bluuebunny/crossref_metadata_2025 huggingface dataset:
Viscoelastic Studies of Surface Layers of Heparin-Fibrinogen Systems and the Antithrombotic Action of Heparin Other than Anticoagulation
Authors: Copley Alfred L. , King Robert G.
Published: May 2019

Decreased rigidity of torque values of surface layers (SL) of heparin plasma was reported by us (Thromb. Res. 1, 1, 1972). In subsequent viscoelasticity studies of SL of fibrinosen (Fg) systems we found lowering of viscosity (nM) and of elasticity (α M) with some heparin prepnrations (HP). No relationship was found between these decreased values and the anti thrombin activity (AA) of the HP, Certain HP, whether fractionated or not, markedly decreased nM and αM,. although their A A was very low. The antithrombotic action (ATA) of HP is therefore not necessarily dependent upon their AA. Since Copley & Robb discovered that HP can clump platelets in vitro & in vivo(Ani.J. Clin. Path. 12,; 416, 563 ,1942), the ATA of HP could hitherto be explained only by AA. Our new findings arc explained by Copley’s concept on initial thrombus formation (Biorheol. 8,79, 1971), in which Fg forms gels without thrombin action (Copley & King, Thromb. Res. 8, Suppl. II. 393. 1976). Such production of Fg gels can no longer be doubted, as found by Tooney & Cohen (J. Molec. Biol, 110, l6l, 1977). Conclusion. The ATA of HP is twofold:(l)further growth of formed thrombi is inhibited by AA;(2) formation of new thrombi is prevented by inhibition of Fg gelation due to constituent, if present, of HP.
Protein Concentration Dependence of Calcium Binding to Prothrombin Fragment-1
Authors: Brenckle G. M. , Peng C. W. , Jackson C.M.
Published: May 2019

Calcium binds to prothrombin and prothrombin fragment-1 with apparent positive cooperativity. It has previously been suggested that the cooperativity is induced by a conformation change in the protein. Another process which can induce cooperativity is ligand induced self-association. Calcium has been shown to induce dimerization in prothrombin and fragment-1. If such an intermolecular process is responsible for the apparent cooperativity, the calcium binding should become non-cooperative at the limit of zero protein concentration. The protein concentration dependence of calcium binding to fragment-1 was investigated using ultraviolet difference spectroscopy and the Hummel-Dryer equilibrium binding technique. A maximum of 10 sites was found on fragment-1 at high calcium concentration (10 μM). The maximum in the Scatchard plot was observed to decrease at fragment-l concentrations less than 10 μM. The difference spectral data was analyzed utilizing the equilibrium binding data. The apparent dissociation constant ob tained from difference spectral titrations is protein concentration dependent. At low protein concentration (1-2 μM) the maximal spectral change occurs with 2 or less Caions bound, while at higher concentrations (50-100 μM), the maximum is reached after ~5 sites are filled.

(Supported by HL14l47, Specialized Center of Research on Thrombosis, NHLBI).
Immunochemical Studies of Bovine Platelet Factor V
Authors: Ittyerah T.R. , Rawala R. , Colman R.W.
Published: May 2019

Utilizing new knowledge of the sub unit structure of bovine plasma factor V, we studied factor V derived from bovine platelets. Platelets were freed of plasma factor V by gel filtration and solubilized in 0.2% Trit on X-100. Incubation of this extract with a specific antiserum against purified plasma factor V rapidly inactivated platelet factor V. Antiserum absorbed with platelet extract failed to inactivate plasma factor V, further indicating that platelet factor V is antigeni cally related to plasmafactor V. Platelet factor V in the extract increased 4.7 fold when activated with thrombin while factor V released from gel filtered platelets by collagen was activated 7.5 fold. In contrast, an 18 fold increase inactivity was observed when diluted plasma or purified plasma V was incubated with thrombin suggesting that plrttelet factor V is partially activated during release. On immunoelectrophoresis, factor V released by collagen or extracted with Triton appeared as a single line close to the origin in contrast to plasma factor V which migrated towardth, anode. Incubation of the collagen releasate with antiserum to plasma factor V resulted in an immunoprec ipitate which when electrophoresed in SDS gave a single band with Mr=270,000, corresponding closely to the h chain of bovine plasma V, Mr =290,000. The alteration in platelet fact or V during release may facilitate its role in prothrombin conversion.
Antithrombin III Replacement Therapy in Patients with Congenital and Acquired Antithrombin III Deficiency
Authors: Thaler E. , Niessner H. , Kleinberger G. , Gaßner A.
Published: May 2019

In 2 patients with congenital (CATD) and 5 with acquired antithrombin III (AT111) deficiency (AATD) replacement therapy with ATI II concentrate (ATC){Behringwerke AG) was per-formed. One CATD patient undergoing neurosurgery received the ATC prophylactically and one for treatment of acute deep venous thrombosis with laboratory evidence of DIG, In the AATD group 1 patient suffered from cerebral falciparum rial aria,3 from septicaemia and 1 from Hodgkin lymphoma complicated by DIC, ATC therapy in AATD was aimed to prevent irreversible tissue damage by fibrin microthrombi, ATI 11 activity (ATI I If} was measured with a heparin cgfactor assay (initial AT111f values of the 7 patients ranging from 20 to 69% of normal controls),Intravenous infusion of the ATC produced no unfavourable side effects. An increase of 1% of normal ATIIIf activity was obtained by infusion of 0.67+0.19 U/kg body weight in the CATD and of 1.14±0.48 U/kg in the AATD group. Peak ATIIIf activities decreased with a biological half life (T/2) of 17.5 to 26.5 hrs in the CATD patients and of 13.3 to 11U.4 hrs in those with AATD, long T/2 values probably indicating reappearance of endogenous ATIIIf. As an indicator of increased sensitivity to heparin a transient prolongation of the thrombin clotting time was found in 4 of 5 patients who received heparin and ATC simultaineously. Though shock was present in 4 of the 5 patients with AATO, irreversible kidney failure did not occur, As the number of patients studied is rather small. a final judgement of the therapeutic value of ATIII replacement cannot be presented.
β-Thromboglobulin and Platelet Survival in Patients with Rheumatic Heart Disease and Prosthetic Cardiac Vaives and their Treatment with Sulphinpyrazone
Authors: Ludlam C.A. , Allen N. , Blandford R.B. , Dowdle R. , Bentley N. , Bloom A.L.
Published: May 2019

The plasma β-thromboglobulin (βtg) concentration has been measured in 202 patients with rheumatic heort disease and prosthetic cardiac volves who were already receiving warforin. In B4 of these patients the plasma βtg concentration was greater than 50ng/ml on at least one occasion . In 9 of these individuals who had persistently elevated plasma βtg concentrations the 51Cr-platelet mean lifespan (PMLS) and plasma and platelet βtg levels were measured before and after treatment with sulphinpyrazone 800mg/day. The results demonstrated that prior to treatment, patients with raised plasma βtg concentrotions hod short PMLS. Treatment with sulphinpyrazone resulted in lengthening of PMLS but there was no change in the plasma or platelet ~tg concentration. It is concluded that in these patients a high plasma βtg level is associated with 0 reduced PMLS and that although sulphinpyrazane increases the PMLS it foils to reduce the plasma βtg concentration. The results suggest that the PMLS is 0 more sensitive measure of the therapeutic effect of sulphinpyrazone than the plasma βtg concentration.
Does Recovery from Aspirin Effect Measure Platelet Survival?
Authors: Catalano P. , Murphy S. , Smith J.B.
Published: May 2019

Recovery from inhibition by aspirin (ASA) of platelet cyclooxygenase has been proposed as a non-isotopic label for determining platelet survival. Reports of a 24 to 48 hour delay in recovery have been sited as evidence that megakaryocyte cyclooxygenase is also inhibited by ASA, an effect that could compromise the usefulness of this technique. Using cotorimetry and radioimmunoassay, we have measured malondialdehyde (MDA) and thromboxane B2 (TXB2) production by platelets obtained at intervals after ingestion of S00 mg of ASA by normal volunteers. The production of both MDA and TXB2 by washed platelets stimulated with NEM or thrombin (TB) returned in a linear fashion with time after ASA. 9.5±1% (M±SEM) return of activity was demonstrated at 24 hours, and 24±1.25 at 48 hours. Using both stimulants and measuring both metabolites, recovery with time closely paralleled normal platelet survival. However, when TXB2 was measured in serum obtained by clotting whole blood with T8 at 37° Only 0.6±1% recovery was present at 24 hours and 5±.5% at 48 hours, suggesting a megakaryocyte effect. To ascertain that this delay in recovery was not an artifact induced by the presence of ASA-treated plasma or cells, a mixture of one volume of normal blood and nine volumes of blood from an ASA-treated donor was stimulated with TB, and the expected production of TXB2 by the 10% normal platelets present could be detected. When washed platelets were resuspended in either platelet-poor plasma or 4% albumin, the delay in recovery at 24 hours was again found. The interpretation of these results is uncertain at present. However, the coincidence of recovery from ASA with normal platelet survival was seen only when protein was excluded from the medium in which platelets were suspended for stimulation.
Prenatal Diagnosis of Haemophilia by Assay of Fetal Factors VIIIC and IX and Immunoradiometric Assay of Factor VIII Clotting Antigen (VIIICAg)
Authors: Mibashan R.S. , Rodeck C.H. , Thumpston J.K. , Peake I.R. , Bloom A.L.
Published: May 2019

Initial studies in adults measured foetor VIII clotting antigen (VIIICAg) in normal serum or plasma by on immunaradiometric assay (IRMA) and showed it grossly reduced or absent in 29 of 30 haemophilic kindreds. Pure fetal blood has been obtained (C.H.R.) at 15-22 weeks by direct fetoscopy with realtime ultra-sound in 64 (86%) of 74 patients studied diagnostically or pre-abortion. At 18-22 weeks 48 of 51 fetoscopies (94%) yielded pure fetal blood. In 22 consecutive somples of fetal citrate-plasma VIIIC was 32-79 (

50) u/ dl, and FIX 8-18 (

12) u/dl. VIIICAg in 9 fetoscopic plasmas ranged from 11-29 (

20) u/ dl, and in 8 su ch sera VIIICAg was 4-14 (

9) u/dl. VIIICAg levels in amniotic fluid were < 0.1 u/dl. Blood was obtained from 6 mole fetuses of obligate hoemophilia A carriers: In 3 only post-termination serum was available, and in one of these VIIICAg of <0.1 u/dl was found. In another both fetoscopic plasma and abortus serum hod < 0.1 u/dl VIIICAg. In the other 2, fetal blood-sampling gave plasma VIIIC levels of 49 and 63 u/dl, and VIIICAg of 13 and 16 u/dl; since these values were within their normal ranges, the pregnancies continued and their neonatal status is still awaited (Feb .79). The results suggest that prenatal diagnosis of haemophilia is possible by combining modified coagulant assays or IRMA with meticulous fetal blood sampling. Further definition of their normal ranges, relative predictive value and added value of VIIIRAg assay is needed.
Platelet Factor 4, β-Thromboglobulin, Malondialdehyde Formation and Blood Lipids in Patients With Diabetes Mellitus
Authors: Zahavi J. , Jenes N.A.G. , Betteridge D.J. , Leyton J. , Galton D. J. , Clark S.E. , Kakkar V.V.
Published: May 2019

Plasma β.-thromboglobulin (βTG) was measured by radioimmnunoassay (RIA) in 126 patients (mean age 54.2) with diabetes mellitus (DM). Their results were compared to age and sex matched healthy subjects and correlated to the incidence of retinopathy and to 5 fractions of serum lipids . In 46 of the patients, platelet factor 4 (PF4) measured by RIA and malondialdehyde (MDA) formation in washed platelets were also determined. Plasma βTG (mean 73.5mg/ml range 19-220), PF4 (mean 35. 7 ng/ml, range 7.6-100 and MDA (mean 12.9 88nM/109 platelets), were significantly elevated in the patients than controls (p < 0.0005). Results were higher in 27 patients with reti nopathy compared to other DM patients. There was a significant correlation between βTG and PF4 (r=0. 4943) suggesting that both proteins are released from the same pool, as well as between βTG and MDA (r=0.5551) or total serum trigl yceride (Tg) level (r=0.3854). These results indicate that in-vivo Platelet “reaction” and apparently prostaglandin (Pg) synthetase activity (measured by MDA), are significantly increased in DM patients, the former resulting mostly from activation of Pg pathways. This abnormal platelet (unction might reflect a pre-thrombotic state. In addition, plasma factors, such as elevated Tg levels, seems to contribute to the abnormal platelet behaviour.
Reology of Blood Flow at Branches
Authors: Einav Shmuel , Berman Herbert
Published: May 2019

Fluid mechanical factors are considered to be possibly responsible for wall changes, veneration of thrombosis and formation of atherosclerosis at branches. The local flow fields in branches would be exnected to derend on the geometry of the branch. In the present study, in vivi flow visualization and velocity nrofile measurements in the plane of the arteriolar and venular branch (T branch) were obtained in the everted, transill-uminsted cheek pouch of the anaesthetized hamster (Mesocritus Auratus). Use was made of our Laser Dorpler Microscope Anemometer (LDHA) which allowed as many as 22 local velocity measurements to be obtained across a 120 μm blood vessel and as many as 6 local velocity measurements across a 38 μm blood vessel. The measured velocity profiles at the measurement cross section of the arteriolar branch were found to be skewed towards the distal wall of the daughter branch, indicating higher wall shear rate at this site. The shear rate on the proximal wall, on the other hand, was found to be smaller than the shear rate at the wall of the main Teasel, The velocity profiles were more skewed at higher Reynolds numbers and higher flow rates. At the venular T branches, no significant changes in the flow profiles or the wall shear rates were observed, indicating lesser tendency to generation of thrombosis or atherosclerosis at these sites (supported in part by NIH Grants HL 00902 and 09447).
Haemorheological Changes Following Plasmapheresis for Raynaud’s Phenomenon
Authors: Dodds A.J. , O’Reilly M.G. , Yates C.J.P. , Cotton L.C. , Flute P.T. , Dormandy J.A.
Published: May 2019

Eight patients with severe Raynaud’s phenomenon were treated by weekly plasma exchange (moan volume removed 1.9 1.) for 4 weeks using a Haemonctics Model 30 Blood Processor. The mean whole blood viscosity at 0.77 sec-1 adjusted for haematocrit was significantly lower following therapy (p<0.001) and the mean red cell deformabi1 ity index was improved (p<0.005). In the patients studied serially the mean percentage fall in whole blood viscosity at 0.77 sec-1 following a single plasma exchange was 44t, but only 12% at 91 sec-1. Seven of the patients noticed symptomatic improvement including healing of ischacmic digital ulcers. In 6, who were tested, improvement in digital artery patency was measured by directional doppler velocimetry. It is concluded that intensive plasmapheresis is an effective means of haemorheological therapy and may be beneficial in patients with severe digital ischacmia by increasing blood flow 4 and reducing the likelihood of thrombosis.
Plasmapheresis and Platelet Function in Raynaud’s Phenomenon
Authors: O’Reilly M.J.G. , Zahavi J. , Dubiel M. , Cotton L.T. , Kakkar V.V.
Published: May 2019

Sixteen patients (mean age 43.5) suffering from Raynaud’s disease or syndrome have been treated by repeated plasmapheresis (P) at weekly IntervalsThey were compared to another 4 untreated patients and 30 healthy subjects. In each patient platelet aggregation (PA) to adenosine diphosphate (ADP) 0.6 and 4μM, 1-eplnephrlne 6μM, collagen 1μg/ml and thrombin 0.2 u/ml as well as serum inmunoglobulin (Ig) levels were determined previous to and after each plasmapheresis. In 7 Patients plasma β-thromboglobulin (βTG) were also determined by radioimmunoassay. Following plasmapheresis the rate and extent of PA to ADP and collagen, which were Increased In all the patients, were reduced by 407, In 10 patients and to thrombin and 1-epinephrine In 6 patients. This change occurred gradually and was maximal after the 4th plasmapheresis. Clinical improvement appeared gradually, as well, in 9 patients. Plasma TG, Initially very high (mean 118.9ng) decreased significantly and rapidly to 71 (p ≤ 0.05) in all the 7 patients. Platelet function studies In the 4 untreated| patients as well as serum Ig level In all the patients, remained unchanged. These results Indicate that in-vlvo platelet “release reaction” measured by βTG which is enhanced in such patients, decreases significantly following the plasmapheresis. Furthermore, they suggest that platelets might be involved in the eventual course of the disease state and the βTG is a useful aid in monitoring plasmapheresis.
Fibrinolysis in Solid Tumors
Authors: Yuen P. , Kwaan H.C. , Ho F.
Published: May 2019

The fibrinolytic activity of 120 malignant and 25 benign solid tumors from autopsy and biopsy specimens was studied by the fibrin slide technique as described by Kwaan and Astrup. The inhibitory activity against fibrinolysis was graded according to the Lysis time of vascuLar tissues within the tumor. The results show chat all malignant solid tumors, with the exception of carcinoma of prostate demonstrate varying degrees of inhibition of fibrinolysis. Persistently high inhibitory activity was found in squamous cell carcinoma of esophagus, the respiratory tract (including squamous cell carcinoma of sinuses, larynx and lung), cervix uteri and skin; carcinoma of uterus, colo-rectal carcinoma; small cell anaplastic carcinoma of lung; neuroblastoma; carcinoma of bile duct; while malignant tumors of the kidney show a lesser degree of inhibition. In contrast, with the exception of hydatldiform mole, benign solid tumors show-little or no Inhibition. There is a difference in fibrinolytic activity between the peripheral and the central portions of a tumor with distinctly greater degree of activity in the former. The pathophysiologic implications of the presence of such a strong inhibition of fibrinolysis is apparent. Biochemical characterization of this inhibition is currently in progress.
Hypercoagulability and Thrombosis in Patients with Gynecological Cancer Under Radiotherapy
Authors: Hugo R.V. , Hiller K.F. , Hafter R. , Lochmüller H. , Graeff H.
Published: May 2019

Cancer patients often develop thromboembolic episodes and show laboratory signs of hypercoagulability. A randomised, controlled, prospective trial was performed in 30 patients which were treated by combined external ir-radation and intravaginal radium application. The incidence of deep vein thrombosis (DVT) diagnosed by 125 I-fibrinogen leg scanning was 43 % in the control group (age 62,9 years; weight 62,6 kg). The application of 2 χ 7.500 U Calciparin s. c/24 h reduced the incidence of DVT to less than 10% (age 61,9 years; weight 64,7 kg). Patients who developed DVT showed a significant increase in circulating soluble fibrin monomer complexes from 4.1 ± 0,8 to 6,7 ± 1,4 % estimated by agarose gel filtration and an increase of fibrinogen from 379 ± 58 to 444 ± 97 mg%. Raised serum FDP levels were found only in two patients with clinically symptomatic leg vein thrombosis. No correlation of AT III levels and DVT was observed. The study shows the efficiency of 2 χ 7.500 U Calciparin s.c.24/ h for the prevention of thromboembolic complications in patients with gynecological cancer under radiotherapy. Results of a continuing study in which the effect of Calciparin, is compared with the one of a semi-synthetic heparan analogue, a 73025 (SSHA) will be presented.
Activation of Blood Coagulation and Anomalous Behaviour of Fibrinpeptide a to Heparin Treatment in Patients with Malignancy
Authors: Peuscher F.W. , van Aken W.G. , Armstrong A.L. , Stoepman-van Dalen E. A. , van Mourik J.A.
Published: May 2019

Hypercoagulability, known to complicate malignant disease, can be detected by the measurement of fibrinopeptide A (FPA). The aim of the present study was to establish the significance of FPA production in. patients (n = 113) with various types of malignancy, without clinical signs of DIC, venous thromboembolism, not receiving cytostatic, anticoagulant or radiotherapy or bedrest. 8 patients presented laboratory abnormalities compatible with DIC; In this group mean FPA was 9.8 ng/ml (normal 1.4±0.9.2SD) and FPA generation In vitro (ΔFPA) was 15 ng/ml/10 min (normal 0.4±0.8, 230). In the remaining 105 patients, mean FPA was 5.2 ng/ml andFPA was 1.8 ng/ml/10 min. Of the 20 patients with both normal FPA and ΔFPA, 13 were clinically in remission, 5 had malignancies without detectable metastases and only 2 showed metastases. In 2 patients FPA was normal but ΔFPA was slIghtLy accelerated. 50 patients had elevated ΔFPA but normal FPA and in 33 cases both FPA and ΔFPA were increased; in these groups the majority of patients had a metastatic malignancy In 22 patients mean FPA before (4.9 ng/ml) and 20 mln after(4.2 ng/ml) receiving heparin (7500 U, i.v.) was not significantly different. These results suggest that apart from DIC tumor cells or the adjacent inflammatory area of tumors may trigger FPA generation, not being affected by heparin. It Is postulated that “heparin resistant” FPA generation Is potentially a sensitive marker of tumor activity.
Bovine Factor VIII: Purification of the Procoagulant Protein
Authors: Vehar G. A. , Davie E. W.
Published: May 2019

Factor VIII has been purified from bovine plasma approximately 500,000 fold with an overall yield of about 5%. Purification steps included affinity chromatography as well as DEAE-Sephadex and sulfate-Sepharose chromatography and gel filtration on Sephadex G-200. The final product readily corrected the clotting of plasma from patients with hemophilia A or severe von Willebrand disease. The purified preparation had no von Wil1ebrand factor activity, as measured by aggregation of platelet - rich human plasma. The coagulant activity was inhibited by antibodies prepared against a purified von Willebrand/factor VIII preparation and was required for the conversion of factor X to factor Xa in the presence of factor IXa, calcium, and phospholipid. The highly purified preparation appeared as a triplet on urea-SDS polyacrylamide gels with an apparent molecular weight of about 70,000. Preincubation of factor VIII with thrombin or factor Xa resulted in (20-50 fold increase in the coagulant activity, and this increase correlated with a reduction in the molecular weight of the protein as determined by SDS polyacrylamide gel electrophoresis. Activated factor VIII was inhibited by preincubation with DFP, in contrast to the precursor form. These data indicate that factor VIII is a trace protein in plasma in a precursor form and is converted to an activated form during the coagulation process. (Supported by NIH Grant HL 16919 and the National Hemophilia Foundation.)
Fibrinogen Proteolysis and Deep Venous Thrombosis (DVT)
Authors: Nossel H.L.
Published: May 2019

Thrombin cleaves fibrinogen in a two-stage reaction first producing fibrin I (fl) and fibrinopeptide A (FPA) and then fibrin II (fill and fibrinopeptide B (FPU). FI forms thinner strands than fil, a property which may be important in the pathogenesis of thrombosis. In the initial stages of plasmin proteolysis of fibrinogen the C-terminal portion of the Aα chain and the N-terminal portion of the EB chain are removed, leaving a molecule called fragment X (X). Release of the N-termlnal peptide Bβ1-42 serves as an index of X formation, llcnco plasma FPA levels serve as an index of X formation by thrombin action and Bβ1-42 levels as an index of X formation by plasmin action. In normal individuals Bβ1-42 levels were approvimately three times higher than FPA levels. In patients with symptoms of DVT, which was confirmed by venogram, PPA levels were almost invariably elevated. Bβ1-42 levels were not significantly elevated in these patients in the absence of complicating pulmonary embolism. Initial studies on patients at high risk for DVT studied prospectively have shown signigicant elevation of the plasma FPA level and little change in the Bβ1-42 level for several days preceding the onset of DVT as indicated by 125I-fibTinogen scan and confirmed by venogram. Bβ1-42 levels became elevated several days later inassociation either with pulmonary embolism or with resolution of the thrombosis. These data suggest_ that the development of DVT is associated with and preceded by imbalance between thrombin action as indicated by plasma FPA levels and plasmin action as indicated by Bβ1-42 levels.
Multiple Drug Therapy to Prevent Thrombosis
Authors: O’Brien J. R.
Published: May 2019

Trials of single drugs to preuent thrombosis have not been strikingly successful. Thus in the complex evolution of a thrombus involving many systems (coagulation, platelets, fibrinolysis etc.) inhibition Df one step in one system can perhaps be bypassed. Perhaps all known systems should be blocked. A more logical approach suggests the measurement of all those attributes that may predispose to thrombosis and which can be altered by drugs. Patient therapy then aims to correct all abnormal tests. Blood flou is influenced by a high fibrinogen which can be normalized by Clofibrate or ICI 55B97. Antithrombin III if loy and factor VII etc, if high can be treated by giving Warfarin. Fibrinolysis can be stimulated by Stanozolol. “Spontaneous platelet aggregation” can be corrected by aspirin. The heparin neutralizing activity of platelet poor plasma (? PF4-like activity released from platelets) is usually abnormal in high-risk patients and this can be normalized by Clofibrate and ICI 55897. Lipid abnormalities should be (studied and if abnormal corrected by diet and Clofibrate. There may be other demonstrable abnormalities that can also be treated. There is no evidence that such “tailor-made polypharmacy” would help but at least one trial involving the concept of “treating the tests” should be considered before more large single drug trials are launched.
Pathophysiological Changes of Flow Patterns of the Blood in Blood, Vessel Diseases
Authors: Ehrly A.M.
Published: May 2019

Blood flow properties have been found to he impaired as well in arte-ial occlusive diseases as in venous diseases. Increased blood and -yiacna viscosity values are found in arterial and venous diseases. Hed cell deformability is impaired In diabetic and non-diabetic patients with chronic peripheral ischemic diseases. In addition, the hemodynamic situation (poststenotic low flow in arterial diseases, impaired venous flow and Increased venous pressure in venous diseases) locally increases blood viscosity and red cell aggregation. These factors and local hamoconcentration reduce blood flow velocity and may induce thrombosis.
Cancer Cell-Endothelial Reactions: The Microinjury Hypothesis and Localized thrombosis in the Formation of Micrometastases
Authors: Warren B. A.
Published: May 2019

The two important cancer cell-endothelial reactions are the entrance of tumour cells into the blood stream and the exit of tumour cells from vessels, (i) In transplantable melanoma transplants in the hamster cheek poach, tumour cells were found to migrate activily througt the endothelial cell junctions and attenuated endothellunf “giant, capillaries” at the edge of the tumours as early as 3 days after transplantation, (ii) The outcome of multiple episodes of microembolism by tumour cell emboli is dependent, in part, upon the structure of the specific microcirculatory units involved, the state of their endothelial lining and the size and deformability of the tumour cell emboli themselves. The Kawaguchi-Nakamura microinjury hypothesis suggests that the waves of emboli reach a specific microcirculatory bed and result in damage at certain sites in the unit due to prolonged temporary blockage of flow through the unit. This depends upon the presence of tumour emboli of a certain size and stiffness. The damage of the unit causes localized thrombus formation-Subsequent flow over the damaged area brings fresh emboli into contact with the injured region which is then favourably altered for the attachment and initial fixation of the circulating tumour cell emboli. In experiments with Walker 256 carcinoma, mouse thymoma and Hela cells these tumour cells were found to attach more firmly to damaged vessel wall than to regions of intact endothelium.
Platelet Survival and the Development of Coronary Disease in the Young : The Effect of Cigarette Smoking, Family History and Medical Therapy
Authors: Fuster V. , Chesebro J.H. , Byrne J.M.
Published: May 2019

The study comprises 53 consecutive patients (pts) under age 50 with symptomatic coronary artery disease (SCAD) confirmed at arteriography and 53 apparently normal individuals (AN) under age 50. We recorded smoking (≥10 pack-years) or strong family history (FH) (first degree relatives ≤age 50) in 923; of the SCAD pts and in 47% of the AN. Platelet depositton to the damaged endothell urn appears important In the pathogenesis of atherosclerosis ; accordingly, platelet consumption was estimated by platelet survival half life with 51Cr (PS). A shortened PS (≤92 hours) occurred in 64% of the SCAD pts, in 52% of the AN with smoking or FH and in only 14% of the AN without smoking or FH (p<0.01). Lengthening of the shortened PS towards normal was obtained:in 10 out of 13 SCAD pts (p<0.01) by using Dipyridamole (225 mgs daily) plus Aspirin (975 mgs daily) and in 7 out of 9 AN smokers (p<0.01) by discontinuation of smoking. The study suggests:(1) a striking association between smoking or strong FH with the development of SCAD in Che young; (2) the possibility that smoking or FH, when associated with short PS, might predict the AN who is developing the disease; and (3) the possibility that in AN the incidence of coronary disease may be prevented by avoiding smoking and by using platelet inhibitors in those wich FH.
Comparison of the Human Activators of Plasminogen
Authors: Mackie M. , Booth N. , Bennett B.
Published: May 2019

Detailed information is available on the molecular characteristics of plasminogen and plasmin. In contrast, few data have been obtained on the number and nature of agents which activate plasminogen in the body. We describe the purification and characteristics of (1) a plasminogen activator present in venous blood after venous occlusion - “circulating activator”, (2) a plasminogen activator generated from plasma by contact with kaolin - “Hageman factor dependent activator”, (3) a plasminogen activator precipitated from plasma by dextran sulphate - “dextran sulphate dependent activator”, (4) a plasminogen activator obtained from cadaveric vascular endothelium - “cadaveric endothelial activator and (5) a plasminogen activator extracted from human heart tissue - “tissue activator”. The properties of these activators are compared with one another and «ich urokinase obtained commercially. The physical properties of activators 1-5 appeared similar though on Sephadex G200 the dextran sulphate dependent activator eluted at a position suggesting a larger molecular size than the other activators. The susceptibility of these activators to enzyme inhibitors were identical but differed from that of urokinase. Anti-sera to urokinase did not inhibit the activity of these activators. Antisera to the cadaveric endothelial activator did not inhibit urokinase or the Hageman factor dependent activator but did inhibit the activity of activators 1,3,4 and 5.
Binding of Allo- and Heteroantibodies to Human Factor IX (F. IX)
Authors: Reisner H.M. , Lewis R.M. , Strand E.A. , Chung K.S. , Roberts H.R.
Published: May 2019

Study of the binding properties of human alloantisera to F.IX is made difficult by the nonprecipitating nature of the antibodies. Most studies have relied on F.IX coagulation neutralization to characterize such immune inhibitors. We have directly measured the binding of antibodies to highly purified radiolabelled F.IX (125 I F.IX) by using either differential precipitation [employing ammonium sulfate (AS) or polyethyleneglycol (PEG)] or column chromatography to separate antigen-antibody complexes from free 125I-F.IX. The two heteroantisera and four immune alloantisera studied all form stabile immune complexes with 125I F.IX in vitro. The various antibodies require different levels of AS or PEG for optimal precipitation of complexes. Precipitability is not related to antibody titer as measured by radioimmunoassay or coagulation inhibition but may be related to the size of the immune complex. Using column chromatography on agarose A-5m, two alloantisera studied form complexes with molecular weights of less than 5 x 105 daltons. The low molecular weight of such complexes suggests that a limited number of antigenic sites on the F.IX molecule are recognized by alloantisera.
Activation of Factor IX by Polymorphonuclear Leukocytes from a Patient with Chronic Granulocytic Leukemia, and from a Patient with Factor XI Deficiency
Authors: Kingdon H.S. , Herion J.C.
Published: May 2019

We have previously reported that preparations of polymorphonuclear leukocytes (PMNs) from normal human blood are capable of activating Factor IX in the presence of calcium ion (Thrombosis Research, 13, 501-507, 1978). In the outrent studies, we demonstrated that cell suspensions subjected to freezing and thawing wete as active or more active than the suspensions subjected to ultrasonic disruption which were used previously. Furthermore, the Factor IX activating activity survived prolonged storage of the frozen cell suspensions. PMNs from a patient with chronic granulocytic leukemia contained potent Factor IX activator, as did PMNs from a patient with severe factor XI deficiency (plasma Factor XI < 1%). Preliminary experiments suggest that the Factor IX activator may reside in the granule fraction of the cells, since Factor IX activation was not observed when strontium was substituted for calcium, we conclude that Factor XIa> is not involved in this cellular activation phenomenon. Furthermore, the presence of an alternate mechanism for activating Factor IX in the blood of the Factor XI deficient patient may explain why the bleeding tendency in this deficiency state is so much milder than in Factor IX or Factor VIII deficiencies.(Supported in part by USPHS grants HL-16633, and DE-0 2668).
Inhibition of Intimal Proliferation of Rabbit Aorta by Ticlopidine
Authors: Reece A. H. , Walton P.L.
Published: May 2019

Proliferation of smooth muscle cells in the intima of arteries at sites adjacent to damaged endothelium may be stimulated by growth factors released from platelets. We have studied the effect of treatment with two inhibitors of platelet function, Ticlopidine and Sulphlnpyrazone, on the rate and extent of intimal proliferation In rabbit abdominal aorta following endarterectomy using a balloon catheter. Aortas were removed at Intervals after surgery, fixed, sectioned and the extent of intimal proliferation was measured using an Image analyser (Quantlmet 720). The arc a of Intimal growth as a percentage of total area of aorta was 42.4±2.8,22.3±1.2 and 50.9±2.0, 33.9±2.0 in control and Ticlopidine treated animals 30 and 60 days after endarterectomy, respectively. Thus treatment with Ticlopidine (50mg/kg/day P. O.) produced a highly significant (p<.001) reduction In the extent of intimai growth In these animals (c 50%), where as Sulphlnpyrazone (50mg/kg/day P. O.) had no effect. ADP Induced platelet aggregation was reduced (c 30%) during the treatment with Ticlopidine but not during treatment with Sulphlnpyrazone.

These findings indicate that Ticlopidine treatment limits intimai proliferation In rabbits following endothelial cell Injury and may be of value in limiting this process In man.
Bay g 6575, An Anti Thrombotic Compound that Stimulates Prostacyclin Release from the Vessel Wall
Authors: Chamone D.A.F. , Vermylen J. , Verstraete M.
Published: May 2019

Oral administration of Bay g 6575 (1-(2-(6-naphtyloxy)ethyl)-3-methyl-2-pyrazolin-5-one) has been shown to reduce or prevent subsequent experimental thrombus formation in arteries or veins of rabbits and rats (I). In these animals, no effect on blood coagulation, fibrinolysis or platelet aggregation could be demonstrated (I). Ingestion of 1.2 g Bay g 6575 daily for one week by six healthy volunteers had no effec t on parameters of blood coagulation, fibrinolysis or platelet aggregation “ex vivo”, but seemed to inhibit platelet aggregation “in vivo” (less decrease of the platelet aggregate ratio after venous occlusion, ref. (2). Plasma drawn from five volunteers after a single oral dose of 1.2 g of the same compound stimulated prostacyclin release from “exhausted” rat aorta slices, while plasma from the same individuals before intake of the substance did not. Whereas administration of Bay g 6575 or dipyridamole alone had no effect on platelet aggregation “ex vivo”, combined administration resulted in a striking and prolonged inhibition of ADP-induced platelet aggregation. It is proposed that the previously described anti thrombotic properties of Bay g 6575 in animals (I) are a consequence of stimulation of prostacyclin release from the vessel wall and that this effect is also demonstrable in humans. (I) Seuter, F. et al. Arzneimittelforschung, 1979, in press. (2) Chamone, D.A.F. et al. Thrombos.Haemostas., 1977, 38, 132.
Fragmentation of Actin by Thrombin-Like Snake Venom Proteases
Authors: Hauck M. , Muszbek L.
Published: May 2019

It has been demonstrated that thrombin can split skeletal muscle actin (Muszbek and Laki, PNAS 1974,71,2208). In the present paper the effect of thrombin-like snake venom proteases (Ancrod and Batroxobins of Bothrops moojeni and Bothrops marajoensis) on actin was studied and compared to the thrombic cleavage of this protein. Only EDTA pretreated G and F actin were split by Ancrod, while, Batroxobins hydrolized native G actin, too. The time course of digestion was followed by SDS PAGE. A split product of 37500 m.w. appeared first which was cleaved further resulting in three lower m.w. fragments. The SDS gel pattern of thrombic fragmentation was well distinguishable from those caused by Ancrod and Batroxobins. The first split products of Batroxobin digestion were isolated and by estimating their N-, and C-terminal end groups and amino acid compositions the peptide bond hydrolyzed first was located in the primary structure of actin. It was established that while thrombin split off two actinopeptides (at Arg ( 28)-Ala(29) and Arg ( 39)-His ( 40) from the N-terminal end of the molecule only Arg ( 39)-His ( 40) was cleaved by Batroxobins.
Effect of Endotoxin on Kinetics of Antithrombin III
Authors: Tanaka H. , Kobayashi N. , Takeuchi T. , Takada M. , Haekawa T.
Published: May 2019

The kinetics of antithrombin III(AT III) in dogs were studied using I-125-labelled AT III and Se-75-methionine. As reported at the last meeting of ISH, the plasma half-life of AT III was 1.7±0.2 days in normal 5 control dogs. The double i.v. administration of 200 ug/ltg of endotoxin and the single i.v. administration of 1 mg/kg of endotoxin resulted in 30% decrease of plasma AT III, 60% decrease of coagulation Factors (I, II, V, VII, VIII, IX), the shortening of plasma half-life of AT III to 1.4 days and the increase of J3u values, suggesting increase of the synthesis rate of AT III. Then, the synthesis of AT III was studied directly using Se-75-methionine. After i.v. injection of Se-75-methionine, blood samples were obtained. One ml of sample plasma was incubated for 24 hrs with 1 ml of anti-AT III antiserum, which was produied in rabbits and the radioactivity of the precipitates were determined. About 80% of AT III was recovered in the precipitates by this method. The maximum radioactivity was obtained 18 hrs after injection of Se-75-methionine, and 0.27 % of total injected Se-75-methionine were utilized to the production of AT III.

These results indicates that; 1. Endotoxin accelerates the metabolism of AT III. 2. The analysis of AT III production is possible using Se-75-methionine as a tracer.
Acquired Antithrombin III (AT III) - Deficiency in Septicaemia
Authors: Deutsch E. , Thaler E.
Published: May 2019

AT III was measured in 34 patients with clinical and bacteriological evidence of septicaemia using a heparin cofactor assay. Based on the results of positive blood cultures gram-negative septicaemia (G-S) was diagnosed in 10 (group 1) and gram positive septicaemia (G+S) in 9 patients (group 2). From the remaining 15 patients {group 3) blood cultures before onset of antibiotic therapy were not obtained and gave negative results throughout the observation period. Based on bacteria] cultures from other sites than venous blood or bacteriological examination of spinal fluid G-S was assumed in 13 and G+S in 2 patients.

In all but one patient of group 1 and one of group 2 AT III activities were decreased below 2 SO of normal controls (n = 91, x = 99.6, SD-8.4) already at the time of the first coagulation screening (patients: n=34,

=58.4, SD-16.6). Analysis of var-ance showed no significant difference between the mean values of the three groups at the c per cent (%) level. The minimal AT III activities during the course of the disease were below the norma] range in all patients studied [n=34,

=51.2, SD=13.6).

Thus AT III deficiency appears to be a constant and early finding in G-S and G+S, causing insufficient inhibition of blood coagulation, and hereby may contribute to irreversible tissue damage caused by microthrombi in septic shock. This deficiency may be an important factor in the failure of heparin therapy to reduce mortality from septic shock.
Antithrombin III Levels in Patients on Long Term Oral Anticoagulants
Authors: Bull Helen , Hackle I.J. , Brozovic Milica , Woodings D.
Published: May 2019

The plasma levels of antithrombin III (AT-III) in patients with congenital AT-III deficiency increaae after treatment with oral anticoagulants (Marciniak et al, 1974). This increase has also been observed in patients on oral antico, gulants for treatment or prophylaxis of thromboembolic diseases (O’Brien and Etherington, 1977). We have studied 27 patients stabilized on long term varfarin 3 months or more after the last thromboembolic episode, in parallel vith 18 healthy controls and 20 patients attending a hypertension clinic, who had never been on oral anticoagulants. Thre, methods vere usedl total progressive antithrombin (TAT); immunoassay by Laurell rocket method (AT-IIIAg); and chromogenic assay (AT-IIIBiol) using S 2238 (Kabi-Vitrum). The values obtained verel TAT 102.6 ± 14.6 and 104.0 ± 7.7; AT-IIIAg 96.9 ± 7.2 and 98.8 ± 9.0; AT-IIIBiol 106.9 ± 12.7 and 101.0 ± 12.51 mean ± SD, for healthy controls and patients on varfarin respectively. There Vere no significant differences between the two groups.

Only one technique, TAT, vas used in hypertenaive patients 1 the mean value vas 104.1 ± 6.8 and vas not significantly different from the other two groups. Our observations did not confirm that the long term oral anticoagulant treatment vas associated with a significant increaae in AT-Ill levels.
Antithrombin III in Contraceptive Treatments with Oestroprogestagens or Progestagen -Only Pills
Authors: Conard J. , Horellou M.H. , Cazenave B. , Samama M. , Zorn J. R. , Neau C.
Published: May 2019

Antithrombin III (AT III) has been investigated in women before tatting the pill and after 3 months Or more, between the 22nd and the 26th day of the menstrual Cycle. The oral contraceptives consistedinacombination of a progestogen with 30 to 50 ug of oestrogen, or in a progestogen - only pill.

AT III was measured by the Hancini method in plasma an serum and the antithrombin activity in serum by the von Kaulla method.

The results were obtained from 120 women before taking the pill, 189 taking an oestro-progestagen : containing 50 μg oestrogenin 96 cases, 30 μg in 65 cases, 30 to 40 μg according to the day of the cycle in 28 cases, and 20 women taking a progestagen-only pill.

The results show a statistically significant decrease in AT III in plasma and serum, and in antithrombin activity with all the oestroprogestagens. No difference is found between 50 and 30 fig oestrogen content. With the progestogen-only pills, no decrease in AT III is observed.
Subunit S and A Ratio in Congenital and Acquired Deficiency of Factor XIII
Authors: Barbui T. , Rodeghiero F. , Dini E.
Published: May 2019

Factor XIII contains two subunits :subunit A, the biologically active component, and subunit S, the “carrier” protein. In 7 homozygotes of factor XIII deficiency, the subunit A was immunologically undetectable while subunit S was on average 32%. The 29 heterozygotes had an average concentration of subunit A 35%, while subunit S was on average 66%. In the heterozygote group, the ratio S/A was 1.93.

In 32 patients with acute myeloblastic leukemia the ratio S/A was found impaired. The group of patients with severe disseminated intravascular coagulation (DIC) had a ratio 2.08 and the cases without a manifest 0IC had a ratio 2.12.

These findings seem to support the view that subunit S plasma concentration drops in accordance to subunit A, even though the reduction of this active componentis due to different mechanisms.
Control of Heparin Treatment with three Different Methods. Behaviour of Antithrombin III
Authors: Bounameaux H. , Marbet G.A. , Airenne H. , Grossmann E. , Stanojevic B. , Eichlisberger R. , Lämmle B. , Duckert F.
Published: May 2019

In 63 plasma samples from patients under heparin we determined the heparin concentration using the chromogenic substrats S-2222 (COATEST Heparin). Thrombin time (TT), activated partial thromboplastin time (APTT), antithrombin III activity (ATIIIact) and concentration (ATIIIimm) were also measured. A good correlation was found between heparin concentration and TT (r= .850, p< .001), heparin concentration and APTT (r =669, p < .001) while the correlation coefficient r between TT and APLT was .896 (p< .001).

We found a statilttically significant reduction of ATIIIact with increasing APTT (p < .05. The ATIIIact and ATIIIimm values were also lower (p < .001) in the overanticoaculated group (n=ll) than in the group with insufficient heparinisation (0.18). The mean (±SO) heparin concentration in 12 plasmas with both TT and APTT in the therapeutic range was .54 (±.15) USP-U/al, very similar to that of 13 plasma (.68 ± .46 U/al) insufficiently heparinised accordirig to the APTT. However, the TT recognised then as correctly anticoagulated. Regarding these findings and our good experience without complication by monitoring heparin therapy with TT we assume that TT is more accurate than APTT for this aim.
A Comparison of Anticoagulation Activity of a Potent Heparin Preparation in Different Test
Authors: Bhargava A.S. , Heinick J. , Schöbel Chr. , Günzel P.
Published: May 2019

The anticoagulant effect of a new potent heparin preparation was compared with a commercially available heparin in vivo after intravenous application in beagle dogs. The anticoagulant activity was determined using thrombin time, activated partial thromboplastin time and whole blood clotting time after 5, 10 and 30 minutes of application. The relative potency of the new heparin preparation (Scherinq) was found to be 1.62 to 2.52 times higher than heparin used for comparison (150 USP units/mg, Dio-synth). The anticoagulant properties of both preparations were also studied in vitro using dog and human plasma. The relative potencies in vitro correlated well with those obtained in vivo. Further characterization with amidolytic method using chromogenic substrate for factor Xa and thrombin (S-2222 and S-2238 from KABI, Stockholm) showed that heparin (Schering) contains 243 to 378 USP units/raq depending upon the test systems used to assay the anticoagulation activity and in addition, proves the validity of the amidolytic method.