bluuebunny/crossref_metadata_2025 huggingface dataset:
Elevated Platelet Antibodies in Septicemic Thrombocytopenia
Authors: Helton J.G. , Neame P. B. , Hirsh J.
Published: May 2019

Thrombocytopenia often complicates bacterial septicemia in the absence of evidence of DIC. Evidence is provided that thrombocytopenia in these patients is caused by adsorption of IgG to platelets (immune thrombocytopenia) which occurs either as a consequence of bacterial injury or adsorption of immune complexes consisting of bacteria or bacterial products and their specific antibodies. Platelet associated IgG (PAIgG) (assayed by inhibition of lysis) was determined in 44 patients with bacterial septicemia. PAIgG was elevated in 8 of 11 thrombocytopenic patients with gram negative septicemia (mean 47 tg IgG/platelet, p<0.0001), 8 of 10 thrombocytopenic gram positive septicemic patients (mean 55 fg IgG/platelet, p<0.0001) and in one of the 31 septicemic patients with normal platelet count (mean 5 fg IgG/platelet). In vitro studies of serum and platelets obtained after 2 patients had recovered demonstrated that PAIgG was elevated to 100 fg IgG/ platelet when their respective bacteria were incubated first with their serum and then with their platelets. In contrast, when bacteria were added to the platelets before Incubation with serum, there was only a slight elevation in PAIgG. Results indicate that PAIgG is frequently elevated In septicemic patients with thrombocytopenia and that in some of these patients the thrombocytopenia is caused by adsorption of immune complexes to the platelets.
Immune Injury to Human Platelets Mediated By IgG Fc Receptor is Prevented by Prostacyclin
Authors: Hawiger J. , Parkinson S.K. , Timmons S. , Glick A.D.
Published: May 2019

Immune injury to human platelets by drugs, bacteria, and viruses which form antigenantibody complexes is mediated by the IgG Fc receptor on human platelets and results in thrombocytopenia. We studied whether immune injury to human platelets mediated by the IgG Fc receptor can be prevented by prostacyclin (Prostagland in 12, PGI2), a novel prost glandin generated by the blood vessel wall. Immune injury to human platelets in whole plasma was elicited by Protein A-bearing staphylococci. Protein A induces binding of I to the human platelet Fc receptor, which results in platelet aggregation and 3H-seroton release in whole plasma. Excess of isolated Fc fragment inhibits aggregation and serotonin release in this model of immune injury. Synthetic PGI2 protected human platelets from this IgG Fc fragment-mediated immune injury in whole plasma. Inhibition was prompt (1 to 5 min) and dose dependent, reaching maximum at 10-6M of PGI2. Removal of plasma proteins and use of IgG-coated cells did not change the inhibitory potency of PGI2, which was at least 1000-fold more active than 6-keto PGF1α. Electron microscopy revealed than PGI2 prevented binding of IgG- coated cel is to human platelet membrane. From comparison with anti-inflammatory steroids (methylprednisolone) and nonsteroidal prostaglandin synthetase inhibitors (ASA), it appears that prostacyclin is the most active agent known the date to protect human platelets from IgG Fc receptor-mediated immune injury in vitro.
Experimental Arterial Thromboembolism in Baboons: Mechanism, Quantitation and Pharmacologic Prevention
Authors: Herker L.A. , Hanson S.R.
Published: May 2019

A quantitative primate model of arterial thromboembolism has been developed that involves the kinetic measurements of 51Cr-platelets and 125I-fibrinogen by femoral arteriovenous cannulae in chaired baboons. Cannula platelet consumption (CPC) correlated directly with exposed cannular area for irradiated Silastic (IS) end polyurethane (correlation coefficients of 0.940 and 0.901 respectively; p < 0.001) and remained steady state for months. Fibrinogen was not measurably destroyed. CPC was independent of cannula flow rate, platelet count, heparin anticoagulation and ancrod defibrinogenation

In-platelet imaging of IS eannulae demonstrated luminal accumulation and subsequent embolization of irregular platelet masses that progressively occluded glomerular vessels with non-fibrin containing platelet thromboemboli.

Oral administration of dipyridamole and sulfinpyrazone decreased cannula platelet consomption in a dose dependent manner with complete interruption at 20 and 250 u mol/kg body wt/day (in three divided doses) respectively, whereas oral acetylsalicylic acid (10-330 umol/kg/day) had no measurable effect on cannula platelet consumption. We conclude that this primate model simulates arterial thrombotic processes in man and that this model is suitable for the in vivo evaluation of biomaterials and of drugs that modify platelet behavior.
Who’s Programme in Thrombosis and Haemostasis Research
Authors: Pisa Dr Z.
Published: May 2019

The Limited success in decreasing the mortality of coronary heart disease (CHD) in general and of sudden death in particular, stresses the needs and importance of systematic research into the etiology and pathogenesis of atherosclerosis and CHD. Prevention seems to be the only effective approach. WHO has included in its CTD long-term programme activities related to these problems. The network of collaborating centres with good epidemiological skills carrying out studies in well-defined populations could provide opportunities for special studies on the role or haemostasis in the etiology and pathogenesis of atherosclerosis and acute MI. In collaboration with ETRO and ISFC methods are being reviewed and sought which are capable of detecting a tendency to arterial thrombosis and atherogenesis. The present position is being discussed.
The Epidemiology of Arterial Thrombosis
Authors: Meade T.W.
Published: May 2019

The epidemiology of clinically manifest ischaemic heart disease (IHD) is not the same as that of atheroma. For example, the rise in IHD incidence that has taken place since early this century has probably not been paralleled by a similar rise in atheroma. The National Necropsy Survey in the United Kingdom found no relationship between physical activity at work and atheroma, but a clear association between activity, on the one hand, and lumen-occlusive changes and myocardial scarring on the other. Later studies have shown an association between occupational and leisure activity and clinical IHD. These and other observations strongly suggest that some process (or processes) other than, or in addition to, the formation of atheroma causes clinical IHD. However, the pathology of the different syndromes usually considered as IHD is not homogeneous. Evidence of recent thrombosis is often found in those dying of myocardial infarction; it is less often found in “sudden death”, though advanced atheroma is a usual finding. Sudden deaths may be due to small, unrecognized platelet aggregates. Clearly, atheroma and thrombogenesis are both involved in the pathogenesis of clinical IHD, and quite possibly to different degrees in different neoole. But thrombogenesis has not been adequately investigated in population-based studies. The epidemiological evidence suggests the need to study this process as contributing not only to infarction and sudden death, but also possibly to the formation of atheroma itself.
Replacement Therapy in Von Willebrand’s Disease
Authors: Sultan Y.
Published: May 2019

Replacement therapy in patients with VWD for bleeding episodes or surgical procedures is not as well defined as treatment of patients with Hemophilia. In severe forms of the disease although it is aesy to obtain high levels of FVIII procoagulant activity after transfusion with FVIII concentrate it is more difficult to achieve a normal bleeding time (B.T.) and moreover to keep it normal throughout surgical procedures and healing.

The normalization of B.T. is dependent on the kind of FVIII concentrates used. To demonstrate this, several commercial preparations of FVIII have been tested for their content in FVIII R Ag and FVIII RWF in comparison with the procoagulant activity ; they have also been tested in vivo for their ability to correct B.T.. shortening of B.T. is also dependent on the quantities injected ; it is more efficient to infuse large doses once a day or every other day, than fractionated doses F It is also dependent on the severity of the disease and the number of transfusions the patient previously received. Some problems remain unsolved, specially when all the parameters of the disease reach normal values in the circulating blood although B.T. is still prolonged.
Factor VIII With Monophasic Degradation, Produced by a New Fastthawing Method
Authors: Ezoan M. , Hansen J.F. , Gormsen J.
Published: May 2019

A new method has been developed for the large-scale production of cryopre-cipitate with a high yield of factor VIII. The freeze-dried factor VIII product has a solubility comparable to that of intermediate purified products and snows a remarkable in vivo stability. These improvements have been mainly accomplished by using a fast thawing method based on energy transfer through microwaves. Deep-frozen plasma bags (200 ml) are thawed in less than 10 min. with the temperature no where exceeding 4°C. In our routine production of freeze-dried, small-pool cryoprecipitate the yield of factor VIII is about 450 Units/liter of plasma. The concentration of the dissolved product is high being at least 2.0 Units/ml. This factor VIII concentrate has been used for home treatment for about one year, Clinical studies have snown a mean factor VIII recovery of 99.7% and a loss in activity of only 25% during the first 5 hours. The degradation follows a single exponential decay with time. This result indicates the absence of a labile component of factor VI VIII, which is usually observed along with the more stable component.
Platelet adhesion by Subendothelium-Bound Factor VIII-VWF:Kinetics and Molecular Weight Dependence
Authors: Bolhuis P. A. , Sakariassen K. S. , Sixma J. J.
Published: May 2019

Platelet adhesion to human subendothelium was determined by perfusions with albumin solutions containing 51Cr-labeled, aspirin-treated platelets and washed red cells (hematocrit 40%) at 37° and a flow rate of 135 ml/min. Adherence was similar with Von Willebrand plasma instead of albumin solution and addition of purified FVIII-VWF caused adhesion similar to that from normal plasma. Incubation of subendotheliurn with FVIII-VWF resulted of binding of FVIII-VWF at the surface and in subsequent perfusions a surface concentration of, FVII-VWF/cm2 was shown to correct the platelet adhesion in albumin solutions towards normal. The kinetics of binding of FVIII-VWF and platelets to the subendothelium confirm the role of bound FVIII-VWF in adhesion. Binding of FVII-VWF occurs rapidly in the first minute of perfusion to about 4 x 10-4U/cm2 and then increases further to about 10-3 u/cm2 in 5 min. Platelet adhesion is similar for perfusates with and without FVIII-WF in the first minute; then the presence of FVIII-VWF results in a two-fold increase of adhesion at 5 min. Reduced adhesion was found with the high-molecular weight component of FVIII-VWF obtained by high iconic strength dissociation. Also, the activity of glycin precipitated FVIII-VWF (e.g. Hemofil FVIII-concentrate) is impaired, cross-electrophoresis of FVIII-VWF from cryoprecipitate and FVIII-VWF after glycin precipitation showed an increased mobility or the latter, indicating a reduced molecular siie. From these experiments we conclude tnat platelet adhesion is mediated by subendothelium-bound FVIII-WWF. The degree of adhesion may depend on the molecular weight of the FVIII-VWF.
The Primary Structure of Horseshoe Crab (Tachypleus tridentatus) Coagulogen and its Homologies with Platelet Factor 4
Authors: Takagi T. , Nakamura S. , Hokama Y. , Miyata T. , Niwa M. , Iwanaga S.
Published: May 2019

Horseshoe crab coagulogen consists of a single basic polypeptide chain having the total 175 amino acid residues. Upon gelation of this protein, a large peptide, named peptide C, which has 28 amino acid residues, was released, and the resulting gel protein consisted of A and B chains, bridged by two disulfide linkages. The sequence studies on peptide C, A chain and B chain provided that their sequences have a significant homology partly with primate fibrinopeptide B and largely with human platelet factor 4. The facts suggest that these are derived from a common ancester or that the coagulogen is a prototype of platelet factor 4. The whole sequence of coagulogen was as follows:Ala-Asp-Thr-Asn-Ala-Pro-Ile-Cys-Leu-Cys-Asp-Glu-Pro-Gly-Val-Leu-Gly-Arg-Thr-Gln-Ile-Val-Thr-Thr-Glu-Ile-Lys-Asp-Lys-Ue-Glu-Lys-Ala-Val-Clu-Ala-Val-Ala-Gln-Glu-Ser-Gly-Val-Ser-Gly-Arg-Cly-Fhe-Ser-Ile-Phe-Ser-His-His-Pro-Val-Phe-Arg-Glu-Cys-Gly-Lys-Tyr-Glu-Cys-Arg-Thr-Val-Arg-Fro-Glu-Hls-Ser-Arg-Cys-Tyr-Asn-Phe-Pro-Pro-Phe-Thr-His-Phe-Lys-Leu-Glu-Cys-Pro-Val-Ser-Thr-Arg-Asp-Cys-Glu-Pro-Va1-Phe-Gly-Tyr-Thr-Va1-Ala-Gly-Glu-Phe-Arg-Va1-1le-Va1-GIn-Ala-Pro-Arg-Ala-Cly-Phe-Arg-GIn-Cys-Val-Trp-Cin-His-Lys-Cys-Arg-Phe-Giy-Ser-Asn-Ser-Cys-Gly-Tyr-Asn-Gly-Arg-Cys-Thr-Gln-Gln-Arg-Ser-Val-Val-Arg-Leu-Val-Thr-Tyr-Asn-Leu-Clu-Lys-Asp-Cly-Phe-Leu-Cys-Glu-Ser-Phe-Arg-Thr-Cys-Gys-Cly-Cys-Pro-Cys-Arg-Ser-Phe,
The Influence of Red Blood Cell Hemolysate (RBCH) on Platelet Aggregation (PA) and Release of Serotonin and β-Thromboglobulin (β-TG) in Heparinized Platelet Rich Plasma (PRP)
Authors: Wurzinger L.J. , Blasberg P. , Jüngling E. , Schmid-Schönbein H.
Published: May 2019

As hemolysis occurs preferentially in artificial internal organs (AIO), and thromboembolic complications are frequently seen in AIO, the influence of liberated RBC-contents on PA was investigated. To perform the experiments under the most physiological conditions, heparin was used as anticoagulant and the platelets were kept at 37°C during withdrawal, preparation, storage and aggregometry. The lysed RBC were freed of membranes and stored at 0°C until use. PA was quantitatively monitored in a defined homogenous shear field in a Couette-chamber by turbidimetry. Dense granule release was assayed using 14C-Serotonin labelled platelets. Alpha granule release was assessed via β-TG radioimmunoassay. PRP samples with added RBCH were compared with samples with ADP added in doses equivalent to that of the samples with RBCH. PA, Serotonin, and β-TG release were found to be positively correlated to the concentration of RBCH In PRP, An increase of PA and release as compared to control samples was observed above RBCH-concentratlons of ea. 0.3 g/L hemoglobin, a concentration frequently found during use of AIO. The addioion of ADP in hemolysate-equivalent doses proved to be less effective. Chromatographic assays showed that ATP, which is found in concentration about tenfold of ADP In the RBC, is broken down to ADP in PRP, thus augmenting the ADP-pool acting on platelets.
Does Fibrin Fragment D-Dimer Cross-Link With Fibrin Monomer?
Authors: Franks J.J. , Kao Betty C.
Published: May 2019

When 2 to 4 ng of radioiodinated human D-dimer (D-D) and 0.2 mg of carrier rabbit D-D in two ml of buffer were added to a .7x10 cm column containing 4 ml of agarose-linked rabbit, or human fibrin monomer, D-D was tightly bound. After a neutral buffer wash D-D was quantitatively eluted (97 to 99% recovery) with 6M urea (pH 8.6). Comparable recovery of unlabeled human D-D, 500 to 8000 ng, was achieved. Human D-D concentration was measured by radioimmunoassay with rabbit anti-human D-D serum. When labeled or unlabeled D-D was mixed with 2ml of serum or when serum from patients with high D-D levels was chromatographed, 40 to 70% of D-D was retained and could not be eluted even by washing the column with alkaline urea for 24 to 48 hours. Irreversible binding of D-D could not be prevented by adding DFP or Tween 20 or by pre-heating the carrier serum to 60°, but binding was completely prevented by 5 mM EDTA in the serum and eluting solutions. Let’s than 1% of labeled D-D remained on the affinity agarose. We have previously suggested that only one of two reciprocal gamma chain cross-linking sites is used when fibrinogen cross-linking takes place (S. Afr. J. Sci., 74:202, 1978). Activated factor XIII could utilize the spare site to form epsilon (gamma glutamyl)lysine bonds between insolubilized fibrin monomer and D-D.
A New Inhibitor of Factor XIII and Platelet Aggregation. Influence of Fibrin Stabilization on Thrombolysis
Authors: Ryckewaert J.J. , Soria J. , Soria C. , Samama M. , Maamer M. , Aurousseau M. , Otero A.M.
Published: May 2019

Antifactor XIII and antiplatelet aggregation activities were discovered in vitro for a synthetic compound with a benzothiophen keton structure. It was found to inhibit the stabiliiation of fibrin as shown by S.D.S. Polyacrylamide gel electrophoresis of a clot previously dissociated by urea and mercaptoethanol, among other tests. The mechanism of action was studied using dansyl cadaverin incorporation into casein : the inhibitor effect was not competitive, related to the inactivation of factor XIIIa, and independant of calcium concentrations. It was much more pronounced in a purified system than in plasma, due to its binding to albumin. We therefore studied its possible displacement from albumin by various drugs.

The thrombolytic activity of urokinase only slightly increased when clot stabilization was inhibited by the studied compound.
Comparative studies on fetal fibrinogen from full term and premature infants
Authors: Galanakis D.K. , Mosesson M. W. , Bannerjee D.
Published: May 2019

Cord plasma or fibrinogen from full term (FT) infants, when compared with that from adults, displays delayed fibrin aggregation under conditions of high ionic strength (0.29); this phenomenon serves as an indicator of a fetal form of fibrinogen. In studies to be reported we compared adult fibrinogen with that from pooled premature (P) (24-35 weeks gestation) or FT (38-42 weeks) infant plasma. The thrombin and arvin clotting times of P fibrinogen (fraction 1-2) were even more prolonged than those of FT fibrinogen. Dodecyl sulfate gel electrophoresis
disclosed no significant differences in the amounts of coagulable fibrinogen catabolites in P, FT, or adult plasma. The thrombin time of P or FT plasma did not change following incubation at 37° for 8 hours. Furthermore, there was no change in the thrombin (citrated) or Reptilase (heparinized) clotting time of P or FT plasma to which had been added sufficient adult fibrinogen to normalize the fetal fibrinogen effect prior to incubation at 370. Thus, functional differences between adult and fetal fibrinogen cannot be attributed to the fetal plasma environment per se. The results suggest that plasma from infants born during the early part of the third trimester of pregnancy is relatively enriched with fetal fibrinogen; that from FT infants, by comparison, contains sufficient amounts of adult fibrinogen to result in shortening of the clotting times.
Effect of Sulfinpyrazone on Experimental Atherosclerosis in Rats
Authors: Horsch A.K. , Klaiber P. , Dennig K. , Hofmann W.
Published: May 2019

Early atherosclerotic lesions in rat aortas were produced by synergy of cholesterol feeding (4 g cholesterol and 2 g cholate in Ioo g pellets) and immunological injury (5 repeated immunizations with 1 mg of horse-radishperoxidase). Cholesterol feeding and immunization was started the same day and resulted after 6 weeks in endothelial injury, intimal thickening and edema, in proliferation of intimal smooth muscle cells and in deposition of lipids and immune complexes in the arterial wall.

100 rats were studied and no lesions were detected in the control group, minimal changes were seen in the animals receiving cholesterol or immunization alone, but lesions were constantly present in the group receiving both regimens. One group of cholesterol fed and immunized rats was given sulfinpyrazone from the beginning of the experiment (160 mg/kg/d) over the 6 weeks and a significant reduction in lesion formation was observed compared to the non treated animals. These results suggest a protection of the arterial wall by sulfinpyrazone in experimental atherosclerosis.
Retroperfusion of the Ischemic Myocardium with Methysergide and Dipyridamole
Authors: Kordenat R.K.
Published: May 2019

In four groups of 6 dogs each, an occluding thrombus was produced in the LAD coronary artery by placing a thrombogenic wire into the vessel, using a catheter technique (under x-ray control). One-haIf hour after occlusion was confirmed (angiographically) in Group I, methysergide (100 micrograms /kg, in 20 ml normal saline) was infused, retrograde, as a single bolus, into the ischemic myocardium, via the great cardiac vein, using a Swann-Gantz balloon catheter. In Group II, dipyridamole (2 mg/kg/ in 20 ml normal saline) was used and in Group III both drugs were combined. Group IV mean heart rate (HR), stroke volume (SV), cardiac output (CO) decreased from baseline (p<0.01, p<0.01 and p<0.001) at the third post-occlusion hour and total peripheral resistance (TPR increased (p<0.001). Few and Less significant changes and some improvement in these and other variables occurred in I and II over the observed period. There were no significant changes in any variable in III. CO, TPR and SV of Groups III and IV were significantly different (p<0.05 to p<0.01), when their mean values were compared at the first and third post-occlusion hours. It appeared that both drugs offer a protective hemodynamic effect during acute myocardial infarction and there may be a positive synergistic action between them.
In Vitro and In Vivo Inhibitory Effects of Alcohol on Platelet Functions of Rats
Fed Dietary Saturated Fats.
Authors: McGregor L. , Renaud S.
Published: May 2019

In the in vitro experiment, alcohol diluted in complete tyrode wae added, at 37°c, 2min before aggregation tests, to platelet-rich plasma (final dilution in plasma : 0.00016 and 0.00032%) from male, Sprague-Dawley rats. These animals were fed either laboratory chow or a high fat (40%) purified diet rich in either polyunsaturated fatty acids (22% corn oil) or in long chain saturated fatty acids (38% with 2% corn oil). Aggregation to thrombin but not to ADP was significantly reduced (50%) in all 3 groups of rats with 0.00032% alcohol, even in hyperaggregable animals fed saturated fats. Addition of 0.00016% alcohol slightly reduced platelet response to thrombin. The in vivo experiment consisted of feeding 48 weanling male Sprague-Dawley rats with purified diets, as mentioned above, rich in either polyunsaturated fatty acids or saturated fatty acids for at least 7 months. Morever, half of these animals had 6% alcohol in their drinking water for at least 2 months. Addition of alcohol, in drinking water, significantly prolonged platelet-rich plasma clotting time of saturated fat (101vs 136 sec) and in polyunsaturated fat group of animals (130vs 145 sec). Platelet maximal response of aggregation to thrombin (7.2vs 4.0 cm) and to ADP (9.0vs 5.7 cm) were significantly reduced by alcohol. Alcohol, in drinking water, appears to markedly inhibit platelet functions in rat. This seems to result from a direct effect on blood platelets since it can be partly reproduced by adding alcohol co platelet-rich plasma in vitro.
Occurence and Evolution of in Vivo Platelet Release Phenomenon. Study in Severe Burns.
Authors: Hourdillé P. , Boisseau M.R. , Bernard Ph. , Sanchez P.
Published: May 2019

During the first post burn period (first and second week) the most severe cases showed evidence of a platelet storage pool deficiency, i.e. : no aggregation with ADP, decreased nucleotide content, reduced number of granules loaded with mepacrine (presence of non granulated platelets) and increase in the percentage of light platelets. During the second post burn period (3rd to 5th week) on the same patients and for the least severe cases, the abnormalities were found heterogeneous, suggesting evidence of a partial release phenomenon, Altough the platelet nucleotide content was reduced, quite a few “empty” platelets were found, there was no significant increase of light platelets.

These data indicate, with regard to the platelets, the presence of a partial release phenomenon during extensive peripheral destruction of platelets which can sometimes reach the state of a Storage Pool Dasease.
Thrombin Enhancement of Human Low Molecular Weight Factor VIII Procoagulant (LMW)
Authors: Horowitz B. , Lippin A. , Woods K.R.
Published: May 2019

LMW was isolated by affinity chromatography on FVIII-Sepharose and characterized with regard to its reaction with thrombin (0.05 u/ml). In contrast to enhancement of the procoagulant activity (FVIII:C) observed with undissociated FVIII, enhancement of FVIII:C of LMW proceeds more slowly and to a lesser extent. Furthermore, thrombin-enhanced FVIII:C of LMW is relatively stable, frequently for periods up to 6 hours. Neither increasing thrombin to 0.5 u/ml nor adding HMW prior to thrombin alters this stability. Thrombin-enhanced LMW was inhibited by addition of human FVIII inhibitor. Measurement of thrombin esterolytic activity revealed the presence of a thrombin inhibitory activity in the LMW but not in the purified, undissociated FVIII preparation. It is not currently known whether this activity is due to a contaminant. The addition of heparin had no effect on the kinetics of inactivation of thrombin indicating the inhibitor is not antithrombin III. These data suggest that LMW may exhibit thrombin inhibitory activity or that FVIII may bind an inhibitor of thrombin.

Advantage was taken of the stability of thrombin-enhanced LMW to determine if it is a serine protease as recently suggested by Vehar and Davie (Science 197: 374, 1977) for bovine FVIII. Human LMW reacted with thrombin was not sensitive to addition of diisopropyl fluorophosphate, indicating that human FVIII is not a serine protease
Production of Two Immunologically Distinct Plasminogen Activators by Human Tissue in Culture.
Authors: Bernik M.B. , Rijken D.C. , Wijngards G.
Published: May 2019

Plasminogen activator is known to be produced by various human tissues in culture, the kidney yielding activator similar to that in urine, urokinase. Little is known, however, about the identity and properties of activator(s) from other tissues. The present study examines activator activity accumulating in serum-free supernates of cultures from lung, kidney, intestine, renal blood vessels, and a cell line from tumor (HeLa). Immunologic properties were examined by quenching assays, performed on fibrin plates using antisera specific for urokinase as well as antiserum to purified uterine tissue activator which showed immunologic relationship to activator from perfused cadaver limbs (vascular activator) but not to urokinase. In these assays, the activity from cultures of intestine and HeLa was not affected by antisera to urokinase but was quenched by antibodies to tissue activator. Lung and blood vessels yielded activity quenched partly (40-70%) by antiserum to urokinase end partly by antibodies to tissue activator. In studies by SDS-polycrylamide gel electrophoresis and immunoassays of gel slices on fibrin plates, urokinase-related activity from lung was located in the region of activator from kidney cultures and of high molecular weight urokinase from urine (Mr.55,000). Tissue activator-re la ted activity from lung and intestine were at approximately 65,000 and that from HeLa at 75,000 molecular weight substances. These studies indicate the production in human tissues of distinct types of plasminogen activators - one corresponding to urokinase, and other(s) related to tissue/vascular activators.
Platelet and Plasma Serotonin in Glomerulonephritis
Authors: Parbtani A. , Frampton G. , Cameron J.S.
Published: May 2019

Intra-platelet and free plasma serotonin levels can be measured easily on casually collected samples of ACD-blood. Platelet serotonin levels were depressed and plasma serotonin levels were raised in progressive immune-complex mediated glomerulonephritis (i.e. patients with disease resulting in renal failure), but were normal in non-progressive forms of glomerulonephritis. Platelet serotonin levels correlated well with the immunological activity of the disease and to a lesser extent with renal function in the groups and in individual patients studied serially, particularly in SLE nephritis. Platelet serotonin levels also correlated with serum-platelet aggregating material (believed to be immune complexes) and with Raji cel1-detected circulating immune complexes. Uptake of 14C-serotonin by platelets in vitro did not correlate with platelet serotonin levels in progressive primary glomerulonephritits, but did so in SLE nephritis.

Platelet serotonin assay provides further evidence of platelet involvement in the pathogenesis of glomerular injury, suggests platelet activation in the circulation, and provides an easy measure of this involvement in relation to the clinical and the immunological disease activity.
Spin Probe Spin Labeled Studies of Heparins and Derivatives
Authors: Lasker S.E. , Pullman I. , Pechauskas R. , Patel C.
Published: May 2019

The polyelectrolyte heparins have a multiplicity of activities in vivo and in vitro including interactions in the coagulation cascade. The stereochemical factors or functional groups specifically associated with these activities have not been clarified. In order to further study the structure-function relationships we used a probe technique, labeling the carboxyl groups of heparins, derivatives and chondroitins with ATEMPO. Electron spin resonance and biological activity studies of nitroxide labeled heparins, and derivatives reveal a small correlation time (Ic) indicating that the label has free rotation when complexed to the carboxyl group. The labeled compound retains 85% of its anticoagulant activity and the complex binds antithrombin III to the same extent as heparin. Nuclear magnetic resonance spectra of the derivatives indicate that the complexing agent, carbodiamide is differentially displaced by ATEMPO at some of the carboxyl groups. The technique permits study of the stereochemistry of the carboxyl groups present in the polysaccharide.
A Heparin Analogue that Releases Endogenous Heparan Sulphate
Authors: Thomas D.P. , Barrowcliffe T.W. , Johnson E.A. , Stocks J. , Dawes J. , Pepper D.S.
Published: May 2019

Heparan sulphate (HS), a near-relative of heparin, but with much less anticoagulant activity in vitro, is bound to cell surfaces. We examined HS isolated from porcine intestinal mucosa, and found that although the material had low anticoagulant activity by APTT, it had a marked effect in anti-Factor Xa clotting assays, giving anti-Xa/APTT ratios of approximately 4:1 in terms of specific activity. In crossed immunoelectrophoresis experiments, HS bound to antithrombin III, but at higher concentrations than heparin. A heparin analogue (a polysulphated chondroitin), while virtually inactive in vitro, nevertheless when administered s.c. to man potentiated the effect of anti-Factor Xa to an extent comparable to that produced by low-dose heparin, but with an anti-Xa/APTT ratio of 4:1. The analogue also produced a marked release of lipoprotein lipase and a four-fold increase in the level of circulating PF4, as measured by radioimmunoassay. The anti-Xa and APTT effects of HS in vitro are very similar to those produced by the analogue in vivo, and it is suggested that the analogue releases not only lipoprotein lipase and PF4, but also HS. The drug-induced release of an endogenous glycosamino-glycan (probably from the endothelial lining of vessels) with anti-Xa activity may represent a fruitful approach to the prophylaxis of venous thrombosis.
Studies on the Organisation of Glycoproteins and Proteins in Human Platelet Membranes
Authors: Nurden A.T. , Dupuis D. , de la Baume H. , Caen J.P.
Published: May 2019

Addition of wheat germ agglutinin (WGA) (50 ug/ml) to washed human platelets (3 x 108/ml) resulted in platelet activation and the release of l4C-5HT within the same time scale as 0.05 units/ml thrombin. In contrast, succinyl-WGA (100 ug/ml) induced no platelet response. The increased valency of WGA (4) compared with succinyl-WGA (2) suggests that the activation is induced through the cross-linking (immobilisation ?) of closely associated receptors in the membrane. This finding induced us to attempt to cross-link and thereby identify adjacent molecules in the membrane by “near-neighbour” analysis. Constituent -SH groups were oxidised employing Cu2+/phenanthroline or diamide as catalysts, and polymers formed as a result of intermolecular -S-S- formation between adjacent molecules were identified by SDS-PAGE. Although previous reports have shown that the major human platelet membrane glycoproteins contain -SH groups, no apparent cross-linking of the glycoproteins was located following the incubation of either washed platelets or isolated membranes with Cu2+/phenanthroline or diamide. However bidimensional SDS-PAGE (1st dimension non-reduced, 2nd dimension reduced) showed the presence of several protein polymers including complexes formed by the cross-linking of 3 large polypeptides of M. Wt. 250 000, 220 000 and 200 000. These components were easily eluted from membrane vesicles at pH 10 and may represent closely associated constituents at the cytoplasmic surface of the plasma membrane.
Selective Absence of Large Forms of Factor VIII/Von Willebrand Factor in Acouired Von Willebrano’s Syndrome. Response to Transfusion
Authors: Meyer D. , Frommel D. , Larrieu M.J. , Zimmerman T.S.
Published: May 2019

A previously healthy elderly man with sudden onset of mucocutaneous bleeding demonstrated a monoclonal IgG gammapathy associated with severe vWD (VIII:C, VIIIR:Ag and VIIIR:RCo <10%). This syndrome has been stable over a ten year period. Associated qualitative abnormalities of F. VIII/vWF were demonstrated by immunoradiometric assay (decreased maximal antibody binding capacity) and radio-crossed immunoelectrophoresis using 125-I-radio-labeled, affinity purified, specific rabbit anti-VIIIR:Ag antibody (lack of the large, slow-moving, forms of VIIIR:Ag). The late clinical onset, negative family history and existence of quantitative as well as qualitative abnormalities of F. VIII/vWF are consistent with an acquired form of variant of vWD. However the abnormality of bleeding time (15 min.) and the hemorrhagic syndrome were less severe than in congenital variants of vWD. Following transfusion of cryoprecicitate (containing all molecular forms of F. VIII/vWF), the recovery of F. VIII/vWF was lower than expected. There was a rapid removal of the large forms of VIMR:Ag, paralleled by a decay of VIIIR:RCo and absence of sustained rise of VIII:C. Although no inhibitor to VIII:C, VIIIR:RCo and VIIIR:Ag was demonstrable when testing the patient’s plasma or IgG fraction, this study indirectly supports the existence of an inhibitor preferentially interacting with the larqe F. VIII/vWF polymers.
Apparent ‘Dominant’ and ‘Recessive’ Inheritance of Von Willebrand’s Disease (VWD) Within the Same Kindreds, Possible Biochemical Mechanisms
Authors: Bloom A.L. , Peake I.R.
Published: May 2019

In each of two vWd kindreds the inheritance pattern exhibited both ‘recessive’ and ‘dominant’ features In one family a severely affected patient had factor VIII clotting activity (VIIIC) 0-lu/dl, Factor VIII related antigen (VIIIRAg) <0.0lu/dl, ristocetin co-factor (VIIIRiCoF)<3u/dl, and bleeding time (BT) >15 min. He developed an inhibitor to VIIIRiCoF during cryoprecipitate (cryo) treatment. His VIIIC response to cryo (? lorge aggregate VIIIRAg) was much less than to Hemofil (? smaller aggregate VIIIRAg) and the antibody inhibited cryo-VIIIRiCoF more than Hemofil-VIIIRiCoF. He had symptomless related parents both of whom had slightly reduced levels of VIIIRAg ond he apparently hod homozygous recessive disease. However another affected Family member had apparently typlcol heterozygous vWd (VIIIC Vdl, VIIIRAg 3u/dl, VIIIRiCoF <3u/dl, BT >15 min). Similarly in the second kindred both a severe(VIIIRAg <0.0lu/dl) and also moderately oFFected patients with classical intermediate vWd were present (eg VIIIRAg 20u/dl). Consanguinity was not demonstrable in the extrerl patient’s symptomless parents but their levels of VIIIRAg were slightly reduced. It is suggested that this mixed inheritance may be associated with the fact that VIIIRAg/RiCoF exists as a series of homologous oligomers of which the largest exhibit greatest platelet-related activities. The variable inheritance pattern may be due to interaction between maternally and paternally derived protomers or related biosynthetic pathways so that various series of oligomers are produced in the offspring.
Suitability of the Factor Sensitive Thromboplastin FS for The Control of Oral Anticoagulation
Authors: Duckert F. , Marbet G. A. , Beeser H. , Gouault-Heilmann Michèle , Loeliger E. A.
Published: May 2019

A new rabbit brain thromboplastin, Thromboplastin FS (Dade) was tested in 4 centers involving 997 patients including 606 coumarin treated patients. It was compared in each center with another rabbit brain preparation (Thromboplastin C, Dade) and with the routinely used thromboplastin (human brain thromboplastin, Thrombotest, Thromborel and Thromboplastin Stago). Tested on plasmas deficient in factor II, V, VII and X the sensitivity of Thromboplastin FS was comparable to human brain preparation and better than Thromboplastin C, Thromborel and Thromboplastin Stago. The calibration constants (cc) between the thromboplastins calculated according to WHO recommendations confirmed the outstanding sensitivity of Thromboplastin FS. It was comparable to human brain thromboplastin (cc=0.986), less sensitve than Thrombotest (cc=0.663), much more sensitive than Thromboplastin C (cc=2.115-2.377), Thromborel (cc=2.323) and Thromboplastin Stago (cc-1.896). The new Thromboplastin FS may be very promising for the control of oral anticoagulation.
Plasminogen Activators in Human Seminal Plasma
Authors: Rijken D.C. , Wijngaards G.
Published: May 2019

The presence of fibrinolytic activity in seminal plasma has been known for many years. However, the identity of this activity has not yet been clarified satisfactorily. Recently, by using antisera against urokinase and tissue activator purified from human uterus, we demonstrated that these activators are distinct proteins. In this study both antisera were used to identify the fibrinolytic activity in human seminal plasma. Identification was achieved on fibrin plates by measuring to what extent the fibrinolytic activity of the seminal plasma was quenched by the IgG fraction of the antisera. It appeared that tissue activator antibodies as well as urokinase antibodies partly quenched the activity. A mixture of both antibodies totally quenched the activity. This indicates, that the fibrinolytic activity of seminal plasma should be ascribed to plasminogen activators and that they can be differentiated on account of their immunological relationship with tissue activator or urokinase.

In contrast to the tissue activator related activity, the urokinase related activity of seminal plasma was found to be inhibited by added Cl inhibitor. However, under similar conditions, the activity of a reference urokinase solution could hardly be inhibited by Cl inhibitor. This discrepancy could be explained by the presence in seminal plasma of a precursor of urokinase which cannot be activated in the presence of Cl inhibitor.
Effects of Low Molecular Weight Fibrinogen Degradation Products on Lymphocytes, Macrophages and Kidney Cells in Cultures
Authors: Kopeć M. , Rosazkowski W. , Szmigielski S. , Gerdin B. , Saldeen T.
Published: May 2019

Dialysable peptides of mw 2200 to 3500 D(LMW-FDP) were isolated from products of exhaustive proteolysis of fibrinogen by plasmin. LMW-FDP inhibited in a dose dependent manner incorporation of 3H-thymidme into cultures of human blood lymphocytes simulated with PHA, concanavalin A or bacterial lipopolisaccharide. These effects were inversely related to stimulation indices. Phagocytosis of 3H-thymidine labelled Staphylococcus aureus by mouse peritoneal macrophages was also inhibited by LMW-FDP. These peptides induced a cytotoxic effect on continuous line of rabbit kidney cells in cultures as manifested by inhibition of 86Rb and 3H-incorporation as well as by releese of 86Rb and 61Cr from prelabelled cells. Eight fractions obtained by molecular sieving of LMW - FDP on Bio-Gel P-6 columns were found to differ pronouncedly in biological activities.
L-Lysine: A Modulator for the Relative Activity of High Molecular Weight (HMW) and Low Molecular Weight(LHW) Urokinase(UK)
Authors: Shen L.L. , Holleman W.H.
Published: May 2019

L-Lysine(Lys), in a concentration dependent manner, progressively inhibited UK-activated lysis of human plasma clots as demonstrated by Ploug test-tube method and elastometric measurements. Lys was more effective with HMW UK than LMW UK, and the effect of Lys with LMW UK from tissue culture and urine sources was the same. Epsilon amino caproic acid(EACA) and tranexamic acid(TXA) were stronger inhibitors but inhibited HMW and LMW UK-induced lysis to the same degree. Elastometric measurements showed that Lys inhibition was not due to its interference with the initial clotting process nor to the reduction of clot rigidity. Amidolytic assays using chromogenic substrates showed that Lys had no direct effect, on UK, and that Lys enhanced the activation of the native Glu-plasminogen(Pg) by LMW UK, but not the activation by HMW UK. When the substrate was human fibrin clots, Lys enhanced the lysis induced by LMW UK while the lysis induced by HMW UK was inhibited; however, the extent of enhancement and inhibition was limited. We concluded that the mode of Lys action is not identical to that of EACA or TXA, and that the stronger Lys inhibition of plasma clot lysis as compared to fibrin clot lysis is due to the potentiation of plasma fibrinolytic inhibitors by Lys. The difference In effect of Lys on HMW and LMW UK-induced lyels is likely due to a partial conformation change of Glu-Pg molecule upon Lys binding. The relatively moderate interaction of Lys with Glu-Fg results In a mildly modified UK substrate which reacts preferentially with the enzyme smaller in size.
Tocainide in the Prevention of Deep-Vein Thrombosis Following Elective Hip-Surgery (Charnley Arthroplasty)
Authors: Cooke E.D. , Bowcock Sally A.
Published: May 2019

Lignocaine is effective in the prophylaxis of deep-vein thrombosis (DVT) after hip surgery. However the drug must be administered intravenously. Tocainide is an oral analogue of lignocaine with similar cardiac anti-arrhythmic effects to the parent drug. This investigation reports the results using the drug in the prevention of DVT in 32 patients following hip surgery using 400mg. 8 hourly commencing 36 hours before surgery.

In the first 12 patients, phlebography was performed on the 7th and 14th postoperative days; in the remaining patients phlebography was performed on the 14th postoperative day. The total incidence of DVT was 34 per cent (expected incidence 70-80%) However many of the thrombi in the calf were minute; the incidence of thigh vein thrombi was 3% (expected incidence 50%). These results indicate the necessity for a double-blind randomised trial of tocainide in the prophylaxis of DVT following elective hip surgery.
The Effect of I-Deamino-8-D-Arginine-Vasopressin (DDAVP) on Von Willebrand (vWF)-and Antihemophilic Factor (AHF) in Hemophilics and Healthy Men
Authors: Blättler W. , Muller M. , Jacky E. , Graf M. , Frick P.
Published: May 2019

A rise of vWF and AHF is brought about infusing DDAVP. Since vWF and AHF kinetics form the basis of its use in classical hemophilia the rise ond disappearance rote of the elicited AHF ond its response to repeated doses were studied. 13 hemophilics {AHF <.01 - .21 U/ml) ond 5 healthy men were given DDAVP (.4 >μg/kg) on 30 occosions. Blood was sampled through on infusion sef at 5 min to 10 h postinfusion. Upon DDAVP vWF increased 2.6 (±.9)-fold within 2 h in all but 1 probands. The AHF increase in probands with o measurable preinfusion activity was 3.5 (± 1.0)-fold and lineoily correlated with the preinfusion volue (r=.97). The disappearance of the elicited AHF followed first order kinetics. In hemophilics, survival time was the longer the higher the rise (r=.87). Upon repeated doses the vWF and AHF response was the smaller the shorter the interval. At 2-th-intervals the AHF rise was 2.0-fold. The study procedure, as checked infusing the solvent into healthy men, increased fibrinopeptide A (FPA) and betathromboglobulin (BTG) levels. In hemophilics, the FPA/BTG generation upon DDAVP was strictly reloted to the AHF rise. Heparin (40 or 120 U/kg) blocked the FPA/BTG-but not the vWF/AHF-increase.
Factor VIII and Fibrinolytic Response to DDAVP and Adrenaline in Normal Subjects and Dissociate Response in Von Willebrand’s Disease
Authors: Ludlam C. A. , Peake I.R. , Allen N. , Davies B.L. , Furlong R.A. , Bloom A.L.
Published: May 2019

The effect of intravenous administration of deamina-D-arginine vasopressin (DDAVP) and adrenaline an levels of factor VIII and plasminogen activator (PA) was studied. In normal subjects and mild haemophiliacs DDAVP caused a rise of plasma levels of PA, procoagulant factor VIII (V IIIC) and its antigen (VIIICAg), factor VIII related antigen (VIIIRAg) and ristacetin co-factor activity (VIIIRiCoF). Similarly in most patients with vWd (including those with VIIIRAg of increased electrophoretic mobility) an increase of PA and all FVIII activities was observed. In one patient with vWd (VIIIC 9u/dl, VIIICAg 13u/dl, VIIIRAg 3u/dl; VIIIRiCoF <3u/dl, bleeding time > 15 min.) in whom an inhibitor of VIIIRiCoF was excluded neither DDAVP nor adrenaline caused the expected rise of PA, VIIIRAg or VIIIRiCoF although VIIIC and VIIICAg were markedly increased. The bleeding times did not shorten. No rise of PA or VIII-related activities was observed after venous occlusion. The results suggest that in some patients with vWd there is defective synthesis/release of both VIIIRAg/VIIIRiCoF and PA. They are consistent with the presence of a primary endothelial cell defectin this disorder.
The Relationship Between Initial Intravascular Coagulation and Later Infectious Complications in Patients with Severe Closed Brain
Authors: Noormaa U. , Tikk A.
Published: May 2019

Survived from initial brain injury(BI), patients may later succumb to infectious complications. The present study evaluates the incidence of infections complications following proven intravascular coagulation(IC) in BI. Data of haemostasis are as follows; platelet counts, intravascular platelet aggregation, fibrinogen concentration, factor xiii activity, paracoagulation tests for soluble fibrin, fibrinolytic activity (FA) and thxomboelastography in systemic arterial and venous and in cerebral venous blood (CVB) were carried out in 62 patients, 50 of them with severe BI. The severity of IC was more clear in cases of severe BI and in CVB samples conpared with arterial ones in the first day of BI, of 43 patients nith slight evidence of IC i.e. hypercoagulation, nobody doveloped later infectious complications, of 23 patients with moderate ic i.e. consumption coagulopathy, 4 developed later pneumonia and 12-urinary tract infection and of 16 patients with severe 10 i.e. hypocoagulation with increased FA, all devoloped later pneucionia and urinary tract infectious and 9 of then - sepsis. So the relationship between the severity of initial IC and later infectious complications in BI excists.