scholarly journals Complex Formation of Covalently Crosslinked Fibrin Oligomers with Agarose Coupled Fibrinogen and Fibrin

1977 ◽  
Author(s):  
R. von Hugo ◽  
R. Hafter ◽  
A. Stemberger ◽  
H. Graeff
Keyword(s):  
Molecular Weight ◽  
Complex Formation ◽  
Affinity Chromatography ◽  
High Molecular Weight ◽  
Calcium Chloride ◽  
Gel Filtration ◽  
Void Volume ◽  
Soluble Fibrin ◽  
Derivatives Of

Crosslinked high molecular weight derivatives of fibrin (fibrinoligomers) were observed during intravascular coagulation. It was the purpose of this study to investigate the complex formation of fibrin oligomers with fibrinogen and fibrinmonomer. Fibrinogen coupled to agarose (Fg-ag) as well as fi-brinmonomer coupled to agarose (Fm-ag) was used as substrate. Soluble oligomers of fibrin were produced by incubating fibrinogen with thrombin, calcium-chloride, cystein and F XIII. They were separated from fibrinogen by gel filtration. Γ-dimers were demonstrated in fractions from the void volume and the shoulder prior to the fibrinogen peak. These fractions were subjected to affinity chromatography. Crosslinked oligomers of fibrin were not adsorbed on Fg-ag, yet adsorption occured on Fm-ag. This indicates that fibrin oligomers have no affinity to fibrinogen, yet readily form complexes with fibrin. This could mean that in vivo they compete with fibrinogen for the fibrinmonomer part of soluble fibrin monomer complexes, and hence have a tendency to increase in size.

Gel Filtration of Plasma from Rabbits Injected with Honomeric DES-AB 125I-Fibrin and 131I–Fibrinogen

1979 ◽  
Author(s):  
I. Kröhnke ◽  
I. Hahn ◽  
W. Krell ◽  
G. Müller-Berghaus
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Void Volume ◽  
Molecular Configuration ◽  
Chromatographic Behaviour ◽  
Fibrin Monomer ◽  
Deutsche Forschungsgemeinschaft

We intended to study the chromatographic behaviour of soluble des-AB fibrin prepared in vitro and injected into rabbits. To prepare des-AB 1251-fibrin, purified rabbit 125I-fibrinogen was clotted by thrombin and the formed clot dissolved in Tris-buffered 3 M urea. Gel filtration of des-AB fibrin in urea through sepharose-CL 6B columns equilibrated with buffered 3 M. urea revealed monomeric fibrin. Rabbits received 131I-fibrinogen and 5 min later monomeric des-AB 125I-fibrin in urea. 30 min after injection blood was drawn and the plasma obtained filtered through sepharose-CL 6B columns eguilibrated with buffered plasma. At 20°C as well as at 37°C des-AB 125I-fibrin was eluted in the void volume in front of the 131I-fibrinogen peak. The data demonstrate that monomeric des-AB 125I-fibrin in urea injected into rabbits remains soluble in plasma. Possibly, monomerJ fibrin is converted to circulating soluble high-molecular weight fibrin aggregates or fibrin monomer changes its molecular configuration, thus showing a different gel filtration behaviour.(Supported by the Deutsche Forschungsgemeinschaft).

In Vitro and in Vivo Antigen-Induced Release of High-Molecular Weight Neutrophil Chemotactic Activity from Human Nasal Tissue

1988 ◽  
Vol 2 (1) ◽  
pp. 7-11 ◽  
Author(s):  
T. Nagakura ◽  
T. Onda ◽  
Y. likura ◽  
T. Endo ◽  
H. Nagakura ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Nasal Polyps ◽  
Gel Filtration ◽  
Chemotactic Activity ◽  
Nasal Tissue ◽  
Nasal Secretions ◽  
Neutrophil Chemotactic Activity

High molecular weight neutrophil chemotactic activity has been identified in resected human nasal polyps, inferior turbinates, and nasal secretions following antigen challenge. The estimated molecular weight, by gel filtration chromatography, was approximately 600,000. However, a heterogeneity of molecular weight in some patients was recognized. Our results suggest a possible role for high molecular weight-neutrophil chemotactic activity in the pathogenesis of hypersensitivity in the human nasal cavity.

scholarly journals High Molecular Weight Fibrinogen Complexes in Patients with History of Myocardial Infarction and Cerebrovascular Disorders

1977 ◽  
Author(s):  
G.G. Neri Serneri ◽  
G.F. Gensini ◽  
R. Abbate ◽  
D. Prisco ◽  
C. Mugnaini ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Molecular Weight ◽  
High Molecular Weight ◽  
Cerebrovascular Disorders ◽  
Gel Filtration ◽  
Globular Proteins ◽  
Void Volume ◽  
Beta Alanine ◽  
Cross Linkage ◽  
History Of

The increased turnover of fibrinogen and decreased platelet survival observed in many patients with history of myocardial infarction (MIP) and in patients with chronic cerebrovascular disorders (CVP) (Neri Serneri et al 1970, Harker and Slichter 1972) could suggest a hypercoagulable state. We investigated 28 MIP, 23 CVP and 31 controls for circulating high molecular weight fibrinogen complexes (HMWFC) by gel-filtration (agarose 4%, 100–200 m, column 1.5 × 90 cm, buffer Tris-Cl-citrate pH 7.6, flow 13 ml/hour, recording of OD at 280 um) of plasma beta-alanine precipitate. HMWFC are eluted in a peak at an alution volume corresponding to the void volume of the column, at which volume globular proteins of MW over 1 million are eluted. HMWFC concentration was in the controls 2.98±1.52% of the fibrinogen eluted, in MIP 8.27±2.9 % (P<0.0l) and in CVP 7.48±1.9 % (P<0.01). When HMWFC concentration was higher than 6-7%, PAA electrophoresis of the eluted complexes (after mercaptoethanol reduction) allowed to detect gamma-gamma dimers, so indicating the cross-linkage of HMWFC. Heparin treatment (12,500 U × 2) markedly lowered the concentration of HMWFC and made gamma-gamma dimers undetectable. These results indicate that in MIP and in CVP a hypercoagulability frequently exists.

Characterization Of Soluble DES-A And DES-AB Fibrin In Human Plasma At 37°C By Agarose-Gel Filtration

1981 ◽  
Author(s):  
G Müller-Berghaus ◽  
J-C Bernhard ◽  
I Mahn
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Void Volume ◽  
Agarose Gel ◽  
High Molecular Weight Material ◽  
Different Temperatures ◽  
Lower Temperature

In two consecutive steps, thrombin cleaves the fibrinopeptides A and B from fibrinogen producing des-A fibrin and des-AB fibrin. Labeled des-A fibrin was prepared by batroxobin and labeled des-AB fibrin by clotting of 125I-fibrinogen with thrombin. Fibrin solubilized in buffered urea was mixed with plasma containing 131I-fibrinogen (fibrin:fibrinogen ratio = 1:20). These fibrinfibrinogen mixtures were applied to sepharose CL- 6B columns eq ui librated with buffered plasma (0.0025 M EDTA, 0.1 M NaCl, 0.05 M tris, 0.005 M EACA, 2 AT U hirudin/ml, 500 KIU a protinin/ml, 0.003 M NaN3, pH 7.4). Plasma was used as an equilibration and elution medium to prevent precipitation of fibrin in the columns. At 20°C, labeled des-A fibrin as well as des-AB fibrin were eluted in the void volume as high-molecular weight aggregates (peak A) and separated from m onomeric labeled fibrinogen (peak B). At 37°C, however, des- A fibrin was eluted at the same position as monomeric fibrinogen (peak B), whereas des-AB fibrin was eluted in the void volume as at 20°C. Rechromatography of isolated fractions of peak A and peak B at different temperatures showed that monomeric fibrin isolated at 37°C formed high molecular weight material at 20°C, and high-molecular weight fibrin isolated at 20°C dissociated at 37 ° C. The results suggest that des-A fibrin solubilized in plasma in the absence of calcium ismonomeric at 37°C but forms high-molecular weight aggregates at lower temperature, whereas des-AB fibrin is oligomeric at 20°C as well as at 37°C.

Distribution of radioactive silver in the subcellular fractions of various tissues of the rat and its binding to low molecular weight proteins

1983 ◽  
Vol 61 (11) ◽  
pp. 1391-1395 ◽  
Author(s):  
Yousef Matuk
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Total Radioactivity ◽  
Subcellular Fractions ◽  
Low Molecular Weight ◽  
Electron Microscopic ◽  
Molecular Weight Peak

In view of the electron microscopic evidence that silver does not penetrate cellular barriers, the distribution of radioactive silver in rat blood and subcellular fractions of liver, kidneys, spleen, and forebrain was studied. It was found that 24 h after a single intraperitoneal injection high levels of radioactivity were reached which decreased at different rates in the various tissues studied. In plasma, liver, and kidneys there was an initial rapid loss of radioactivity which was followed by a slower rate of loss. In the blood, forebrain, and spleen the loss of radioactivity was linear and somewhat slower than in the other three tissues. The cytosols of the liver and kidneys contained 60% while those of the forebrain and spleen contained 30% of the total radioactivity found in the tissue homogenates. Gel filtration on Sephadex G-75 showed that all cytosols contained two peaks of radioactivity; a high molecular weight peak which eluted just after the void volume and a low molecular weight peak. The amount of radioactivity in both peaks was, however, much lower in the chromatographic peaks of the forebrain and spleen than that found in those of the liver and kidneys. Furthermore, the spleen had a comparatively very small low molecular weight radioactive peak. In vitro experiments with liver cytosol showed similar results to those found in vivo in that the high molecular weight radioactive peak could be removed by heat. It is concluded that silver does enter cells and that silver thionein exists in the cytosols of forebrain, spleen, kidney, and liver.

scholarly journals Demonstration of kallikrein-like protease activity in nonactivated plasma of patients with Cooley's anemia

Blood ◽  
1983 ◽  
Vol 61 (2) ◽  
pp. 232-237
Author(s):  
M Andrew ◽  
M Manno ◽  
M Karpatkin
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Trypsin Inhibitor ◽  
Gel Filtration ◽  
Lima Bean ◽  
Thrombin Time ◽  
Diisopropyl Fluorophosphate ◽  
Bean Trypsin Inhibitor ◽  
Cooley's Anemia

Routine evaluation of 12 children with Cooley's anemia revealed that each one had a prolonged partial thromboplastin time. However, prothrombin time and thrombin time were within the normal range. Specific assays demonstrated low levels of the four contact factors: factors XI, XII, prekallikrein, and high molecular weight kininogen. Further investigation revealed activity against para-nitroanilide peptide substrates in unactivated plasma from all 12 patients. Following gel filtration on Sephadex G200, the activity emerged in one peak in the void volume, indicating a molecular weight of greater the 500,000. Activity was greatest against H-D-Pro-Phe-Arg-pNA, the substrate for plasma kallikrein, and was inhibited by diisopropyl fluorophosphate and trasolyl. It was unaffected by hirudin, soy bean trypsin inhibitor, and lima bean trypsin inhibitor. It was destroyed by heating at 56 degrees C. Specific antisera against human prekallikrein and human alpha-macroglobulin did not reduce the activity. It is concluded that a high molecular weight kallikrein-like protease, is present in the plasma of these patients. It is postulated that it is released into the circulation from tissue as a result of damage due to iron overload. It is further postulated that this protease brings about in vivo activation of the contrast factors, resulting in a fall in their circulating levels.

scholarly journals Enhancement of blood coagulation by soluble fibrin complexes.

1975 ◽  
Vol 141 (5) ◽  
pp. 944-961 ◽  
Author(s):  
M S Hansen ◽  
N U Bang ◽  
R D Barton ◽  
L E Mattler
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Biological Properties ◽  
Major Surgery ◽  
Gel Chromatography ◽  
Soluble Fibrin ◽  
Enhanced Sensitivity ◽  
Intravascular Thrombosis ◽  
Normal Plasma

We have detected a species of soluble fibrin complexes with significant biological properties. Agarose gel chromatography of normal plasma or purified fibrinogen previously incubated with small amounts of thrombin revealed the presence of a species of high molecular weight soluble fibrin complexes, which contained only small quantities of fibrinogen by immunological assays but which exhibited enhanced sensitivity to thrombin. In addition, these complexes substantially shortened the thrombin-clotting time of normal plasma and enhanced the resistance of normal plasma to heparin action. Similar thrombin-sensitive soluble fibrin complexes were demonstrated in vivo in rabbits for up to 10 min after the infusion of 50 U of thrombin. Thrombin-sensitive soluble fibrin complexes were also demonstrated in 3 of 12 patients with documented thromboembolic disease and in 2 of 20 patients after major surgery. High molecular weight soluble fibrin complexes, which exhibit enhanced thrombin sensitivity and which are capable of increasing the rate of normal fibrinogen-to-fibrin conversion by thrombin, may appear consequent to clinical thrombosis and situations involving trauma (e.g., major surgery). Such soluble complexes, although they have no proven role in the primary pathogenesis of intravascular thrombosis, may contribute to a temporary "hypercoagulable state" and may accelerate the build-up and extension of already existing thrombotic deposits.

scholarly journals High Molecular Weight Fibrin(OGEN) Complexes

1977 ◽  
Author(s):  
N. Alkjaersig
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Complex Form ◽  
Plasma Fibrinogen ◽  
Fibrin Deposition ◽  
Exclusion Chromatography ◽  
The Individual ◽  
Immunologic Assay ◽  
Derivatives Of

Plasma fibrinogen, because of catabolism of fibrinogen in vivo, exhibits biochemical heterogeneity, manifested by molecular weight and other differences between the various thrombin clottable moieties. These comprise high molecular weight fibrin(ogen) complexes (HMWFC) ranging in molecular weight from 400,000 to 1,000,000, native fibrinogen (m.w. 330,000) and derivatives of fibrinogen smaller than the native protein (m.s. 260,000 or less). These moieties may be assayed, in plasma as well as in the purified system, by gel exclusion chromatography with immunologic assay of chromatographic effluents by the Technicon Immunoprecipi-tator and mathematical analysis of elution profiles. The coefficient of assay variation for the individual moieties is 7%.Normal control subjects show (mean ± 1 s.d.) 7.7 ± 6.2% (18.4 ± 21 mg%) of plasma fibrinogen in higher molecular weight complex form, 67 ± 10.3% monomeric fibrinogen and 25.8 ± 6.4% fibrinogen first derivative. Values for plasma HMWFC exceeding mean + 2 s.d. of normal (20% or 71 mg%) are classed as abnormal and regarded as demonstrative of enhanced fibrin formation. The hypothesis that enhanced fibrin deposition is associated with the presence of demonstrable pathology, extravascular or intravascular fibrin deposistion or thrombosis, has been validated in several human disease models.

Regulation of intestinal goblet cell secretion. I. Role of parasympathetic stimulation

1982 ◽  
Vol 242 (4) ◽  
pp. G370-G379 ◽  
Author(s):  
R. D. Specian ◽  
M. R. Neutra
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Goblet Cell ◽  
Gel Filtration ◽  
Goblet Cells ◽  
Mucus Secretion ◽  
Cell Secretion ◽  
Intestinal Mucus ◽  
Compound Exocytosis

The in vivo effects of the parasympathomimetic drug pilocarpine on rat intestinal goblet cells were analyzed by autoradiography, light microscopy (LM), and electron microscopy (EM). Pilocarpine accelerated the release of mucus by compound exocytosis from crypt (but not surface) goblet cells throughout the small and large intestine. Pilocarpine-induced mucus secretion was blocked by atropine alone in ileum and colon, but total inhibition in proximal small intestine required a combination of atropine and tubocurarine. The sensitivity of morphological-autoradiographic methods for detection of goblet cell secretion was compared with that of a biochemical detection method, separation of labeled high-molecular-weight glycoproteins by Sepharose 4B gel filtration of luminal washings. Even when secretion of labeled mucus by compound exocytosis was clearly demonstrated by LM, EM, and autoradiography, gel filtration assay of luminal washings from pilocarpine-injected rats failed to reveal an increase in labeled high-molecular-weight glycoproteins. Autoradiographs of mucosal tissue after luminal washing showed that newly secreted, labeled mucus was retained in the crypts and was thus unavailable to the biochemical assay. Thus, direct observation of exocytosis in individual goblet cells provides a qualitative, but sensitive, assay for short-term acceleration of intestinal mucus secretion.

Tumor Uptake of High and Low Molecular Weight Fractions from 67Ga-Citrate Labelled Blood Plasma

1984 ◽  
Vol 23 (02) ◽  
pp. 59-61 ◽  
Author(s):  
M. C. Crone ◽  
P. Thouvenot ◽  
F. Brunotte ◽  
C. Marchai ◽  
J. Robert ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Blood Plasma ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Weight Fraction ◽  
Blood Protein ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
67Ga Citrate

SummaryBlood plasma from tumor-bearing rats was incubated with 67Ga-citrate, and two fractions of high molecular weight (proteins) and low molecular weight were isolated by dialysis and by gel-filtration chromatography. Both fractions showed a different in vivo uptake by DS-sarcoma-bearing animals, the high molecular weight fraction being accumulated to a lesser extent. Compared to 67Ga-citrate the low molecular weight fraction showed a different uptake which for most tissues was significatively higher. This behavior suggests the presence of 67Ga in chemical forms other than citrate in the low molecular weight fraction. The lower uptake of the blood protein fraction is discussed.

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