bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Application of Urikinase to Nephropathy. (Report of Group Study)
Authors: Abe T.
Published: May 2019

Coagulation and fibrinolysis system have been imagined to have some pathogenic influence on various kinds of nephropathy. Our study group on the clinical effect of urokinase (UK) administration to nephropathies applied this material to 106 patients with acute and chronic glomerulonephritis, nephrotic syndreome with different causes and renal failure together with 25 control subjects. From 120×104 to 900×104 units of UK was given in ten to twenty-two days solely or together with corticosteroid, dipyridamole, Clofibrate, furosemide, spironolactone, -methyl-dopa, indomethacine, cyclophosphamide, azathiopurine and so on. Heparin was also employed to some control or comparison cases in daily doses of five to fifteen thousand units.

Daily amounts of urine, urinary protein, phenolsulphon-phthalein excretion, creatinine clearance were found responsive to UK administration and their beneficial changes were realized statistically in cases of chronic glomerulonephritis (22 cases in 47), rapidly progressive glomerulonephritis (2 in 5), nephrotic syndrome (5 in 35) and some others showed only decrease of urinary protein. As for the cases of renal failure, no definite coclusion was drawn. Only a few cases gave some clues of side effect such as headache, nausea and aggravated hematuria. Histological classification was also tried to find that the proliferative type was most responsive to this treatment.
The Levels of Fibrin Stabilizing Factor Activity in the Course of Myocardial Infarcation
Authors: Morishita Takeshi , Shindo Toru , Hasegawa Susumu , Hirai Junichi , Suzuki Shinichiro , Amanuma Mitsuru , Koizumi Nichizo , Koya Niroku , Shirai Tatsuo
Published: May 2019

It is known that coronary thrombosis is found in most cases of myocardial infarction. There is surely relationship between the infarction and thrombosis. Platelet aggregation and fibrin formation mechanism are of course concerned with the thrombosis, but the purpose of our research is to know how fibrin stabilizing factor is involved in that. From this respect, we observed the behavior of F.S.F. in myocardial infarction. The typical changing pattern of plasma F.S.F. in the myocardial infarction is as follows; it holds high level just after the onset, rapidly decreases right after, goes down to the minimum at 5th - 7th day, gradually increases thereafter, returns to normal level at about 14th day. This profile was almost same as that in case of experimental myocardial infarction in dogs and was very specific in heart tissue, but was not observed in the control surgical experiment, and in the patients with brain bleeding and thrombosis. The experiment of Master’s two-step test to see the effect on plasma F.S.F. level gave an interesting result. The tests of about 100 normal plasma showed the age dependency which means the increasing F.S.F. concentration in plasma related on age. The mean of F.S.F. in adult was 21±3 (μM MDC/10 min).
Effect of Mini-Dose of Chlorpromazine on the Bleeding time and Platelet Aggregation
Authors: Zahavi J. , Schwartz G.
Published: May 2019

The impairment, of hemostasis by ehlorpromazine (c) at concentrations below those which inhibit platelet release reaction, has been evaluated in 24 normal subjects. In each individual, modidied Ivy bleeding time and platelet aggregations (PAg) induced by adenosine diphosphate (ADP) 0,6 MM and 4 MM collagen 0,003% and 1-epinephrine 3MM well as platelet serotonin concentration, were performed prior to and 20 min. after intramuscular administration of (C) or saline. Normal saline was injected to 6 subjects and 5, 12,5 and 20 mg. of (c) to another 8, 5 and 5 subjects respectively. There was a slight but significant prolongation of the bleeding time and a decrease in the initial wave of aggregation to ADP 0,6 MM in the subjects on (C) compared to those on-saline. Platelet serotonin, however, and (PAg) to collagen, 1-epinephrine and ADP 4 MM, were the same in both groups. The impaired hemostasis by mini-dose of (C) might be attributed to the prevention of leakage of ADP from intact erythrocytes in-vivo and, if so, may signify an important new application of this drug in the treatment of thrombotic disorders.
The in Vitro Bleeding Time
Authors: Blakely J.A. , Glynn M. F. X. , Prchal J.
Published: May 2019

The observation by Didisheim of platelet-fibrin hemostatic plugs occluding puncture sites in teflon AV shunts prompted an attempt to produce similar plugs in vitro using a pump to circulate shed blood. Various anticoagulants, tubing types, flow rates, pressures and methods of producing suitable holes have been investigated. Satisfactory results have been obtained with drilled holes 0.013 - 0.018” diameter in tygon or teflon tubing using heparinized or native, but not EDTA or citrated whole blood. Holes are occluded by platelet fibrin thrombus. Red cells are necessary for plugs to form but are not a component of the plug. Time required for plugs to form correlate with the template bleeding time and are prolonged by thrombocytopenia, aspirin administration, and Von Willebrand’s disease. Possible applications are being explored.
Lacking Schistocytosis in Experimental Disseminated Intravascular Coagulation after Defibrination by Plasmin
Authors: Heves H. , Slijepcevic B.
Published: May 2019

A disseminated intravascular coagulation (DIC) was induced in rats by infusion of a subthreshold dose of thrombin after producing a hypercoagulable state by i.v, injection of ellagic acid. After pretreatment with a 90 min.-infusion of 50 cas.-u. of plasmin (= plasmin group) or normal saline (= control group) 20 mg AMCHA were applied i.v. for stopping fibrinolysis. Blood parameters of the control group revealed a typical constellation for DIC; additionally, it was observed a highly accellerated 131-I-fibrinogen cata-bolism and an accumulation of 131-I-radioactivity in lungs and kidneys accompanied by haemoglobinaemia and schistocytosis. In spite of the previous defibrination by plasmin, the plasmin group showed partly similar results; the deposition of radioactivity was not so impressive, though plasma haemoglobin levels were neither increased nor schistocytosis observed.

Considering the complexity of the presented experimental model, these results show that red cells are not fragmented to schistocytes in absence of fibrinogen because of missing fibrin strands, which probably cause the destruction of red cells in the small blood vessels.
Inhibitory Effect of Aprotinin on the Activation Process of Contact Factors and Platelet Aggregation
Authors: Yamaguchi I. , Yano Y. , Kitahara T. , Yatabe M. , Ukita M. , Fujimaki M. , Fukutake K.
Published: May 2019

It has been reported by several investigators that aprotinin demonstrates inhibitory action on the process of intrinsic blood thromboplastin generation. In this paper the authors will present the facts that on the hydrolytic action of plasma kallikrein, and on the activation process of plasma prekallikrein as well, the inhibitory action of aprotinin is indicated by the use of chromozym PK assay and the latter process might be caused by the inhibition of factor XII activated with Kaolin, and as to platelet aggregation induced by thrombin, aprotinin also plays the inhibitory effect which is more sensitive than the antithrombin activity of aprotinin measured by thrombin clotting time. However, the inhibitory effect of aprotinin on platelets might be caused not only by antithrombic action of this enzyme inhibitor, but also other influence of the inhibitor on release mechanism of the platelets, because epinephrine induced platelet aggregation can be disturbed by the addition of this inhibitor. In summary it mighy be suggested in this study that the clinical use of aprotinin preparation might be one way for anticoagulant thrapy on thrombotic deseases associated with intrinsic hypercoagulability of plasma factors and platelets.
Fibrinolytic Activity of Gel Filtered Platelet
Authors: Watada M. , Nakagawa M. , Nishizawa A. , Ijichi H.
Published: May 2019

Too many factors in the plasma has made it complicated to investigate the functions of platelets, and gel filtration of the platelet rich plasma was proved to be the beneficial method to separate platelets undamaged from plasma protein. Avoiding the influences of the surrounding plasma factors, this gel filtered platelets (GFP) were considered to be applicable for the various platelet research. Using this GFP, the fibrinolytic activities were measured by means of the plasminogen free fibrin plates and of the caseinolytic techniques on the separated platelets and disrupted platelets. The inhibitors like alpha-1 antitrypsin or alpha-2 macroglobulin were also assayed. The lytic area on the fibrin plate was observed when GFP was placed while platelet rich plasma did not lyze, however the lytic area was increased in diameter when GFP was placed with plasmin, comparing with plasmin or GFP alone. In case of the ultra sonically disrupted GFP, it showed the greater lytic area. Caseinolytic activity of GFP supported also the above mentioned results. These observation proved that the platelet has fibrinolytic activity which seems to be masked by inhibitors in the plasma circumstance.
Fibrinolysis in the Upper Gastrointestinal Tract
Authors: Low J. , Dodds A.J. , Biggs J.C.
Published: May 2019

Local fibrinolytic activity can contribute towards the continuation of haemorrhage from many tissues. Fibrinolysis has been documented in the gastro-duodenal mucosa, gastric venous drainage and gastric secretions and a recent double blind trial indicates a beneficial effect of a fibrinolytic inhibitor in patients with gastrointestinal haemorrhage. In the present study, the fibrinolytic role of acid, pepsin, trypsin, chymotrypsin and other components of the gastro-duodenal and biliary secretions has been studied in control subjects and patients with a variety of diseases using a labelled clot lysis method. The effect of inhibitors and pH changes on the various systems has been assessed. In addition, the fibrin degradation products have been followed by acrylamide gel electrophoresis. In general, gastrointestinal fibrinolysis can be inhibited by pH changes or by various inhibitors and this may have therapeutic implications.
Effects of Suloctidil on Circulating Platelet Aggregates Formation in Rats
Authors: Roba J. , Claeys M. , Lambelin G.
Published: May 2019

In a search for new models allowing testing of antithrombotic agent in vivo, we have adapted the Wu and Hoak technique for measuring circulating platelet aggregates in rats. The pattern of reversible and irreversible platelet aggregates is studied by sampling blood in vena cava and/or orbital sinus following a) i.v. ADP injection; b) implantation of an aortic catheter; c) adjuvant induced arthritis.

Suloctidil (p.o.) can prevent formation of circulating aggregates. Its effects are compared with those of other antiplatelet agents.
Measuring Antithrombin III with a Chromogenic Substrate(Chromozym-th): Which Buffer System?
Authors: Troger N. , Spycher P. , Furlan M. , Beck E.A.
Published: May 2019

Synthetic oligopeptides, coupled with a chromophore, may be used for measurement of thrombin as well as of thrombin inhibitors. This principle was applied to a recently synthesised tripeptide derivative (Tos-Gly-Pro-Arg-PNA, commercial designation: Chromozym-Th). Following partial activation of antithrombin III with heparin we found a marked dependence of the activation kinetics on some common buffer systems. At constant pH (8.2), molarity (0.15) and heparin concentration (0.5 NIH units/ml), thrombin inhibition was as follows: Tris > glycine > imidazole >> triethanol-amine. Following full activation of antithrombin III with excess amounts of heparin these differences were less apparent. Since partial activation of antithrombin III might also be expressed as biologic activity of heparin, careful attention should be given to the selection of an appropriate buffer system before such tests are introduced into the clinical laboratory.
Platelet Function Changes During Hemodialysis
Authors: Levin Robert D. , Ivanovich Peter , Kwaan Hau C.
Published: May 2019

Platelet function during hemodialysis was studied in 27 patients during 62 dialyses on a Cor-dis-Dow Hollow Fiber Artificial Kidney CDAK-4 with anticoagulation by porcine mucosal heparin. Platelet levels fell from 2.08 + 0.52 × 109/1 (Mean ± S.D.) prior to dialysis to 1.74 ± 0.49 × 109/1 after 1/2 hour of dialysis (P<0.01), and remained at depressed levels throughout the dialysis period. Platelet adhesiveness in arterial blood did not significantly vary during dialysis. By aggregometry, the fraction of aggregable platelets decreased from 69 ± 13% prior to dialysis to 51 ± 15% after 1/2 hour (P<0.001) and afterwards remained depressed. The ratio of aggregation after 1.2 /μM ADP to that after 4.7 /μM ADP elevated from 87 ± 15% prior to dialysis to 93 ± 7% after 1/2 hour (P<0.05). Platelets from the venous return limb of the hemodialyzer were poorly aggregable to 50/aM ADP or 10 /iM epinephrine.

The findings are compatible with an enhanced reactivity of platelets and the concomitant adherence of aggregable platelets upon the dialyzer fibers, leaving an increased fraction of poorly functioning platelets in the circulation. The enhanced reactivity is apparently induced by heparin, increased levels of ionized calcium, and other factors.
Absorption of Different Salts of Heparin Following Subcutaneous Injection
Authors: Cella G. , Lane D.A. , MacGregor I. , Kakkar V.V.
Published: May 2019

Two recent studies (1,2) have demonstrated that the MW of heparin and nature of the inorganic ion associated with it influence the heparin level in blood following subcutaneous injection. In the present study we have studied the absorption of heparins containing different amounts of Na and Ca and yet all having similar MW’s when characterized by gel filtration upon a polyacrylamide-agarose matrix (3). Four heparins from the same source material containing approximately 11% Na, 2.5% Ca, 7%, 10.5% Ca together with two Na salts and two Ca salts that are commercially available have been studied in a double-blind crossover trial in six normal volunteers. It was found that mean heparn levels circulating 1, 3, 5 and 7 hours post injection fell as the% of Ca associated with the heparins increased. The differences between the mean levels obtained following injection of the Na salts and the 10-11% Ca salts were highly significant at 1, 3 and 5 hours, while differences between Na salt and 7.3% Ca were significant only at 1 hour post injection. In one individual with small amounts of subcutaneous tissue inconsistent results were obtained, with two Ca salts (7.3% and 11%) producing twice the levels obtained following subcutaneous injection of the Na salts and another Ca salt (10.5%) producing characteristically low levels.

1. Johnson, E.A. et al. Thrombosis and Haemostasis 35: 586, 1976.

2. Thomas, D.P. et al. Thrombosis Research 9: 241, 1976.

3. Johnson, E.A. and Mulloy, B. Carbohydrate Research 51: 119, 1976.
Effect of Drugs and Membrane Poisons on Platelet Adhesion to a Metal
Authors: Lucas T. , Bonavita F. , Ramasamy N. , Sawyer P.N.
Published: May 2019

The effect of anticoagulants and membrane poisons on the adhesion of human platelets to a platinum electrode was studied in an in-vitro electrochemical optical test system. The potential of the electrode was varied from -600 mv vs NHE to +600 mv vs NHE in 100 mv steps. Platelets were resuspended in phosphate buffered saline. The cell concentration was 3 × 105 cells/ml. Experiments were conducted at 22°C. Thirty samples were examined. The number of platelets adhering to the electrode at each potential was observed and indirect ADP assays were conducted. The results were the following; (i) anodic potentials equivalent to injury potentials+200mv vs NHE enhanced platelet adhesion, (ii) Heparin, ASA and 3, 5 dichloro-salicylic acid reduced platelet adhesion, (iii) protamine sulfate and chlorpromazine enhanced adhesion, (iv) sulfhydryl (SH) blocking agents (mercuric acetate and mercuric benzoate) reduced platelet adhesion 65%, (v) iodoacetic acid (carboxyl and thiol group reducing agents) reduced platelet adhesion by 987o. The simple test system can be employed to evaluate the physical-chemical nature of platelet recognition and adhesion to a prosthetic surface. Based on this study platelet adhesion to a metallic surface with known electrochemical characteristics appears to be mediated by a number of surface reactive groups. Alteration or neutralization of these active groups leads to platelet inactivation and blocks adhesion.
A Colourimetric Technique for Monitoring Variations in the Composition of F VIII Reagents
Authors: Seghatchian M.J. , Miller-Andersson M.
Published: May 2019

An inter-laboratory estimation of F VIII potency has shown that the reagent was the main source of variation. The complicated nature of the F VIII reagent dictates the “plateau of the optimal activation” to be determined experimentally. Thus it is essential to design a direct method to surmount problems due to the reagent variability and stability. S2222 is a highly sensitive substrate for F Xthat has been successfully used instead of plasma substrate for measuring F generation. The rate of cleavage was found to be strongly influenced by small changes in the volume of serum and phospholipid. This allows the possibility of “tuning” the reactants to obtain a specific rate of reaction. The optimal F Xa activation also depend upon the form in which the functionary active reactants (Ca2+, phospholipid, serum dilution) are added. Some commercially combined reagents gave very poor rate of cleavage and lack of slope for various dilutions of reference plasma. This makes the results invalid. The colourimetric test using S2222 is quick and sensitive. The resultsare reproducible. Thus the method fulfils the requirements for an acceptable test for monitoring and standardizing the composition of factor VIII reagents.
The Application of the Pyrimidopyrimidine RA 233 for Platelet Transfusion
Authors: Reuter H. , Borberq H. , Linker H.
Published: May 2019

As an alternative for the buffy-coat preparation of platelets a combination of the continuous flow centrifugation with the single batch processing technique was examined. The use of the membrane stabilizing agent RA 233 has shown this procedure to be sufficiently useful for clinical application. The use of RA 233 made available platelet yields of about platelets per blood donor. The drug was given in an amount of 100 mg i.v. to the blood donor and added in doses of 80 mg to each 500 ml plasma collection bag containing 10% ACD-A solution as an anticoagulant. From the platelet rich plasma the platelets were obtained by further concentration in a Christ IV KS blood bank centrifuge with 2.600 × g. As could be shown by transfusion of RA 233 prepared platelets to leukemic patients the drug is reversibly bound to the platelet membrane. While during preparation platelet adhesion and aggregation are slightly inhibited by RA 233, the functional activity of the platelets is regained after transfusion. The survival of transfused 51Cr-labeled platelets prepared in the presence of RA 233 was found to be in the normal range.
Kinetic Studies of Fibrinogen in Pregnant Rabbits
Authors: Moeller R.-M. , Mahn I. , Müller-Berqhaus G.
Published: May 2019

During gestation increased concentrations of fibrin (ogen)derivatives are observed indicative of intravascular action of thrombin. The aim of this study was to elucidate the kinetics of fibrinogen during gestation. The elimination of homologous 1-125-fibrinogen was studied in 14 pregnant rabbits during the first as well as during the third trimesters of gestation. Control studies were performed with 10 non-pregnant rabbits.

The mean distribution volume of labeled fibrinogen did not significantly differ between pregnant and non-pregnant rabbits. During the third trimester pregnant rabbits demonstrated a pronounced shortening of T 1/2 of labeled fibrinogen from a mean of 55.3 hr during the first to a mean of 29.7 hr during the third trimester. The experiments showed a significant increase in the fractional catabolic rate from 45.0 to 69.9% per day in the course of gestation. The shortening of T 1/2 of labeled fibrinogen correlated to the number of fetusses per litter.

This study indicates an average acceleration of the fibrinogen turnover during gestation of about 50%. These direct quantitative measurements demonstrate that fibrinogen catabolism is pronouncedly accelerated during pregnancy.
Measurement of the Electrical Impedance of Blood During Clotting
Authors: Theiss W. , Ulmer A.
Published: May 2019

Characteristicchanges of the electrical impedance are observed during blood clotting, the measurement of which can be used as a screening test for hemorrhagic diatheses (L. Arnhold, W. Theiss, J. Tiemann: Deutsche Medizinische Wochenschrift 100:1657-1663, 1975). Large scale use of the test was hampered by the technically delicate experimental set-up of a matched pair of precision measuring cells arranged in a bridge circuit and by the necessity to perform measurements immediately on native blood samples.

Comparative measurements on blood samples of 30 healthy volunteers in a conventional bridge circuit with a pair of measuring cells and in a modified circuit with a single measuring cell revealed good results also with the latter method. Experiments with recalcification of citrated whole blood showed satisfactory correlation with measurements carried out on native whole blood.

This newly developped straight forward procedure of measuring in a single cell and the possibility to work with citrated blood are two major advances in making the measurement of the electrical impedance of clotting blood amenable for use on a larger scale in further research work and eventual routine use.
Structural Features in “Spontaneous” Focal Endothelial Cell Desquamation of Rabbit Aorta
Authors: Svendsen E. , Jørgensen L.
Published: May 2019

Rabbit aortas were examined by light, scanning and transmission electron microscopy to observe the morphological steps in focal endothelial cell desquamation. The aortas were not subjected to any instrumentation prior to fixation by either immersion or perfusion. In areas where boundary layer separation of flow with eddy formation is known to occur, many elongated protruding injured cells were observed, some obviously partly loosened. Some of the latter cells appeared to be suspended in two ends and were twisted longitudinally. Single cells, or even sheets of cells, were completely detached; the breaks seemed to have taken place close to, and parallel with the intercellular junctions, but not within these structures. Platelets had reacted both with injured cells and the denuded intimai surface. Thus, the sequence appears to be protrusion of cell body, partly loosening of cells through breaks near the periphery, twisting of cells, and complete detachment.
Inhibitory Effect of Diamines and Polyamines on Human Platelet Aggregation and [14C]-Serotonin Release Reaction
Authors: Subbarao K. , Forestier F.
Published: May 2019

Physiological diamines and polyamines occur in high concentrations in various parts of animal tissues. These amines are known to interact with and stabilize nucleic acids, membranes and ribosomes (Tabor and Tabor, Pharmac. Rev., 16, 245). The effect of putrescine, cadaverine, spermidine and spermine on platelet function is not yet fully explored. We studied the effect of these reagents on in vitro aggregation of human platelet rich plasma (PRP) induced by the addition of ADP, thrombin, collagen and serotonin. Cadaverine, spermidine and spermine at concentrations from 2-5 μM strongly inhibited the aggregation of platelets and the [14C]-serotonin release reaction induced by ADP and thrombin in a concentration dependent manner, but did not show any effect on aggregation induced by other agents. Putrescine, on the other hand, failed to produce any effect on the aggregation of platelets and [14C]-serotonin release reaction. Studies on the binding of purified human thrombin treated with [14C]-diisopropylfluoro-phosphate (DFP) to washed human platelets indicated that cadaverine (1-5 μmoles) increased the binding of total [14C]-DFP-thrombin to platelets by 30%. The data suggest that the alteration of platelet function by diamines and polyamines was probably achieved by their binding to platelet membranes.
Two Dimensional Immunoelectrophoretic Patterns of Alpha2 Macroglobulin in Subjects with and Without Thrombotic Tendencies
Authors: Stevenson Mabel M. , Davidson Ann G. , Tarnay Thomas J.
Published: May 2019

In an attempt to determine whether compensatory changes in alpha2 macroglobulin occur in patients with thrombotic tendencies, studies were performed by the two dimensional immunoelec-trophoretic technique of Laurell on the plasmas of healthy laboratory workers and patients with possible thrombotic tendencies. A single, broad, asymmetrical peak was noted in 16 of 18 laboratory workers. A slower moving discrete second spike of lesser magnitude was found in 2 of the 18. A pattern with two peaks appeared in 11 of 14 patients on warfarin therapy for recurrent thrombophlebitis (8 of 8), for prophylaxis after artificial heart valve replacement (3 of 3), and coronary revascularization (1 of 3). Antithrombin III patterns studied according to Sas were normal in all but 2 of these patients. A volunteer ingesting warfarin was normal. A single patient with liver disease showed two peaks.

Preliminary in vitro incubation of plasma with thrombin, plasmin, or streptokinase failed to create more than one peak.

These preliminary data suggest changes in alpha2 macroglobulin may represent in vivo coagulation and might provide another means to identify thrombosis prone individuals.
Serum Antithrombin III in Cerebrovascular Stroke
Authors: Takamatsu S. , Sakuta S. , Henmi K. , Tamada Y. , Mizuno S.
Published: May 2019

The purpose of this study is to evaluate the significance of serum antithrombin ILE (AT III) in acute cerebrovascular stroke. AT III levies were determined by means of single radial immunodiffusion method in 73 serum specimens of 19 patients with cerebral hemorrhage (CH.), 129 of 24 patients with cerebral infarction (C.I.) and 82 of 82 healthy controls. AT III levels of patients with C.H. and with C.I. on the 1st day of the attacks were significantly higher than the value of controls. The high levels maintained from the 1st to the 25th day exculding 7th and 8th days in hemorrhagic patients. In patients with C.I., the levels from the 6th to the 25 day were significantly higher than that from the 1st to the 5th day. No difference was found in the types of the diseases. In 150 serum specimens obtained from the 1st to the 7th day, AT III levels were significantly lower in the patients with high fever, disturbance of consciousness and abnormal ECG findings than those in patients without these clinical findings. In serum specimens obtained from the 5th to the 7th day, AT III level of survivals were significantly higher than that of the dead.AT III levels in patients with high FDP levels were higher than those in patients with normal FDP levels. In C.I., AT III levels were directly proportional to prothormbin and plasminogen levels, and were inversely proportional to α1-antitrypsin levels. It is supposed that high AT III levels in acute stroke patients are caused by the protective effects on excess thrombus formation occurring parimarily in C.I. and secondarily in C.H. The high levels in both types of strokes and the close relation to outcome of patients indicate usefulness in the clinical evaluation of patient’s prognosis.
Thrombogenic Characteristics of Activated and Standard Concentrates of Prothrombin Complex
Authors: Penner J.A. , Leach K. , Rohwedder J.
Published: May 2019

The composition of a variety of prothrombin complex concentrates(PCs), including an “activated” product, was examined to determine thrombogenic potential. Small amounts of Xa activity were found to be presentin all products surveyed, although much greater quantities were present in the activated one. Valuesfor prothrombin factors VII, IX, and X activity varied over a limited range, from one productto another, and small quantities of factors XI and XII were identified in several concentrates. Animal recipients of each product, receiving a dose of 100 u/kg developed intravascular coagulation. Prior incubation of the products with Antithrombin III to eliminate activated factors did not alter the coagulopathy pattern.

It was concluded that the presence of factor Xa was not responsible for the observed intravascular coagulation syndrome, and that other factors, such as increased procoagulant levels in the circulation, or the presence of other prothrombin intermediates must be involved.
Session: Hemophilia Therapy: A Challenge to the World Blood Resources” Topic: “How can the Problem be Solved? Is Blood a World Resource?”
Authors: Bidwell E.
Published: May 2019

The production of factor VIII is only one of many interdependent commitments of the Blood Transfusion Service, and is unlikely to be the final determining factor in limiting the total commitment. The real challenge of Haemophilia Therapy cannot be met by any amount of effort by plasma fractionators - it lies in the gaining and retaining of the willing co-operation of the volunteer donor, and of the ordinary citizen who directly or indirectly supports the haemophiliac by his contribution to national wealth. There must be a comprehensive programme to utilise human blood and blood products in which haemophilia therapy is only a part. I do not think this challenge can be met by regarding blood as a World Resource. It is a Community or National resource. The practice of moving surplus blood or blood products from one affluent country to another is of doubtful wisdom, in the long term, for either participant.
The Interaction of the Platelet and von Willebrand Factor
Authors: Fass D.N. , Booyse F. , Lewis J.C. , Bowie E. J. W.
Published: May 2019

A culture of pig aortic endothelial cells was used for experiments to investigate the interaction between the platelet and von Willebrand factor. An antibody was raised in rabbits to purified porcine von Willebrand factor. A semi-confluent culture of pig endothelial cells was stained immunofluorescently by the sandwich technique using anti-Willebrand factor IgG. An extensive extracellular meshwork of microfilaments was revealed. In endothelial cell cultures from von Willebrand pigs, no immunoreactive microfilaments were found. Immunoelectronmicro-scopy with peroxidase linked antibody has been used to identify similar filaments in normal pig endothelial cells. Washed platelets were shown to adhere to semiconfluent or damaged normal endothelial cell cultures. If the cultures had been previously incubated with anti-Willebrand factor IgG, the washed platelets did not adhere. There was no adherence of platelets when they were added to semiconfluent or damaged von Willebrand endothelial cells.
Platelets, Endothelial Cells and von Willebrand Factor
Authors: Ruggeri Z.M.
Published: May 2019

In plasma and platelets (PI) from 10 patients (pts) with “severe” recessive von Willebrand’s disease (vWD) factor VIII-related antigen (VIIIR:AG) was unmeasurable in 7 and extremely low in 3 using a very sensitive immunoradiometric assay (IRMA); ristocetin cofactor (VIIIR:RCo) was always unmeasurable and VIM:AG was not detectable on immunofluorescence on endothelial cells (EC) of the vessel wall. In 12 pts with “classical”, dominant vWD characterized by reduced plasma level of VIIIR:AG and VI I IR:RCo, normal values were found in PI and the mobility of PI VIIIR:AG on crossed Immunoelectrophoresis was not different from that of normal controls; VIIIRr AG was normal on EC. In 7 pts showing a faster electrophoretic mobility of plasma VIIIR:AG (variant: vWD), the same abnormality was found in PI. VIIIR:AG concentration was normal both in plasma and in PI when measured by electroimmunodiffusion, whereas IRMA gave lower values and VI I IR:RCo was decreased; VIIIR:AG was normal on EC. These findings show that “severe” vWD is the expression of a marked reduction of factor VIII synthesis fully expressed in EC, PI and plasma. In “classical” vWD the plasma defects are not reflected in the cellular compartments, suggesting a defective transfer from EC and PI to plasma. Patients with “variant” vWD are the expression of a qualitative alteration of the factor VIII molecule, functionally defective both in PI and in plasma.
Factor VIII Associated Antigen, Platelet Retention and Platelet Aggregation in Patients with Vascular Diseases
Authors: Scharrer I. , Pander U.
Published: May 2019

Factor VIII associated Antigen (F.VIII ASS AG) or Factor VIII related protein (FVIIIRP) was found in the vessel wall by several investi gators. Breddin and co-workers described enhanced platelet aggregation in patients with vascular diseases. Therefore we investigated the level of F. VII lassAG in patients with vascular diseases and the relation of it to platelet aggregation and platelet retention.. The following methods were used: Quantitative immunelectrophoresis in antiserum containing agarosegel according to Laurel 1, platelet retention test by application of the adeplat-S-pump system and platelet aggregation test III according to Breddin. We compared the results of 50 healthy persons and 53 patients with vascular diseases. We found a significant correlation of F.VIII assAG to the age of 50 healthy males. The Spearman rang correlation coefficient was r = 0.4188. We could also demonstrate a relation of the increased level of F.assAGs to different vascular diseases. The patients with myocardial infarction (18), with peripheral arterial occlusion (18), with cardiac valve replacement (9) and with peripheral venous thrombosis (8) showed a distinctly increased F.assAG. We found neither a correlation of F.VIIIassAG to the aggregation nor to the retention.
Mutant forms of Factor IX
Authors: Roberts H.R. , Chung K.S. , Goldsmith J.C.
Published: May 2019

Evidence for the heterogeneity of hemophilia B is based on the observation that differing abnormal Factor IX molecules can be isolated and characterized from certain patients with hemophilia B. The abnormal molecules, so far studied, are present in virtually normal amounts when measured by immunological techniques, but have reduced clotting activity. Other hemophilia B patients have reduced concentrations of antigenically detectable Factor IX as well as reduced clotting activity. Still other hemophilia B patients have no detectable Factor IX, either by immunological techniques or by functional assays.

The purpose of this report is to describe another kindred with two affected members who have an abnormal Factor IX molecule. The affected members have a bleeding disorder typical of moderate hemophilia B. However they have a normal prothrombin time (PT) and an only slightly prolonged partial thromboplastin time (PTT). The PTT becomes markedly prolonged when the lipid employed in the assay is diluted. Factor IX can be detected in the affected patients by immunological techniques and has recently been isolated by purification procedures involving, as a final step, heparin agarose affinity chromatography. This abnormal Factor IX, tentatively named Factor IXAlabama, differs from Factor IXBM (prolonged PT and PTT) and Factor IXChapel Hill (prolonged PTT and normal PT). Thus, the heterogeneous nature of Factor IX is being elucidated and clarified.
Results of Liver Biopsies, Rate of Icteric Hepatitis, and Frequency of Anti-HBs and Hbs-Antigen in Patients of the Heidelberg Hemophilia Center
Authors: Schimpf Kl. , Zimmermann K. , Rüdel J. , Thamer G. , Zeltsch P.
Published: May 2019

With the opening of the hemophilia center in Oct. 1972 an intensive replacement therapy with clotting factor concentrates was started the first time for many patients. Concerning the induced liver damages, three years later the following résumée could be drawn. From 113 examined patients 17(19%) acquired an icteric hepatitis(i.h.). Excluding 2 cases the interval between beginning of intensive clotting factor substitution and outbreak of i.h. was found to be 46 days in the average (13-117).1o 827 u (1 8oo-27 99o) of clotting factor concentrates had been injected until to this moment.(The average consumption by the 113 examined patients from Oct.1972 to Oct. 1975 was 57 186 u per patient and year.) In Oct.197 5 the distribution of Anti-HBs and HBs-Ag positive patients(radio immune assay) was: Frequently transfused cases (more than 15 000 u per year): Anti-HBs 86%, HBs-Ag 8%(total 94%); less frequently transfused cases”: Anti-HBs 71,5%, HBs-Ag 9,5%(total 81%). In 18 patients(not identical to those with i.h.) liver biopsies could be performed with the. following results: chronic hepatitis: 9, subsided hepatitis without activity: 4, alcoholic liver damages: 3, acute dystrophia: 1, cholanqiolitis: 1.
Liver Disease in Hemophilia
Authors: Spero Joel A. , Lewis Jessica H. , Hasiba Ute
Published: May 2019

Abnormal liver function tests (LFTs) in patients with Hemophilia A and B who have been treated with commercial factor concentrates have been observed since 1973. One hundred and seventeen of these had had LFTs performed at least twice and at least six months apart. Of these, 13 were chronically HbsAg positive and the rest anti-Hbs positive. Forty percent of the anti-Hbs group had persistently abnormal LFTs for greater than 6 months, the majority for more than two years. Owing to the uncertain relationship of this “transaminitis” with true liver pathology, especially in the anti-body positive patients, seven closed liver biopsies were performed without incident under high coverage with factor VIII replacement. All patients were asymptomatic at the time of biopsy. One had a prior episode of illness probably related to subclinical hepatitis. An eighth patient underwent an open liver biopsy as part of a staging laparotomy for Hodgkins Disease. Six biopsies showed varying degrees of chronic persistent hepatitis (CPH). One was not evaluable owing to changes related to a retrospectively obtained history of ETOH intake. Finally, the individual with previous clinical liver disease had post-necrotic cirrhosis. Fluorescent studies with anti-Hbs and-anti-Hbc as well as electron microscopy were done on the seven closed biopsy specimens. The biopsy results suggest that the “transaminitis” found in transfused hemophiliacs represents histologic liver disease. In the majority of patients studied, CPH, a relatively benign entity, was present; and this does not justify withdrawal of concentrate therapy at this time.
Hemophilic Arthropathy in the Home Care Setting
Authors: Levine P.H. , McVerry B.A. , O’Connell F.D. , Zimbler S.
Published: May 2019

Thirty-four severe and moderately severe hemophiliacs on a formal comprehensive self-therapy program were prospectively evaluated as to the presence of hemophilic arthropathy, and as to progression or regression of arthropathy over a two-year period. Significant baseline deficits were noted in: flexion and extension of the knee (43 and 25 percent of joints, respectively), dorsiflexion and plantar flexion of the ankle (62 and 81 percent), and flexion and extension of the elbow (74 and 47 percent). Abnormalities of the hips and shoulders were unexpectedly common. In spite of the early and intensive use of plasma products and conservative supportive measures, there was considerable progression of arthropathy during the study period, especially in the older patients, and in patients with significant baseline arthropathy. These data indicate the need for continued aggressive management of arthropathy in patients on home care programs.
Psycho-Social Status of Patients on Home Care-6 Year Follow UP.
Authors: Granich B. , Hilgartner M.W.
Published: May 2019

Hemophiliacs on home care are more concerned with the daily management of their illness than with the more global considerations of longer life and better health. In a study of the total pop ulation of patients on home care at the New York Hospital, and of their families, the obvious advantages of greater personal freedom, better management of pain and stress, great saving of time, and control of and responsibility for their own lives was affirmed by the respondents. This has led the patients to seek to and largely succeed in giving up the sick role, which requires close dependence on medical supervision. Patients now tend to see themselves as partners in the management of their illness rather than as passive recipients of medical care. However, they recognize a continuing need for information and psychological support and for involvement in organizations which will supply these services. They recognize that they are receiving excellent care. In general, It seems that for this group of patients, hemophilia has become less a disaster and more a problem to be managed. The study was done in July-September, 1976, under the auspices of the New York Hospital and the Metropolitan Chapter of the Hemophilia Foundation. The information was gath ered in 146 face to face interviews with patients, parents and spouses, by trained social workers from Fordham University. The study population was largely middle class, white, suburban and 50% Catholic. By contrast, a study of a similar population from the same hospital, done before the advent of home care, revealed a principal concern to be complete dependence on hospital care and the physical and emotional burden that is imposed.
An Artificial ‘Haemophilic’ Plasma for One-Stage Factor VIII Assay
Authors: Chantarangkul V. , Ingram G.I.C. , Darby S.
Published: May 2019

An ‘artificial’ haemophilic plasma for one-stage factor-VIII assays is made by incubating human plasma with EDTA, Na2 to destroy factor VIII, and afterwards removing the anticoagulant by dialysis. Bovine factor V is then added to a given level. In the assay, contact activation is controlled by adding contact product.

It was confirmed that factor-VIII activity was destroyed and that the EDTA, Na2 was subsequently removed. The fibrinogen in the treated plasma clotted normally with thrombin. Likely variation in factor-V activity would not be critical. The concentration of fibrinogen and other factors was adequate. Variation between batches was small. The artificial plasma yielded assay results closely comparable to haemophilic plasma in samples with factor-VIII activities in the range 0.01-20.0 iu/ml.
Experimental Studies on Factor VIII Inhibitor Bypassing Activity
Authors: Vinazzer H.
Published: May 2019

The exact action of factor VIII inhibitor bypassing activity (FEIBA) is still unclear. For this reason, a series of experimental studies was carried out. Procoagulant activities were examined by standard one-stage methods while factor Xa and thrombin were measured by chromogenic substrates. Activities of factors II, VII, IX, and X were similar to PPSB fractions. In addition, low factor V activity and a phospholipid were detected. No activated factor X was present in FEIBA but there was a trace amount of 2.1 NIH units of thrombin per 100 FEIBA units. On addition of calcium chloride slow thrombin formation could be observed which however, reached 1100 NIH units of thrombin per 100 FEIBA units within an incubation time of 10 min. The velocity of thrombin formation was greatly enhanced by addition of a PTT reagent and of thromboplastin respectively. Factor Xa on the other hand, was neither formed after addition of calcium chloride nor by a PTT reagent. Tissue thromboplastin however, activated Xa from FEIBA in the same manner as a PTT reagent plus barium sulfate plasma. From these results, the conclusion could be drawn that thrombin could readily be made available from FEIBA while activation of Xa either needed the complete endogenous pathway or the presence of tissue thromboplastin. The procoagulant activity of FEIBA therefore, could be attributed to direct thrombin formation. By this process, an activation of the clotting mechanism in plasmas deficient in endogenous coagulation factors, and a complete independence from the presence or absence of a specific antibody could be explained.
The Aspirin Bleeding Time - A Screening Test for Evaluation of von Willebrand’s Disease
Authors: Stuart Marie J. , Miller Merrill , Wolk Joel , Davey Fredrick
Published: May 2019

In an attempt to elucidate the usefulness of the Bleeding Time (BT) post Aspirin (ASA) ingestion this test was done with other tests of coagulation in 30 controls and 77 patients evaluated for a possible bleeding diathesis. Coagulation studies included PT, PTT, TT, VIII AHF & AGN, IX, XI, XIII platelet retention and aggregation, and modified Ivy BTs pre and 2 hrs. post 600 mgms ASA. The mean control BT in 25 normals was 3.6'±3.2' (3SD). Following ASA the BT was 6.4'±4.1' (3SD). 5/30 “controls” without bleeding histories had abnormal BTs post ASA. In this group, 4 were proven to have unrecognized von Willebrand’s disease (VWD) and one a platelet defect. Of the 74 patients studied, 28 had initial BTs that were abnormal (23 with VWD and 5 with platelet defects). Of the remaining 46 with initially normal BTs, 26 had abnormal BTs post ASA. 13/26 had VWD at their first evaluation. In 9/26, however, the abnormal BT post ASA was the only abnormality at initial evaluation. On repeated testing, these patients were also shown to have VWD. 4/26 with abnormal BTs post ASA were found to have platelet abnormalities. The remaining 20 patients had normal BTs pre and post ASA. 16/20 revealed no hemostatic abnormality. In 4 eventually proven to have VWD, the BT post ASA was normal. The use of the BT post ASA raised the sensitivity of the BT as a screening test from 48% to 93% in the abnormal patient population. VWD appears to be the most common symptomatic/ asymptomatic bleeding disorder. The BT post ASA is a valuable screen in the evaluation of hemostatic defects i.e., mainly VWD, as also the occasional patient with a thrombocytopathy.
Comparison of Factor VIII Levels After Adrenalin IN Hemophilia a and von Willebrand’s Disease (VWD)
Authors: Eyster M.E. , Ballard J.O. , Prager D.
Published: May 2019

Two-fold or greater increases of Factor VIII procoagulant activity (VIII AHF) have been described following the administration of adrenalin in normal individuals and in some patients with VWD.VIII AHF and Factor VIII-like antigen (VIII AGN) levels were measured pre and one hour post the s. q. injection of adrenalin 0.35 cc. in 4 adult VWD patients and in 10 adult hemophiliacs with VIII AHF levels ranging from 1-29%, normal VIII AGN levels and normal VIII VWF levels as measured by the washed platelet ristocetin assay. Eighteen normal adults served as controls. The four patients with VWD showed two-fold or greater increases of VIII AHF with variable VIII AGN responses. All 10 hemophiliacs showed no change in VIII AHF with significant increases in VIII AGN when analyzed by the paired t-test.

The fixed rate of VIII AHF production, release or activation observed in hemophiliacs may help to explain the constancy of VIII AHF levels in individual hemophiliacs and their affected family members. Furthermore, this simple provocative test may be helpful in distinguishing certain VWD patients from those with hemophilia.
Femoro-Patellar Syndrome in Hemophilic Knee
Authors: Caffinière J.Y. De La , Allain J.P. , Laurian Y. , Larrieu M.J.
Published: May 2019

A high incidence of anatomical alterations of the patellar cartilage is observed in the chronic knee arthropathy of hemophiliacs. A systematic investigation of the role of an abnormal femoro-patellar joint in this knee arthropathy was undertaken. Fifty five knee joints in 38 children and 33 knee joints in 17 adults were studied. Clinical evidence of a femoro-patellar dysfunction was present in 21 children and 6 adults. Pain was the major symptom and this pain often made physiotherapy impossible. A permanent flexion deformity of the knee was frequent. The natural history of this syndrome was documented by regular X-Rays and was correlated with age, number of hemarthrosis, and associated anatomical abnormalities. Surgical procedures were performed in 5 cases in order to improve the knee function and relieve pain : 2 patellectomies, one section of the external patellar ligament and 2 condylar remodelling. After a follow up period of 6 months to 2 years, the clinical and functional results remained questionnable.