Influence of Serum Incubation Conditions on Asolectin Thromboplastin Generation Results

1960 ◽  
Vol 4 (01) ◽  
pp. 083-092
Author(s):  
Arthur J. Seaman ◽  
Karen-Marie Karlsen
Keyword(s):  
Sodium Citrate ◽  
Normal Serum ◽  
Hemophilia B ◽  
Good Separation ◽  
Minimal Time ◽  
Serum Activity ◽  
Normal Populations ◽  
Soybean Phosphatide ◽  
Generation Test ◽  
Oxalated Plasma

SummaryThromboplastin generation in a system employing Asolectin, a soybean phosphatide, suspension as a platelet substitute is greatly influenced by the time and temperature of serum incubation.Reproducible results and good separation of populations of normal and PTC deficient sera were obtained in the asolectin TGT system when test sera were incubated for 1 hour at 20° C prior to the thromboplastin generation test. Longer time or higher temperatures of serum incubation resulted in progressively less reliable, separation of pathologic and normal populations in asolectin TGT systems than in fresh platelet TGT systems. Cephalin suspension was shown in a smaller number of tests to behave similarly to asolectin suspension when used as a platelet substitute in TGT systems.The 0.14 M sodium citrate eluate of citrated normal or Hemophilia B sera (incubated 1 hour at 37° C) had the ability to shorten the prolonged asolectin TGT minimal time of “over-incubated” normal sera to values obtained with the same sera in fresh platelet TGT systems. This correction, however, was probably non-specific since Hemophilia B sera results were sometimes similarly normalized. Citrated (but not oxalated) plasma incubated at 37° C for 24 hours, then recalcified to manufacture serum, and incubated an additional hour at 20° C showed no loss of the fresh 1 hour 20° C serum activity in asolectin thromboplastin generation, further suggesting that the thermolabile component of normal serum influencing asolectin TGT is a reaction product evolved during clotting.

Action of Erythrocyte Extract on the Prothrombin Consumption Test

1958 ◽  
Vol 194 (3) ◽  
pp. 527-530 ◽  
Author(s):  
Feliks Stanski ◽  
Armand J. Quick
Keyword(s):  
Whole Blood ◽  
Platelet Rich Plasma ◽  
Normal Serum ◽  
Human Erythrocytes ◽  
Hemophilia B ◽  
Clotting Factor ◽  
Serum Factor ◽  
Rabbit Plasma

The prothrombin consumption time for human and rabbit whole blood and for platelet-rich plasma is approximately the same. The addition of an extract of human erythrocytes to the blood or plasma before clotting increases prothrombin consumption significantly except in platelet-poor rabbit plasma. If a mixture of aged normal serum and an extract of erythrocytes is added to the latter, the consumption of prothrombin is greatly increased. This potentiating activity of normal aged serum is also present in aged hemophilic serum, but is diminished in the serum from a patient with hemophilia B (PTC deficiency). The serum factor is destroyed at 60°C and is adsorbed with Ca3(PO4)2. The findings suggest that the cofactor of erythrocytin, the clotting factor in the erythrocyte, may be related to the agent lacking in hemophilia B.

scholarly journals Circulating serum lectins of patients with IgA nephropathy stimulate IL-6 release from mesangial cells.

1997 ◽  
Vol 8 (2) ◽  
pp. 208-213
Author(s):  
C Libetta ◽  
T Rampino ◽  
G Palumbo ◽  
C Esposito ◽  
A Dal Canton
Keyword(s):  
Affinity Chromatography ◽  
Iga Nephropathy ◽  
Mesangial Cells ◽  
Lectin Binding ◽  
Normal Serum ◽  
In Vitro Studies ◽  
Human Mesangial Cells ◽  
Serum Activity ◽  
Peripheral Leukocytes

Previously, the authors reported that the serum of patients with immunoglobulin (Ig) A nephropathy stimulated peripheral leukocytes, and this effect was inhibited by nominal haptens for lectins. In vitro studies have shown that lectins can bind to rat mesangial cells and cause their activation. This study was performed to investigate whether the serum of IgA nephropathy patients contains lectins that activate mesangial cells, i.e., induce release of interleukin (IL)-6, a nephritogenic cytokine. The serum of patients was adsorbed by affinity chromatography on resins loaded with lectin-binding sugars. After adsorption, serum supernatant was collected and the resins were then eluted. Human mesangial cells were conditioned with native serum, post-adsorption supernatant, and eluate (all three at 10%) for 24 h, and the release of IL-6 was determined by ELISA. Normal serum was used as control. Incubation of mesangial cells with IgA nephropathy patients serum raised average IL-6 release from 8.5 pg/mL to 274.1 pg/mL. Adsorption in beta-D-glucose and N-acetyl-D-glucosamine caused a fall in the activity of patients' serum, to 17.0 and 63.7 pg/mL, respectively, and the activity lost was recovered in the eluate (185.2 and 142.7 pg/mL, respectively). Neither adsorption on N-acetyl-D-galactosamine nor on fucosylamine was associated with any effect on serum activity; accordingly, no activity was found in the eluates. Serum of patients with non-IgA mesangiocapillary nephritis did not stimulate mesangial cells. These results show that the serum of IgA nephropathy patients contains specific lectins that stimulate IL-6 nephropathy by mesangial cells and are, therefore, potential nephritogenic.

The Effect of Hirudin, a Potent Antithrombin, on the Thromboplastin Generation Test

1967 ◽  
Vol 17 (01/02) ◽  
pp. 256-263 ◽  
Author(s):  
M. J Caldwell
Keyword(s):  
Factor Viii ◽  
Inhibitory Effect ◽  
Normal Serum ◽  
Factor Ix ◽  
Thrombin Inhibitor ◽  
Generation Test ◽  
Dependent Interaction

SummaryThe presence of hirudin at the time adsorbed plasma and serum were combined in TGT inhibited the formation of the prothrombin converting activity. Hirudin, a potent antithrombin, probably exerted its inhibitory effect by neutralizing any thrombin which might form. If adsorbed plasma and serum were incubated together for a period as short as 1 to 3 min before the addition of hirudin, calcium and lipid, hirudin no longer exerted the same inhibitory effect, but, in order to exert the same inhibitory effect, had to be present in higher concentration. Preincubation of factor VIII deficient plasma with serum and calcium was not effective in overcoming hirudin inhibition, but preincubation with factor IX deficient serum with adsorbed plasma and calcium was able to overcome inhibition due to hirudin equally as well as normal serum.Once again the importance of traces of thrombin in the formation of activity capable of rapidly clotting substrate plasma in TGT has been shown by using a specific thrombin inhibitor. A thrombin dependent interaction appears to take place between adsorbed plasma and serum and also between product I and phospholipid.

Cryofibrinogen: formation and inhibition in heparinized plasma

1963 ◽  
Vol 204 (3) ◽  
pp. 419-422 ◽  
Author(s):  
R. A. Heinrich ◽  
E. C. Vonder ◽  
A. R. W. Climie
Keyword(s):  
Sodium Citrate ◽  
Heart Surgery ◽  
Soluble Fraction ◽  
Cell Mass ◽  
Sodium Oxalate ◽  
Heparin Concentration ◽  
Red Cell Mass ◽  
Considerable Problem ◽  
Cardiac Surgeon ◽  
Generation Test

Cold precipitable proteins (cryoglobulins, cryofibrinogen, and heparin-precipitable fractions) have presented a considerable problem to the clinician in treatment of patients who are discovered to have such fractions and to the cardiac surgeon who is confronted with clogged filters in heart surgery with hypothermia. When heparin, 0.04 mg/ml blood, is added as an anticoagulant, fibrin threads or clots may be observed in the red cell mass. The plasma of such blood, when exposed to temperatures of 5 C for 16 hr will show two precipitable fractions, one of which is soluble at 37 C whereas the other is not. As the heparin concentration is increased, there is a serial disappearance of each fraction. The heat-soluble fraction is the last to disappear, and this occurs when the heparin concentration reaches 3.0 mg/ml blood. Normal blood similarly treated with sodium oxalate or sodium citrate shows no cold precipitation. Normal plasma, after addition of thrombin, shows both cold-precipitable fractions. Twelve patients who had varying amounts of cryofibrinogen were studied by employing a modification of the thromboplastin generation test. Eight of the twelve patients showed a hypercoagulable state.

The blood coagulation mechanisms in thrombocytopenic blood

1957 ◽  
Vol 01 (03/04) ◽  
pp. 433-444 ◽  
Author(s):  
Shirley A. Johnson ◽  
M. June Caldwell ◽  
Raymond W. Monto
Keyword(s):  
Blood Coagulation ◽  
Sodium Citrate ◽  
Normal Serum ◽  
Normal Blood ◽  
Blood Clots ◽  
Consumption Value

SummaryThrombocytopenic serum supports a short prothrombin consumption time, however, the same serum contains as little prothrombin as normal serum. The prothrombin consumption test of thrombocytopenic serum can be prolonged by adsorption on BaCO3 or reduced again by the addition of the sodium citrate eluate. The antihemophilic activity of thrombocytopenic blood disappears when the blood clots as in normal blood and autoprothrombin I is present in much smaller amounts than in normal serum. The authors suggest that a new factor, a possible derivative of prothrombin, is responsible for the short prothrombin consumption value in thrombocytopenic blood.

scholarly journals Study of the Normal Serum Activity Sensitizing Stabilized Fibrin to Urea Dispersion by Sephadex G. 200 Chromatography

1977 ◽  
Author(s):  
F. de Cataldo ◽  
F. Baudo ◽  
E. Mussini
Keyword(s):  
Ammonium Sulphate ◽  
Normal Serum ◽  
Final Concentration ◽  
Distilled Water ◽  
Platelet Poor Plasma ◽  
Serum Activity ◽  
Two Peaks ◽  
Phosphate Buffered Saline ◽  
Stabilized Fibrin ◽  
Second Peak

Clots from human normal native platelet-poor plasma are dispersed by the addition of urea at a final concentration of 2.5 M. The same clots, washed in buffered saline (pH 7.4), are not dispersed by urea, but are rendered susceptible to its dispersing action by prior incubation in normal undiluted plasma and serum or 33% ammonium sulphate fractions. The serum fraction was dissolved in and dialyzed against pH. 7.4 phosphate-buffered saline and analyzed by Sephadex G. 200 chromatography. Two peaks were obtained; the relative materials were dialyzed against pH. 7.4 phosphate-buffered distilled water, lyophilized and dissolved in saline. The activity sensitizing the stabilized fibrin to urea dispersion was recovered in the second peak material.

scholarly journals A LABILE COMPONENT OF NORMAL SERUM WHICH COMBINES WITH VARIOUS VIRUSES. NEUTRALIZATION OF INFECTIVITY AND INHIBITION OF HEMAGGLUTINATION BY THE COMPONENT

1949 ◽  
Vol 90 (5) ◽  
pp. 475-495 ◽  
Author(s):  
Harold S. Ginsberg ◽  
Frank L. Horsfall
Keyword(s):  
Calcium Ions ◽  
Newcastle Disease ◽  
Guinea Pigs ◽  
Influenza A ◽  
Sodium Citrate ◽  
Normal Serum ◽  
Mouse Serum ◽  
Human Beings ◽  
Serum Component ◽  
Labile Component

A labile component present in the serum of human beings, guinea pigs, and rabbits neutralizes the infectivity of mumps, Newcastle disease, influenza A and B viruses. The labile component of these sera and of mouse serum also inhibits hemagglutination of chicken RBC by these viruses. The component is inactivated by heating at 56°C. for 30 minutes and upon storage at 4°C. for periods longer than 2 weeks. The virus-neutralizing and hemagglutination-inhibiting properties result from serum component-virus combination in the presence of calcium. The combination is stable, and does not undergo spontaneous dissociation. Partial separation of virus can be brought about by heating mixtures held for 24 hours or by removal of calcium ions with sodium citrate. The labile serum component appears to be distinct from hemolytic complement.

Observations on Factor VIII (Antihaemophilic A Factor) Stability in Rabbit and Rat Plasma Estimation of Factor VIII in Rat Blood by a New Method

1965 ◽  
Vol 14 (03/04) ◽  
pp. 374-386 ◽  
Author(s):  
D Nilsson ◽  
B. A Waaler
Keyword(s):  
Factor Viii ◽  
Animal Species ◽  
Rat Plasma ◽  
Test System ◽  
Test Material ◽  
Rabbit Plasma ◽  
Factor Viii Activity ◽  
Generation Test ◽  
Rat Tail ◽  
Oxalated Plasma

Summary1. Factor VIII activity of rat and rabbit plasma can be measured with a modification of the thromboplastin generation test.2. Using this method it can be shown that factor VIII activity is unstable on storage and on repeated freezings and thawings in oxalated plasma from the two animal species. The factor VIII activity is especially unstable in oxalated rat plasma, from which it disappears almost completely by four or more repeated freezings and thawings.3. A rat oxalated plasma, which had been frozen and thawed ten times, could be used as a factor VIII-free reagent in a simple one-stage test system for measurements of this factor in rat or rabbit plasma.4. The method developed may be used with rat tail capillary blood as test material, making repeated observations on rats possible. The use of the method is discussed.

A “New” Family with Stuart-Prower Deficiency

1959 ◽  
Vol 03 (01) ◽  
pp. 059-076 ◽  
Author(s):  
J Roos ◽  
C van Arkel ◽  
M. C Verloop ◽  
F. L. J Jordan
Keyword(s):  
Normal Serum ◽  
One Stage ◽  
New Family ◽  
Haemorrhagic Diathesis ◽  
Generation Test ◽  
Mode Of Inheritance

SummaryA description is given of the coagulation disturbances in six patients with a haemorrhagic diathesis due to Stuart-Prower deficiency. Relatives of these patients, if heterozygous for this deficiency, showed no significant haemorrhagic diathesis; in the laboratory, their one-stage “prothrombin” times showed only a slight prolongation. Thromboplastin formation in these relatives was sufficient, but determination of the Stuart-Prower factor revealed lower values. The relatives in question were incapable of giving the same correction of the thromboplastin generation test in their bleeder relatives as normal serum does.The relation between Stuart-Prower deficiency and Christmas factor is discussed.One patient is described who combined heterozygotism for Stuart-Prower deficiency with alcaptonuria.The mode of inheritance of the Stuart-Prower deficiency is discussed.

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