Maternal Circulating Levels of Activin A, Inhibin A, sFlt-1 and Endoglin at Parturition in Normal Pregnancy and Pre-Eclampsia

Background: Maternal circulating levels of anti-angiogenic factors such as soluble fms-like tyrosine kinase-1 (sFlt-1), endoglin (sEng) and placental proteins like activin A and inhibin A are increased before the onset of pre-eclampsia. There is evidence for oxidative stress in pre eclampsia. Recently it was shown that placental oxygen concentration is related to sFlt-1 and inhibin A. In addition it is reported that oxidative stress markers are increased in placental tissue delivered after labour. Therefore, the objective of this study is to investigate if these proteins are altered in maternal circulation of labouring preeclampsia and normal pregnancies. Methodology: To assess the effects of labour, samples were taken from 10 normal pregnant (NP) and 10 pre-eclamptic (PE) women pre-labour, full dilation, placental delivery and 24 h. To assess the effects of placental delivery, plasma samples were taken from 10NP and 10PE women undergoing elective Caesarean section, pre-delivery, placental delivery and 10 min, 60 min and 24 h post delivery. SFlt-1 and sEng and activin A and inhibin A were measured using commercial and in house ELISA’s respectively. Results: The levels of sFlt-1 and sEng were significantly higher in PE compared to NP women in both groups. In labour, sFlt- 1 levels increased significantly at full dilatation in PE women, before declining by 24 hr. However there was no significant rise in sEng levels in labour. Activin A and inhibin A levels declined rapidly with placental delivery in NP and PE pregnancies. There was a significant rise in activin A levels during labour in PE compared to pre labour, but inhibin levels did not increase. Conclusion: Labour in pre-eclamptic women increases the levels of sFlt-1 and activin A. This pilot data suggests that increase in the maternal levels of these factors in labour could predict and/or contribute to the maternal syndrome postpartum.Citation: Reddy, A. et al. (2009). 'Maternal circulating levels of activin A, inhibin A, sFlt-1 and endoglin at parturition in normal pregnancy and pre-eclampsia', PLoS ONE 4(2): e4453. [Available at http://www.plosone.org]. © 2009 Reddy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Measurement of refractive index by nanoparticle tracking analysis reveals heterogeneity in extracellular vesicles

Introduction: Optical techniques are routinely used to size and count extracellular vesicles (EV). For comparison of data from different methods and laboratories, suitable calibrators are essential. A suitable calibrator must have a refractive index (RI) as close to that of EV as possible but the RI of EV is currently unknown. To measure EV, RI requires accurate knowledge of size and light scattering. These are difficult to measure as most EVs cannot be resolved by light microscopy and their diameter is smaller than the wavelength of visible light. However, nanoparticle tracking analysis (NTA) provides both size and relative light scattering intensity (rLSI) values. We therefore sought to determine whether it was possible to use NTA to measure the RI of individual EVs. Methods: NTA was used to measure the rLSI and size of polystyrene and silica microspheres of known size and RI (1.470 and 1.633, respectively) and of EV isolated from a wide range of cells. We developed software, based on Mie scattering code, to calculate particle RI from the rLSI data. This modelled theoretical scattering intensities for polystyrene and silica microspheres of known size (100 and 200 nm) and RI. The model was verified using data from the polystyrene and silica microspheres. Size and rLSI data for each vesicle were processed by the software to generate RI values. Results: The following modal RI measurements were obtained: fresh urinary EV 1.374, lyophilised urinary EV 1.367, neuroblastoma EV 1.393, blood EV 1.398, EV from activated platelets 1.390, small placental EV 1.364–1.375 and 1.398–1.414 for large placental EV (>200 nm). Large placental EV had a significantly higher RI than small placental EV (p<0.0001). The spread of RI values was narrower for small EV than for the more heterogeneous large EV. Discussion: Using NTA and Mie scattering theory, we have demonstrated that it is possible to estimate the RI of sub-micron EV using NTA data. EV typically had a modal RI of 1.37–1.39, whereas values of >1.40 were observed for some large (>200 nm) microvesicles. Conclusion: This method for measuring EV RI will be useful for developing appropriate calibrators for EV measurement

Astronomy below the survey threshold in the SKA era

Astronomy at or below the 'survey threshold' has expanded significantly since the publication of the original 'Science with the Square Kilometer Array' in 1999 and its update in 2004. The techniques in this regime may be broadly (but far from exclusively) defined as 'confusion' or 'P(D)' analyses (analyses of one-point statistics), and 'stacking', accounting for the flux-density distribution of noise-limited images co-added at the positions of objects detected/isolated in a different waveband. Here we discuss the relevant issues, present some examples of recent analyses, and consider some of the consequences for the design and use of surveys with the SKA and its pathfinders

CO Luminosity Density at High-z (COLDz) Survey: A Sensitive, Large-area Blind Search for Low-J CO Emission from Cold Gas in the Early Universe with the Karl G. Jansky Very Large Array

We describe the CO Luminosity Density at High-z (COLDz) survey, the first spectral line deep field targeting CO(1–0) emission from galaxies at z = 1.95–2.85 and CO(2–1) at z = 4.91–6.70. The main goal of COLDz is to constrain the cosmic density of molecular gas at the peak epoch of cosmic star formation. By targeting both a wide (~51 arcmin2) and a deep (~9 arcmin^2) area, the survey is designed to robustly constrain the bright end and the characteristic luminosity of the CO(1–0) luminosity function. An extensive analysis of the reliability of our line candidates and new techniques provide detailed completeness and statistical corrections as necessary to determine the best constraints to date on the CO luminosity function. Our blind search for CO(1–0) uniformly selects starbursts and massive main-sequence galaxies based on their cold molecular gas masses. Our search also detects CO(2–1) line emission from optically dark, dusty star-forming galaxies at z > 5. We find a range of spatial sizes for the CO-traced gas reservoirs up to ~40 kpc, suggesting that spatially extended cold molecular gas reservoirs may be common in massive, gas-rich galaxies at z ~ 2. Through CO line stacking, we constrain the gas mass fraction in previously known typical star-forming galaxies at z = 2–3. The stacked CO detection suggests lower molecular gas mass fractions than expected for massive main-sequence galaxies by a factor of ~3–6. We find total CO line brightness at ~34 GHz of 0.45 ± 0.2 μK, which constrains future line intensity mapping and CMB experiments

Syncytiotrophoblast extracellular vesicles from pre-eclampsia placentas differentially affect platelet function

Pre-eclampsia (PE) complicates around 3% of all pregnancies and is one of the most common causes of maternal mortality worldwide. The pathophysiology of PE remains unclear however its underlying cause originates from the placenta and manifests as raised blood pressure, proteinuria, vascular or systemic inflammation and hypercoagulation in the mother. Women who develop PE are also at significantly higher risk of subsequently developing cardiovascular (CV) disease. In PE, the failing endoplasmic reticulum, oxidative and inflammatory stressed syncytiotrophoblast layer of the placenta sheds increased numbers of syncytiotrophoblast extracellular vesicles (STBEV) into the maternal circulation. Platelet reactivity, size and concentration are also known to be altered in some women who develop PE, although the underlying reasons for this have not been determined. In this study we show that STBEV from disease free placenta isolated ex vivo by dual placental perfusion associate rapidly with platelets. We provide evidence that STBEV isolated from normal placentas cause platelet activation and that this is increased with STBEV from PE pregnancies. Furthermore, treatment of platelets with aspirin, currently prescribed for women at high risk of PE to reduce platelet aggregation, also inhibits STBEV-induced reversible aggregation of washed platelets. Increased platelet reactivity as a result of exposure to PE placenta derived STBEVs correlates with increased thrombotic risk associated with PE. These observations establish a possible direct link between the clotting disturbances of PE and dysfunction of the placenta, as well as the known increased risk of thromboembolism associated with this condition

New mechanism for Notch signaling to endothelium at a distance by Delta-like 4 incorporation into exosomes.

Notch signaling is an evolutionary conserved pathway that is mediated by cell-cell contact. It is involved in a variety of developmental processes and has an essential role in vascular development and angiogenesis. Delta-like 4 (Dll4) is a Notch ligand that is up-regulated during angiogenesis. It is expressed in endothelial cells and regulates the differentiation between tip cells and stalk cells of neovasculature. Here, we present evidence that Dll4 is incorporated into endothelial exosomes. It can also be incorporated into the exosomes of tumor cells that overexpress Dll4. These exosomes can transfer the Dll4 protein to other endothelial cells and incorporate it into their cell membrane, which results in an inhibition of Notch signaling and a loss of Notch receptor. Transfer of Dll4 was also shown in vivo from tumor cells to host endothelium. Addition of Dll4 exosomes confers a tip cell phenotype on the endothelial cell, which results in a high Dll4/Notch-receptor ratio, low Notch signaling, and filopodia formation. This was further evidenced by increased branching in a tube-formation assay and in vivo. This reversal in phenotype appears to enhance vessel formation and is a new form of signaling for Notch ligands that expands their signaling potential beyond cell-cell contact

The Evolution of the Baryons Associated with Galaxies Averaged over Cosmic Time and Space

We combine the recent determination of the evolution of the cosmic density of molecular gas (H2) using deep, volumetric surveys, with previous estimates of the cosmic density of stellar mass, star formation rate and atomic gas (H i), to constrain the evolution of baryons associated with galaxies averaged over cosmic time and space. The cosmic H i and H2 densities are roughly equal at z ~ 1.5. The H2 density then decreases by a factor ${6}_{-2}^{+3}$ to today's value, whereas the H i density stays approximately constant. The stellar mass density is increasing continuously with time and surpasses that of the total gas density (H i and H2) at redshift z ~ 1.5. The growth in stellar mass cannot be accounted for by the decrease in cosmic H2 density, necessitating significant accretion of additional gas onto galaxies. With the new H2 constraints, we postulate and put observational constraints on a two-step gas accretion process: (i) a net infall of ionized gas from the intergalactic/circumgalactic medium to refuel the extended H i reservoirs, and (ii) a net inflow of H i and subsequent conversion to H2 in the galaxy centers. Both the infall and inflow rate densities have decreased by almost an order of magnitude since z ~ 2. Assuming that the current trends continue, the cosmic molecular gas density will further decrease by about a factor of two over the next 5 Gyr, the stellar mass will increase by approximately 10%, and cosmic star formation activity will decline steadily toward zero, as the gas infall and accretion shut down

ST2 and IL-33 in Pregnancy and Pre-Eclampsia

Normal pregnancy is associated with a mild systemic inflammatory response and an immune bias towards type 2 cytokine production, whereas pre-eclampsia is characterized by a more intense inflammatory response, associated with endothelial dysfunction and a type 1 cytokine dominance. Interleukin (IL)-33 is a newly described member of the IL-1 family, which binds its receptor ST2L to induce type 2 cytokines. A soluble variant of ST2 (sST2) acts as a decoy receptor to regulate the activity of IL-33. In this study circulating IL-33 and sST2 were measured in each trimester of normal pregnancy and in women with pre-eclampsia. While IL-33 did not change throughout normal pregnancy, or between non-pregnant, normal pregnant or pre-eclamptic women, sST2 was significantly altered. sST2 was increased in the third trimester of normal pregnancy (p<0.001) and was further increased in pre-eclampsia (p<0.001). This increase was seen prior to the onset of disease (p<0.01). Pre-eclampsia is a disease caused by placental derived factors, and we show that IL-33 and ST2 can be detected in lysates from both normal and pre-eclampsia placentas. ST2, but not IL-33, was identified on the syncytiotrophoblast layer, whereas IL-33 was expressed on perivascular tissue. In an in vitro placental perfusion model, sST2 was secreted by the placenta into the ‘maternal’ eluate, and placental explants treated with pro-inflammatory cytokines or subjected to hypoxia/reperfusion injury release more sST2, suggesting the origin of at least some of the increased amounts of circulating sST2 in pre-eclamptic women is the placenta. These results suggest that sST2 may play a significant role in pregnancies complicated by pre-eclampsia and increased sST2 could contribute to the type 1 bias seen in this disorder