142 research outputs found
Die plazentare Syndecan-1 Expression im 1. und frßhen 2. Trimenon: Ein prädiktiver Faktor fßr das kindliche Schwangerschaftsoutcome?
Syndecanâ1 ist ein Proteoglykan, das bei Zell-Kontakten, -proliferation, und -differenzierung mitwirkt. Der Nachweis von Syndecan in frĂźhen humanen Abort- und Mausplazenten legt eine Funktion in der embryonalen Entwicklung nahe. 203 Chorionzottenbiopsien wurden immunhistochemisch auf Syndecan-1 gefärbt. Die Ergebnisse der mikroskopischen Auswertung wurden mit den Daten des Kollektives aus der PIA-Fetal Database korreliert. Wir konnten die Mikrovilli als Ort der intensivsten Syndecanexpression identifizieren. Es zeigte sich ein hochsignifikanter Unterschied in der Syndecanfärbeintensität zwischen groĂen und kleinen Chorionzotten und zwischen dem Nikotinabusus der Mutter und der erniedrigten Syndecanexpression. Eine erniedrigte Expression ist auch assoziiert mit FrĂźhgeburtlichkeit, eine vermehrte Expression mit Lebendgeburten und normalem Karyotyp. Unsere Ergebnisse zeigen erstmalig, dass Syndecan-1 die Liste der 1. Trimester-Prädiktoren von Schwangerschaftskomplikationen erweitern kann
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Human platelet protein ubiquitylation and changes following GPVI activation
Platelet activators stimulate post-translational modification of signalling proteins to change their
activity or their molecular interactions leading to signal propagation. One covalent modification is attachment of the small protein ubiquitin to lysine residues in target proteins. Modification by ubiquitin can either target proteins for degradation by the proteasome or act as a scaffold for other proteins. Pharmacological inhibition of deubiquitylases or the proteasome inhibits platelet
activation by collagen, demonstrating a role for ubiquitylation, but relatively few substrates for
ubiquitin have been identified and the molecular basis of inhibition is not established. Here we report the ubiquitome of human platelets and changes in ubiquitylated proteins following stimulation by collagen related peptide (CRP-XL). Using platelets from six individuals over three
independent experiments, we identified 1634 ubiquitylated peptides derived from 691 proteins, revealing extensive ubiquitylation in resting platelets. 925 of these peptides show an increase of more than 2-fold following stimulation with CRP-XL. Multiple sites of ubiquitylation were 16 identified on a number of proteins including Syk, filamin and integrin heterodimer subunits. This work reveals extensive protein ubiquitylation during activation of human platelets and opens the possibility of novel therapeutic interventions targeting the ubiquitin machinery
Molecular Recognition of Glycan-Bearing Glycomacromolecules Presented at Membrane Surfaces by Lectins: An NMR View
Lectinâglycan interactions are at the heart of a multitude of biological events. Glycans are usually presented in a multivalent manner on the cell surface as part of the so-called glycocalyx, where they interact with other entities. This multivalent presentation allows us to overcome the typical low affinities found for individual glycanâlectin interactions. Indeed, the presentation of glycans may drastically impact their binding by lectins, highly affecting the corresponding binding affinity and even selectivity. In this context, we herein present the study of the interaction of a variety of homo- and heteromultivalent lactose-functionalized glycomacromolecules and their lipid conjugates with two human galectins. We have employed as ligands the glycomacromolecules, as well as liposomes decorated with those structures, to evaluate their interactions in a cell-mimicking environment. Key details of the interaction have been unravelled by NMR experiments, both from the ligand and receptor perspectives, complemented by cryo-electron microscopy methods and molecular dynamics simulations.M.H. and L.H. thank the DFG for support through the ViroCarb research consortium (HA5950/5-2) and the CeMSA@HHU (Center for Molecular and Structural Analytics @ Heinrich-Heine University) for recording the mass spectrometric and the NMR-spectroscopic data for the structural conformation of the glycomacromolecules and their lipid conjugates. The CIC bioGUNE EM platform is also thanked for infrastructural support during cryo-EM data collection. The group in Spain thank the European Research Council (RECGLYCANMR, Advanced grant no. 788143), MCIN/AEI/10.13039/501100011033 for grants PDI2021-1237810B-C21, PID2021-126130OB-I00, CEX2021-001136-S, and CIBERES, an initiative of Instituto de Salud Carlos III (ISCIII), Madrid, Spain, for generous funding
Colorectal cancer liver metastatic growth depends on PAD4-driven citrullination of the extracellular matrix
Citrullination of proteins, a post-translational conversion of arginine residues to citrulline, is recognized in rheumatoid arthritis, but largely undocumented in cancer. Here we show that citrullination of the extracellular matrix by cancer cell derived peptidylarginine deiminase 4 (PAD4) is essential for the growth of liver metastases from colorectal cancer (CRC). Using proteomics, we demonstrate that liver metastases exhibit higher levels of citrullination and PAD4 than unaffected liver, primary CRC or adjacent colonic mucosa. Functional significance for citrullination in metastatic growth is evident in murine models where inhibition of citrullination substantially reduces liver metastatic burden. Additionally, citrullination of a key matrix component collagen type I promotes greater adhesion and decreased migration of CRC cells along with increased expression of characteristic epithelial markers, suggesting a role for citrullination in promoting mesenchymal-to-epithelial transition and liver metastasis. Overall, our study reveals the potential for PAD4-dependant citrullination to drive the progression of CRC liver metastasis
Male reproductive aging arises via multifaceted mating-dependent sperm and seminal proteome declines, but is postponable in Drosophila
I.S. and S.W. were supported by a Biotechnology and Biological Sciences Research Council (BBSRC) Fellowship to S.W. (BB/K014544/1) and S.W. additionally by a Dresden Senior Fellowship. B.M.K., P.D.C., and R.F. were supported by the Kennedy Trust and John Fell Funds. R.D. was supported by Marie Curie Actions (Grant 655392). B.R.H. was funded by the EP Abraham Cephalosporin-Oxford Graduate Scholarship with additional support from the BBSRC Doctoral Training Programme. M.F.W. was supported by a NIH Grant R01HD038921. Work in the J.S. Laboratory was supported by NIH Grant R15HD080511.Declining ejaculate performance with male age is taxonomically widespread and has broad fitness consequences. Ejaculate success requires fully functional germline (sperm) and soma (seminal fluid) components. However, some aging theories predict that resources should be preferentially diverted to the germline at the expense of the soma, suggesting differential impacts of aging on sperm and seminal fluid and trade-offs between them or, more broadly, be-tween reproduction and lifespan. While harmful effects of male age on sperm are well known, we do not know how much seminal fluid deteriorates in comparison. Moreover, given the predicted trade-offs, it remains unclear whether systemic lifespan-extending inter-ventions could ameliorate the declining performance of the ejacu-late as a whole. Here, we address these problems using Drosophila melanogaster. We demonstrate that seminal fluid deterioration con-tributes to male reproductive decline via mating-dependent mech-anisms that include posttranslational modifications to seminal proteins and altered seminal proteome composition and transfer. Additionally, we find that sperm production declines chronologically with age, invariant to mating activity such that older multiply mated males become infertile principally via reduced sperm transfer and viability. Our data, therefore, support the idea that both germline and soma components of the ejaculate contribute to male reproduc-tive aging but reveal a mismatch in their aging patterns. Our data do not generally support the idea that the germline is prioritized over soma, at least, within the ejaculate. Moreover, we find that lifespan-extending systemic down-regulation of insulin signaling re-sults in improved late-life ejaculate performance, indicating simul-taneous amelioration of both somatic and reproductive aging.Publisher PDFPeer reviewe
Identification of a Phytase Gene in Barley (Hordeum vulgare L.)
Background: Endogenous phytase plays a crucial role in phytate degradation and is thus closely related to nutrient efficiency in barley products. The understanding of genetic information of phytase in barley can provide a useful tool for breeding new barley varieties with high phytase activity. Methodology/Principal Findings: Quantitative trait loci (QTL) analysis for phytase activity was conducted using a doubled haploid population. Phytase protein was purified and identified by the LC-ESI MS/MS Shotgun method. Purple acid phosphatase (PAP) gene was sequenced and the position was compared with the QTL controlling phytase activity. A major QTL for phytase activity was mapped to chromosome 5 H in barley. The gene controlling phytase activity in the region was named as mqPhy. The gene HvPAP a was mapped to the same position as mqPhy, supporting the colinearity between HvPAP a and mqPhy. Conclusions/Significance: It is the first report on QTLs for phytase activity and the results showed that HvPAP a, which shares a same position with the QTL, is a major phytase gene in barley grains
Myosin V regulates synaptopodin clustering and localization in the dendrites of hippocampal neurons
The spine apparatus (SA) is an endoplasmic reticulum-related
organelle that is present in a subset of dendritic spines in cortical
and pyramidal neurons, and plays an important role in Ca2+
homeostasis and dendritic spine plasticity. The protein
synaptopodin is essential for the formation of the SA and is widely
used as a maker for this organelle. However, it is still unclear which
factors contribute to its localization at selected synapses, and how it
triggers local SA formation. In this study, we characterized
development, localization and mobility of synaptopodin clusters in
hippocampal primary neurons, as well as the molecular dynamics
within these clusters. Interestingly, synaptopodin at the shaftassociated clusters is less dynamic than at spinous clusters. We
identify the actin-based motor proteins myosin V (herein referring to
both the myosin Va and Vb forms) and VI as novel interaction partners
of synaptopodin, and demonstrate that myosin V is important for the
formation and/or maintenance of the SA. We found no evidence of
active microtubule-based transport of synaptopodin. Instead, new
clusters emerge inside spines, which we interpret as the SA being
assembled on-site
Molecular Recognition of Glycan-Bearing Glycomacromolecules Presented at Membrane Surfaces by Lectins: An NMR View
Age influence on effectiveness of a novel 3-phytase in barley-wheat based diets for pigs from 12 to 108 kg under commercial conditions
[EN] The main objective of this study was to evaluate the influence of pig's age on the effectiveness of a new microbial 3-phytase, produced by Komagataella phaffii, under commercial conditions in barley-wheat based diets. Two experiments were conducted in weaned, growing and finishing pigs; firstly, to determine phytase efficacy on dry matter, organic matter, energy, protein and mineral (phosphorus, P and calcium, Ca) digestibility (n = 48; Experiment 1), and secondly, to evaluate the effect of phytase on growth performance and bone mineralization (n = 312; Experiment 2). In each experiment, three barley-wheat based diets were formulated following the recommendations for each animal age, of which two versions were manufactured, including 0 and 1000 phytase units (FTU)/kg of feed of the new 3-phytase to be tested. Results showed the new phytase had the potential to increase the digestibility of Ca and P (on av. + 0.05 and +0.06, respectively; P < 0.01), especially P digestibility in growing pigs (+0.10; P < 0.001), consequently decreasing P and Ca excretion. Digestible energy (DE) of the diet increased with the addition of phytase in weaned pigs (+0.69 MJ/kg of dry matter (DM); P < 0.001). Dietary inclusion of new 3-phytase enhanced average daily gain from 46 to 94 days of age (+0.07 kg/d; P < 0.05) and decreased feed conversion ratio from 46 to 154 days of age (on av. -0.13; P < 0.05), although no significant effect was observed from 154 to 185 days of age. Addition of the new 3-phytase also promoted bone mineralization, increasing the weight of the bones (+3.99 and +3.64 g of tibia at 95 days and metacarpus at 100 days of age, respectively; P < 0.05) and the ash, Ca and P content in these bones (e.g. + 0.46 and +0.33 g of P in tibia at 95 days and metacarpus at 100 days of age, respectively; P < 0.001). In conclusion, pig age affected the efficacy of a new 3-phytase on P and Ca digestibility both in weaned and growing diets and DE content of the weaned diets, which also resulted in improvements in growth, feed conversion and bone development until 154 days of age. These effects seem to be reduced during the finishing period, although the advantages of the new 3-phytase on bone mineralization were maintained until 185 days of age.We thank the technical staff at the experimental farms of the Research and Technology Animal Centre (CITA-IVIA), the Institute of
Animal Science and Technology (Universitat Politècnica de Valencia) and Javier Gómez (Crianzas Campovivo) for expert technical
assistance and experimental support.Cambra LĂłpez, M.; Cerisuelo, A.; Ferrer, P.; RĂłdenas MartĂnez, L.; AliguĂŠ, R.; Moset, V.; Pascual AmorĂłs, JJ. (2020). Age influence on effectiveness of a novel 3-phytase in barley-wheat
based diets for pigs from 12 to 108 kg under commercial conditions. Animal Feed Science and Technology. 267:1-13. https://doi.org/10.1016/j.anifeedsci.2020.114549S113267Adeola, O., & Cowieson, A. J. (2011). BOARD-INVITED REVIEW: Opportunities and challenges in using exogenous enzymes to improve nonruminant animal production. Journal of Animal Science, 89(10), 3189-3218. doi:10.2527/jas.2010-3715Almeida, F. N., Sulabo, R. C., & Stein, H. H. (2013). Effects of a novel bacterial phytase expressed in Aspergillus Oryzae on digestibility of calcium and phosphorus in diets fed to weanling or growing pigs. Journal of Animal Science and Biotechnology, 4(1). doi:10.1186/2049-1891-4-8Arredondo, M. A., Casas, G. A., & Stein, H. H. (2019). Increasing levels of microbial phytase increases the digestibility of energy and minerals in diets fed to pigs. Animal Feed Science and Technology, 248, 27-36. doi:10.1016/j.anifeedsci.2019.01.001Atakora, J. K. A., Moehn, S., Sands, J. S., & Ball, R. O. (2011). Effects of dietary crude protein and phytaseâxylanase supplementation of wheat grain based diets on energy metabolism and enteric methane in growing finishing pigs. Animal Feed Science and Technology, 166-167, 422-429. doi:10.1016/j.anifeedsci.2011.04.030Blaabjerg, K., Nørgaard, J. V., & Poulsen, H. D. (2012). Effect of microbial phytase on phosphorus digestibility in non-heat-treated and heat-treated wheatâbarley pig diets1. Journal of Animal Science, 90(suppl_4), 206-208. doi:10.2527/jas.53920Brady, S., Callan, J., Cowan, D., McGrane, M., & OâDoherty, J. (2002). Effect of phytase inclusion and calcium/phosphorus ratio on the performance and nutrient retention of grower-finisher pigs fed barley/wheat/soya bean meal-based diets. Journal of the Science of Food and Agriculture, 82(15), 1780-1790. doi:10.1002/jsfa.1262BranĚa, D. V., Ellis, M., CastanĚeda, E. O., Sands, J. S., & Baker, D. H. (2006). 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Bioavailability of Iron, Zinc, Phytate and Phytase Activity during Soaking and Germination of White Sorghum Varieties
The changes in phytate, phytase activity and in vitro bioavailability of iron and zinc during soaking and germination of three white sorghum varieties (Sorghum bicolor L. Moench), named Dorado, Shandweel-6, and Giza-15 were investigated. Sorghum varieties were soaked for 20 h and germinated for 72 h after soaking for 20 h to reduce phytate content and increase iron and zinc in vitro bioavailability. The results revealed that iron and zinc content was significantly reduced from 28.16 to 32.16% and 13.78 to 26.69% for soaking treatment and 38.43 to 39.18% and 21.80 to 31.27% for germination treatments, respectively. Phytate content was significantly reduced from 23.59 to 32.40% for soaking treatment and 24.92 to 35.27% for germination treatments, respectively. Phytase enzymes will be activated during drying in equal form in all varieties. The results proved that the main distinct point is the change of phytase activity as well as specific activity during different treatment which showed no significant differences between the varieties used. The in vitro bioavailability of iron and zinc were significantly improved as a result of soaking and germination treatments
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