bluuebunny/crossref_metadata_2025 huggingface dataset:
Structural Characteristics of Prothrombin
Authors: Walz Daniel A. , Seegers Walter H.
Published: May 2019

The amino acid sequences of human and bovine prothrombin fragments 1 and 2 have been completed. In addition, partial sequence information has been obtained on the respective fragments from chicken prothrombin. Fragment 1 from human (155 amino acids), bovine (156 amino acids) and chicken (approximately 161 amino acids) contains the vitamin K-dependent calcium binding region (residues 1-32) and is a highly conserved region of the fragment. Fragment 2 from human (118 amino acids), bovine (118 amino acids) and chicken (113 amino acids) is apparently undergoing a more rapid rate of nucleotide substitution than is fragment 1. The A-chain of the enzyme from human (36+13 amino acids), bovine (49 amino acids) and chicken (49 amino acids) have a highly variable amino-terminal portion (residues 1-13) and a conserved portion (residues 14-49). Calculations for fragment 1 indicate the presence of 3α-helical regions (23.2%), 9β-sheet regions (34.2%) and 10β-turns (25.8%). The calcium binding region is an α-helical region of the molecule. Fragment 2 has 5α-helical regions (38.1%), 4β-sheet regions (19.5%) and 9 β-turns (30.5%). The differing rates of substitution found for fragments 1 and 2 are having no apparent effect on the calculated α-helical or β-sheet regions within prothrombin and may account for the limited, conserved activation bonds available within the prothrombin molecule. (Supported in part by a grant from the National Institutes of Health, HL-03424-19 and the McGregor Foundation of Detroit).
Assay for Prekallikrein Levels and Prekallikrein Activation in Human Plasma Using Dextran Sulphate as a Soluble Activator
Authors: Kluft C.
Published: May 2019

Incubation of human citrated plasma with dextran sulphate at 0°C was found to generate kallikrein activity as determined with the chromogenic synthetic tripeptide α-N-Bz-Pro-Phe-Arg-p. nitroanilide (0.16 mM). The possible use of the synthetic soluble dextran sulphate (MW 500,000) instead of the commonly used insoluble kaolin for activating the plasma kallikrein system was explored. The rate of activation of prekallikrein to kallikrein was dependent on: a) the concentration of dextran sulphate until a plateau was reached at 12.5 μg/ml; and b) the concentration of factor XII in the entire range up to 100%, as varied by mixing deficient and normal plasma. Activation was only slightly influenced by increased levels of C1-inactivator. In comparing dextran sulphate with kaolin, it was noticed that activation by means of dextran sulphate was slower (optimum at 7 and 2 min, respectively) and could be stopped by dilution. With kaolin (or glass), the activation continued even at high dilution (1:162) in the cuvet, preventing a proper study of the activation process. Dextran sulphate also did not interfere with the spectrophoto-metric assay, in contrast to the light-scattering properties of the insoluble kaolin. Optimum conversion of prekallikrein to kallikrein is reached in normal pooled plasma in 7 min at 0°C with 25 μg/ml dextran sulphate. The same is true for other plasma samples, if not liable to unwanted preactivation or incomplete activation, as revealed by control experiments. In 15 apparently healthy individuals, a mean activity of 476 ± 58 (SD) mU/ml was found. It is concluded that dextran sulphate can be used advantageously in the assay of prekallikrein activation and prekallikrein levels.
Further Physicochemical and Immunological Data Concerning the D-Dimer Complex
Authors: Gaffney P.J. , Brasher M. , Joe F.
Published: May 2019

It has been suggested that a sensitive assay of D-dimer, a fragment found in plasmin-mediated crosslinked fibrin digests, might contribute to our understanding of natural and induced fibrinolysis in man (P.J. Gaffney, Lancet (1972) ii, 1422). Towards this end, further characterisation of the heterogeneity of the D-dimer complex was attempted using gel electrophoretic techniques, while some attempts to raise rabbit antisera to D-dimer were also assessed.

It was confirmed that D-dimer, following plasmin-mediated release from crosslinked fibrin, was non-covalently linked to fragment E. The E fragment was released from the D-dimer/E complex during progressive digestion. The earliest digests examined showed that more than 50% of the total E fragment (immunologically measured) was complexed with D-dimer while some E remained in the complexed form even after intensive digestion. During examination of various timed digests it was observed that the electrophoretic properties of the various D-dimer complexes changed while the detergent gel patterns remained consistent. Antisera raised in rabbits to the D-dimer/E complex, following absorption with fragments D and E, reacted with purified fibrinogen and suggests that these antibodies may be recognising the dimeric D structure in fibrinogen.

It was concluded that studies of the immunological properties of the D-dimer/E complex, this being the earliest form of soluble crosslinked fibrin fragments, might be the most useful in eventually establishing an assay which would further our understanding of natural and induced fibrinolysis.
Platelet Plasma Membrane Glycoproteins in Normal and Genetically Abnormal Platelets
Authors: Phillips David R.
Published: May 2019

One approach to determine what specialized structures on the platelet membrane surface perform platelet specific functions is to compare the surface of normal to genetically abnormal platelets. We have used three techniques; lactoperoxidase-catalyzed iodination, neuraminidase/galactose oxidase/(3H)-NaBH4 labeling, and periodate/(3H)-NaBH4 labeling, to investigate the molecular organization of the proteins in these membranes. Several generalizations can be made about the normal membrane: (i) all of the major proteins exposed on the surface (12 have been identified thus far), are glycosylated, (ii) some of these glycoproteins are trans-membrane (i.e., they span the thickness of the membrane), (iii) the glycosylated segments are exposed to the outside of the cell, (iv) many disulfides (both inter- and intramolecular) are present in the platelet membrane glycoproteins, and (v) most of these glycoproteins contain sialic acid.

Analysis of the glycoprotein composition of the platelets from 16 individuals with Glanzmann’s thrombasthenia (characterized by a lack of aggregation) showed a decreased concentration of glycoproteins IIb and III. In contrast, platelets from an individual with Bernard-Soulier syndrome (characterized by absent adhesion) had primarily a decrease in glycoprotein Ib. The data demonstrate that both genetic abnormalities are caused by different defects in platelet membrane glycoproteins and suggest molecular entities which may be involved in specific platelet functions. Supported by NIH Career Development Award HL-0080 and Grant HL-15616.
Absence of the 145000 Molecular Weight, Soluble Platelet Glycoprotein - Lack of Platelet Agglutination
Authors: Solum N.O. , Hagen I. , Peterka M. , Gjemdal T.
Published: May 2019

The agglutination of human platelets by bovine factor VIII related protein has been studied in relation to the presence or absence of the 145ooo molecular weight, soluble platelet glycoprotein (GPS) using the presence of a granule-located glycoprotein as a positive control. GPS is found in the soluble fraction after subcellular fractionation of platelet homogenates on sucrose density gradients, and is probably identical to the soluble membrane protein glycocalicin The granule and the soluble glycoproteins separate into two bands on SDS-polyacrylamide gels both in reduced and unreduced samples when the gels contain urea and EDTA and are stained with periodic acid-Schiff reagent (PAS). A concomitant loss, or specific elution, of GPS and loss of agglutination was observed when platelets were stored in Tris-buffered saline, pH 7.4 (TS)at 4°C for 7 days, or when freshly prepared platelets were frozen and thawed when suspended in TS. When suspended in an EDTA-containing buffer during freezing and thawing, the resulting platelet ghosts still contained GPS and still agglutinated. The soluble fraction obtained by subcellular fractionation of giant platelets(Bernard-Soulier)did not show any PAS bands, whereas soluble fraction of these platelets prepared by freezing and thawing in TS clearly showed the presence of the granule protein but not the GPS band. The data indicate an association between GPS and the agglutination phenomenon.
Effects of Anticoagulant Therapy on Levels of Plasma Antithrombin III
Authors: Matsuda T. , Ogawara M. , Hirabayashi N. , Seki T. , Yokouchi M. , Murakami M.
Published: May 2019

Levels of plasma antithrombin III (AT-III) are frequently decreased in acquired disorders which predispose to thrombosis. The purpose of this study is to elucidate the mechanism underlying the decreased concentration of AT-III in thrombophilic states. Levels of AT-III were determined before and after institution of anticoagulant therapy with warfarin in nine cases. These patients consisted of three cases with ischaemic heart diseases, three with mitral stenosis with atrial fibrillation, one with cardiac aneurysm, one with thoracic and abdominal aneurysm and one with transient ischaemic attack. In seven cases, AT-III increased significantly following the institution of anticoagulant therapy. In two cases, AT-III decreased again after cessation of the anticoagulant therapy. Except for two cases of aneurysm, in whom coagulation analysis revealed presence of chronic consumption coagulopathy, plasma fibrinogen content did not increase following the anticoagulant treatment. α2-macroglobulin was unaffected by anticoagulant therapy in all cases. Levels of AT-III were inversely correlated with thrombotest values in the case of cardiac aneurysm in whom warfarin was continued for 14 months. From these results, it is concluded that AT-III is consumed even in cases in whom intravascular coagulation is not apparent and that the consumption of AT-III isaffected by levels of vitamin K dependent factors.
Clinical Use of Platelet Adhesion
Authors: Bick R.L. , Fekete L.F. , Wilson W.L.
Published: May 2019

Platelet adhesion as a measurement of platelet function was performed in 50 normal and 100 abnormal patients. A Bowie-type glass bead column and a constant perfusion technique was used. Template bleeding times (TBT) and platelet aggregation was also obtained in abnormals. The abnormal population consisted of patients referred with a disorder of hemostasis thought to be due to platelet dysfunction. Results on the normal population revealed “normal” adhesion to be defined as 89% to 100% retention, with an average of 94.8 ± 2.7% and a standard error of 0.41 (entire range for normals = 88% - 98%). In all 100 abnormals sent for evaluation, platelet adhesion correlated very well with aggregation, and more importantly, with clinical bleeding. In several cases abnormal adhesion and normal aggregation was seen; these 6 patients were classical von Willebrands, In 4 patients normal adhesion and abnormal aggregation was noted; this was due to aspirin use. Four patients had borderline adhesion and aggregation, and in all others the pattern was either normal adhesion and aggregation or abnormal adhesion and aggregation. In the vast majority of abnormals there was absolute correlation between adhesion and the TBT; in those 6 patients where correlation was lacking, the TBT was normal and adhesion was abnormal. These 6 patients had a malignant paraprotein disorder with paraprotein-induced platelet dysfunction. The results of this study suggest platelet adhesion, when done in a standardized fashion, to be a highly reproducible modality to assess platelet function. Using this methodology the normal range is narrow and excellent correlation is noted between abnormal adhesion, abnormal aggregation and clinical bleedability.
Therapy with Factor IX Concentrate Resulting in D.I.C. and Thromboembolic Phenomena
Authors: Campbell E.W. , Neff Sandra , Bowdler A.J.
Published: May 2019

The development of subclinical D.I.C. in a factor IX hemophiliac receiving concentrate replacement therapy during surgery is discussed with respect to pertinent laboratory features. The case is unique in that factor IX levels were followed carefully and corresponded inversely to the intensity of the D.I.C. Of additional significance is the failure of heparin failure given in conjunction with factor IX concentrate to prevent D.I.C. The value of adequate laboratory monitoring during factor IX replacement therapy is apparent.
The Identification of Accelerated Coagulability
Authors: Caprini J.A. , Zuckerman L. , Cohen E. , Vagher J.P. , Lipp V.
Published: May 2019

This report describes a new method for comparing overall clotting characteristics between normal individuals and those with proven malignancy using thrombelastographic (TEG) comparison of native and celite-activated specimens. Native whole blood TEG, celite-activated TEG, and standard coagulation tests were performed on the same blood collection in 90 normals and 90 patients with new malignancies and the results were used to derive a discriminate equation This equation classified correctly all 90 normals and 88 out of 90 cancer patients. The formulation was verified with an additional 82 patients with only one incorrect classification in the 31 cancer subjects. The standard coagulation tests did not discriminate without significant overlap between cancer patients and normals. The high degree of discrimination obtained with our TEG analysis provides a new tool to compare individual clotting differences that may have important clinical applications. However, the results cannot be extrapolated to selectively identify those with malignancy among the general population since many other stimuli may produce accelerated coagulability. This analysis does provide identification of accelerated coagulability in association with new malignancies.
Procoagulant Activity of Thrombosthenin Preparations
Authors: Muszbek L. , Fésüs L. , Szabó T. , Hársfalvi J.
Published: May 2019

3 × precipitated bovine thrombosthenin preparation exerted a powerful procoagulant effect similar to platelet factor 3 activity. In the presence of thrombosthenin the recalcification time, the kaolin clotting time and the Stypven time considerably shortened, the partial thromboplastin time only slightly altered and the activated partial thromboplastin time did not change at all. Thrombosthenin highly accelerated the generation of intrinsic thromboplastin but it was not active in a factor IXa generation test. It neither possessed tissue factor like property nor influenced the activity of tissue factor preparation or the action of thrombin on purified fibrinogen. Thrombosthenin contained 109 μg phospholipid/mg protein, the composition of which was quantitatively determined by thin layer chromatography. The procoagulant activity as well as the phospholipid content were retained after agarose gel filtration. It is suggested that thrombosthenin having an associated phospholipid component significantly contributes to the clot promoting effect of activated platelets.
Blood Coagulation, Fibrinolytic Activity, Platelet Function and Immunoglobulins in Frostbite
Authors: Chohan I.S. , Singh I.
Published: May 2019

Fifteen males, 19-45 years old, stationed between altitudes 3690 and 5540 m in the Western Himalayas who were frostbitten were studied within 24 hours of the injury and then 4 weeks and 1 year after for blood coagulation defects. The following disturbances were found: fibrinogen degradation products and factor VIII-related antigen were increased; fibrinogen, platelet counts and haematocrit were decreased; platelet adhesiveness was increased; euglobulin lysis time was prolonged; antithrombin III,α-1 antitrypsin and α-2 macroglobulin were markedly decreased; IgG and IgA immunoglobulins and cryoglobulins were increased; serum albumin was decreased and IgM immunoglobulin consumption was increased.

These abnormalities increase platelet adhesiveness and diminish fibrinolytic activity and promote intravascular thrombosis.

Furosemide increases fibrinolytic activity and suppresses platelet adhesiveness in vivo (I. Singh and I.S. Chohan, Int. J. Biometeor. 17, 73, 1973). Its use in the prevention of frostbite is under investigation.
The Isolation of a Potent Plasma Kallikrein with Weak Plasminogen Activator Activity
Authors: Gallimore M.J. , Fareid E. , Stormorken H.
Published: May 2019

Kallikrein was isolated from human plasma by the following procedures: removal of euglobulins at pH 5.3; QAE Sephadex chromatography: gel filtration on Sephadex G-150 and G-100. The partially purified preparation was then freed of contaminants by running it down a column of Sepharose-4B to which had been linked antibodies to IgG and pre-PTA. Pre-kallikrein and kallikrein activities were monitored during the fractionation procedures using a new synthetic chromogenic substrate for plasma kallikrein (Chromozyme-PK, Penta-pharm, Basle, Switzerland). 5 mg of enzyme was obtained with a specific activity of 3.75 Chromozyme PK (CPK) units/mg at 22° (yield = 9.5%: purification factor = 6250) and the yield of kinin from heated plasma was 1.79 μg/CPK unit/min. The kallikrein exhibited very weak plasminogen activator activity when tested on fibrin plates and in fibrin clot lysis assays(1 CPK unit =0.133 CTA units urokinase). Some other properties of the enzyme are discussed.
The Effect of Procoagulative Phospholipids on the Synthetic Substrate Determined Thrombin Generation in Vitro
Authors: Schulte F. , Klug O. , Roth Ursula
Published: May 2019

The effect of procoagulative phospholipids (Procops) with platelet factor-3 like activity on the generation of thrombin in plasma can be determined in vitro with the aid of synthetic chromogenic substrates. Standard citrated plasma of two manufacturers and from different batches incubated with amounts of 2-10 meg Procops as 1:100 - 1:500 diluted Tachostyptan (micellar Procops in form of a pharmaceutical speciality) was activated. The generated amount of thrombin activity catalyses the hydrolysis of chromogenic substrate to a tripeptide and to p-nitroaniline which was measured kinetically with a spectrophotometer at 405 nm. At optimum concentrations of Procops, activities adequate to 80 (SD ± 7, VK 8. 8%) - 115 i. u. (SD ± 2. 5, VK 2.2%) thrombin per ml plasma were measured depending on the conditions of incubation and activation. Having made the basis of appropriate procedures for incubation and activation the in vitro effect of Procops on the thrombin generation in plasma is reproducibly measurable with the aid of chromogenic substrate.
The Action of Urokinases on Plasm Inogens
Authors: Lormeau J.C. , Goulay J. , Vairel E.G. , Choay J.
Published: May 2019

The enzymic activity of the two forms of urokinase (I, MW 33,000, and II, MW 54,000) was studied, using different assay systems and towards glutamyl plasminogen, lysyl plasminogen and a synthetic substract. This study indicates that a significant difference exists between the two forms of urokinase : urokinase II is about 2.5-fold more active than urokinase I towards glutamyl plasminogen, the plasminogen circulating form. An hypothesis is proposed.
Platelet Function, Ultrastructure and Management in Glanzmann’s Thrombasthenia
Authors: Lusher J.M. , Barnhart M.I. , Pullen J. , Warrier A.I.
Published: May 2019

This exhibit will demonstrate laboratory abnormalities found in longitudinal studies on six patients with Glanzmann’s thrombasthenia. All have had mucous membrane bleeding (predominantly epistaxis) since infancy. In addition to prolonged bleeding times, absent clot retraction and little or no platelet retention in a glass bead column, all six have defective platelet factor 3 release with Kaolin and their platelets do not aggregate with ADP, epinephrine or collagen. The platelets of all six do aggregate with ristocetin, however. Transmission electron microscopy (TEM) of thin sectioned platelets revealed normal ultrastructure. However, in the TEM platelet function test (Thromb. Diath. Haemorrh., Suppl. 42:321-344, 1970) there was difficulty in capturing the thrombasthenic platelets even on the formvar surface. Those platelets which did adhere to the formvar exhibited normal percentages of dendritic and spread forms although pseudopod detail was unusual. Inspection by scanning electron microscopy (SEM) of surfaces of platelets responding to contact activation and captured on companion formvar coverslips revealed surface membrane features consistent with inhibition of membrane activation. Platelets were swollen with convoluted membranes while pseudopod outgrowth from discrete regions was not prominent. Suppression of membrane activation was also noted by SEM of platelets obtained by cytocentrifugation of buffy coat preparations. Upon exposure to collagen platelet adhesion and morphologic features of aggregation were abnormal under our test conditions.

In addition to local measures, recommended management of severe bleeding episodes is infusion of single donor platelet concentrates, obtained from an HLA matched donor by plateletpheresis.
Heparin Concentrations in Plasma During Surgery in Heparin Low-Dose
Authors: Kruse-Blinkenberg H. O. , Gormsen J.
Published: May 2019

The concentration of heparin in plasma was measured by the method of Denson & Bonnar and by a method using a chromogenic substrate for activated factor X (S-2222, Bofors, Sweden), pre-, per- and postoperatively in 126 patients undergoing major abdominal-thoracic, gynaecologic or orthopedic surgery. The patients received heparin (Heparin (R), Novo) 5,000 units 2 hours preoperatively and every 8 hours postoperatively for 1 week. No significant correlation was found between concentrations of heparin, sex, age, presence of malignant diseases, the type of operation or frequency or extent of per- and postoperative bleeding. Zero values were found immediately postoperatively in 41 of 126 patients, most frequently after abdominal surgery.

The correlation between the values found by the 2 methods was satisfactory.
Morphologic Platelet Changes in Vitro and During Thrombus Formation in the Rat
Authors: Wiedemann R. , Breddin K. , Grun H. , Weichert W.
Published: May 2019

Using high power Nomarski optics the shape change of thrombocytes can be shown. Platelets appear in their native shape as flat discs, less than 25% show pseudopodes. Depending on the time after blood sampling and on incubation temperature platelets in PRP or citrated blood swell and form tentacles. The addition of ADP to PRP induces the formation of aggregates. Single platelets form large vesicles rupturing and releasing granulated material. The remaining platelet material fuses. Bencyclan affects platelet morphology by inducing a spherical transformation, which is paralleled by the inhibition of platelet adhesiveness, spreading and aggregation. Observations in small mesenteric vessels of the rat show platelets in their native shape under stasis conditions. Vascular lesions are produced with a focused laserbeam (Hadron 513 biolaser). Immediately after the lesion platelets stick to the site of the microburn. Within seconds these platelets swell and form protrusions. After 3 - 10 min the vessel is occluded by a thrombus, of platelets, which undergo further swelling. Later the thrombus is partially or completely swept away and the vessel is recanalized. Irreversible fusion of platelets is rarely observed. These morphologic platelet changes differ markedly from those observed during in vitro aggregation. Injection of a new antithrombotic substance (Bay G 6575) diminishes the adhesion of platelets on the vessel lesion. The morphologic changes of single platelets (primary shape change) probably represent basic processes in hemostasis and thrombus formation.
The Microembolism Syndrome
Authors: Saldeen Tom
Published: May 2019

Clinical, autopsy and experimental investigations in parallel have revieled the existence of a microembolism syndrome where respiratory insufficiency is the main finding. In the early, usually benign form of the syndrome, transient pulmonary microemboli give rise to transitory pulmonary dysfunction. There is a phase of bronchiolar constriction, probably due to platelet release of prostaglandins. In the delayed, often malignant, form of the syndrome persistent pulmonary microemboli result in a progressive respiratory insufficiency. Three important factors in the pathogenesis of the delayed syndrome are fibrin, a delayed fibrin elimination from the lungs and a fibrin degradation product (FDP) which causes an increased vascular permeability. The delayed fibrin elimination is mainly caused by a fibrinolysis inhibitor, which primarily inhibits plasminogen activation. The inhibitor has been isolated in serum from patients with the delayed syndrome by affinity chromatography. The liver synthesis of this inhibitor can be prevented by nicotinic acid, probably due to a decreased release of free fatty acids. Dextran decreases the effect of this inhibitor. The contact time between the permeability increasing FDP and the pulmonary endothelium is much longer in a state of delayed fibrin elimination than during normal fibrin elimination where the FDP are rapidly swept away from the lungs. During the last years there has been a drop in the number of deaths in this syndrome, probably due to new prophylactic measures, especially those decreasing the degree of intravascular coagulation and inhibition of fibrinolysis.
Alteration of Streptokinase-Induced Fibrinolysis Under Clinically Relevant Conditions
Authors: Sutor A.H. , Wünsch-Macholz G. , Künzer W.
Published: May 2019

Testing streptokinase-induced fibrinolysis in human plasma we obtained the following results: Optimal fibrinolysis takes part at pH-values between 6.8 and 7.25, at NaCl-concentrations between 0.04 and 0.08 g/1, at temperatures between 36° and 42°, and at streptokinase-concentrations between 0.05 and 2 U/ml plasma. Outside of these ranges fibrinolytic activity is reduced. No correlation was found between streptokinase-induced fibrinolysis and hemoglobin concentration, but a strongly positive correlation between fibrinolysis and erythrocyte stromata concentration.
On the Nature of Fibrinolytic Activity of Tissues
Authors: Donner L. , Klener P. , Vodáková L.
Published: May 2019

Plasminogen activator of the vessel wall of thoracic, abdominal aortas, coronary, renal and carotic arteries was examined by means of the Todd’s histochemical method.

Activator was removed from sections by treatment for one hour with kalium thiocyanate. After saline wash possible proactivator was converted with streptokinase or urokinase solution and tested again with the histochemical method. Sections treated with thiocyanate showed no or very low fibrinolytic activity. After streptokinase or urokinase treatment a local fibrinolysis appeared again. It is suggested that thiocyanate despite its power to remove activator is unable to extract proactivator from tissue. No differences were found in different types of arteries examined neither in atheroclerotic or normal arteries.
Prophylaxis for Hemophilic Arthropathy
Authors: Ross Mackay S.
Published: May 2019

Hemophilia patients at the Rochester, New York, Hemophilia Center Clinic on three different modes of transfusion therapy were reviewed. It was found that those patients receiving prophylactic transfusion required less material over twelve months than those on home infusion or out-patient therapy. It was concluded that many hemorrhages may have a psychomotor origin and that prophylactic transfusion suppressed hemorrhage on a psychomotor basis.
Platelet Functions in Relation to Dietary Fats in two Regions of France
Authors: Renaud S. , Dumont E. , Godsey F. , Supplisson A.
Published: May 2019

To evaluate whether dietary fats might affect platelet functions in man as observed in animals, platelet aggregation (to thrombin, ADP, collagen, epinephrine) and clotting activity of PRP, PPP and of washed platelets, were studied in a mobile-laboratory in 50 healthy male farmers (40-45 years) from the Var and the Moselle, in relation to lipemia, glycemia, anthropometric determinations, dietary fats and other nutriments, as well as platelet phospholipid fatty acid composition. The main difference in the diet of the two regions was the percentage of calories from saturated fats (19% in Moselle, 12% in Var). In the Moselle subjects, the platelet clotting activity of PRP and of washed platelets (but not of PPP) and the platelet aggregation to thrombin and ADP, but not to collagen and epinephrine, was highly significantly (p < 0.001) increased as compared to those of the Var. In addition, the platelet clotting activity (p < 0.001) and the platelet aggregation (p < 0.01) were significantly correlated on an individual basis, with the diet content (calories) in long chain saturated fatty acids (palmitic/2 + stearic acids). The platelet clotting activity was also significantly (p < 0.001) correlated with the fatty acid composition of certain platelet phospholipids (PS + Pi) but not to plasma cholesterol which was identical in the two regions. Results of this pilot study in man appear to duplicate those in animals which indicate that it is the long chain saturated fatty acids in the dietary fats which increase the clotting and aggregating properties of platelets, in relation to thrombosis. The enhanced platelet functions in Moselle might also be related to the increased incidence of coronary heart disease in that region.
Some Studies on the Coagulation and Fibrinolytic Activity in Sickle Cell Patients
Authors: Osamo N.O. , Famodu A.A.
Published: May 2019

Basic studies on the activity of the coagulation and fibrinolytic systems have been carried out on two hundred sickle cell patients and the results compared to another two hundred AA subjects covering a wide spectrum of socio-economic classes. Euglobulin lysis times (ELT) Fibrinogen, degradation products (FDP), Fibrinogen levels, and levels of Factors V and VIII were measured using standard techniques with proven reproducibility. These estimations were performed in some of the sickle cell patients who arrived promptly at the onset of crisis and the results compared to the measurements obtained during their quiescent asymptomatic periods before and after crisis. It was found that there was no significant difference in the fibrinolytic activity among patients with sickle cell disease during their asymptomatic periods and normal AA subjects. At the onset of crisis, however, FDP levels in sickle cell patients appeared to drop while ELT was shortened. Factor VIII levels were consistently higher in the sickle cell patients than in their normal counterparts while Factor V levels were more variable. The significance of these findings in relation to the natural history of the disease and the general management of these patients will be discussed.
Activity and Concentration of Antithrombin III in Plasma and Serum
Authors: Róka L. , Bleyl H.
Published: May 2019

The concentration and activity of antithrombin III contained in plasma and serum of a single individual were compared with each other. The concentration of antithrombin III was determined by means of the rockett technique according to Laurell, antithrombin activity was measured by thrombin neutralization, residual thrombin activity was quantified using the chromogenic substrate Chromozym TH. The patients’ plasmas examined could be divided into different groups according to their antithrombin III specific activity. Most of the samples contained about 34 U/mg, the specific activities of two other groups were about 20 u/mg and 15 U/mg respectively. Only 3 samples contained more than 45 U/mg. In plasma samples with low specific antithrombin III activity the simultaneous occurrence of free antithrombin and an antithrombin-thrombin complex formed in vivo has been demonstrated by means of the two-dimensional Immunoelectrophoresis with heparin added to the agarose-gel medium. (Sas et al., 1975). The antithrombin-thrombin complex could be separated from free antithrombin by adsorption to a heparin-agarose column and fractionated elution, using the different heparin affinity of complex-bound and free antithrombin.
Role of Streptokinase in Treatment of Venous Thrombosis
Authors: Persson A.V. , Foti C.E. , Pierce L.E. , Sower N.D.
Published: May 2019

Fifty-four patients with deep venous thrombosis diagnosed on the basis of venographic studies were treated by four physicians using the same protocol but acting independently. In the early part of the study all patients were treated with streptokinase; in the latter part patients were randomized and treatment was either heparin or streptokinase. Of 40 patients who. received streptokinase, 22 were randomized and 18 were nonrandomized. Both drugs were administered by continuous IV infusion for 72 hours. Patients treated with streptokinase in the early stages of disease did very well; 90% of patients treated within five days of the onset of clinical symptoms and 75% treated within two weeks of onset showed marked improvement on post-treatment venograms. Only one patient who was treated with heparin showed a good result. Clinical results were similar in both groups. The incidence of important complications, such as significant bleeding and pulmonary emboli, was the same with both modalities. Fever of more than 101 F developed in about 50% of patients treated with streptokinase but in only one patient treated with heparin. We conclude that streptokinase is very effective in treating patients with acute deep venous thrombosis.
Study of the Normal Serum Activity Sensitizing Stabilized Fibrin to Urea Dispersion by Sephadex G. 200 Chromatography
Authors: Cataldo F. de , Baudo F. , Mussini E.
Published: May 2019

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.
Determination of Antithrombin, Heparin Co-Factor and Heparin in Plasma Using the Chromogenic Substrate S2238
Authors: Scully M.F. , Kakkar V.V.
Published: May 2019

Antithrombin, the thrombin inhibiting capacity of human plasma, was measured in normal plasma and also as a partially purified protein with the synthetic substrate, H-D-Phe-Pip-Arg-p-nitroanilide-2HC1. (S2238, KabiVitrum, London). A method was developed which was sensitive in the range 0.05 to 0.25 NIH units of thrombin in a total volume of 0.5 ml. Under these conditions a change in optical density of 0.36 per minute per 0.1 units of thrombin was observed. The substrate is, therefore, 5 to 6 times more sensitive than the previously used B2-L-Phe-Val-Arg-p-nitroanilide (S2160, KabiVitrum, London). Using this procedure antithrombin levels in up to 10 pis of plasma were measured without preliminary defibrination.

The method has been adapted for the automated determination of thrombin (0.025 to 0.4 units/ incubate and also antithrombin levels in 5-50μls of plasma using an LKB 2086 reaction rate ana-lzer. Triplicate measurements of 25 μl aliquots of 15 random samples of normal human plasma agreed within each sample set to ± 3% (1 standard deviation). A correlation was shown with antithrombin III levels measured by a clotting method. A modification of the method produced a more sensitive assay measuring in the range 80-l007o plasma antithrombin. Heparin cofactor was measured in 10 μl aliquots of heparinized plasma (NIH/ml). An adaption allowed automated measurement of heparin levels in the range 0.2 to 1.0 units/ml of plasma.

Preliminary experience indicates that with the use of this substrate various clotting tests may be substituted with a method which is automated and readily reproducible.
Problems of Streptokinase Therapy in Childhood
Authors: Sutor A.H. , Künzer V.
Published: May 2019

A generally accepted dosage regimen of streptokinase (SK) therapy is not available for the pediatric age group. In our experience with 35 children we found the determination of proactivator plasminogen (PP) by a capillary blood method very helpful for guidance of the initial and maintainance SK-therapy. Children with shock and consumption coagulopathy (due to gram negative sepsis, due to hypovolemia or due to metabolic disorders) have low PP-values and need an initial SK-dose of 4000 U/kg, followed continuously by 1000 U/kg/h for at least 24 hrs. Some children with local thromboses and the hemolytic-uremic-syndrome have high PP values which require an initial SK dose of 6000 U/kg and a maintenance dose of 1500 U/kg/h. PP-values during adequate SK-therapy drop to less than 3 %. Higher PP-values during SK-therapy indicate an underdosage of SK and require an increase of the SK-dose to prevent hyperplasminemia which may be followed by bleeding complications.
Complex Formation of Covalently Crosslinked Fibrin Oligomers with Agarose Coupled Fibrinogen and Fibrin
Authors: Hugo R. von , Hafter R. , Stemberger A. , Graeff H.
Published: May 2019

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.
Antithrombin-III as a Diagnostic AID in Disseminated Intravascular Coagulation
Authors: Bick R.L. , Dukes M.L. , Wilson W.L. , Fekete L.F.
Published: May 2019

Antithrombin-III (AT-III )/heparin cofactor is now recognized as a major inhibitor of thrombin and other serine proteases in coagulation. Since the reaction between AT-III and serine proteases is irreversable.AT-III consumption should be expected in pathological intravascular coagulation and attendent generation of thrombin and other serine proteases. Using a new AT-III assay system, unaffected by heparin or fibrino(geno)lytic degradation products, AT-III was monitored in 17 patients with DIC, It was found that early and significant decreases occured in all pts. It was further noted that monitoring of AT-III during therapy for DIC reflected a cessation of AT-III consumption and, thus, appeared to reflect efficacy of therapy in stopping the clotting process. Only 1 of 17 patients failed to show an increase in AT-III with initiation of therapy. In this group of patients, mini-heparin therapy appeared to be as efficacious as large doses of heparin in correcting AT-III consumption and other laboratory abnormalities of acute DIC, and in controlling hemorrhage of acute DIC. Four patients had chronic DIC with malignancy; the use of ASA and dipyridamole corrected AT-III consumption and clinical manifestations in these patients, although the response required more time than with heparin or mini-heparin. These findings suggest that the monitoring of DIC with AT-III levels may be useful in both confirming the diagnosis and, more importantly, in monitoring efficacy of therapy. Significant rises in AT-III were noted in all but 1 patient after initiating therapy, presumably reflecting cessation of consumption. In addition, mini-heparin appeared to be as efficacious as large heparin doses in stopping acute DIC and antiplatelet therapy appeared to stop AT-III consumption and clinical manifestations in patients with chronic DIC associated with malignancy.
Exchange Between Intra- and Extravascularly Injected Radioiodinated Fibrinogen
Authors: Rüegg R. , Straub P.W.
Published: May 2019

To test the hypothesis that in certain diseases plasma fibrinogen may be degraded extra-vascular ly and that circulating fibrin/-ogen degradation products (FDP) may stem from extra-vascular ly degraded fibrin/-ogen, 8 patients with pleural or peritoneal effusions (4 transudate, 4 exudate) were given 125I-fibrinogen i.v. and simultaneously 131I-fibrinogen intraca-vitarily. Measurements of plasma and effusion volumes allowed quantitation of the exchange of fibrinogen and its protein-bound derivatives. Plasma t/2 was shortened in 6/7 patients. At 48 hrs 7.6 ± 3.9% of i.v. injected radioactivity was found in the effusion (18% of it clottable, 24% protein-bound but unclottable), 4.8% ± 2.2% of intracavitarily injected radioactivity was found in plasma (29% clottable and 28% protein-bound unclottable). Iramunoreactive serum FDP were elevated in 7/8, plasma euglobulin lysis time normal in all 7/7 patients. Effusion fluids were fibrinolytically active, usually showing higher FDP concentrations than serum. It is concluded that 1) the shortened plasma half-life of fibrinogen is due in part to loss into the effusion, 2) fibrinogen is degraded in effusions and 3) plasma FDP in patients with ‘effusions may stem partly from extravascular fibrinogen proteolysis. The findings suggest cautious interpretation of FDP levels and fibrinogen turnover results in patients with suspected DIC and effusions.
Spontaneous Platelet Aggregation and Sulfinpyrazone Influence in Patients with Diseases of the Kidneys and Hypertension
Authors: Kirch W. , Köhler H.
Published: May 2019

In some diseases, particularly in diseases of the vessels, there is a spontaneous aggregation of platelets. So it was of interest, whether in patients with diseases of the kidneys and essential hypertension there is a frequent spontaneous aggregation as well. In addition, the effect of the antithrombotic substance sulfinpyrazone on platelet aggregation, on plasmatic coagulation and on clinical parameters, for instance on proteinuria, was to be investigated. In 47 patients with diseases of the kidneys or essential hypertension spontaneous platelet aggregation, thrombin time, partial thromboplastin time, fibrinogen and factor VIII were studied. Patients with spontaneous platelet aggregation were given 800 mg sulfinpyrazone daily for I to 6 months. In 15 of 16 patients with chronic glomerulonephritis (serum creatinine 0,7-2,0 mg%) and 13 of 14 patients with a chronic pyelonephritis (serum creatinine 0,8–1,7 mg%) spontaneous platelet aggregation was found as well. So in almost all patients with chronic glomerulonephritis and pyelonephritis a spontaneous aggregation was seen. The frequency of spontaneous platelet aggregation in patients with essential hypertension is likely to depend on the stage of hypertension. Spontaneous platelet aggregation did not return to normal in any of our patients on sulfinpyrazone and the substance had no influence on proteinuria, haematuria and leukocyturia in patients with diseases of the kidneys.
Arachidonic Acid Metabolism of Platelet in Diabetes
Authors: Kurosawa T. , Tojima T. , Funayama H. , Takahashi Y. , Shiokawa Y.
Published: May 2019

Recent reports have indicated that platelet aggregation is enhanced in some diabetics who have proliferative retinopathy and that platelet function is a altered by glucose loading. But the mechanism is not clarified yet. Arachidonic acid, the precursor of prostaglandin endoperoxide, plays a major role on platelet aggregation. Blood samples were collected with sodium citrate at 0, 30, 60, 120 and 300 minutes after 100 g glucose loading. Platelet-rich plasma was obtained by centrifugation and platelet aggregation was studied photometrically adding ADP. Platelet was obtained by further centrifugation and was kept freeze-dried. Platelet samples were extracted and transesterificated and separated by gas liquid chromatography. The quantitative regulation of arachidonic acid in platelets was measured by the composition ratio of arachidonic acid (C20-4)/linoleic acid (C18-2)=AL index. The results of platelet aggregation after glucose loading were as follows; platelet aggregation was not changed remarkably in normal subjects, but was enhanced at 30 and 60 and suppressed at 120 minutes in diabetics.AL index is as fol lows:prior to glucose loading, AL index of diabetics (4.6 ± 1.2) was higher than that of normal subjects (3.5 ± 0.5). After glucose loading, no significant change was observed in normal subjects, but AL index was increased at 30 (4.8 ± 1.4) and 60 (4.9 ± 1.4) and decreased at 120 minutes (4.1 ± 0.9) in diabetics. The results indicates that there is a certain relationship between quantitative regulation of arachidonic acid in platelet and platelet aggregation and that hyperaggregation may be induced by abnormal prostaglandin metabolism in diabetes.
Platelet Survival and Antiplatelet Drugs in Diabetes Mellitus Disease
Authors: Bremer G. , Jacobi E.
Published: May 2019

Platelet survival and platelet adhesiveness was measured in patients with long time diabetes mellitus in need of insulin. They are characterized by the complications of diabetic gangrene, neuropathy, nephropathy and retinopathy. Compared to a control group of healthy persons, patients with diabetes show a significant decrease in platelet survival (normal persons t/2 = 4.1; diabetics t/2 = 3.24).SH 1117, a dipyridamole derivative, alone in doses of 500 mg three times per day, has little modifying effect on platelet consumption. By the application of a combination of 75 mg dipyridamole and 330 mg acetylsalicylic acid, a normalization of platelet survival and platelet adhesiveness is achieved.
In Vivo and In Vitro Comparison of Various Factor VIII Concentrates
Authors: Nilsson I.M. , Hedner U.
Published: May 2019

Five different factor VIII concentrates, AHF-Kabi(=fraction 1-0), Krynativ-Kabi(=cryoprecipitate), Hemofil-Hyland, AHF-Profilate-Abbott, Kryobulin-Immuno, available in Sweden for treatment of haemophiliacs were compared with respect to in vivo recovery of F VIII:C and survival time and in vitro properties. The parameters studied were F VIII:C, F VIIIR:AG, crossed Immunoelectrophoresis, F VIII:Rcof, fibrinogen content and F XIII activity. All the preparations had higher values for F VIIIR:AG than for F VIII:C. The quotient was highest for Hemofil, Krynativ-Kabi and Kryobulin and varied between 4 and 7. The lowest quotient, 1.3 to 4, showed AHF-Kabi. The units of F VIII:Rcof were almost the same as the units of F VIII:C. AHF-Kabi had the highest fibrinogen content and was the only preparation with high amounts of F XIII. In cross Immunoelectrophoresis AHF-Kabi showed a similar pattern to that of normal plasma. The other preparation had a different pattern suggesting less hetero-genicity of the molecule. The in vivo recovery was about the same for all the concentrates but AHF-Kabi had a significantly longer half-life (18-26 hrs); the corresponding figures for Hemofil were 8-16 hrs when given to the same patients. Only AHF-Kabi was able to completely normalize the defect in von Willebrand’s disease.