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S. Gillis B. C. Furie B. Furie H. Patel M. C. Huberty M. Switzer W. B. Foster H. A. Scoble M. D. Bond 《Protein science : a publication of the Protein Society》1997,6(1):185-196
The gamma-carboxyglutamic acid (Gla) domains of the vitamin K-dependent blood coagulation proteins contain 10 highly conserved Gla residues within the first 33 residues, but factor IX is unique in possessing 2 additional Gla residues at positions 36 and 40. To determine their importance, factor IX species lacking these Gla residues were isolated from heterologously expressed human factor IX. Using ion-exchange chromatography, peptide mapping, mass spectrometry, and N-terminal sequencing, we have purified and identified two partially carboxylated recombinant factor IX species; factor IX/gamma 40E is uncarboxylated at residue 40 and factor IX/gamma 36,40E is uncarboxylated at both residues 36 and 40. These species were compared with the fully gamma-carboxylated recombinant factor IX, unfractionated recombinant factor IX, and plasma-derived factor IX. As monitored by anti-factor IX:Ca (II)-specific antibodies and by the quenching of intrinsic fluorescence, all these factor IX species underwent the Ca(II)-induced conformational transition required for phospholipid membrane binding and bound equivalently to phospholipid vesicles composed of phosphatidylserine, phosphatidylcholine, and phosphatidylethanolamine. Endothelial cell binding was also similar in all species, with half-maximal inhibition of the binding of 125I-labeled plasma-derived factor IX at concentrations of 2-6 nM. Functionally, factor IX/gamma 36,40E and factor IX/gamma 40E were similar to fully gamma-carboxylated recombinant factor IX and plasma-derived factor IX in their coagulant activity and in their ability to participate in the activation of factor X in the tenase complex both with synthetic phospholipid vesicles and activated platelets. However, Gla 36 and Gla 40 represent part of the epitope targeted by anti-factor IX:Mg(II)-specific antibodies because these antibodies bound factor IX preferentially to factor IX/gamma 36,40E and factor IX/gamma 40E. These results demonstrate that the gamma-carboxylation of glutamic acid residues 36 and 40 in human factor IX is not required for any function of factor IX examined. 相似文献
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Robin Heinen S. Emilia Hannula Jonathan R. De Long Martine Huberty Renske Jongen Anna Kielak Katja Steinauer Feng Zhu T. Martijn Bezemer 《Ecology letters》2020,23(6):973-982
Soil legacy effects are commonly highlighted as drivers of plant community dynamics and species co‐existence. However, experimental evidence for soil legacy effects of conditioning plant communities on responding plant communities under natural conditions is lacking. We conditioned 192 grassland plots using six different plant communities with different ratios of grasses and forbs and for different durations. Soil microbial legacies were evident for soil fungi, but not for soil bacteria, while soil abiotic parameters did not significantly change in response to conditioning. The soil legacies affected the composition of the succeeding vegetation. Plant communities with different ratios of grasses and forbs left soil legacies that negatively affected succeeding plants of the same functional type. We conclude that fungal‐mediated soil legacy effects play a significant role in vegetation assembly of natural plant communities. 相似文献
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Abstract Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals. 相似文献
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Salame MY More RS Verheye S Leimbach ME Iii SB Chronos NA 《International journal of cardiovascular interventions》1999,2(4):207-215
Glycoprotein IIb/IIIa receptor inhibitors represent a relatively new therapeutic approach in the field of antiplatelet therapy. Following the development of abciximab a number of small molecule GPIIb/IIIa inhibitors have been introduced such as tirofiban and eptifibatide. In this fast-moving field the interventional cardiologist needs a framework to guide decision-making for the individual patient. This review covers the efficacy and safety data from the clinical trials of GPIIb/IIIa inhibitors in the context of patients undergoing percutaneous coronary intervention for unstable angina/non-Q-wave myocardial infarction. There is an increasing body of evidence to support the efficacy of GPIIb/IIIa inhibitors in reducing the risk of adverse ischemic events in high and low risk patients undergoing percutaneous coronary intervention. A number of unresolved efficacy and safety issues remain, including the duration of treatment before and after intervention; whether a reduction in the heparin dose would further decrease the risk of hemorrhage without affecting the periprocedural thrombotic rate in patients undergoing PTCA with adjunctive GPIIb/IIIa inhibitors; and the cost-effectiveness of this therapy. When a thorough analysis of cost-effectiveness has been made, it will be easier to advocate the widespread use of these agents in all patients undergoing coronary intervention. 相似文献
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Michel Batista Fabricio K Marchini Paola AF Celedon Stenio P Fragoso Christian M Probst Henrique Preti Luiz S Ozaki Gregory A Buck Samuel Goldenberg Marco A Krieger 《BMC microbiology》2010,10(1):259
Background
The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches. 相似文献7.
In population ecology, dispersal plays a fundamental role, but is potentially costly. Traditionally, studies of phenotypic trade-offs involving dispersal focus on resource allocation differences between flight and reproduction. However, investments in dispersal may also result in reduced allocation to other “third-party traits” (e.g. compensatory feeding) that are not directly associated with reproduction. Such traits remain largely uninvestigated for any phytophagous insect despite their importance for performance and survival. Using two wing-dimorphic, phloem-feeding planthoppers, Prokelisia dolus and Prokelisia marginata that differ dramatically in dispersal abilities, we sought evidence for a trade-off between investments in dispersal (flight apparatus) and ingestion capability (allocation to the esophageal musculature governing ingestion). Dispersal allows species to meet nutrient demands by moving to higher-quality resources. In contrast, enhanced investment in esophageal musculature increases ingestion capacity and allows phloem feeders to compensate for deteriorating plant nutrition on site. Our objectives were to compare differences in flight and feeding investment between P. dolus and P. marginata and between the wing forms of both species, and to compare ingestion capacity between the two species and wing forms. Morphometric and gravimetric measures of investment in flight versus feeding indicate that the sedentary P. dolus allocates more muscle mass to feeding whereas P. marginata invests more heavily in flight. Likewise, brachypters invest more in feeding and less in flight than macropters. The greater esophageal investment in P. dolus is associated with enhanced ingestion capacity compared to P. marginata. As a consequence, P. dolus is better equipped to meet on-site nutrient demands when faced with deteriorating plant quality than P. marginata, which must migrate elsewhere to do so. Notably, such third-party trade-offs place constraints on how insect herbivores cope with changing resources and set the stage for fundamental differences in population dynamics. 相似文献
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Eric AF Simoes 《Respiratory research》2002,3(1):6
In sarcoidosis, host genetic factors are discussed as contributing to disease susceptibility and course. Since tumor necrosis factor (TNF)-α is a central mediator of granuloma formation and since elevated TNF-α levels are found during active phases of sarcoidosis, genetic polymorphisms correlating with influences on TNF-α levels are of special interest. The complete sequencing of the MHC region and the increase in the number of identified gene polymorphisms in this locus associated with TNF-α production offer the opportunity of detecting new genes associated with sarcoidosis and perhaps of defining disease-associated haplotypes that bear the potential of serving as predictive markers for this disease. 相似文献
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Hanneke Vlaming Tibor van Welsem Erik L de Graaf David Ontoso AF Maarten Altelaar Pedro A San-Segundo Albert JR Heck Fred van Leeuwen 《EMBO reports》2014,15(10):1077-1084
Histone H2B ubiquitination is a dynamic modification that promotes methylation of histone H3K79 and H3K4. This crosstalk is important for the DNA damage response and has been implicated in cancer. Here, we show that in engineered yeast strains, ubiquitins tethered to every nucleosome promote H3K79 and H3K4 methylation from a proximal as well as a more distal site, but only if in a correct orientation. This plasticity indicates that the exact location of the attachment site, the native ubiquitin-lysine linkage and ubiquitination cycles are not critical for trans-histone crosstalk in vivo. The flexibility in crosstalk also indicates that other ubiquitination events may promote H3 methylation. 相似文献