bluuebunny/crossref_metadata_2025 huggingface dataset:
Kinetics and In Vivo Distribution of In-111-Labelled Platelets and Platelet Function in Familial Hypercholesterolaemia
Authors: Wessels P , Heyns A du P , Esterhuysen A J , Badenhorst P N , Lötter M G , Pieters H , Kotzè H F
Published: June 2018

The kinetics, in vivo distribution and sites of sequestration of autologous In-111-labelled platelets and other platelet function parameters were studied in ten patients with type IIa or IIb familial hypercholesterolaemia and thrombotic complications of atherosclerosis. The in vitro platelet aggregation response to ADP (P = 0.50) and collagen (P = 0.46); binding of fibrinogen to platelets (P = 0.61); and plasma beta-thromboglobulin levels (P = 0.42) of the patients and normal reference subjects did not differ significantly. The in vivo distribution of In-111-labelled platelets at equilibrium was within normal limits, and at the end of platelet life-span the sequestration pattern of labelled platelets in the reticuloendothelial system was also normal (spleen P = 0.31; liver P = 0.54). There was minimal evidence of in vivo platelet activation: only mean platelet lifespan (MPLS), 195±57 hours (difference between mean MPLS of patients and controls was 25 hours, with a 95% confidence interval from 23 to 31 hours; P = 0.02); mean platelet platelet turnover, 2298±824 platelets/μl/hour (P = 0.005); plasma platelet factor 4 (P = 0.02); and the mean circulating platelet aggregate ratio, 0.8±0.1 (P = 0.02); differed significantly from normal. These results suggest that abnormalities of platelet function and kinetics observed in type II hyperlipoproteinaemia cannot be ascribed wholly to the hyperlipidaemia, but may be induced by the associated atherosclerosis.
Tissue Plasminogen Activator and Urokinase in Normal, Dysplastic and Cancerous Squamous Epithelium of the Uterine Cervix
Authors: Larsson G , Larsson Å , Åstedt B
Published: June 2018

The distribution of tissue plasminogen activator (t-PA) and urokinase (u-PA) was investigated immunohistochemically in squamous epithelium of the uterine cervix, in normal and dysplastic conditions as well as in preinvasive and invasive carcinomas. Normal epithelium showed presence of both t PA and u-PA immunoreactivity only in the superficial cellular layer, whereas in preinvasive lesions they were present in all layers. In sections of normal specimens as well as preinvasive lesions the UK immunoreactivity was weaker compared to that of t-PA. Tnvnsivc lesions showed a patchy distribution of t-PA and u-PA immunoreactivity. Furthermore, it could be demonstrated that stromal cells close to the infiltrating malignant squamous epithelium contained t-PA immunoreactivity and also u-PA immunoreactivity. Further immunohistochemical studies disclosed that these cells were macrophages.
Does Increased Platelet Release Normalize During Anti-Hypertensive Treatment?
Authors: Lande Knut , Kjeldsen Sverre Erik , Eide Ivar , Leren Paul , Gjesdal Knut
Published: June 2018

Blood platelet function was evaluated in 10 men, all 50 years old, with untreated, mild hypertension. Each patient was examined four times: At the beginning of the study, after 5 weeks on placebo treatment, after the following 5 weeks on propranolol 160 mg daily, and finally after a second period of 5 weeks on placebo. At baseline the plasma level of the platelet release product (β-thromboglobulin (BTG) was 41.6 (30.5-57.0) μg/l (median and 95% confidence interval). During the first placebo period BTG was normalized to 21.0 (14.1-25.9) μg/l. While systolic blood pressure and heart rate fell during β-adrenergic receptor blockade, BTG remained unchanged throughout the rest of the observation periods. Platelet size increased significantly during treatment with β-blocker. The present study indicates that the normalization of elevated platelet function which previously has been reported to occur during anti-hypertensive drug therapy, may be explained by patient adaptation to the blood sampling procedure.
Experimental Studies on the Relative Efficacy of Dermatan Sulphate and Heparin as Antithrombotic Agents
Authors: Merton R E , Thomas D P
Published: June 2018

In this study, the anticoagulant and antithrombotic properties of unfractionated heparin (UFH) and dermatan sulphate (DS) were compared. The ability of UFH and DS to impair thrombin generation in vitro and in ex vivo plasma samples was also studied. DS has minimal anticoagulant activity by conventional assays but impairs thrombin generation both in vitro and in ex vivo plasma samples. However, thrombin generation could not be suppressed below about 35% of control values at all doses of DS studied. While this was sufficient to impair experimental venous thrombosis during 10 minutes’ stasis, DS was ineffective in preventing thrombosis following 20 minutes’ stasis in doses up to 1.25 mg/kg. In contrast, 1 μg/ml of UFH completely suppressed thrombin generation in vitro, and 150 μg/kg prevented throm- bogenesis over a period of 20 minutes’ stasis. Neither drug prolonged the bleeding time (BT) at effective antithrombotic doses, but 2.5 mg/kg UFH significantly increased the BT, whereas DS did not. While DS has antithrombotic activity, it is less effective than UFH in inhibiting thrombin generation, and as an antithrombotic agent.
The Influence of Low Molecular Weight Heparin in Combination with Dihydroergotamine on Experimental Thrombosis and Haemostasis
Authors: Bergqvist David , Nilsson Bodil
Published: June 2018

The influence of low molecular weight heparin in combination with dihydroergotamine (DHE) on thrombus formation and primary haemostasis was investigated in rabbit models. Conventional heparin in the dose of 60 units anti Xa activity/kg b. w. effectively prevented thrombus formation but the same dose of a low molecular weight fragment only gave a marginal decrease of the frequency of thrombosis. The thrombus weight was, however, significantly reduced. Doubling the dose of the heparin fragment totally abolished thrombus formation as did the combination of 60 units with DHE. With DHE it was also possible to diminish the low molecular weight heparin dose to 30 units with a good prophylactic effect. There was a small but significant increase of haemostatic plug formation time in all treatment groups except the one with low molecular weight heparin fragment 30 units anti Xa activity combined with DHE. Thus, by combining low molecular weight heparin in a low dose with DHE it is possible to prevent thrombus formation without influencing primary haemostasis in rabbits.
Human Gamma-Thrombin: Lack of Correlation Between a Platelet Functional Response and Glycoprotein V Hydrolysis
Authors: Jandrot-Perrus Martine , Guillin Marie-Claude , Nurden Alan T
Published: June 2018

The ability of purified human gamma-thrombin to stimulate platelet function was related to its capacity to degrade GP V. Compared to alpha-thrombin, much greater amounts of gamma- thrombin were required to induce platelet aggregation; and this also applied to secretion from dense bodies, alpha-granules and lysosomal granules. Platelet stimulation by gamma-thrombin was additionally characterized by the presence of a lag-phase. Platelets with 3H-labelled surface glycoproteins showed the same functional response to both alpha- and gamma-thrombin as unlabelled platelets. But while threshold levels of alpha-thrombin induced little GP V hydrolysis confirming McGowan et al. (1), amounts of gamma-thrombin which induced substantial degradation (e. g. 8.3 nM degraded 36% of platelet GP V in 3 min) were unable to sustain either platelet aggregation or secretion. These results suggest that protein-binding regions remote from the catalytic site of alpha-thrombin are more important for platelet activation than GP V hydrolysis. They also provide further support to the argument that GP V hydrolysis may not be the essential trigger of platelet activation by thrombin.
Pharmacokinetics of Heparin and Low Molecular Weight Heparin Fragment (Fragmin®) in Rabbits with Impaired Renal or Metabolic Clearance
Authors: Palm M , Mattsson Ch
Published: June 2018

The kinetics and tissue distribution of 3H-heparin and a 3H- labelled low molecular weight heparin fragment were compared in normal rabbits as well as in rabbits with blocked renal function or reticuloendothelial system (RES). Radioactivity in plasma, urine, liver and kidneys, as well as anti-FXa activity in plasma were determined. The plasma elimination of heparin was, when compared to normal controls, prolonged both in rabbits with renal dysfunction as well as in rabbits with blocked RES, while renal dysfunction was the only parameter that significantly prolonged the plasma half-life of Fragmin. Studies on tissue distribution in normal rabbits revealed that about 60 per cent of the radioactive heparin dose accumulated in the liver and kidney three hours after the injection, whereas the corresponding value was less than 10 per cent for the Fragmin-derived radioactivity. The recovery of radioactivity in urine within three hours was 5 and 35 per cent of the dose, respectively, for 3H-heparin and 3H- Fragmin. It is concluded from the present study that the rapid plasma elimination of heparin in the rabbit (tl/2 = 17 minutes) is mainly due to a high tissue distribution (liver and kidney) while the plasma elimination of Fragmin (t1/2 = 28 minutes) is mainly caused by renal excretion.
Redistribution of Platelet Glycoproteins Induced by Allo- and Autoantibodies
Authors: Santoso S , Kiefel V , Mueller-Eckhardt C
Published: June 2018

The redistribution of platelet glycoproteins (GP) Ib, IIb and IIIa in response to stimulation with human platelet specific alloantibodies, autoantibodies and a quinidine-dependent antibody was investigated using immunofluorescence and a quantitative radioimmunoassay. The platelet GPs carrying the corresponding epitopes were determined using immunoblot technique or radioimmunoprecipitation. In accordance with previous results obtained with murine monoclonal platelet specific antibodies, redistribution upon stimulation with human platelet reactive antibodies was observed only due to reactions with epitopes on GP IIb, IIIa, respectively, but not on GP Ib. We therefore believe that membrane redistribution of human platelets induced by various antibodies is a function of the GP recognized by those antibodies rather than the source of the platelet reactive antibody.
Defective Fibrinolytic Response in Atherosclerotic Patients – Effect of lloprost and Its Possible Mechanism of Action
Authors: Bertelé Vittorio , Mussoni Luciana , Rosso Gianfranco del , Pintucci Giuseppe , Carriero Maria Rita , Merati Maria Grazia , Libretti Arnaldo , Gaetano Giovanni de
Published: June 2018

Plasma fibrinolytic activity and tissue-type plasminogen activator (t-PA) were defective in response to venous stasis in five out of ten patients with peripheral occlusive artery disease. Discontinuous infusions of iloprost, a stable synthetic analogue of prostacyclin, restored a normal fibrinolytic response in all five patients but did not induce a parallel increase of plasma t-PA. These findings suggest that in addition to the possible benefits due to its vasodilatory and antiplatelet activity, iloprost may improve the fibrinolytic activity in patients with atherosclerotic disease, providing them with further antithrombotic protection. The profibrinolytic effect of iloprost seems not to depend on its ability to induce vascular t-PA release. Rather, it might be related to its inhibitory effect on PAI release from platelets, endothelial cells and/or hepatocytes. Venous occlusion test represents an easy diagnostic approach to fibrinolytic defects, even if related to arterial disease, and may help select patients who need therapeutic intervention.
Correlation Between the Enzymatic Activity, Anticoagulant and Antiplatelet Effects of Phospholipase A2 Isoenzymes from Naja nigricollis Venom
Authors: Manjunatha Kini R , Evans Herbert J
Published: June 2018

The three phospholipase A2 isoenzymes from Naja nigricollis crawshawii snake venom inhibit both blood coagulation and platelet aggregation. To investigate the correlation between phospholipid splitting ability of these enzymes and their inhibitory activities, the effects of various preincubation times and the inclusion of EDTA were examined. Preincubation of plasma and thromboplastin with the phospholipase isoenzymes resulted in an increase in Ca2+-initiated clotting time with time of preincubation. Incubation of the isoenzymes with EDTA before their addition to the plasma-thromboplastin mixture reduced the anticoagulant effect. These results indicate that the catalytic activity contributes at least partially to the anticoagulant effect. However, inhibition of platelet aggregation appears to be independent of enzymatic activity since there is no increase in inhibition with time of preincubation of platelets and phospholipases, and inclusion of EDTA has no significant effect on inhibition of aggregation. All three enzymes appear to belong to class B of the platelet affector PLA2 enzymes as determined by platelet effects, since they do not initiate platelet aggregation.
Glycoprotein Ib Has a Partial Role in Platelet-von Willebrand Factor Collagen Interaction
Authors: Aihara Morio , Tamura Ken , Kawarada Ryuko , Okawa Keizou , Yoshida Yutaka
Published: June 2018

The adhesion of human fixed washed platelets (FWP) to collagen was decreased after treatment with Serratia marcescens protease (SP), which removed 95% of the glycocalicin from platelet membrane glycoprotein (GP) lb. However, the diminished adhesion of SP treated FWP to collagen could still be increased in the presence of purified von Willebrand factor (vWF). This ability of vWF to increase FWP adhesion to collagen is defined as collagen cofactor (CCo). The adhesion of FWP to collagen was not affected by a monoclonal antibody (MAb) to GP Ilb/IIIa (10E5), that inhibits ADP and collagen induced platelet aggregation. On the other hand, it was decreased by 50% by a MAb to GP lb (6D1), that inhibits ristocetin induced platelet aggregation. Adhesion of FWP in buffer to collagen was completely inhibited by Ricinus communis agglutinin I or concanavalin A, while Lens culinalis agglutinin and wheat germ agglutinin showed 50% inhibition. The FWP adhesion to collagen in the presence of vWF (normal plasma) was unaffected by MAbs to GP Ilb/IIIa (10E5, P2, HPL1) but was decreased to 32-38% by MAbs to GP lb (6D1, AN51, HPL11). A MAb to vWF (CLB-RAg 35), that inhibits ristocetin induced binding of vWF to platelets, decreased the CCo of normal plasma by 70%. The MAb, CLB-RAg 201, that inhibits the binding of vWF to collagen, completely inhibited the CCo of normal plasma. In conclusion, our data suggest that (1) GP lb has a partial role in FWP adhesion to collagen; (2) the binding of vWF to collagen is required for the expression of CCo; (3) CCo is partly mediated through GP lb; but (4) other platelet membrane protein(s) besides GP lb or GP Ilb/IIIa must also be involved in FWP-vWF-collagen interactions.
Decreased Platelet-Free Dopamine and Unchanged Noradrenaline and Adrenaline in Essential Hypertension
Authors: Kjeldsen S E , Gjesdal K , Leren P , Eide I K
Published: June 2018

The content of free-catecholamines in blood platelets is much higher than in plasma and platelet catecholamines must be taken up from plasma, since platelets lack the enzymes for catecholamine synthesis. There is some evidence that platelet catecholamine content under certain circumstances may be an integrated measure of plasma catecholamine concentrations over time. Platelet-free catecholamines were therefore assayed in 18 untreated patients with essential hypertension and in 16 normotensive control subjects. Mean platelet-free dopamine in the hypertensive group was 3.7 ± 0.4 pg/mg platelet weight, i.e. significantly less than the 6.5 ± 0.9 pg/mg found in the normotensive (p <0.005). Platelet contents of noradrenaline and adrenaline did not differ. Decreased platelet-free dopamine and unchanged platelet noradrenaline and adrenaline persisted after adjustment for increased body weight in the hypertensive group. Although the reasons for decreased platelet-free dopamine in the hypertensive group remain unknown, this finding may add to previous result showing facilitated release of granular contents from blood platelets in patients with essential hypertension. Our data do not support platelet levels of free-catecholamines to be a marker of increased sympathetic tone in essential hypertension.
The Influence of Carbohydrate Structure on the Clearance of Recombinant Tissue-Type Plasminogen Activator
Authors: Hotchkiss A , Refino C J , Leonard C K , O'Connor J V , Crowley C , McCabe J , Tate K , Nakamura G , Powers D , Levinson A , Mohler M , Spellman M W
Published: June 2018

Modification of the carbohydrate structures of recombinant tissue-type plasminogen activator (rt-PA) can increase or decrease its rate of clearance in rabbits. When rt-PA was treated with sodium periodate to oxidize carbohydrate residues, the rate of clearance was decreased from 9.6 ± 1.9 ml min−1 kg−1 to 3.5 ± 0.6 ml min−1 kg−1 (mean ± SD, n = 5). A similar change in the clearance of rt-PA was introduced by the use of endo-β-N-acetyl- glucosaminidase H (Endo-H), which selectively removes high mannose asparagine-linked oligosaccharides; the clearance of Endo-H-treated rt-PA was 5.0 ± 0.5 ml min−1 kg−1. A mutant of rt-PA was produced with an amino acid substitution at position 117 (Asn replaced with Gin) to remove a potential glycosylation site that normally contains a high mannose structure. The clearance of this material was also decreased, similar to the periodate and Endo-H-treated rt-PA. Conversely, when rt-PA was produced in the CHO 15B cell line, which can produce only high mannose oligosaccharide structures on glycoproteins, the clearance was increased by a factor of 1.8. These results demonstrate that the removal of rt-PA from the blood depends significantly upon the nature of its oligosaccharide structures.
The Anticoagulant Properties of a Modified Form of Protein S
Authors: Mitchell C A , Kelemen S M , Salem H H
Published: June 2018

Protein S (PS) is a vitamin K-dependent anticoagulant that acts as a cofactor to activated protein C (APC). To date PS has not been shown to possess anticoagulant activity in the absence of APC.

In this study, we have developed monoclonal antibody to protein S and used to purify the protein to homogeneity from plasma. Affinity purified protein S (PSM), although identical to the conventionally purified protein as judged by SDS-PAGE, had significant anticoagulant activity in the absence of APC when measured in a factor Xa recalcification time. Using SDS-PAGE we have demonstrated that prothrombin cleavage by factor X awas inhibited in the presence of PSM. Kinetic analysis of the reaction revealed that PSM competitively inhibited factor X amediated cleavage of prothrombin. PS preincubated with the monoclonal antibody, acquired similar anticoagulant properties. These results suggest that the interaction of the monoclonal antibody with PS results in an alteration in the protein exposing sites that mediate the observed anticoagulant effect. Support that the protein was altered was derived from the observation that PSM was eight fold more sensitive to cleavage by thrombin and human neutrophil elastase than conventionally purified protein S.

These observations suggest that PS can be modified in vitro to a protein with APC-independent anticoagulant activity and raise the possibility that a similar alteration could occur in vivo through the binding protein S to a cellular or plasma protein.
Antithrombin III Geneva: A Hereditary Abnormal AT III with Defective Heparin Cofactor Activity
Authors: de Moerloose P A , Reber G , Vernet Ph , Minazio Ph , Bouvier C A
Published: June 2018

A 43-year-old man presented a pulmonary embolism. The unusual circumstances of apparition, the age and the increased heparin requirements suggested an antithrombin III (AT III) deficiency. AT III activity was low in the propositus and seven other members of his family (mean 55%), but immunologic levels were normal (mean 110%). Crossed immunoelectrophoresis in absence of heparin showed a normal pattern, but in presence of heparin showed an abnormal peak as compared with controls. Kinetics experiments showed a normal inhibition of thrombin and Xa in absence of heparin, but abnormal in presence of heparin. Affinity chromatography on heparin-Sepharose revealed two populations of AT III, one of which was devoid of heparin cofactor activity. The toponym AT III Geneva is proposed for this new familial abnormal AT III with defective heparin cofactor activity.
The Interaction of Activated Protein C and Thrombin with the Plasminogen Activator Inhibitor Released from Human Endothelial Cells
Authors: de Fouw N J , van Hinsbergh V W M , de Jong Y F , Haverkate F , Bertina R M
Published: June 2018

The effects of human activated protein C (APC) and thrombin on plasminogen activator inhibitor (PAI-1) released from cultured human umbilical endothelial cells, grown in serum-free 35S-methionine containing medium, were studied in two ways: 1) measurement of PAI-1 activity with an amidolytic assay, and 2) immunoprécipitation of the medium with anti-PAI-1 IgG, antiprotein C IgG or anti-thrombin IgG followed by SDS-PAGE and autoradiography.

Addition of APC or thrombin to the endothelial cell conditioned medium results in a time and concentration dependent loss of PAI-1 activity and in the degradation of PAI-1 from 46 kD into a 42 kD product.

After incubation of the medium with APC in the presence of cells, an additional band of 95 kD was found, which could be immunoprecipitated with both anti-PAI-1 IgG and anti-protein C IgG, indicating the formation of an APC-PAI-1 complex before degradation occurs. No complex could be detected after incubation of the medium with thrombin in the presence of endothelial cells.

Blocking the active sites of APC and thrombin prevented both the formation of APC-PAI-1 complexes and the inactivation and degradation of PAI-1. After removal of the active PAI-1 from the medium, no degradation of the inactive PAI-1 by APC or thrombin could be found.

It is concluded that both APC and thrombin react with the active PAI-1, resulting in inactivation and degradation of PAI-1.
Evaluation of Monoclonal Antibodies to vWf Antigen for Use in Autoradiographing vWf Multimer Analysis
Authors: Enayat M S , Hill F G H , Robinson W , Prowse C V , Hornsey V S
Published: June 2018

Nine monoclonal antibodies (MAb, coded ESvWF 1-5, 7-10) to human von Willebrand’s factor (vWf) have been studied for their labelling characteristics with 125I and their ability to demonstrate vWf multimers by autoradiography after discontinuous SDS electrophoresis on agarose and agarose/acrylamide gels in plasma from normal and von Willebrand’s disease (vWD) patients. Of ESvWF 1, 2, 4, 7, 8 and 9, ESvWF 2 gave autoradiographs most similar to those obtained with a polyclonal antibody. The others gave much fainter staining of all bands suggesting that the epitope detected by ESvWF 2 is either more common or, less easily altered by SDS than the epitopes identified by the other MAb’s. ESvWF 2 gave a different staining pattern with a IIC vWD patient than that obtained using a polyclonal antibody. The most striking result, however, was shown by ESvWF 3, 5 and 10 which failed to stain the lower band of the lower molecular weight multimer. This suggests that the lower triplet band, unlike the central and upper bands of the triplet, does not contain the epitope identified by these three MAb’s. With these reagents it has been possible to show, for the first time, that epitope differences do exist between the constituent bands of the vWf triplet.
Responses of Platelets to Strains of Streptococcus sanguis: Findings in Healthy Subjects, Bernard-Soulier, Glanzmann’s, and Collagen-Unresponsive Patients
Authors: Soberay A H , Herzberg M C , Rudney J D , Nieuwenhuis H K , Sixma J J , Seligsohn U
Published: June 2018

The ability of endocarditis and dental strains of Streptococcus sanguis to induce platelet aggregation in plasma (PRP) from normal subjects were examined and compared to responses of PRP with known platelet membrane glycoprotein (GP) and response defects.

S. sanguis strains differed in their ability to induce normal PRPs to aggregate. Strains that induced PRP aggregation in more than 60% of donors were significantly faster agonists (mean lag times to onset of aggregation less than 6 min) than those strains inducing response in PRPs of fewer than 60% of donors.

Platelets from patients with Bernard-Soulier syndrome aggregated in response to strains of S. sanguis. In contrast, platelets from patients with Glanzmann’s thrombasthenia and from a patient with a specific defect in response to collagen were unresponsive to S. sanguis. These observations show that GPIb and V are not essential, but GPIIb-IIIa and GPIa are important in the platelet response mechanism to S. sanguis. Indeed, the data suggests that the platelet interaction mechanisms of S. sanguis and collagen may be similar.
Eine In-Vitro-Methode zur Bestimmung der Thrombozytenadhäsivität
Authors: Jacobi E , Hagemann G , Poliwoda H
Published: June 2018

Es wird über eine neue In-vitro-Methode zur Bestimmung der Thrombozyten-adhäsivität berichtet. Aus strömendem Zitratblut scheiden sich an Kunstfasern Plättchen ab, deren Menge mit Hilfe des Folin-Reagenz bestimmt wird. Dabei müssen folgende Parameter konstant gehalten werden: Blutströmungsgeschwindigkeit, Temperatur und Tageszeit der Blutabnahme. Als besonders vorteilhaft haben sich Fasern mit rauher Oberfläche erwiesen. Die Adhäsivität als Funktion der Thrombozytendichte (n) stellt sich in nächster Annäherung als 0,1 √n heraus, welches eine Umrechnung auf eine normale Thrombozytendichte bei unterschiedlich gemessenen Dichten erlaubt. Die Standardabweichung der Meßwerte wurde mit ± 9,1%, der Meßbereichsumfang mit wenigstens dem 30fachen der Standardabweichung bestimmt.
Appearance of Thrombogenic Tendency Induced by Adrenaline and Its Prevention by β-Adrenergic Blocking Agent, Nialamide and Pyridinolcarbamate
Authors: Yamazaki H , Sano T , Odakura T , Takeuchi K , Matsumura T , Hosaki S , Shimamoto T
Published: June 2018

In 52 men and 33 women, the total, adhesive and nonadhesive platelet counts, onestage prothrombin time, euglobulinlysis time, and plasma nonesterified fatty acid were measured before and after an intravenous injection with 0.1 μg/kg of adrenaline or 1-noradrenaline. Three hours before the injection, placebo, 30 mg of phenoxybenzamine, 300 mg of pronethalol, 100 mg of nialamide or 100 mg of pyridinolcarbamate was administered orally under the double blind technique. Five minutes after the injection with adrenaline in 15 cases pretreated with placebo, a decrease in adhesive platelet count, and a shortening of one-stage prothrombin time and euglobulin lysis time were observed with statistical significance (P ˂ 0.01 ∼ 0.05). These changes were not observed in 15 cases injected with saline or noradrenaline. The changes were not observed in 10 cases pretreated with pronethalol, but were observed in 10 cases pretreated with phenoxybenzamine. So the effect of adrenaline may be due to stimulation of ß-adrenergic receptor sites. Meanwhile, the decrease in adhesive platelet count and the enhancement of blood coagulability were prevented by nialamide and pyridinolcarbamate, while the enhancement of fibrinolysis was not. The preventive effect of nialamide was less than that of pyridinolcarbamate. Phenoxybenzamine and nialamide prevented an increase in systolic blood pressure induced by adrenaline, and pronethalol prevented tachycardia by adrenaline, while pyridinolcarbamate did not affect these changes. Changes in plasma nonesterified fatty acid showed no relationship to the above hematological changes.
The Effect of Stress and Warfarin on the Adrenal Gland in Relation to Spontaneous Hemorrhage, as Judged by Measurement of Adrenal Ascorbic Acid and Serum Corticosterone
Authors: Chattopadhyay S , Johnson D. D , Millar G. J , Jaques L. B
Published: June 2018

Rats were subjected to the following procedures: No treatment, Stressor (10% NaCl i.p.), Warfarin for 7 days, Stressor followed by Warfarin; and groups were sacrificed at intervals for assessment of spontaneous hemorrhage and of adrenal ascorbic acid concentration. There was no hemorrhage in the no treatment and stressor groups; some hemorrhage in the warfarin group; profound hemorrhage with Warfarin + Stressor. The adrenal ascorbic acid concentration was found to be lower, 8 h and again 5 days after stress, and remained lower in the warfarin + stress animals. Warfarin had no effect on adrenal ascorbic acid level.

In another series of experiments in which the stress consisted of an electric current to the cage floor for 6 sec over 15 min, rats were sacrificed daily for determination of serum corticosterone concentration and occurrence of spontaneous hemorrhage. There was a statistically significant increase of serum corticosterone concentration with stress, warfarin and combined warfarin and stress treatments (P< 0.001 for all three variables). There was a significant correlation (r = 0.96 and 0.89, P< 0.01) for serum corticosterone concentration with hemorrhage score and incidence of hemorrhage in stressed rats receiving warfarin, but not in those receiving only warfarin. The results indicate an activation, rather than an exhaustion, of the pituitary-adrenal axis during the combined action of anticoagulant and stress, which results in the development of spontaneous hemorrhage.
Quantitative Determination of Intravascular Coagulation in Vivo in Various Organs in the Dog
Authors: Busch Ch , Lundqvist H , Saldeen T
Published: June 2018

A technique for determination of intravascular coagulation in different organs by means of continuous external detection of 125I-fibrinogen in the dog is described.

It is suggested that this technique would be of value in human beings as a complement to the present, often unsatisfactory methods for determination of microembolism.
The Estimation of Blood Platelet Survival
Authors: Murphy E. A
Published: June 2018

A study has been made of the distortion introduced by using the multiple-hit (gamma) model for the construction of population curves where some of the underlying assumptions are violated. The three violations, over which, because of physiological evidence, there is greatest occasion for concern, are considered in some detail. These distortions are that a latent interval exists between hits so that true survival would follow a gamma distribution shifted to the right; that the probability of responding to an environmental insult may change with the age of the platelet so that survival would follow an Erlangian distribution; and that platelet destruction may be governed by competing processes of differing relative importances. Under fairly general conditions, the discrepancies between these models (if true) and the “best fit” (minimax) gamma function of integral order are small (less than 3%) especially when compared with the kind of experimental errors usually encountered.

It seems then that until such time as very precisely measured data are available, the multiple-hit model may be used with confidence at least as a device for estimating the mean.

It is pointed out, however, that the estimation of the number of hits to destruction, or the mean waiting time between them separately, is likely to be imprecise. Nevertheless, the estimation of the mean (which is the ratio of the number of hits to the reciprocal of the waiting time) may still be much more satisfactory than estimates not explicity defended by rational models. The latter consideration is statistical and will be dealt with in a later paper in this series.
The Leukocyte Digestion of Fibrinogen Degradation Products (FDP)
Authors: Gibiński K , Lipiński B , Trusz-Gluza M
Published: June 2018

While the native fibrinogen is not digested by the leucocyte proteases both the early and late FDP are digestible without any denaturating reagent. Thus, this reaction may occur in vivo indicating an unknown role of granulocytes in paracoagulation.
A Comparative Study of Adult and Fetal Sheep Fibrinogen, Sulf-Fibrinogen and Fibrinogen Degradation Products
Authors: Loly W , Israels L. G , Bishop A. J , Israels E. D
Published: June 2018

The results of these comparative studies of adult and fetal sheep fibrinogen are as follows:

1. Adult D fragments and sulf-fibrinogen are identical to their fetal counterparts when examined by acid alkaline polyacrylamide gel electrophoresis.

2. Adult and fetal E fragments show identical mobility on acid and alkaline polyacrylamide gel electrophoresis, but in the alkaline system the adult E piece consistently shows 3 separate bands while the fetal E peice, with one exception, is one diffuse band. The pattern of banding of the two E fragments is identical in the acid gel system.

3. Adult D and E fragments and sulf-fibrinogen are immunologically identical to their fetal counterparts.

4. Thrombin times on fetal plasmas are, on the average, prolonged over those on adult plasmas ; this is probably due to the lower absolute fibrinogen concentration of fetal plasma.

5. Adult and fetal fibrinogen showed no differences in the pH dependency of their thrombin times, indicating that the polymerization process is probably the same in both.

6. It is essential, in studies of fibrinogen structure and function, to work with very pure fibrinogen preparations, free from fibrin monomers and fibrinogen degradation products.
Corrective Effect of Sialic Acid on the Clotting of Factor VIII Deficient Blood
Authors: Rubin H , Fernbach T , Ritz N. D
Published: June 2018

Optimal concentrations of N-acetyl neuramic acid (NANA) accelerated the clotting of factor VIII deficient whole blood and plasma. There was no effect on normal plasma and certain other factor deficient plasmas.

NANA increased thromboplastin formation by factor VIII deficient plasma, as measured by the Thromboplastin Generation Test.

NANA possesses an inhibitory effect on rabbit brain thromboplastin and cephalin in recalcification tests, over the values of its controls, but-in sharp contrast, enhanced the effect of Russell Viper Venom (RVV) and RVV-cephalin.

The amino sugar N-acetyl mannosamine, possessing common functional groups with NANA, also shortened the clotting time of factor VIII deficient whole blood and plasma.

It is suggested that NANA acts as a cofactor to factor VIII which is present in trace amounts in hemophilic plasma.
The Adsorption of Coagulation Factors onto Phospholipids
Authors: Hemker H. C , Kahn M. J. P , Devilee P. P
Published: June 2018

It is shown that factors VIII and IX react with phospholipid in a way fully analogous to the reaction of factors V and X. From this it is concluded that a complex of factors VIII and IX adsorbed onto a phospholipid micelle may be the entity that activates factor X in the intrinsic pathway. The implications of this finding for the concepts of the reaction mechanism of blood coagulation are discussed. It is shown that instead of the original 7-step cascade of Macfarlane, a 4-step cascade consisting partly of complex enzymes is more probable.
Assay of Progressive Antithrombin in Plasma
Authors: Abildgaard U , Gravem K , Godal H. C
Published: June 2018

A simple two-stage assay of progressive antithrombin activity in plasma is described. After heat defibrination, plasma is diluted VlO in buffer (pH 8.0) and incubated with thrombin for 3 min at 37° C. After incubation, residual thrombin activity is determined with standard substrate (antithrombin clotting time). By substituting buffer for diluted plasma, the buffer clotting time is determined. The antithrombin activity may be calculated from these two clotting times or read from a standard curve. It is expressed in per cent of pooled normal plasma. The range in 36 normal plasmas was 74-116%, the S. D. 11%, and the S. E. M. 5.7%. The combination of simplicity and accuracy makes the present method compare favourably with earlier methods.
A Simple Technique for the Measurement of Plasma Heparin Concentration during Anticoagulant Therapy
Authors: Marder V.J
Published: June 2018

Blood is incoagulable for several hours after therapeutic doses of intravenous heparin when tested by the glass clotting time (CT) or activated partialthromboplastin test (APTT). A simple modification of the APTT is described which allows the quantitative measurement of plasma heparin levels at any time during therapy. In principal the test plasma is diluted in normal plasma before assay, thereby reducing the heparin concentration to a range which can be measured by the APTT. The corresponding plasma heparin concentration is estimated by reference to a single standard curve which applies for concentrationsfrom 0.1 to 10 I.U./ml of plasma. Since all samples are diluted in normal plasma, storage of blood at 4° or even 37° for up to 48 h does not significantly alter the results. The assay is applied in the study of plasma heparin disappearance time and in the control of anticoagulant therapy forthromboembolic disease.
Antithrombin- und Antiplasminwirkung von 4-Amidinophenylbrenztraubensäure (APPA) in vivo
Authors: Markwardt F , Klöcking H.-P , Nowak G
Published: June 2018

Es wurde die Antithrombin- und Antiplasmin Wirkung der 4-Amidinophenylbrenztraubensäure (APPA) an Kaninchen untersucht. Dabei hat sich gezeigt, daß die in biochemischen Untersuchungen als kompetitiver Hemmstoff der Gerinnungsfermente wirkende Verbindung auch in vivo antikoagulierende und antifibrinolytische Wirkungen entfaltet, die sich sowohl nach intravenöser als auch nach oraler Gabe des Inhibitors messend verfolgen lassen.

Die Wirkung von Thrombininfusionen oder Streptokinaseinfusionen wurde bei Gegenwart des Inhibitors im Blut unterbunden.

Die Resorptionsquote der APPA nach oraler Applikation beträgt 45% und die Halbwertzeit 100 min. Ein Teil der applizierten Inhibitormenge wird in unveränderter Form mit dem Urin ausgeschieden.

APPA stellt ein pharmakologisches Modell für eine neue Klasse von Stoffen zur medikamentösen Beeinflussung der Blutgerinnungs- und Fibrinolyse Vorgänge dar.
Blood Clotting, Plasma Kinins and Fibrinolysis
Authors: Gjønnæss H , Stokmorken H
Published: June 2018

In plasma of women on oral contraception a marked activation of factor VII takes place, when stored overnight at low temperatures. This activation is dependent on factor XII, and evidence is presented that it may be mediated by the plasma kinin system.
Human Prothrombin
Authors: Derleth Th. R , Penner J. A
Published: June 2018

A method is presented for the chromatographic separation of human prothrombin from variable volumes of plasma. The procedure is simple, consisting of chromatography on ECTEOLA which provides a potent product containing factor IX and X activity sufficient for therapeutic use. Additional chromatography on DEAE or Sephadex G200 will produce a prothrombin of high specific activity which appears homogeneous by ultracentrifugation and electrophoresis on cellulose acetate, although multiple protein fractions can be identified on starch gel electrophoresis.

Human prothrombin prepared by this method migrates as an alpha 2-globulin and does not appear to be altered electrophoretically or antigenically from its original state in the plasma. The characteristics of human prothrombin obtained from patients with deficiencies of factor VII or IX do not differ from normal.
Immunological Identification of Three Urokinases
Authors: Day Eugene D. , Ball Anne P.
Published: June 2018

A third urokinase has been detected by immunochemical tests in two clinical preparations of urokinase, one from Abbott and one from Sterling-Winthrop. It is component V in a series of components found in Sterling-Winthrop clinical urokinase. Components I and II are confined to Sterling-Winthrop urokinase whereas components III, IV, and V are present also in Abbott urokinase. Evidence suggests but does not prove that V is the SI component of Abbott; that IV is the S2 component of Abbott as well as the crystalline component of Sterling -Winthrop ; and that III is a higher molecular weight species of urokinase, unrelated to either IV or V, which may correspond to the 104,000 MW uroprotein of Doleschel and Auerswald. It is proposed that III be referred to as S3.
A Simple Test for Quantitation of Platelet Adhesion to Glass
Authors: Mason R. G , Gilkey J. M
Published: June 2018

A simple method for quantitating pletelet adhesiveness to glass is described. The use of minimally manipulated native whole blood and direct counting of adherent platelets are principal features of the method. Normal values for platelet adhesiveness to glass both in the absence and presence of added ADP are given. Without ADP, abnormally low platelet adhesiveness values were observed in afibrinogenemia, thrombasthenia, uremia, homocystinuria and in some patients with von Willebrand’s disease or Hageman trait. Under the same test condition, abnormally elevated values were found in some patients with von Willebrand’s disease or diabetes mellitus. With added ADP, which induced aggregation of some platelets in the blood, abnormally low platelet adhesiveness values were observed in thrombasthenia and one diabetic patient, while abnormally high values were found in von Willebrand’s disease, and in one patient with homocystinuria. The test for platelet adhesiveness performed with added ADP may have diagnostic value for patients with von Willebrand’s disease.
Untersuchungen über die durch Kollagen ausgelöste 5-HT-Freisetzung aus Blutplättchen
Authors: Barthel W , Markwardt F
Published: June 2018

Bei Einwirkung von Sehnenextrakt auf Kaninchenblutplättchen in Tyrodelösung kommt es zu einer Freisetzung des gespeicherten 5-HT. Die 5-HT-Freisetzung ist abhängig von der zugesetzten Sehnenextraktmenge und läuft mit großer Geschwindigkeit ab. Glukose- und Kalziumentzug sowie die Vorbehandlung des Sehnenextrakte mit Kollagenase hemmen die Freisetzungsreaktion. Papaverin, NEM und das Isochinolinderivat B30 hemmen die durch Sehnenextrakt ausgelöste 5-HT-Freisetzung in dosisabhängiger Weise. Adenosin und Azetylsalizylsäure hatten unter den verwendeten Versuchsbedingungen keinen Einfluß auf die 5-HT-Freisetzung.
Morphologische, biochemische und eiweißanalytische Untersuchungen bei einem Fall von Riesenplättchen-Thrombozytopathie mit Bence-Jones-Proteinurie
Authors: Schneider W , Hübner G , Scheurlen P. G , Joist J , Kübler W , Lechler E , Hellriegel K. P , Gross R
Published: June 2018

Die Ergebnisse feinstruktureller, biochemischer, immunologischer und chromosomenanalytischer Untersuchungen bei einem Patienten mit Riesenplättchenthrombozytopathie werden berichtet und Übereinstimmungen und Unterschiede zu den Befunden an anderen genetisch bedingten, thrombozytopenischen Thrombozytopathien aufgezeigt.
Über den Reaktionsmechanismus der Veränderungen von Kaninchenfibrinogen durch Kobalt(II)-Ionen
Authors: Taube Chr , Fiedler H
Published: June 2018

Bei einigen Tieren und beim Menschen führen Kobalt(II)-Ionen durch Änderung der Konformation des Fibrinogenmoleküls zu einer Gerinnungsstörung. Die Artspezifität des Fibrinogenmoleküls spielt dabei keine Rolle. Der Effekt ist von der Anwesenheit eines hochmolekularen Plasmafaktors abhängig, der bei einigen Tierarten nicht oder in ungenügender Konzentration vorhanden ist. Kobalt-58 wird an Fibrinogen angelagert, ohne daß es dadurch in jedem Fall zu einer Gerinnungsstörung kommt. Die Inkubation eines Gemisches aus Fibrinogen und defibriniertem, mit Kobalt(II) vorinkubiertem Plasma führt selbst dann zu einer Störung der Gerinnungs-fähigkeit des Fibrinogens, wenn die freien Kobalt(II)-Ionen nach Zugabe des Plasmas als Chelatkomplex gebunden werden.
Activation of the Fibrinolytic Enzyme System in Laboratory Animals and in Man
Authors: Baillie A. J , Sim A. K
Published: June 2018

The activation characteristics of the fibrinolytic system of several species have been evaluated in vitro and compared with the human enzyme system. The cat, dog and baboon would appear to be the most suitable species for further study of fibrinolytic activation. The present findings have been found to parallel the species differences in platelet behaviour which have been reported separately from this laboratory.
Platelet Sensitivity to Aggregation in Normal and Diseased Groups
Authors: Sano T , Boxer M. G. J , Boxer L. A , Yokoyama M
Published: June 2018

A method to estimate the sensitivity of platelets to aggregation by ADP has been described. Platelet-rich plasma was obtained from 8% trisodium citrate anticoagulated blood. ADP in trishydroxymethylaminomethane buffered saline was prepared by serial two-fold dilutions in concentrations of 2−1 through 2−18 mg/ml. Fifty μl each of platelet-rich plasma and ADP solution were mixed in a plastic test tube and agitated for 1 min. Aggregation was observed utilizing a light microscope. All procedures were performed at room temperature. The coefficient of variation in one sample was 1.4% or less and the daily variation was slight in healthy male volunteers. An advantage of this method is that it requires only a small amount of blood for the performance of many tests.

In 105 healthy Japanese males, the sensitivity of platelets to ADP-induced aggregation was positively correlated with age with a correlation coefficient of 0.824.

In the acute stage of thromboembolism of myocardial infarction or stroke, value of platelet sensitivity to ADP-induced aggregation was significantly higher at the 0.05 to 0.01 confidence limits than in age-, and sex-matched controls.
Aggregation of Fibrinogen and Its Degradation Products by Basic Proteins
Authors: Stewart G. J , Niewiarowski S
Published: June 2018

Fibrinogen and early fibrinogen degradation products (FDP) of plasmin digestion containing Fragment XfP gave highly ordered polymers of limited branching with protamines and synthetic polylysine (molecular weight 175,000). Histones, polylysine of molecular weight 2,500, granulocyte cationic protein and platelet factor 4 formed amorphous precipitates with fibrinogen and early FDP. Fibrin degradation products (fdp) containing complexes of Fragment X° formed highly ordered polymers of extensive branching with these basic proteins.

The splitting off of fibrinopeptides from fibrinogen and Fragment XfP was not essential for the formation of highly ordered polymers. However, in the presence of fibrinopeptides, high specificity of polymerizing agents such as basic protein is required. In the absence of fibrinopeptides, fibrin monomer or Fragment X° could polymerize after dissociation from complexes with other degradation products by any of the basic proteins.

It has been suggested that the polymerization of fibrinogen and its derivatives by basic proteins may be an important factor contributing to the formation and deposition of fibrin-like material in clinical conditions connected with the destruction of tissues (release of histones), damage of granulocytes (release of lysosomal cationic protein) and intravascular formation of platelet aggregates (release of platelet factor 4).
Influence of Climate on the Homeopathic Prescription
Authors: van Woensel Erik
Published: June 2018

The different types of climate our patients are sensitive to and their physical generals are frequently overlooked in case-taking; yet, this particular source of information provides us with the key to unlock the door to the correct homeopathic remedy.
The Papaveraceae: Benevolence—Opium, Chelidonium and Sanguinaria
Authors: Leupen Alex
Published: June 2018

Alex Leupen has worked in the Homeopathic Doctors Centre in Utrecht, the Netherlands, since 1990. Eight colleagues work in this centre, including Jan Scholten. Alex is an experienced teacher (the Netherlands, Belgium, France and Germany) on the underlying systems in the plant families, animal kingdom, sarcodes, nosodes, fungi and the periodic table of the elements pertaining to homeopathic practice. In this column, he shares clinical tips.