Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

It is shown that scanning force microscopy (SFM), operated in the attractive mode, can be used to obtain high resolution pictures of adsorbed fibrinogen molecules on solid surfaces, without the need for staining or special microscope grids. SFM also reveals the three-dimensional structure of the adsorbed molecules. Two forms of adsorbed fibrinogen are demonstrated on hydrophobic silicone dioxide surfaces: a trinodular about 60 nm long and a globular with about a 40 nm diameter. Polymeric networks formed after storage of the surface with adsorbed fibrinogen in PBS for 11 days are also shown. The SFM-results for the trinodular structure suggest the existence of loops or peptide chains extending outside the basic structure of the fibrinogen molecule.
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PMID:Structure of adsorbed fibrinogen obtained by scanning force microscopy. 184 83

Rouleau formation by back scattered light and flow behaviour by viscometry of heat-treated (48.4, 48.8 and 49.5 degrees C) and normal (37 degrees C) human red blood cells (RBC) were investigated. Washed RBC were treated in PBS and afterwards resuspended in their own plasma. It was found that the time behaviour of the fibrinogen mediated weak RBC-RBC interaction is influenced (decreased) already at an incubating temperature of 48.4 degrees C. Kinetic measurements are more sensitive than a static aggregation characterization. Beside the decreased deformability of the RBC also a heat-altered structure of the RBC-glycocalyx and the aggregating energy of the macromolecules have to be considered.
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PMID:Moderate heat treatment of only red blood cells (RBC) slows down the rate of RBC-RBC aggregation in plasma. 646 8

Potent and novel fibrinolytic enzymes (lumbrokinase [LK]) were extracted from the earthworm, Lumbricus rubellus. These enzymes were very stable and showed greater antithrombotic activity than other currently used fibrinolytic proteins. An LK fraction showing the most potent fibrinolytic activity was immobilized onto a polyurethane (PU) surface to investigate its enzymatic activity and antithrombotic activity. A methanol-extracted PU surface was coated with 3% (wt/vol) maleic anhydride methylvinyl ether copolymer (MAMEC)/tetrahydrofuran (THF) solution, and the surface was incubated in an LK solution/phosphate-buffered saline (PBS, pH 7.4). The surface properties were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA), and dynamic contact angle. The stability of immobilized LK was determined by caseinolytic activity assay and the specificity of immobilized LK on fibrinogen/fibrin was observed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The antithrombotic activity of immobilized LK was evaluated using an ex vivo rabbit A-A shunt experiment. LK immobilization was confirmed by ATR-FTIR and ESCA. Immobilized LK demonstrated stable proteolytic activity during various incubation periods. Immobilized LK proteolyzed fibrinogen and fibrin almost specifically, while it hardly hydrolyzed other plasma proteins including plasminogen and albumin. In the ex vivo A-A shunt experiment, the LK-immobilized surface significantly prolonged occlusion time over control surfaces. This is primarily due to the high thrombolytic activity of immobilized LK. In this work, a highly efficient surface modification method on the PU surface was developed, and this LK immobilization technique will be very useful in improving the blood compatibility of blood-contacting devices.
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PMID:Surface characteristics and properties of lumbrokinase-immobilized polyurethane. 761 90

Six fractions of strong and novel fibrinolytic enzymes (lumbrokinase, LK) were extracted from the earthworm Lumbricus rubellus. The enzymes in these fractions appeared to be very stable and showed greater antithrombotic activity than other currently used antithrombotics. The authors immobilized an LK fraction that shows the most potent fibrinolytic activity on a polyurethane (PU) surface to investigate its enzymatic and antithrombotic activity. The methanol extracted PU surface was treated with a 3% (wt/vol) maleic anhydride methylvinyl ether copolymer (MAMEC) solution and finally incubated in an LK solution in PBS (pH 7.4). The immobilized LK activity was estimated by the fibrin plate method and caseinolytic activity assay. The antithrombotic activity was evaluated by in vitro 125I-fibrinogen adsorption in fresh whole blood and 99mTc platelet adhesion tests. In addition, the occlusion time was determined through ex vivo rabbit A-A shunt experiments. The content and unit activity of immobilized LK were found to be 24 micrograms/cm2 and 18 IU/cm2, respectively. The relative activity ratio of immobilized LK to soluble LK was found to be approximately 34%. Immobilized LK was stable within a various pH range and resistant to inhibitors and thermal inactivation. Less fibrinogen was adsorbed and fewer platelets adhered on an LK-immobilized surface than on PU and PU-MAMEC controls. The ex vivo occlusion time of untreated PU and PU-MAMEC surfaces were only 32 and 42 minutes, respectively. But that of LK-immobilized PU was extended to 140 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Antithrombotic activity of a lumbrokinase immobilized polyurethane surface. 826 50

Plasma protein adsorption onto an artificial surface is strongly influenced by not only the surface characteristics of materials, but also by the fluid dynamics inside the blood pump, and it would influence subsequent platelet adhesion or activation, which plays a major role in the initiation of thrombus formation at the blood-material interface in vivo. In vitro flow visualization of an electrohydraulic LVAD was performed by a video camera (CCD, Hitachi) and an image processor (PC VISION PLUS) with an IBM PC. The electrohydraulic LVADs were implanted in mongrel dogs of approximately 20 kg. The authors sectioned the blood contacted ventricle after animal death according to the level of shear rate. Because analysis of adsorbed protein might be influenced by the size of the ventricle segment, the number of segments was limited to eight per ventricle. Platelet adhesion and its morphology were observed by scanning electron microscopy (SEM). Adsorbed plasma proteins (fibrinogen, albumin, and IgG) on each segment were quantified by enzyme linked immunosorbent assay (ELISA). The specimens were soaked in 2% (wt/vol) SDS/PBS for 2 days and the released protein concentration assessed. A well developed large vortex was observed at the center of the artificial ventricle. Polyurethane blood pumps displayed different degrees of protein adsorption and subsequent platelet adhesion on each segment.
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PMID:The fluid dynamic effect on protein adsorption in left ventricular assist devices. 826 54

As inhaled nitric oxide (iNO) may differently increase bleeding time (BT) and inhibit platelet aggregation in normal and lung-injured patients or experimental models, we studied the effects of iNO on hemostasis in presence and absence of an endotoxic lung injury in the rat. Eight hours after intratracheal administration of endotoxin (lipopolysaccharide [LPS]) or its solvent (phosphate-buffered solution [PBS]), four groups of rats were randomized according to the presence or absence of 15 ppm iNO added for an additional 10 h. We measured BT, ex vivo platelet aggregation, plasma fibrinogen, euglobulin clot lysis time (ECLT), and platelet and aortic cyclic guanosine 5'-monophosphate (cGMP) contents. Acute lung inflammation did not influence BT, but increased platelet aggregability, fibrinogen levels, and platelet and aortic cGMP. In control and endotoxic rats, iNO increased BT, reduced platelet aggregability, and increased platelet cGMP. iNO increased aortic cGMP only in healthy rats. ECLT was increased by LPS and unchanged with iNO. These results suggest that the extrapulmonary "systemic" effects induced by iNO on hemostasis were not strictly similar in healthy and LPS rats, inflammation inducing proper changes in coagulation parameters. However, iNO attenuated the procoagulant activity induced by acute lung inflammation, suggesting a potentially beneficial effect of this therapy.
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PMID:Impact of inhaled nitric oxide on platelet aggregation and fibrinolysis in rats with endotoxic lung injury. Role of cyclic guanosine 5'-monophosphate. 973 Oct 13

Interleukin-1beta (IL-1beta) was isolated from LPS-stimulated brushtail possum alveolar macrophages using PCR primers based on conserved regions of mammalian IL-1beta. The complete cDNA was cloned by 5' and 3' rapid amplification of cDNA ends (RACE). The predicted protein of 269 amino acids shared 4346% identity with several mammalian IL-1beta proteins. Constructs were made to express the mature IL-1beta in Escherichia coli and two recombinant IL-1beta proteins, rpIL-1beta1 and rpIL-1beta2, which differed in length by four amino acids at the N-terminus, were produced. Both proteins induced a weak proliferative response in a possum thymocyte assay. Possums injected intravenously with 100 microg of rpIL-1beta1 or rpIL-1beta2 showed profound changes in body temperature and numbers of circulating leukocytes. A sharp decrease in temperature occurred within 2 h of administration followed by an elevation of temperature peaking at 24 h. The smaller rpIL-1beta1 protein had a greater effect on temperature than rpIL-1beta2. Both rpIL-1beta proteins caused a marked decrease in number of neutrophils and lymphocytes at 2-6 h after injection. At 24 h after injection, neutrophil and lymphocyte numbers were elevated 6.0-fold and 2.6-fold, respectively in the possums injected with rpIL-1beta1 and 3.9-fold and 1.5-fold, respectively in the possums injected with rpIL-1beta2. Fibrinogen levels were elevated at 24 and 72 h after injection with both proteins. In comparison, neither recombinant bovine IL-1beta (rbIL-1beta) nor PBS had significant effects on body temperature or blood haematology. The studies have shown that the two recombinant forms of IL-1beta were biologically active in possums and that the IL-1beta with four fewer amino acids at the N-terminus was the more active.
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PMID:Molecular cloning and physiological effects of brushtail possum interleukin-1beta. 1020 3

Red blood cell (RBC) aggregation is of prime importance in vivo and in vitro for low flow rates. It may be estimated by rheometrical measurements at low shear rates, but these are perturbed by slip and migrational effects which have already been highlighted in the past. These effects lead to a torque decay with time so that the true value of the stress at low shear rates may be greatly underestimated. Elevated aggregation being associated with different diseases, pathological blood samples show more pronounced perturbing effects and a strong time dependency in low shear rate rheometry. To test the dependence of slip and migrational effects on RBC aggregation, and particularly to determine the way in which they depend upon fibrinogen concentration ([Fb]), a home-made measuring system with roughened internal and external walls (170 microns roughness) was used to study low shear rate rheometry for RBC suspensions in PBS buffer containing albumin (at 50 g/l) and fibrinogen at various concentrations. The influences of hematocrit, shear rate, and fibrinogen concentration were investigated. Particular attention was paid to data acquisition at low shear rates (10(-3) s-1 to 3 x 10(-2) s-1). The combined influence of hematocrit and fibrinogen was investigated by adjusting hematocrit to 44 or 57% and fibrinogen concentration ([Fb]) to 3.0-4.5-6.5 g/l. Microscopic observations of the blood samples at rest were performed. They showed that different structures were formed according to fibrinogen concentration. The rheometrical measurements indicated that torque decay with shearing duration was strongly dependent on fibrinogen concentration and on shear rate at fixed hematocrit. Migrational and slip effects were more pronounced as shear rate decreased, fibrinogen concentration was raised, and hematocrit was lowered. The results have been explained on the basis of the expected microstructure of flowing blood in relation to the microscopic observations at rest.
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PMID:Blood low shear rate rheometry: influence of fibrinogen level and hematocrit on slip and migrational effects. 1047 59

To clarify the release properties of anti-cancer drugs from fibrin glue, a study was performed using several anti-cancer drugs with remarkably different physical properties. Concentrated fibrinogen, fibronectin, and coagulation factor XIII were prepared from healthy human plasma according to the cryoprecipitate method. Fibrin glue containing anti-cancer drugs was prepared as follows; the cryoprecipitate was mixed with each anti-cancer drug and aprotinin, then thrombin was added. These glues were incubated in PBS containing plasminogen and urokinase at 37 degrees C for seven days, and the medium was then sampled several times after centrifugation. The drug concentration in each sample was measured using HPLC. Fibrin glue without aprotinin was quickly hemolyzed and disappeared after 2--4 h. That with aprotinin was only slightly hemolyzed and more than half remained after 7 days. Mitomycin C and fluorouracil were quickly released from the glue regardless of the presence or absence of aprotinin. However, enocitabine was gradually released from glue with aprotinin although quickly released from that without. The rate of release of each drug from the glue with aprotinin correlated well with its hydrophobicity. Thus, to establish a sustained release system using fibrin glue, one should use the more lipophilic anti-cancer drugs and a fibrinolytic enzyme inhibitor.
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PMID:Novel drug delivery system using autologous fibrin glue--release properties of anti-cancer drugs. 1072

The effect of fibrinogen on corrosion behavior of SUS316L and SUS317L stainless steel in artificial blood PBS solution has been investigated with electrochemical technology. The results showed that the corrosion potential (Ec) of stainless steel shifted negatively, the passivated current (ip) became less and the pitting corrosion potential (Eb) shifted negatively with the existence of fibrinogen in PBS. These indicate that samples become more sensitive to corrosion under this circumstance. SEM pictures demonstrated that stainless steel adsorbed fibrinogen on its surface.
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PMID:[Effect of fibrinogen on corrosion behavior of stainless steel in artificial blood solution]. 1179 9


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