator (uPA) and its major physiological uPA inhibitor (PAI)-1 were inhibited. While tyrosine phosphorylation of STAT3 was unaffected by mTOR inhibition and knockdown, serine phosphorylation was diminished. We conclude that mTOR signaling is one major mechanism in a tightly regulated network of intracellular signal pathways including the JAK/STAT system to regulate invasion in human trophoblast cells by secretion of enzymes that remodel the extra-cellular matrix (ECM) such as MMP-2, -9, uPA and PAI-1. Dysregulation of mTOR may contribute to pregnancy-related pathologies caused through impaired trophoblast invasion.

Signal Analysis of Auditory Evoked Cortical Fields in Fetal Magnetoencephalography

Magnetoencephalography (MEG) using auditory evoked cortical fields (AEF) is an absolutely non-invasive method of passive measurement which utilizes magnetic fields caused by specific cortical activity. By applying the exceptionally sensitive SQUID technology to record these fields of dipolar configuration produced by the fetal brain, MEG as an investigational tool could provide new insights into the development of the human brain in utero. The major constraint to this application is a very low signal-to-noise ratio (SNR) that has to be attributed to a variety of factors including the magnetic signals generated by the fetal and maternal hearts which inevitably obscure a straightforward signal analysis. By applying a new algorithm of specific heart artefact reduction based on the relative regularity of the heart signals, we were able to increase the chance of extracting a fetal AEF from the raw data by the means of averaging techniques and principle component analysis. Results from 27 pregnant, healthy women (third trimester of their uncomplicated pregnancy) indicate an improved detection rate and the reproducibility of the fetal MEG. We evaluate and discuss a-priori criteria for signal analyses which will enable us to systematically analyze additional limiting factors, to further enhance the efficiency of this method and to promote the assessment of its possible clinical value in the future.

Changes in Brain Size during the Menstrual Cycle

Background There is increasing evidence for hormone-dependent modification of function and behavior during the menstrual cycle, but little is known about associated short-term structural alterations of the brain. Preliminary studies suggest that a hormone-dependent decline in brain volume occurs in postmenopausal, or women receiving antiestrogens, long term. Advances in serial MR-volumetry have allowed for the accurate detection of small volume changes of the brain. Recently, activity-induced short-term structural plasticity of the brain was demonstrated, challenging the view that the brain is as rigid as formerly believed. Methodology/Principal Findings We used MR-volumetry to investigate short-term brain volume changes across the menstrual cycle in women or a parallel 4 week period in men, respectively. We found a significant grey matter volume peak and CSF loss at the time of ovulation in females. This volume peak did not correlate with estradiol or progesterone hormone levels. Men did not show any significant brain volume alterations. Conclusions/Significance These data give evidence of short-term hormone-dependent structural brain changes during the menstrual cycle, which need to be correlated with functional states and have to be considered in structure-associated functional brain research.

Recommended standards for fetal magnetocardiography

Fetal magnetocardiography (FMCG) is increasingly being used in research and diagnostics of fetal heart function. Currently, FMCG is the only noninvasive procedure available, comparable to postnatal ECG, which can be used to assess cardiac electrophysiology during the second and third trimester of pregnancy. For a reliable evaluation and full clinical acceptance of this new technique, large numbers of patient investigations are required which can only be obtained in multicenter studies. An international standard protocol is needed to allow pooling of sufficient data and to permit the comparison of studies performed in different centers. This article provides recommended standards for FMCG in the fields of data acquisition and data analysis. (PACE 2003; 26:2121–2126)

The application of fetal magnetocardiography (FMCG) to investigate fetal arrhythmias and congenital heart defects (CHD)

Fetal magnetocardiography (FMCG), a new non‐invasive diagnostic tool in the analysis of the electrophysiological changes of the heart, was selectively applied in cases of fetal arrhythmias and congenital heart defect (CHD) to demonstrate its value for diagnosis and prenatal management.

Fetal heart rate variability reveals differential dynamics in the intrauterine development of the sympathetic and parasympathetic branches of the autonomic nervous system

The aim of this study was to investigate the hypothesis that fetal beat-to-beat heart rate variability (fHRV) displays the different time scales of sympatho-vagal development prior to and after 32 weeks of gestation (wks GA). Ninety-two magnetocardiograms of singletons with normal courses of pregnancy between 24 + 1 and 41 + 6 wks GA were studied. Heart rate patterns were either quiet/non-accelerative (fHRP I) or active/accelerative (fHRP II) and recording quality sufficient for fHRV. The sample was divided into the GA groups <32 wks GA/>32 wks GA. Linear parameters of fHRV were calculated: mean heart rate (mHR), SDNN and RMSSD of normal-to-normal interbeat intervals, power in the low (0.04-0.15 Hz) and high frequency range (0.15-0.4 Hz) and the ratios SDNN/RMSSD and LF/HF as markers for sympatho-vagal balance. fHRP I is characterized by decreasing SDNN/RMSSD, LF/HF and mHR. The decrease is more pronounced <32 wks GA. Beyond that GA SDNN/RMSSD is predominantly determined by RMSSD during fHRP I and by SDNN during fHRP II. In contrast to fHRP I, during fHRP II, mHR is positively correlated to SDNN/RMSSD instead of SDNN >32 wks GA. LF/HF increases in fHRP II during the first half of the third trimester. Non-accelerative fHRP are indicative of parasympathetic dominance >32 wks GA. In contrast, the sympathetic accentuation during accelerative fHRP is displayed in the interrelations between mHR, SDNN and SDNN/RMSSD. Prior to 32 wks GA, fHRV reveals the increasing activity of the respective branches of the autonomic nervous system differentiating the types of fHRP.

Microarray analysis of placental tissue in intrauterine growth restriction

Objective  Besides foetal or maternal disorders, placental dysfunction is a major cause of intrauterine growth restriction (IUGR). Although numerous macro‐ and histopathological changes have been described, little is known about the precise aetiology and the contribution of foetal/placental genes in this disorder.

Human fetal heart rate variability-characteristics of autonomic regulation in the third trimester of gestation.

Abstract Aim: To describe developmental aspects of the sympatho-vagal heart rate regulation in the human fetus by applying numerics to visual descriptions of fetal heart rate patterns throughout the third trimester of pregnancy. The focus is to determine potential benefits of this alternative means of assessing the maturation of the fetal autonomic nervous system by analysis of fetal heart rate variability (fHRV). Subjects and methods: The magnetocardiograms of 103 normal fetuses between 24+1 and 41+6 weeks of gestation were studied. Fetal heart beat intervals were determined with a temporal precision of 1 ms. The levels of fetal activity were estimated according to characteristic heart rate patterns (I–III) prior to 32, between 32–35 and beyond 35 (groups 1–3) completed weeks. Mean heart rate (mHR), standard deviation of normal-to-normal beat intervals (SDNN) and root mean square of successive differences of normal beats (RMSSD) served as fHRV indices, mean permutation entropy (PE_Mean) as complexity measure. SDNN/RMSSD was introduced as a potential marker for sympatho-vagal balance. Results: Low activity fHRP (I) were characterized by significantly lower level fHRV indices and higher PE_Mean when compared to fHRP II. We found that SDNN/RMSSD decreases with gestation in fHRP I, which suggests increasing vagal influence. In fHRP III (assigned to active awake fetuses only after 32 weeks), highest level SDNN and mHR are associated with a dramatically reduced complexity. fHRV indices cluster characteristically with the activity levels. Conclusions: We conclude that a combined analysis of fHRV, based on SDNN/RMSSD and PE_Mean, and fHRP is advantageous in the assessment of maturation of the fetal autonomic nervous system.