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1.
Ektoras Hadjipanayi Peer-Hendrik Kuhn Philipp Moog Anna-Theresa Bauer Haydar Kuekrek Lilit Mirzoyan Anja Hummel Katharina Kirchhoff Burak Salgin Sarah Isenburg Ulf Dornseifer Milomir Ninkovic Hans-Günther Machens Arndt F. Schilling 《PloS one》2015,10(8)
Conceptually, premature initiation of post-wound angiogenesis could interfere with hemostasis, as it relies on fibrinolysis. The mechanisms facilitating orchestration of these events remain poorly understood, however, likely due to limitations in discerning the individual contribution of cells and extracellular matrix. Here, we designed an in vitro Hemostatic-Components-Model (HCM) to investigate the role of the fibrin matrix as protein factor-carrier, independent of its cell-scaffold function. After characterizing the proteomic profile of HCM-harvested matrix releasates, we demonstrate that the key pro-/anti-angiogenic factors, VEGF and PF4, are differentially bound by the matrix. Changing matrix fibrin mass consequently alters the balance of releasate factor concentrations, with differential effects on basic endothelial cell (EC) behaviors. While increasing mass, and releasate VEGF levels, promoted EC chemotactic migration, it progressively inhibited tube formation, a response that was dependent on PF4. These results indicate that the clot’s matrix component initially serves as biochemical anti-angiogenic barrier, suggesting that post-hemostatic angiogenesis follows fibrinolysis-mediated angiogenic disinhibition. Beyond their significance towards understanding the spatiotemporal regulation of wound healing, our findings could inform the study of other pathophysiological processes in which coagulation and angiogenesis are prominent features, such as cardiovascular and malignant disease. 相似文献
2.
Tim Schwarting Philipp Lechler Johannes Struewer Marius Ambrock Thomas Manfred Frangen Steffen Ruchholtz Ewgeni Ziring Michael Frink 《PloS one》2015,10(2)
IntroductionSuccessful graft ingrowth following reconstruction of the anterior cruciate ligament is governed by complex biological processes at the tendon-bone interface. The aim of this study was to investigate in an in vitro study the effects of bone morphogenetic protein 7 (BMP-7) on tendon-bone integration.ResultsIn both models, positive effects of BMP-7 on ALP enzyme activity were observed (p<0.001). Additionally, similar results were noted for LDH activity and lactate concentration. BMP-7 stimulation led to a significant increase in OCN expression. Whereas the effects of BMP-7 on tendon monoculture peaked during an early phase of the experiment (p<0.001), the cocultures showed a maximal increase during the later stages (p<0.001). The histological analysis showed a stimulating effect of BMP-7 on extracellular matrix formation. Organized ossification zones and calcium carbonate-like structures were only observed in the BMP-stimulated cell cultures.DiscussionThis study showed the positive effects of BMP-7 on the biological process of tendon-bone integration in vitro. Histological signs of improved mineralization were paralleled by increased rates of osteoblast-specific protein levels in primary bovine osteoblasts and fibroblasts.ConclusionOur findings indicated a role for BMP-7 as an adjuvant therapeutic agent in the treatment of ligamentous injuries, and they emphasized the importance of the transdifferentiation process of tendinous fibroblasts at the tendon-bone interface. 相似文献
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To elucidate how atmospheric CO2 enrichment, enhanced nutrient supply and soil quality interact to affect regrowth of temperate forests, young Fagus sylvatica and Picea abies trees were grown together in large model ecosystems. Identical communities were established on a nutrient-poor acidic and on a more fertile calcareous soil and tree growth, leaf area index, fine root density and soil respiration monitored over four complete growing seasons. Biomass responses to CO2 enrichment and enhanced N supply at the end of the experiment reflected compound interest effects of growth stimulation during the first two to three seasons rather than persistent stimulation over the whole duration of the experiment. Whereas biomass of Picea was enhanced in elevated CO2 on both soils, Fagus responded negatively to CO2 on acidic but positively on calcareous soil. Biomass of both species profited from enhanced N supply on the poor acidic soil only. Leaf area index on both soils was greater in high N supply as a consequence of a stimulation early in the experiment, but was unaffected by CO2 enrichment. Fine root density on acidic soil was increased in high N supply, but this did not stimulate soil respiration rate. In contrast, elevated CO2 stimulated both fine root density and soil CO2 efflux on calcareous soil, especially towards the end of the experiment. Our experiment suggests that future species dominance in beech-spruce forests is likely to change in response to CO2 enrichment, but this response is subject to complex interactions with environmental factors other than CO2, particularly soil type. 相似文献
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Extracellular Enzymes and Nutritional Physiology of Ampelomyces quisqualis Ces., Hyperparasite of Powdery Mildew, in vitro 总被引:1,自引:0,他引:1
W.-D. Philipp 《Journal of Phytopathology》1985,114(3):274-283
In vitro, the powdery’ mildew hyperparasite Ampelomyces quisqualis produced constitutively two groups of extracellular enzymes; β-glucosidase, β-N-acetylglucosaminidase and acid phosphatase possessing unusual high molecular weights (M, about 340 000), and ribonuclease, β-l→3-glucanase and α-l→4-glucanase representing smaller molecules (M, about 15 000–55 000). As can be concluded from adsorption to Concanavalin A-Sepharose 4 B, these enzymes are of glycoprotein nature Furthermore, traces of phospholipase could be detected in all of the isolates tested, whereas only phosphorylated carbohydrate components and nucleotides including cAMP as nutrients. The results of these studies suggest that in the biotrophic phase of parasitization A. qwsqualis interferes with energy metabolism, protein synthesis, cell wall synthesis and, possibly, regulation of the host, whereas proteins and membranes remain nearly unaffected. Thus, depletion of the energy reserves of the powdery mildew may be mainly responsible for the degeneration of host ultrastructure which represents the beginning of the necrotrophic phase of parasitization. 相似文献
7.
R Philipp J O McIntyre B H Robinson H Huth W Trommer S Fleischer 《Biochimica et biophysica acta》1984,790(3):251-258
Calcium efflux and EGTA-induced calcium release from an internal platelet membrane fraction have been studied after the oxalate-supported calcium uptake had reached steady state. Increasing external calcium concentrations stimulate the calcium efflux velocity, with an apparent half-maximal stimulation at about 5 microM outside calcium concentration and a maximal velocity of calcium efflux of 4.66 +/- 2.32 nmol X min-1 X mg-1. Moreover, the ratio of the liberated calcium on the loaded calcium seems to be independent of the increasing external calcium concentration. Increasing the calculated internal calcium concentration by varying the oxalate potassium concentration from 10 mM to 1 mM results in an increase of the liberated calcium from the membrane vesicles from 7.4% to 63%, respectively, without changing the calcium efflux velocity. Similar conclusions can be drawn from the observation of results from the calcium efflux and EGTA-induced calcium release methods. Moreover, calcium pump reversal does not seem to be responsible for the calcium efflux or calcium release. All these different points added to the previously described regulation of calcium efflux by the catalytic subunit of cAMP protein kinase suggest us that the mechanism of calcium liberation by the platelet membranes is different from the calcium uptake. 相似文献
8.
A generalized mathematical model, previously developed and experimentally validated, was modified and used to computer-simulate two dialysate-feed systems for operating a dialysis continuous process for the ammonium lactate fermentation. The simulations predicted that the feeding of substrate into the dialysate circuit and thence into the fermentor circuit via dialysis should greatly improve the production of cell mass and metabolite product. Experiments were conducted to test the system in which the fermentor is operated without an effluent, thus immobilizing the cells. Dried cheese whey ultrafiltrate was rehydrated to contain a normal concentration of lactose (62 mg/ml), supplemented with yeast with an adapted culture of Lactobacillus bulgaricus. The system was operated without interruption for 26 days. Results during steady-state conditions showed that the system is a new and useful way to immobilize living cells for the purpose of producing a metabolite at a high rate for a prolonged time. The substrate consumed by the cells is converted to product via maintenance metabolism only and is sterilized by dialysis. 相似文献
9.
Johana A. Luna Coronell Khulan Sergelen Philipp Hofer István Gyurján Stefanie Brezina Peter Hettegger Gernot Leeb Karl Mach Andrea Gsur Andreas Weinhäusel 《基因组蛋白质组与生物信息学报(英文版)》2018,16(1):73-84
Characterization of the colon cancer immunome and its autoantibody signature from differentially-reactive antigens (DIRAGs) could provide insights into aberrant cellular mechanisms or enriched networks associated with diseases. The purpose of this study was to characterize the antibody profile of plasma samples from 32 colorectal cancer (CRC) patients and 32 controls using proteins isolated from 15,417 human cDNA expression clones on microarrays. 671 unique DIRAGs were identified and 632 were more highly reactive in CRC samples. Bioinformatics analyses reveal that compared to control samples, the immunoproteomic IgG profiling of CRC samples is mainly associated with cell death, survival, and proliferation pathways, especially proteins involved in EIF2 and mTOR signaling. Ribosomal proteins (e.g., RPL7, RPL22, and RPL27A) and CRC-related genes such as APC, AXIN1, E2F4, MSH2, PMS2, and TP53 were highly enriched. In addition, differential pathways were observed between the CRC and control samples. Furthermore, 103 DIRAGs were reported in the SEREX antigen database, demonstrating our ability to identify known and new reactive antigens. We also found an overlap of 7 antigens with 48 “CRC genes.” These data indicate that immunomics profiling on protein microarrays is able to reveal the complexity of immune responses in cancerous diseases and faithfully reflects the underlying pathology. 相似文献
10.
Summary The natural radionuclide content of mineral phosphate fertilizers has been determined gammaspectrometrically. The investigations comprised ca. 70% of the mineral phosphate fertilizers authorized and used in the Federal Republic of Germany (FRG). At maximum, we found specific activities of 62 nCi Unat/kg, 23 nCi226Ra/kg, 1.6 nCi Thnat/kg and 262 nCi40K/kg. The mean values, weighted by the percentual agricultural consumption of the main phosphate fertilizer groups in 1973/74 and related to the phosphate content, amounted to 58, 40, 2, and 584 nCi/kg P2O5 for Unat,226Ra, Thnat, and40K respectively. This resulted in an annual distribution due to phosphate fertilizing of about 3.9 µCi Unat, 2.7 µCi226Ra, 0.1 µCi Thnat, and 39.9 µCi40K per ha of arable or pasture land in 1973/74 on the average. From these values the air dose rates over agricultural areas have been estimated under extreme conservative assumptions resulting in an additional external exposure of members of the population of 0.02 mrd/a on the average and 0.4 mrd/a in the region of highest phosphate fertilizing intensity. If it is assumed that radium contained in phosphate fertilizers were completely accumulated in the soils during the last 80 years, this value would be raised to 0.3 mrd/a on the average. The occupational external radiation exposure due to natural radionuclides contained in phosphate fertilizers was estimated to be 0.1 mrd/a on the average and 2.3 mrd/a at maximum for persons working in agriculture. These estimates show that natural radionuclides in phosphate fertilizers contribute but very little to the mean terrestrial radiation exposure of the population which is 50 to 55 mrd/a in Germany. Only for the small group of persons working in fertilizer production plants or storehouses a significant increase of the external radiation exposure has to be expected which could reach a doubling of the mean natural exposure value.Dedicated to Prof. Dr. H. Muth on the occasion on his 60th birthday 相似文献