The Use of DEAE Cellulose in the Preparation of a Factor VIII (AHG)-free Bovine Fibrinogen Reagent

1961 ◽  
Vol 6 (02) ◽  
pp. 287-307 ◽  
Author(s):  
W. F Stapp
Keyword(s):  
Factor Viii ◽  
Protein Concentration ◽  
Tap Water ◽  
Deae Cellulose ◽  
Fibrinogen Concentration ◽  
Nacl Solution ◽  
Factor Viii Activity ◽  
Bovine Plasma ◽  
Nacl Concentration ◽  
Optimum Ph

SummaryAdequate fibrinogen reactivity is suggested as an additional requirement for specific fibrinogen reagents, to be added to the well known requirements of protein concentration, optimum pH, and optimum ionic strength. Reasons and a method for testing are outlined.A method for the preparation of a specific fibrinogen reagent, deficient in factor VIII, from BaSO4 adsorbed bovine plasma is described in detail. The method described utilizes either fresh or regenerated DEAE cellulose as an adsorbent in a non-column chromatographic adsorption procedure, associated with a dialysis procedure in which the final fibrinogen product is concentrated and precipitated as a “euglobulin”. The “euglobulin” fibrinogen precipitate was then dissolved/suspended in buffered saline at a specific pH and NaCl concentration. Essentially the method can be briefly outlined into 5 separate steps:1. Dialysis of BaSO4 adsorbed bovine plasma against 0.5% NaCl at pH 6.6 in 0.005 M Imidazole-HCl buffer.2. Washing of DEAE cellulose with the same type of buffered NaCl solution to specific conditions.3. Mixing of the dialyzed BaSO4 adsorbed bovine plasma with the washed DEAE cellulose and separation by centrifugation.4. Concentration of the cellulose adsorbed plasma as a “euglobulin” fibrinogen precipitate by dialysis against running tap water.5. Dissolving of the “euglobulin” fibrinogen precipitate in 0.9% NaCl buffered at pH 6.8.Fibrinogen preparations, prepared with the use of regenerated DEAE cellulose were found to be similar to those prepared by fresh DEAE cellulose in fibrinogen reactivity and deficiency in factor VIII activity, but were found to have less fibrinogen concentration than those prepared using fresh DEAE cellulose.Suggestions were also made for possibly increasing the over-all yield of the fibrinogen reagent through use of a “washing-elution” procedure.The procedure that was used for regenerating the DEAE cellulose was also described in detail and commented upon.

Thrombelastography during and after Elective Abdominal Surgery

1978 ◽  
Vol 39 (02) ◽  
pp. 488-495 ◽  
Author(s):  
J M Butler
Keyword(s):  
Abdominal Surgery ◽  
Factor Viii ◽  
Whole Blood ◽  
Dominant Factor ◽  
Fibrinogen Concentration ◽  
Factor Viii Activity ◽  
Elective Abdominal Surgery

SummaryThrombelastography has been performed on recalcified whole blood from 50 patients before, during and after elective abdominal surgery. The characteristic changes of the thrombelastographic indices r, k and mA are described.During operation r and k shortened, but no change in mA was observed. This response was in part associated with an increase in factor VIII activity. Following operation, while r time was somewhat shortened, much more marked changes in k and mA were evident. Increasing fibrinogen concentration was the dominant factor in determining the post-operative changes in the thrombelastograph.

Difference in rehydration process due to salt concentration of drinking water in rats

1988 ◽  
Vol 64 (6) ◽  
pp. 2438-2443 ◽  
Author(s):  
T. Okuno ◽  
T. Yawata ◽  
H. Nose ◽  
T. Morimoto
Keyword(s):  
Tap Water ◽  
Albino Rats ◽  
Fluid Loss ◽  
Salt Appetite ◽  
Nacl Solution ◽  
Mutual Cooperation ◽  
Positive Balance ◽  
Nacl Concentration ◽  
Rehydration Process ◽  
Additional Loss

Albino rats were thermally dehydrated (approximately 8% of body wt), divided into five groups, and given tap water or 0.2, 0.45, 0.9, or 2.0% NaCl solution ad libitum for 16 h. Rats given 0.9 or 0.45% NaCl solution regained fluid loss completely in 3–3.5 h, whereas those given 0.2% solution became fully rehydrated at 10 h. The rats in the tap water and 2.0% NaCl groups were only 78 and 59% rehydrated, respectively, within 16 h. Na balance was positive in the 0.9% NaCl group by about five times the amount of the cations lost during the dehydration period. A positive balance of Na was also observed in the 0.45 (approximately 250%) and 2.0% NaCl groups (300%), whereas the 0.2% NaCl group regained lost water and Na simultaneously at 10 h. With tap water, additional loss of cations was observed. These findings show that for the replacement of water due to thermal dehydration there is a range of NaCl concentration with which the rats can rehydrate with the mutual cooperation of thirst, salt appetite, and kidney function.

Fibrinogen Concentration and Factor VIII Activity in Women with Preeclampsia

2007 ◽  
Vol 26 (4) ◽  
pp. 415-421 ◽  
Author(s):  
Vaughan K. Williams ◽  
Adrian B. M. Griffiths ◽  
Sarah Carbone ◽  
William M. Hague
Keyword(s):  
Factor Viii ◽  
Fibrinogen Concentration ◽  
Factor Viii Activity

Decreased Fibrinolysis in Behçet’s Disease

1973 ◽  
Vol 29 (03) ◽  
pp. 610-618 ◽  
Author(s):  
Tova Chajek ◽  
Eta Aronowski ◽  
G Izak
Keyword(s):  
Factor Viii ◽  
Plasminogen Activator ◽  
Behçet’S Disease ◽  
Fibrinolytic Activity ◽  
Behcet’S Disease ◽  
Behçet's Disease ◽  
Fibrinogen Concentration ◽  
Factor Viii Activity ◽  
Fibrinolytic Agents ◽  
Behcet's Disease

SummaryThe coagulation and fibrinolytic systems have been investigated in nine patients suffering from Behçet’s disease. Six of these had thrombophlebitis.The results of these studies revealed markedly elevated fibrinogen concentration and increased factor VIII activity as well as a pronounced decrease in the spontaneous fibrinolytic activity. This latter finding may have resulted from the presence of a potent antiplasmin activity and that of an inhibitor of the plasminogen activator detected in these patients’ sera.It is suggested that decreased fibrinolysis aggravates the thrombotic episodes frequently encountered in Behçet’s disease; accordingly treatment with fibrinolytic agents was found to offer marked benefits.

“Inhibitor Like” Effects of Hematoporphyrin on Factor VIII (Antihaemophilic Globulin)

1967 ◽  
Vol 17 (03/04) ◽  
pp. 299-306
Author(s):  
R. B Davis ◽  
Rosemary Biggs ◽  
K. W. E Denson
Keyword(s):  
Factor Viii ◽  
Fibrinogen Concentration ◽  
Factor Viii Activity ◽  
Temperature Dependent ◽  
Naturally Occurring ◽  
Progressive Loss ◽  
Rate Of Reaction

SummaryThe incubation of factor VIII with HP resulted in a progressive loss of factor VIII activity which was dependent on the porphyrin concentration. The rate of reaction was retarded by fibrinogen and glyoxaline, but was not accelerated by reduction in fibrinogen concentration. No increase in factor VIII activity was found when fibrinogen was added after destruction of factor VIII by HP, suggesting irreversibility of factor VIII destruction. The reaction was light but not temperature dependent; in other respects it closely resembled the kinetics observed during factor VIII destruction by naturally occurring anticoagulants and antibody inhibitors of factor VIII.

Measurement of factor VIII activity of B-domain deleted recombinant factor VIII

2001 ◽  
Vol 38 (2, Suppl 4) ◽  
pp. 13-23 ◽  
Author(s):  
M. Mikaelsson ◽  
U. Oswaldsson ◽  
M. A. Jankowski
Keyword(s):  
Factor Viii ◽  
Recombinant Factor Viii ◽  
Factor Viii Activity

Methods for the Concentration of Bovine Ac-Globulin (Factor V)

1961 ◽  
Vol 06 (03) ◽  
pp. 435-444 ◽  
Author(s):  
Ricardo H. Landaburu ◽  
Walter H. Seegers
Keyword(s):  
Room Temperature ◽  
Barium Carbonate ◽  
Deae Cellulose ◽  
Reducing Agents ◽  
Factor V ◽  
Isoelectric Precipitation ◽  
Stabilizing Agent ◽  
Bovine Plasma ◽  
The Stability

SummaryAn attempt was made to obtain Ac-globulin from bovine plasma. The concentrates contain mostly protein, and phosphorus is also present. The stability characteristics vary from one preparation to another, but in general there was no loss before 1 month in a deep freeze or before 1 week in an icebox, or before 5 hours at room temperature. Reducing agents destroy the activity rapidly. S-acetylmercaptosuccinic anhydride is an effective stabilizing agent. Greatest stability was at pH 6.0.In the purification bovine plasma is adsorbed with barium carbonate and diluted 6-fold with water. Protein is removed at pH 6.0 and the Ac-globulin is precipitated at pH 5.0. Rivanol and alcohol fractionation is followed by chromatography on Amberlite IRC-50 or DEAE-cellulose. The final product is obtained by isoelectric precipitation.

Use of Factor VIII in the Treatment of Hemophilia

1962 ◽  
Vol 07 (02) ◽  
pp. 230-238 ◽  
Author(s):  
A Pavlovsky ◽  
H Peterson ◽  
G Casillas ◽  
C Simonetti ◽  
A Martinez Canaveri ◽  
...  
Keyword(s):  
Factor Viii ◽  
Tannic Acid ◽  
Deae Cellulose ◽  
Fibrinolytic System ◽  
Purification Method ◽  
Fraction I

SummaryThis publication describes the results obtained by treatment of haemophiliacs with factor VIII preparations isolated from Cohn fraction I by use of tannic acid, FI-O-Ta.The authors stress the rapidity of the disappearance of factor VIII after injection. Transfusions are generally well tolerated. One reaction of the pyrogen type has been observed and also a case of activation of the fibrinolytic system.A second purification method by means of chromatography on DEAE-cellulose is described.

Heparin, Protamine and Polybrene

1965 ◽  
Vol 13 (02) ◽  
pp. 550-560 ◽  
Author(s):  
Anthony Britten
Keyword(s):  
Factor Viii ◽  
Incubation Mixture ◽  
Factor V ◽  
Factor Viii Activity ◽  
Rapid Loss

SummaryThe effects of incubating heparin, protamine or Polybrene with plasma were studied. All three drugs cause rapid loss of factor V from decalcified plasma, while Polybrene also accelerates the loss of factor VIII activity. These changes are related to temperature, the period of incubation and the dose of the drug used, and can be partially prevented by inclusion of neutralizing doses of the appropriate antagonist in the incubation mixture.The implications of these findings are discussed.

The Immunological Properties of Factor VIII

1966 ◽  
Vol 16 (03/04) ◽  
pp. 559-573 ◽  
Author(s):  
L Uszyński
Keyword(s):  
Factor Viii ◽  
Inhibitory Activity ◽  
Hemophilia A ◽  
Severe Form ◽  
Molecular Defect ◽  
Bovine Plasma ◽  
Human Factor Viii ◽  
Antigenic Properties ◽  
Coagulation Tests ◽  
Normal Human

SummaryRabbits immunized against human AHG fibrinogen-free preparations, were shown to produce anti-AHG antibodies. The inhibitory activity of these antibodies was tested by thromboplastin generation test, thrombelastography, and the specific anti-AHG antibodies neutralization test. The latter test permitted quantitative determination of antigenic form of factor VIII. The inhibitory activity of anti-FI-O-Ta serum resulted exclusively from the anti-AHG antibodies which in coagulation tests behaved like circulating anticoagulants directed against factor VIII.The anti-AHG antibodies were neutralizable by normal human serum or plasma even contained only trace of AHG activity after storage. There was no antigenic form of factor VIII in the severely affected patients with hemophilia A, von Willebrand’s disease nor in the normal plasma adsorbed on bentonite. The presented results suggest a molecular defect of factor VIII in patients with hemophilia A. The severe form of this disease depends, probably, on a major impairment of AHG biosynthesis, leading to changes in the antigenic properties of the molecule. The AHG from rabbit, porcine and bovine plasma respectively did not neutralize the anti-AHG antibodies formed in rabbits immunized against human factor VIII preparations.

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