Expression of plasma prekallikrein mRNA in human nonhepatic tissues and cell lineages suggests special local functions of the enzyme

At present it is generally accepted that plasma prekallikrein (PPK) is synthesized in the liver and secreted into the bloodstream. Surprisingly, it has recently been shown that PPK mRNA is present also in RNA from the kidney, adrenal gland and placenta. In spite of its novelty and possible important physiological implications this finding has been neglected. Here we report that PPK mRNA is expressed also in the human brain, heart, lung, trachea, endothelial cells and leukocytes as well as in a variety of fibroblast and epithelial cell lines. Expression of PPK mRNA in fibroblasts, endothelial cells and leukocytes suggests that PPK mRNA detected in RNA preparations from whole tissue may originate solely from these ubiquitously occurring cells. However, PPK mRNA expression in various epithelial cell lines demonstrates that tissue-specific cells also transcribe the PPK gene. The presence of PPK mRNA in nonhepatic tissues and cells indicates that they have the capacity to synthesize the protein. The physiological role of PPK synthesized in extrahepatic tissue is unknown. It may participate in local actions within tissues as well as contributing to the PPK pool in blood plasma. Cultured cells will provide a valuable model for exploring the physiological significance of extrahepatic PPK expression

Neutrino emission characteristics of black hole formation in three-dimensional simulations of stellar collapse

Neutrinos are unique probes of core-collapse supernova dynamics, especially in the case of black hole (BH) forming stellar collapses, where the electromagnetic emission may be faint or absent. By investigating two 3D hydrodynamical simulations of BH-forming stellar collapses of mass 40 and 75 M_sun, we identify the physical processes preceding BH formation through neutrinos, and forecast the neutrino signal expected in the existing IceCube and Super-Kamiokande detectors, as well as in the future generation DUNE facility. Prior to the abrupt termination of the neutrino signal corresponding to BH formation, both models develop episodes of strong and long-lasting activity by the spiral standing accretion shock instability (SASI). We find that the spiral SASI peak in the Fourier power spectrum of the neutrino event rate will be distinguishable at 3 sigma above the detector noise for distances up to O(30) kpc in the most optimistic scenario, with IceCube having the highest sensitivity. Interestingly, given the long duration of the spiral SASI episodes, the spectrograms of the expected neutrino event rate carry clear signs of the evolution of the blue spiral SASI frequency as a function of time, as the shock radius and post-shock fluid velocity evolve. Due to the high accretion luminosity and its large-amplitude SASI-induced modulations, any contribution from asymmetric (dipolar or quadrupolar) neutrino emission associated with the lepton emission self-sustained asymmetry (LESA) is far subdominant in the neutrino signal.Comment: 25 pages, including 19 figures. Discussion on LESA expanded; conclusions unchanged. Matches version accepted for publication in Phys. Rev. D. Animated visualizations available at: https://wwwmpa.mpa-garching.mpg.de/ccsnarchive/data/Walk2019

Stellar Collapse Diversity and the Diffuse Supernova Neutrino Background

The diffuse cosmic supernova neutrino background (DSNB) is observational target of the gadolinium-loaded Super-Kamiokande (SK) detector and the forthcoming JUNO and Hyper-Kamiokande detectors. Current predictions are hampered by our still incomplete understanding of the supernova (SN) explosion mechanism and of the neutron star (NS) equation of state and maximum mass. In our comprehensive study we revisit this problem on grounds of the landscapes of successful and failed SN explosions obtained by Sukhbold et al. and Ertl et al. with parametrized one-dimensional neutrino engines for large sets of single-star and helium-star progenitors, with the latter serving as proxy of binary evolution effects. Besides considering engines of different strengths, leading to different fractions of failed SNe with black-hole (BH) formation, we also vary the NS mass limit, the spectral shape of the neutrino emission, and include contributions from poorly understood alternative NS-formation channels such as accretion-induced or merger-induced collapse events. Since the neutrino signals of our large model sets are approximate, we calibrate the associated degrees of freedom by using state-of-the-art simulations of proto-neutron star cooling. Our predictions are higher than other recent ones because of a large fraction of failed SNe with long delay to BH formation. Our best-guess model predicts a DSNB electron-antineutrino-flux of 28.8^{+24.6}_{-10.9} cm^{-2}s^{-1} with 6.0^{+5.1}_{-2.1} cm^{-2}s^{-1} in the favorable measurement interval of [10,30] MeV, and 1.3^{+1.1}_{-0.4} cm^{-2}s^{-1} with electron-antineutrino energies > 17.3 MeV, which is roughly a factor of two below the current SK limit. The uncertainty range is dominated by the still insufficiently constrained cosmic rate of stellar core-collapse events.Comment: 43 pages, 16 figures, 10 tables; revised version with new Sections 6 & 9, and new Appendix E; accepted by Ap

Production of 44Ti and iron-group nuclei in the ejecta of 3D neutrino-driven supernovae

The radioactive isotopes of 44Ti and 56Ni are important products of explosive nucleosynthesis, which play a key role for supernova (SN) diagnostics and were detected in several nearby young SN remnants. However, most SN models based on non-rotating single stars predict yields of 44Ti that are much lower than the values inferred from observations. We present, for the first time, the nucleosynthesis yields from a self-consistent three-dimensional (3D) SN simulation of an approximately 19 Msun progenitor star that reaches an explosion energy comparable to that of SN 1987A and that covers the evolution of the neutrino-driven explosion until more than 7 seconds after core bounce. We find a significant enhancement of the Ti/Fe yield compared to recent spherically symmetric (1D) models and demonstrate that the long-time evolution is crucial to understand the efficient production of 44Ti due to the non-monotonic temperature and density histories of ejected mass elements. Additionally, we identify characteristic signatures of the nucleosynthesis in proton-rich ejecta, in particular high yields of 45Sc and 64Zn.Comment: 12 pages, 7 Figures, accepted for publication in ApJ

Analysis of Oversulfation in a Chondroitin Sulfate Oligosaccharide Fraction from Bovine Aorta by Nanoelectrospray Ionization Quadrupole Time-of-Flight and Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry

A combination of negative ion nano-electrospray ionization Fourier-transform ion cyclotron resonance and quadrupole time-of-flight mass spectrometry was applied to analysis of oversulfation in glycosaminoglycan oligosaccharides of the chondroitin sulfate type from bovine aorta. Taking advantage of the high-resolution and high mass accuracy provided by the FT-ICR instrument, a direct compositional assignment of all species present in the mixture can be obtained. An oligosaccharide fraction containing mainly hexasaccharides exhibited different levels of sulfation, indicated by the presence of species with regular sulfation pattern as well as oversulfated oligosaccharides with one additional sulfate group. Oversulfation can be directly identified from the high-resolution/high mass accuracy FT-ICR mass spectra according to their specific isotopic fine structure. Location of sulfate groups was analyzed by Q-TOF MS and low-energy CID MS/MS. Tetrasulfated hexasaccharides were analyzed by use of collision-induced dissociation at variable collision energy for an unambiguous assignment of the attachment site of the sulfate groups by minimizing unspecific neutral losses. Cleavage of glycosidic bonds gave rise to B- and C-type ions and their respective complementary Y- and Z-type fragment ions

Small proteoglycans of normal adult human kidney: Distinct expression patterns of decorin, biglycan, fibromodulin, and lumican

Small proteoglycans of normal adult human kidney: Distinct expression patterns of decorin, biglycan, fibromodulin, and lumican.BackgroundAmong the members of the small leucine-rich proteoglycan family, decorin, biglycan, and fibromodulin have been proposed to be potent modulators of transforming growth factor-β (TGF-β) activity, thereby playing an important role in the pathogenesis of fibrotic kidney diseases. Furthermore, decorin expression influences the expression of p21WAF1/CIP1, which has been related to kidney hypertrophy and hyperplasia. However, none of the members of this proteoglycan family have been investigated in normal adult human kidney cortex, thus making it impossible to correlate disease-mediated alterations of their expression with the normal situation in vivo.MethodsThe chondroitin/dermatan sulfate proteoglycans, decorin and biglycan, and the keratan sulfate proteoglycans, fibromodulin and lumican, were investigated in normal human adult renal cortex by immunohistochemistry on the light and electron microscopic level and by in situ hybridization. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) methods were used to get an estimate of their expression in isolated glomeruli. Decorin excretion with the urine was measured by Western blotting.ResultsTwo bands of decorin and a single band of biglycan mRNA were identified in Northern blots of isolated glomeruli. Amplification by RT-PCR was required to detect the signals for fibromodulin and lumican. All four proteoglycans were preferentially expressed in the renal interstitium with accumulations around tubules. Weak expression was found in the mesangial matrix. Biglycan was expressed by glomerular endothelial cells and, together with fibromodulin, was synthesized and deposited in distal tubular cells and collecting ducts. Immunogold labeling indicated the presence of the proteoglycans in the glomerular basement membrane, which was interpreted as a result of glomerular filtration. Indirect evidence suggested tubular reuptake of decorin after glomerular filtration.ConclusionThe data indicate that the different cells of the adult human kidney are characterized by a distinct expression pattern of the four small proteoglycans. It is suggested that these proteoglycans may have distinct pathophysiological roles depending upon whether they are expressed by mesangial cells, endothelial cells, epithelial cells, or cells of the tubulointerstitium

A Novel 110-kDa Receptor Protein is Involved in Endocytic Uptake of Decorin by Human Skin Fibroblasts

The small leucine-rich proteoglycan (SLRP) decorin is efficiently internalized by a variety of cultured cells. A 51-kDa protein has previously been described as a receptor mediating endocytosis of decorin and of the structurally related SLRP biglycan. Recent findings suggest that endocytosis of SLRPs may also be mediated by additional receptors. The class-A scavenger receptor, the endocytic mannose receptor, the epidermal growth factor receptor, and insulin-like growth factor-I receptor have emerged as candidates. We used a combined approach of immunoprecipitation and photoactivated cross-linking to identify endocytosis receptors for decorin in human skin fibroblasts. Decorin was purified by HPLC-DEAE-ion exchange chromatography from the secretions of human skin fibroblasts under nondenaturing conditions. Confocal immunofluorescence microscopy revealed that both biotinylated decorin and decorin conjugated to the heterobifunctional cross-linker sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1-3'-dithiopropionate (SASD) were endocytosed with equal efficiency. SASD-conjugated decorin was added to [35S]-methionine-labeled fibroblasts and cross-linked intracellularly to receptor molecules by photoactivation on endocytic uptake. Cross-linked decorin-receptor complexes were purified from the extracts of trypsin-treated fibroblasts by anion exchange chromatography and immunoprecipitation with a decorin-specific antiserum. Analysis by 2D electrophoresis and autoradiography revealed that decorin was specifically cross-linked to a protein of 110 kDa, which exhibited an isoelectric point of 5.5. In a second approach, unlabeled fibroblasts were subjected to decorin endocytosis and photoactivated cross-linking followed by Western blotting of DEAE-purified cell extracts. A shift of biotinylated decorin immunoreactivity from 165 kDa (decorin-receptor complex) to 54 kDa (SASD-conjugated biotinylated decorin) was noted on reductive cleavage of the cross-linker, representing a difference in molecular weight of approximately 110 kDa. The identification of a 110-kDa protein as a novel endocytosis receptor for decorin provides further support for the emerging concept of a redundancy of receptor molecules in the endocytosis of SLRP

The endothelial glycocalyx: composition, functions, and visualization

This review aims at presenting state-of-the-art knowledge on the composition and functions of the endothelial glycocalyx. The endothelial glycocalyx is a network of membrane-bound proteoglycans and glycoproteins, covering the endothelium luminally. Both endothelium- and plasma-derived soluble molecules integrate into this mesh. Over the past decade, insight has been gained into the role of the glycocalyx in vascular physiology and pathology, including mechanotransduction, hemostasis, signaling, and blood cell–vessel wall interactions. The contribution of the glycocalyx to diabetes, ischemia/reperfusion, and atherosclerosis is also reviewed. Experimental data from the micro- and macrocirculation alludes at a vasculoprotective role for the glycocalyx. Assessing this possible role of the endothelial glycocalyx requires reliable visualization of this delicate layer, which is a great challenge. An overview is given of the various ways in which the endothelial glycocalyx has been visualized up to now, including first data from two-photon microscopic imaging