Pre-eclampsia and intrauterine growth restriction (IUGR) are common complications of pregnancy with significant implications for mother and fetus. The exact cause is unknown, but current theories suggest that abnormal trophoblast invasion leads to placental under perfusion and hypoxia with the generation of reactive oxygen species (ROS). A result of this insult is increased trophoblast apoptosis. The cell cycle regulator p53 has been identified in increased amounts in the trophoblast in pre-eclampsia and IUGR. It is possible that modulation of this potent apoptotic signal could improve placental function, potentially offering a therapy for these presently untreatable conditions.In these studies, trophoblast apoptosis was induced with Nutlin-3, a synthetic compound which inhibits the binding of p53 to its inhibitor, Mdm2, therefore acting as a p53 activator. Treatment with Nutlin-3 induced apoptotic cell death in BeWo choriocarcinoma cells, primary trophoblast cells and placental villous explants, suggesting that p53 is intrinsic to trophoblast survival. This increase in apoptosis was associated with an increase in p53, Mdm2, p21 and Puma protein expression in BeWo cells and p21 and Puma mRNA in placental villous explants. No effect was observed in Bax expression. Co-treatment with Nutlin-3 and the p53 inhibitor Pifithrin-α reduced apoptosis to the level of control tissue in BeWo cells, primary trophoblast cells and villous explants, suggesting that reducing p53 activity improves trophoblast function.The use of hypoxia and ROS was associated with an increase in apoptosis and p21 and Puma in placental villous explants. Furthermore, the extent of apoptosis and p21 and Puma expression was reduced by co-treatment with Pifithrin-α, suggesting that the ameliorating effects of Pifithrin-α on trophoblast apoptosis are maintained following an insult from a less specific p53 activating mechanism, further supporting the continued investigation of p53 modulation as a therapeutic manipulation of placental function.Pregnant mice were exposed to Nutlin-3 to ascertain the effect of p53 over-activity on pregnancy outcome. The establishment of a model of placental dysfunction in the mouse would have allowed investigation into the effect of p53 inhibition with Pifithrin-α. Despite clear evidence of p53 modulation in maternal tissues, the decidua, no effect was observed upon pregnancy outcome or upon fetal or placental tissue. These findings suggest that the mouse placenta may not be a good model of p53 over-activity, although the exact mechanism of resistance remains unclear.In conclusion, p53 exaggeration is inducible in trophoblast with Nutlin-3, hypoxia and ROS. Furthermore, Pifithrin-α is able to reduce this effect, suggesting that the p53 pathway may be a realistic target for modulation in the placenta.

Thesis (Ph. D.)--Virginia Commonwealth University, 2007. / Prepared for: Dept. of Obstetrics and Gynecology. Bibliography: leaves 166-192. Also available online via the Internet.

Indiana University-Purdue University Indianapolis (IUPUI) Indiana University School of Dentistry / Background: Pre-eclampsia a potentially life threatening hypertensive disorder occurring in 3-14% of pregnancies. Its etiology is multifactorial involving the placenta. The only “cure” that currently exists is the delivery of the baby, which is often pre-term. There is no early pregnancy screening test to recognize those at risk. Recently, an altered immune-inflammatory responses at the placental level in response to infectious agents (eg., periodontal pathogens) have been proposed to be etiological for this pregnancy complication. A new class of Pattern Recognition Receptors called Peptidoglycan Recognition Proteins (PGRPs) constituting 4 distinct molecules PGRP 1-4 is emerging as a key player in modulating host responses to peptidoglycan and its breakdown products. A critical knowledge gap exists on the role of PGRPs in the innate immune responses that occur at the maternal-fetal interface in response to pathogens and their components that may be present in maternal circulation secondary to chronic infections. Aim: The aim of this pilot study is to investigate the expression PGRPs in the placenta of pre-eclamptic women. The overall goal is to better understand the association of periodontal disease and adverse pregnancy outcomes. Methods and Materials: This case control study consisted of subjects with: (1) normal term pregnancies (n=7) (2) pre-eclampsia (n=7). Preeclampsia was defined as hypertension (systolic blood pressure of ≥ 140 mm Hg or diastolic blood pressure of ≥ 90 mm Hg on at least 2 occasions, 4 hours to 1 week apart) and proteinuria (≥ 300 mg in a 24-hour urine collection or one dipstick measurement of ≥ 2+). A real time quantitative PCR array was used to analyze the relative mRNA expression of TLR2, TLR4, NOD1, NOD2, PGRP1, PGRP2, PGRP3, and PGRP4. Immunohistochemistry was performed to determine the cell type(s) expressing the PGRP proteins in the placental tissue. Summary statistics (mean, standard deviation, range, 95% confidence interval for the mean) were calculated for PGRP 1-4 expression for each group. Results and conclusions: The PCR data showed the expression of PGRPs 1, 3 and 4 in the placental samples. There was an up-regulation of PGRP-1 (1.4 fold) and down regulation of PGRP-3 (1.3 fold) and PGRP-4 (1.6 fold). TLR2, TLR4 and NOD2 mRNA were also elevated in the placental samples. Immunohistochemistry demonstrated positive staining for PGRPs 3 and 4 in the trophoblasts. The results from this novel research could lead to development of salivary and/or plasmatic biomarkers for early detection of PE and warrants further investigation.

Excessive release of syncytiotrophoblast vesicles (STBM) from the placenta into the maternal circulation may cause the inflammatory response, endothelial dysfunction and activation of the coagulation system characteristic of pre-eclampsia (PE). Consequently, other cell types including platelets, leukocytes, red blood cells (RBC) and endothelium may be activated to release cellular vesicles which exacerbate the disease. This thesis aimed to develop methodology for enumerating and phenotyping STBM and the other vesicle types to determine whether they could be used as biomarkers for PE. In vitro derived STBM and vesicles from the other cells of the vascular compartment were examined to select a suitable panel of antibodies to analyse these same vesicle types in plasma samples from non-pregnant (NonP), normal pregnant (NormP) and PE women. Our flow cytometer was shown to detect microvesicles ≥290nm, hence smaller nanovesicles and exosomes could not be detected by this method. Therefore, a novel technique for analysing both microvesicles and nanovesicles, Nanoparticle Tracking Analysis (NTA), was explored and was found to be able to detect vesicles as small as 70nm. The origins of the vesicles that change in pregnancy are not yet known. Flow cytometry and NTA were used in parallel to determine the size, number and phenotype of STBM and other cellular vesicles in NonP, NormP and PE women. Flow cytometry showed that majority of vesicles were derived from platelets, followed by RBC vesicles, leukocyte vesicles and STBM. NTA showed that the total number of vesicles in plasma was significantly elevated in NormP and late-onset PE women compared to NonP controls, and the vesicles were smaller in size. Similarly, flow cytometry showed differences in the composition of vesicles between pregnant and non-pregnant women, demonstrating that pregnancy affects vesicle release. However, no differences were found between NormP and PE women. This was probably due to the majority of samples studied being from late rather than early-onset PE. Thus, although this is the most comprehensive analysis of circulating vesicles in pregnancy to date, their use as biomarkers for PE remains an open question.

Dissertation for MMed (Obstetrics and Gynaecology) and FCOG Part II / Hypertensive disorders in pregnancy are common and their incidence appears to be on the increase. Preeclampsia is a multi-organ, heterogeneous disorder of pregnancy associated with significant maternal, fetal and neonatal morbidity and mortality. Because preeclampsia is a progressive disorder, invariably delivery remote from term is often necessary to halt disease progression to benefit the mother and fetus. Objectives:  To determine the maternal outcomes in women with severe preeclampsia  To determine fetal and neonatal outcomes of infants born preterm Methods This was a prospective, descriptive study performed in three academic hospitals affiliated to the University of the Witwatersrand in Johannesburg. Data was collected from women with severe preeclampsia, who delivered between gestational ages of 26 weeks and 33 weeks, with a minimum neonatal weight of 500g as determined by sonography. Results: In the sample of 92 patients enrolled, there were two maternal deaths as a result of severe preeclampsia. Eclampsia and HELLP syndrome were the most frequently observed maternal complications at 34% and 49% respectively. Caesarean section was the most frequent method used to expedite delivery in 84% of women. Of the 97 babies delivered, 20% were confirmed intra-uterine fetal deaths, 7% demised during the early neonatal period and a there was a 40% very low birth weight rate. Conclusion: Despite interventions to reduce maternal and neonatal morbidity and mortality in our setting, our outcomes are similar to those observed in other parts of the world.

by Wong Wah-Kwan, Herman. / Thesis (M.Phil.)--Chinese University of Hong Kong. / Includes bibliographical references (leaves 69-79). / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Pregnancy-induced hypertension --- p.2 / Chapter 1.1a --- Definition of pregnancy-induced hypertension --- p.2 / Chapter 1.2 --- A brief history of pregnancy-induced hypertension --- p.5 / Chapter 1.3 --- The epidemiology of pregnancy-induced hypertension --- p.6 / Chapter 1.4 --- Prediction of pregnancy-induced hypertension --- p.8 / Chapter 1.5 --- Erythrocyte sodium-lithium countertransport --- p.14 / Chapter 1.5.1 --- The history and characteristics of SLC --- p.14 / Chapter 1.5.2 --- SLC in essential hypertension --- p.19 / Chapter 1.5.3 --- SLC in pregnancy --- p.26 / Chapter 1.5.4 --- SLC in other diseases --- p.27 / Chapter 1.6 --- Methods used in the study of erythrocyte SLC --- p.32 / Chapter 1.7 --- Aims of the projects --- p.34 / Chapter CHAPTER 2 --- MATERIALS & METHODS --- p.35 / Chapter 2.1 --- Anthropometric measurements --- p.36 / Chapter 2.1.1 --- Blood pressure measurements --- p.36 / Chapter 2.1.2 --- Physical measurements --- p.36 / Chapter 2.1.3 --- Gestational age --- p.36 / Chapter 2.2 --- Materials --- p.37 / Chapter 2.3 --- Method for the measurement of erythrocyte SLC --- p.37 / Chapter CHAPTER 3 --- PRECISION & INTRA-INDIVIDUAL VARIATION OF ERYTHROCYTE SLC IN MALES & FEMALES --- p.42 / Chapter 3.1 --- Assessment of the precision of the methods --- p.43 / Chapter 3.2 --- Method of study --- p.45 / Chapter 3.3 --- Statistics --- p.46 / Results --- p.47 / Chapter CHAPTER 4 --- PREDICTION OF PREGNANCY-INDUCED HYPERTENSION: SLC ACTIVITIES IN SECOND & THIRD TRIMESTERS --- p.54 / Chapter 4.1 --- Method of Study --- p.55 / Chapter 4.2 --- Results --- p.55 / Chapter CHAPTER 5 --- DISCUSSION --- p.63 / REFERENCES --- p.69

Yim Ka Wing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 93-102). / Abstracts in English and Chinese. / Statement --- p.I / Abstract --- p.Ii / Acknowledgements --- p.Iv / Publications and awards --- p.Vii / General abbreviations --- p.Viii / Technical abbreviations --- p.Ix / Abbreviations of chemicals --- p.X / List of figures --- p.Xi / List of tables --- p.Xiii / Table of contents --- p.Xiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Pre-eclampsia --- p.3 / Chapter 2.1 --- Inadequate Uteroplacental Circulation --- p.5 / Chapter 2.2 --- Placenta Ischaemia --- p.7 / Chapter 2.3 --- Oxidative Stress --- p.7 / Chapter 2.4 --- Systemic Inflammatory Response --- p.9 / Chapter 2.5 --- The Continuum Theory of Clinical Syndromes --- p.12 / Chapter 2.6 --- Origin of Stimuli to Inflammatory Response --- p.13 / Chapter Chapter 3 --- Proteomic Analysis --- p.16 / Chapter 3.1 --- Methodology in Proteomic Research --- p.17 / Chapter 3.1.1 --- 2D-PAGE --- p.17 / Chapter 3.1.2 --- Mass Spectrometry --- p.18 / Chapter 3.1.2.1 --- Peptide Mass Fingerprinting --- p.18 / Chapter 3.1.2.2 --- Product-ion Data --- p.19 / Chapter 3.1.2.3 --- Matrix-assisted Laser Desorption/Ionization Tandem Time-of-Flight Mass Spectrometry (MALDI-TOF/TOF MS) --- p.19 / Chapter 3.1.3 --- Bioinformatics Tools --- p.20 / Chapter 3.2 --- Applications of Proteomic Analysis --- p.20 / Chapter Chapter 4 --- Materials and Methods --- p.21 / Chapter 4.1 --- Overview --- p.21 / Chapter 4.2 --- Patients --- p.23 / Chapter 4.2.1 --- Pre-eclampsia --- p.23 / Chapter 4.2.2 --- Recruitment --- p.23 / Chapter 4.3 --- Stage I: Comparative Proteomic Analysis --- p.24 / Chapter 4.3.1 --- Sample Processing --- p.24 / Chapter 4.3.2 --- Modified Bradford's Method --- p.24 / Chapter 4.3.3 --- Albumin Depletion --- p.25 / Chapter 4.3.4. --- Comparative Proteomic Analysis --- p.26 / Chapter 4.3.4.1 --- First Dimension: Isoelectric Focusing (IEF) --- p.28 / Chapter 4.3.4.2 --- Reduction and Alkylation --- p.29 / Chapter 4.3.4.3 --- Second Dimension: SDS-PAGE --- p.30 / Chapter 4.3.4.4 --- Gel Imaging --- p.31 / Chapter 4.3.5 --- 2D-PAGE Data Analysis --- p.32 / Chapter 4.3.5.1 --- Gaussian Spot --- p.32 / Chapter 4.3.5.2 --- Matchset --- p.33 / Chapter 4.3.5.3 --- Normalization --- p.33 / Chapter 4.3.5.4 --- Significance Analysis of Microarrays --- p.34 / Chapter 4.4 --- Stage II: Protein Identification --- p.35 / Chapter 4.4.1 --- Tryptic Peptide Fingerprinting --- p.37 / Chapter 4.4.1.1 --- Removal of Silver Ions --- p.37 / Chapter 4.4.1.2 --- Reduction and Alkylation --- p.38 / Chapter 4.4.1.3 --- In-gel Digestion --- p.38 / Chapter 4.4.1.4 --- Clean up of Peptides --- p.39 / Chapter 4.4.1.5 --- Mass Spectrometric Analysis --- p.39 / Chapter 4.4.2 --- Database search: Profound-Peptide Mapping --- p.40 / Chapter 4.4.3 --- Database search: Mascot-MS/MS Ion Search --- p.40 / Chapter 4.5 --- Stage III: Immunoassays --- p.41 / Chapter 4.6 --- Sample Size and Power Calculations --- p.42 / Chapter 4.7 --- Statistical Analysis --- p.42 / Chapter Chapter 5 --- Results --- p.43 / Chapter 5.1 --- Patients' Demographic Characteristics and Obstetric Outcomes for 2D-PAGE Analysis --- p.43 / Chapter 5.2 --- Plasma Protein Quantity --- p.45 / Chapter 5.3 --- 2D-PAGE --- p.47 / Chapter 5.4 --- SAM Analysis --- p.47 / Chapter 5.5 --- Spot # 5204 and Spot # 6210 --- p.50 / Chapter 5.6 --- Protein Identification --- p.54 / Chapter 5.6.1 --- Identification of Protein Spot # 5204 --- p.54 / Chapter 5.6.2 --- Identification of Protein Spot # 6210 --- p.61 / Chapter 5.7 --- Patients' Demographic Characteristics and Obstetric Outcomes for Immunoassays --- p.68 / Chapter 5.8 --- Immunoassays --- p.70 / Chapter Chapter 6 --- Discussion --- p.81 / Chapter 6.1 --- Role of ficolins and MBL in complement activation pathway --- p.82 / Chapter 6.2 --- Structure of ficolins and MBL --- p.83 / Chapter 6.2.1 --- H-ficolin --- p.85 / Chapter 6.2.2 --- L-ficolin --- p.86 / Chapter 6.3 --- Immune Response and Pre-eclampsia --- p.86 / Chapter 7 --- Conclusion --- p.89 / Appendix --- p.90 / References --- p.93

Pre-eclampsia is a major cause of maternal and perinatal morbidity and mortality worldwide. A predictor of pre-eclampsia would enable intervention, close surveillance and timely delivery, and thereby reduce the negative consequences of the disorder. The overall aim of this thesis was to study potential predictors of pre-eclampsia by biochemical and epidemiological methods. Angiopoietin-1 (Ang-1) and Angiopoietin-2 (Ang-2) are regulators of angiogenesis, which is important for placental development. In a prospective and longitudinal study of a low-risk population the Ang-1/Ang-2 ratio was evaluated. The Ang-1/Ang-2 ratio increased during pregnancy in all women but at gestational week 25 and 28 the ratios were significantly lower in women who later developed pre-eclampsia. The relevance of Histidine-rich glycoprotein (HRG), a protein with angiogenic properties, was furthermore evaluated. HRG levels decreased in all women, with significantly lower levels at gestational week 10, 25 and 28 in women who later developed pre-eclampsia. Thus both Ang-1/Ang-2 ratio and HRG may predict pre-eclampsia. To evaluate the predictive value of HRG in combination with uterine artery Doppler early in pregnancy a study was performed in a high-risk population. The results revealed that the combination was better able to predict preterm pre-eclampsia than each marker individually, with a sensitivity of 91% at a specificity of 62%.  A possible association between hyperemesis gravidarum and pre-eclampsia, as well as other placental dysfunctional disorders, was investigated. Hyperemesis gravidarum may be caused by high levels of human chorionic gonadotrophin (hCG) and increased levels of hCG in the second trimester is associated with later development of pre-eclampsia. A cohort of all pregnancies in the Swedish medical birth register between 1997 and 2009 was studied. After adjustment for confounding factors an association between hyperemesis gravidarum in the second trimester and preterm pre-eclampsia, placental abruption and infants born small for gestational age was demonstrated. In conclusion, the ratio of Ang-1/Ang-2 as well as HRG in plasma may be potential predictors of pre-eclampsia. Combination with uterine artery Doppler further increases the predictive value of HRG for preterm pre-eclampsia. Hyperemesis gravidarum in the second trimester may be considered as a clinical risk predictor of pre-eclampsia and other placental dysfunctional disorders.

Preeclampsia (PE) is a maternal disorder of pregnancy, characterized by late-onset hypertension and proteinuria. It affects roughly 5-7% of all pregnancies worldwide and is a leading cause of maternal and fetal/neonatal morbidity and mortality. Cigarette smoking in pregnancy is associated with a 33% reduction in the incidence of PE, and this is dose dependent. It is hypothesized that carbon monoxide (CO), a combustion product in cigarettes, may confer cytoprotective and regulatory properties leading to the decreased incidence of PE. CO is produced endogenously by the enzyme heme oxygenase (HO), and it is thought that the manipulation of the HO/CO system in pregnancy can ameliorate or reduce the pathophysiologic signs of PE. The exposure of pregnant mice to 250 ppm CO led to an increase in each of the maternal uterine blood flow, vascularity of the placenta and vessel diameter, with a shift towards angiogenesis in the placenta tissue proteins Exposure of human placental villous explants to 250ppm CO led to a decreased production and release of the soluble vascular endothelial growth factor (VEGF) receptor -1 (sFlt-1). This molecule is increased in maternal plasma and placenta tissue of women with PE and it binds with molecules of angiogenesis, limiting their ability to interact with the endothelium. Using an AdsFlt-1 PE-like mouse model, the exposure of mice to 250ppm chronic CO prevented the hypertension, proteinuria and glomerular alterations, supporting the use of CO as a future therapeutic for women with PE. We completed a pilot study to evaluate the exposure of healthy volunteers to two, one hour inhalations of 250ppm CO. We determined the half-life of CO and we provide baseline kinetics data for males and females following CO inhalation. These data are important for future therapeutic studies in order to better establish proper dosing, concentration of CO and method of delivery. The results of this thesis contribute to the understanding of the pathophysiology of PE and provide evidence to support the use of CO as a therapeutic for this disorder. / Thesis (Ph.D, Anatomy & Cell Biology) -- Queen's University, 2014-05-01 14:38:42.584

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.