Wolfgang Holzgreve

Oral misoprostol vs. intravenous oxytocin in reducing blood loss after emergency cesarean delivery

Objective: To compare the effectiveness of oral misoprostol and intravenous oxytocin in reducing blood loss in women undergoing indicated or elective cesarean delivery (CD) under spinal anesthesia.

MALDI-TOF Mass Array Analysis of RASSF1A and SERPINB5 Methylation Patterns in Human Placenta and Plasma

Differences in DNA methylation patterns between placenta and blood cells of pregnant women have been suggested as potential biomarkers for noninvasive prenatal diagnostic strategies, including for common obstetrical complications, such as preeclampsia. New findings in epigenetic origins of fetal or placental disorders may improve our ability for optimal management of these conditions. Using a novel high-throughput mass spectrometry on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass array, we compared the quantitative methylation changes of RASSF1 and SERPINB5 (also known as MASPIN) genes in placenta and plasma samples. We analyzed the methylation status of a total of 3569 CpG dinucleotides on these two genes in 83 different samples: 50 plasma samples (20 from pregnant women and 30 from nonpregnant women) and 33 placenta tissue samples (25 from normal pregnancies and eight from preeclamptic pregnancies). The aim of this study was to assess the utility of epigenetic changes as biomarkers for noninvasive prenatal diagnostic procedures. Using a two-way hierarchical cluster analysis, significantly different methylation levels of the RASSF1 gene were found between placenta (normal and preeclamptic) and plasma samples of pregnant women. Although the SERPINB5 gene was hypomethylated in placenta DNA more than in plasma DNA, it did not demonstrate significant differences between studied groups. The MALDI-TOF mass spectrometry analysis of placenta and plasma DNA methylation patterns may serve as a tool for the study of gender-independent biomarkers in noninvasive prenatal diagnosis.

Circulating cell-free DNA as a potential biomarker for minimal and mild endometriosis

It has recently been reported that high concentrations of circulating cell-free (ccf) nucleic acids in plasma and serum could be used as biomarkers for non-invasive monitoring a wide variety of malignant and benign proliferations and inflammatory conditions. Endometriosis is one of the most common benign gynaecological proliferations with inflammatory activation in premenopausal women. Real-time multiplex polymerase chain reaction was used for synchronized quantification of the glyceraldehyde-3-phosphate dehydrogenase gene sequence in nuclear DNA (nDNA) and the ATP synthase-8 gene sequence in mitochondrial DNA (mtDNA). DNA was extracted from 500 microl serum and plasma of 19 cases with endometriosis to measure the total amount of ccf nDNA and ccf mtDNA. The concentration of ccf nDNA in plasma was significantly higher in the endometriosis group than in the control group (P = 0.046). The cut-off value selected by a receiver operating characteristic curve could provide a sensitivity of 70% and a specificity of 87% to discriminate between the minimal or mild cases and normal controls. The finding of significantly increased concentrations of ccf nDNA in plasma of patients with endometriosis suggests that ccf nDNA might be a potential biomarker for developing non-invasive diagnostic test in endometriosis.

Current Understanding of Mitochondrial DNA in Breast Cancer

Abstract:  The recent surge in mitochondrial research has been driven by the identification of mitochondria‐associated diseases and the role of mitochondria in apoptosis and aging. Mitochondrial DNA (mtDNA) has been proposed to be involved in carcinogenesis because of its high susceptibility to mutations and limited repair mechanisms in comparison to nuclear DNA. As mtDNA lacks introns, it has been suggested that most mutations will occur in coding sequences. The subsequent accumulation of mutations may lead to tumor formation. By virtue of their clonal nature, high copy number and high frequent mutations may provide a powerful molecular biomarker for the detection of cancer. It has been suggested that the extent of mtDNA mutations might be useful in the prognosis of cancer outcome and/or the response to certain therapies. In this review article, we aim to provide a brief summary of our current understanding of mitochondrial genetics and biology, review the mtDNA alterations reported in breast cancer, and offer some perspectives as to the emergence of mtDNA mutations, including their functional consequences in cancer development, diagnostic criteria, and therapeutic implications.

Simultaneous quantitative assessment of circulating cell-free mitochondrial and nuclear DNA by multiplex real-time PCR

Quantification of circulating nucleic acids in plasma and serum could be used as a non-invasive diagnostic tool for monitoring a wide variety of diseases and conditions. We describe here a rapid, simple and accurate multiplex real-time PCR method for direct synchronized analysis of circulating cell-free (ccf) mitochondrial (mtDNA) and nuclear (nDNA) DNA in plasma and serum samples. The method is based on one-step multiplex real-time PCR using a FAM-labeled MGB probe and primers to amplify the mtDNA sequence of the ATP 8 gene, and a VIC-labeled MGB probe and primers to amplify the nDNA sequence of the glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) gene, in plasma and serum samples simultaneously. The efficiencies of the multiplex assays were measured in serial dilutions. Based on the simulation of the PCR reaction kinetics, the relative quantities of ccf mtDNA were calculated using a very simple equation. Using our optimised real-time PCR conditions, close to 100% efficiency was obtained from the two assays. The two assays performed in the dilution series showed very good and reproducible correlation to each other. This optimised multiplex real-time PCR protocol can be widely used for synchronized quantification of mtDNA and nDNA in different samples, with a very high rate of efficiency.

Implications of Feto-maternal Cell Transfer in Normal Pregnancy

Traditionally, the placenta has been thought to be a well-built barrier that separates the genetically different mother and offspring. This has been studied and discussed by K.E. von Baer already in 1828 when he found separate vascular beds in dogs.1 In the following years, it has been suggested that there is communication between the placenta and the host mother, because syncytiotrophoblasts have been found in the lungs of women who died of eclampsia.2 Transplacental fetal “bleeding” was recognized only in cases with a significant placental trauma, e.g., car accident, amniocentesis, chorionic villous sampling, termination of pregnancy, and external version done for breech presentation.

Experiences with In Utero Transplantation of Mesenchymal Stem Cells

In utero stem cell transplantation (IUT) has become a valuable therapeutic option in fetuses with congenital immunologic disorders, such as severe combined immunodeficiency (SCID) or bare lymphocyte syndrome [1, 2]. However, other diseases such as thalassemias, storage defects, or osteogenesis imperfecta have either resulted in no detectable engraftment or microchimerism with uncertain effect on the phenotype. Although IUT was performed as early as the end of first trimester, neither bone marrow nor fetal liver cells resulted in relevant engraftment. It can be postulated that the fetal immune system deletes the allogeneic stem cells since several studies suggest that the fetal thymus is colonized in the first third of gestation [3]. A fetal T-cell-mediated alloresponse is evident as early as the second trimester and has cleared most allogeneic cells by term [4, 5]. In principle, IUT could result in long-term chimerism when performed early enough in pregnancy since, for instance, persistent blood group chimerism has been demonstrated for dizygotic twins [6]. But obviously, also the “transmaternal” traffic of cells from a first born to the next infant in a later pregnancy leads to tolerance induction within the T-cell population [7]. The early presentation of allogeneic cells to the developing fetal thymus results in specific tolerance, whereas later appearance (for instance, due to IUT of allogeneic hematopoietic stem cells at embryonic day 14 post conception/E14) leads to clearance of allogeneic cells from the circulation within months by the recipient’s immune system [8].

Misoprostol for prevention of postpartum hemorrhage: a randomized controlled trial

Women who showed a major collagen reduction (Group A) were treated by Ca+Vit D, while the other (Group B) received oral HRT. Both therapies have been administered once a day, for one year. After six months there was a cross-over between the two different therapies. Results: The collagen thickness has been evaluated at the sixth month and at the end of the study. In both groups there was an increase of collagen thickness. All the oral HRT treated subjects showed during the six months a major increase than the other group, but not very significant. Conclusion: The follow-up, we performed 60 days after therapy discontinuity demonstrated that the effects of both therapies were persistent.