bluuebunny/crossref_metadata_2025 huggingface dataset:
Antithrombins in Childhood Malnutrition
Authors: Jiménez R.A. , Ingram G.I.C. , Vargas W.
Published: May 2019

Antithrombin activities in 30 severely malnourished children and 40 normal children were estimated in clotting tests by thrombin neutralization, as antiXa and by a heparin antithrombin assay;and by immunodiffusion as ∞2-globulin and ∞1-antitrypsin. The patients’ mean ∞2-globulin was severely depressed, and there were less marked depletions in mean values for thrombin neutralization, antiXa and in the heparin antithrombin assay (which showed the flat curve thought to reflect a thrombotic tendency). The ∞1-antitrypsin values were normal. The findings support the concept of antithrombin as the summation of ∞2-Globulin and ∞1-antitrypsin (with ∞2-macroglobulin); and may be related to the high incidence of thrombosis reported in childhood malnutrition.
Factor VII Activity and Antigen in Hemophilia B Variants
Authors: Mazzucconi M.G. , Bertina M. , Orlando D. , Romoli G. , Avvisati G. , Mariani G.
Published: May 2019

In 23 patients with Hemophilia B (variants: II B-, 5 BR and 7 B+) factor VII Activity (VII:C) and Antigen (VII:Ag), in correlation with Thrombotest were measured. Thrombotest was found prolonged in 14 patients (in 8 > x+3SD and in 6 >x+2SD). Factor VII:C was found reduced in 1/11 B-, in 2/5 BR and in 4/7 B+ variants. Factor VII:Ag was normal in all but one patient (a 5 years old boy). The ratio VII:C/VII:Ag was abnormal in 8 patients. In these patients factors II and X activities were always within the normal range. The discrepances between VII:C and VII:Ag may be due to: 1) very minor vitamin K deficiency (presence of small amounts of PIVKA-II), 2) an inhibition of factor VII activation by a factor IX “abnormal” molecule, and 3) the synthesis of an abnormal molecule of factor VII.
Polymerisation Defect of Fibrinogen London
Authors: Lane D.A. , VanRoss M. , Cuddigan B. , Kakkar V.V.
Published: May 2019

In the present study further work has been carried out on fibrinogen that has been isolated from a patient with a coagulation defect tentatively designated fibrinogen London. The dysfibrinogenemia was characterised by normal polypeptide chains on SDS gel electrophoresis, normal release of fibrinopeptide A and a delayed polymerisation of fibrin monomers. The polymerisation defect was pH and ionic strength dependent and partially corrected by protamine sulphate and Ca++ ions. Studies on the NH2 and COOH terminal polymerisation domains were carried out using insolubilised fibrin monomers prepared from patient and normal fibrinogen (Kudryk et al, Thromb. Res, 9,25. 1976). High MW fragment D, prepared by plasmin digestion of normal fibrinogen, bound to both normal and patient fibrin monomer and was eluted with 8M urea indicating no gross malfunction of the NH2 terminal polymerisation domain. Also purified fibrinogens from normal and patient plasma a bound to normal fibrin monomer sepharose, suggesting that the COOH terminal domain is capable of binding to the NH2 terminal domain of normal fibrin. These results suggest that the polymerisation defect of fibrinogen London is different in character to that of fibrinogen Detroit, where the NH2 terminal domain does not bind to the normal COOH terminal domain.
Fibrinogen Logroño. A New Fibrinogen Molecular Variant
Authors: Fernandez J. , Lasierra J. , Narvaiza M.J. , Vilades E. , Palaciosy E. , Rocha E.
Published: May 2019

We present a new molecular variant of fibrinogen. The majority of biological tests were normal except prolonged prothrombin and thrombin times, fibrinogen deficiency as quantified by a colourimetric method, and the polymerization of fibrin’s monomers. Normal thrombin times were obtained by addition of normal plasma, bovine thrombin, CaCl2, and by changes in pH and ionic strength.

The study of the purified fibrinogen showed normal fibrinopeptides’ liberation and sialic acid and hexosamine levels. Fibrin stabilization, coagulability index = 64%, electrophoretic mobility, pI = 5.4 (N = 5.8), and S20,w = 10 (N = 8.2), were abnormal.
Increased Sensitivity to Endotoxin Induced Disseminated Intravascular Coagulation (DIC) in Vitamin E-Deficient Rabbits
Authors: Lipinski B. , Machlin L.J.
Published: May 2019

DIC in rabbits was induced by i.v, injection of E. coli endotoxin (e) (50μg/kg). The levels of fibrinogen (F) and FDP, fibrin monomer (FM), PTT, white blood cells and platelet counts were determined before and 3 hours after e injection. Fibrin (f) deposited in organs was calculated by counting the radioactivity of 125I-F introduced i.v, beforehand. Pretreatment of rabbits, maintained on laboratory chow, with dl-α-tocopheryl acetate (vit. E) (40mg/kg/day i.m. for 3 days) did not protect the animals against U1C induced with 2 doses of e 24 hours apart, and only slightly reduced amounts of f deposited in organs. On the other hand, rabbits maintained on a vit. E-deficient diet were found to be extremely sensitive to e. Single doses of e caused 100% mortality within 24 hours, as compared to 100% survival in a group fed the same diet containing vit. E. Laboratory findings (F depletion, presence of FM, increased FDP and prolonged PTT) and increased f deposition in organs (kidneys, lung, liver and spleen) indicated activation of intravascular coagulation by a single dose of c in vit. E-deficient rabbits. It is possible that normal levels of vit. E protect against cell injury by e and thus prevent a release of procoagulants into the blood stream.
Procoagulant Properties of Ascitic Fluid in Hepatic Cirrhosis
Authors: Phillips L.L. , Rodgers J.B.
Published: May 2019

Re-infusion of ascitic fluid into the vasculature of patients with liver cirrhosis corrects many of the protein, fluid, and electrolyte abnormalities. Such infusion can lead to D.I.C. Three such cases are reported here. One had intermittent drainage and re-infusion, the other two had Le Veen shunts installed. All three patients showed laboratory evidence of D.I.C. Ascitic fluid on one patient showed procoagulant material by the thromboplastin activation test. In the presence of ascites from another patient, plasmas deficient in Factors XI, IX, and VIII gave recalcification and non-activated partial thromboplastin times similar to those of normal plasma. RCT’s and PTT’s of normal heparinized plasma and of plasmas deficient in Factors X and VII-X were shortened to a lesser extent. Clotting times of Factor V deficient plasma were prolonged by ascites. These results indicate that an activator of Factor X is present in the ascitic fluid. Additional tests such as RVV and prothrombin times suggest that some activated Factor X and/or tissue activator (endotoxin and leukocytes) may have been formed in vivo.
Preparation of a Factor VII Concentrate
Authors: MacLeod A.J. , Dickson I.
Published: May 2019

A factor VII concentrate has been prepared from pooled citrated fresh frozen plasma following removal of cryoprecipitate and factors II, IX and X. The method involved batch adsorption on DEAE-Sephadex A-50, fractionation of the subsequent batch eluate by PEG precipitation and passage through a column of DEAE-Sepharose CL-.6B. A phosphate-citrate buffer pH 6.9 was used throughout, this was made 0.2M with NaCl for the batch elution and a 0 - 0.2H NaCl linear gradient was used to elute the components from the column. Factor VII activity was clearly resolved from the bulk of the protein, including caeruloplasmin, and could be recovered as a concentrate at about 20 U FVII/ml with a specific activity of in excess of 1 U FVII/mg of protein and an overall recovery of 40% to 50%
Influence of Platelet Inhibitory Drugs on the Thrombogenicity of Dacron Arterial Grafts Perfused in an Artificial Circulation
Authors: McCollum C.N. , Rajah S.M. , Crow M.J. , Kester R.C.
Published: May 2019

The effect of Acetyl Salicylic Acid (ASA), Dipyridamole (DPM) and in combinations on the response of platelets to woven Dacron perfused in an artificial circulation was investigated. Circuits were perfused by heparinised blood donated by 6 volunteers after each had taken: -1) No drugs 2) ASA 300mg tds 3) DPM 100mg qds 4) ASA 300mg + DPM 75mg tds 5) ASA 80mg + DPM 75mg tds 6) ASA 80mg tds, for 1 week. Bleeding times were performed prior to each donation. Platelet functions were estimated during perfusion. Each grafts surface was examined by scanning electron microscopy (SEM) at the end of perfusion.

Platelet inhibition by low dose ASA + DPM was accompanied by less extension of bleeding time, and we suggest that such combination may protect patency in Dacron grafts at risk.
Platelet Release Reaction in vivo in Patients with Ischemic Heart Disease (IHD) After Exercise and its Prevention with Dipyridamole
Authors: Yamazaki H. , Motomiya T. , Sano T.
Published: May 2019

Although an interaction between platelets and arteriosclerotic vessel wall is thought to be important in thrombus formation, a little information was obtained in clinical subjects. We have reported that platelet aggregation Increased in patients with IHD after exercise. To analyse the mechanism of this phenomenon, changes in platelet sensitivity to ADP aggregation, plasma von Willebrand factor and beta-thromboglobulin level were measured in 30 IHD and 30 healthy controls before and Immediately after an isometric exercise (handgrip of 50% voluntary contraction for 2 min). Platelet sensitivity and vWF were determined by original methods detecting microscopically the highest dilution of serially two-fold diluted ADP or test plasma mixed with ristocetin to give platelet aggregation. Beta-TG was measured by RIA Kit. An effect of anti-platelet drug was also observed in IHD. The patients with IHD were administered with placebo or dipyridamole (400 mg/day for 4 weeks) in a crossover single blind fashion. Under placebo, platelet sensitivity to aggregation, vWF and beta-TG increased immediately after the exercise with a statistical significance in IHD. In the healthy control and IHD under dipyridamole, these increases were not observed. The phenomenon may suggest that platelets circulating in sclerotic vessels tend to release and are enhanced in reactivity with smaller stimuli than those in healthy. Such changes might be prevented with dipyridamole.
A Prospective Trial of Sulfinpyrazone and Survival After Thrombotic Stroke
Authors: Blakely J. A.
Published: May 2019

From Jan. 1974 to Dec. 1977, 1260 patients who had recovered from the acute effects of stroke were identified in chronic care hospitals and stroke rehabilitation units. Stroke had been attributed to thrombosis in 754 patients; those with azotemia, hyperuricemia, anemia, gout, bleeding diathesis, active peptic ulcer, malignancy, severe disability, poor immediate prognosis, or antiplatelet or anticoagulant therapy were excluded. 346 were considered suitable for entry and 290 consented and were randomly allocated to Sulfinpyrazone 800 mgs. daily or placebo. Patients were followed in double-blind fashion to Dec. 1978. Randomization was stratified for age, sex, normal or abnormal ECG and interval of more or less than 6 months since stroke. Suspect drug toxicity was similar in treatment and placebo groups. 107 patients ceased to take medication over the course of the study, 52 in the treatment and 55 in the placebo group. All but 6 were followed to the termination of the study. There were 53 deaths in patients currently taking study medications, 25 in the treated find 28 in the placebo groups, and 44 deaths in patients who had stopped taking study medications at the time of death, 20 in the treatment and 24 in the placebo groups. Further analysis is in progress. The overall mortality rates do not suggest that Sulfinpyrazone improves survival after recovery frcm presumed thrombotic stroke.
Sulfinpyrazone in the Prevention of Cardiac Death: The Anturane Reinfarction Trial
Authors: Margulies E.H.
Published: May 2019

The Anturane Reinfarction Trial is a randomized, double-blind, multicenter clinical trial comparing sulfinpyrazone (200 mg q.i.d.) and placebo in the prevention of cardiac mortality among patients with a recent documented myocardial infarction. Results represent data accumulated on 1475 eligible patients entered 25-35 days after myocardial Infarction and followed for an average of 8.4 months, and have been previously reported. The results indicated sulfinpyrazone was effective in reducing cardiac deaths with the most pronounced effect in reducing sudden cardiac death.

The trial was completed in August, 1978. The final results represent data accumulated on 1528 eligible patients with an average follow-up of 16 months. All 106 deaths were of a cardiovascular nature (62 placebo, 43 sulfinpyrazone). Of the 105 cardiac deaths (62 placebo, 43 sulfinpyrazone), 59 were sudden cardiac deaths (37 placebo, 22 sulfinpyrazone), 35 were myocardial infarctions (18 placebo, 17 sulfinpyrazone) and 11 (7 placebo, 4 sulfinpyrazone) were of other cardiac causes. Various approaches of data analysis employed resulted in a statistically significant difference (p=0.047, Cox’s Method) attributable to sulfinpyrazone in reducing sudden cardiac death.

Sulfinpyrazone was very well tolerated with the most frequently associated newly observed sign and symptom being minor gastrointestinal complaints.
Characteristics of the Major Platelet Membrane Site used in Binding to Collagen
Authors: Meyer F.A. , Frojmovic M.M.
Published: May 2019

In binding to collagen surfaces, platelets seem to recognize the high content of proline and hydroxyproline.1 A site seems to be involved which can be blocked by preincubating platelets with soluble polymers rich in proline or hydroxyproline e.g. polyhydroxyproline (PHP). Binding to this site can also be blocked by interacting the platelets with low concentrations of ADP. Preincubation with 100 μg/ml PHP1 or 0.5 μM ADP produce 60±l0% reduction in adhesion.2 The action of PHP is immediate but that of ADP is delayed. The dose of ADP used produces shape change, PHP does not cause shape change. But shape change is irrelevant to the effect since also adhesion of fixed platelets is reduced by both treatments. Addition of ADP and PHP together does not produce greater inhibition of platelet adhesion than either agent alone showing that the same binding site must be influenced. However, ADP reduces adhesion also on albumin - and globulin-coated surfaces showing that the indirect effect of ADP influences other platelet binding sites as well. AMP (1 mM) blocks the effect of ADP but not of PHP. The receptor with which ADP interacts on the platelet surface is not directly involved in binding but can inactivate binding sites presumably through some local interaction mechanism.

1. Meyer, F.A., Weisman, Z., Thromb. Res. 12, 431 (1978).

2. Meyer, F.A., Weisman, Z. and Frojmovic, M.M. (this volume.
Role of Metabolic Inhibitors on the Adhesion of Rabbit Gel Filtered Platelets to Collagen - Sepharose
Authors: Capobianco J.O. , Holleman W.H.
Published: May 2019

The adhesion of rabbit gel filtered platelets (GFP) to collagen-Sepharose was found to be largely energy independent, since metabolic inhibitors only partially suppressed this process. 51Cr and 14C-serotonin labeled GFP were incubated at 37°C/10 min. with anti-metabolites prior to passage over collagen-Sepharose columns. Adhesion and release responses were determined by 51Cr content of columns and 14C content of the supernatant from column eluents. Antimycin-A (AA, 2μg/ml), or AA plus 2-deoxy-D-glucose (2DG, 32mM) or Rotenone (R, 2μg/ml) produced small reductions in adhesion. All three compounds inhibited release, their effects being additive. 2DG or glucono-§-lactone (GLAC, 10mM) alone had little or no effect on adhesion. 2DG reduced release response by 40%, while GLAC had no effect. AA or R reduced adhesion by 35% and 31%, and release by 51% and 47%, respectively. The combination of 2DG, GLAC, AA and R did not reduce adhesion more than 45%. The Inhibitory effects of AA were time and dose related, and the addition c glucose (0.1%) circumvented these effects. These experiments suggest that adhesion includes energy dependent and independent phases. In addition, spherical platelets (GFP at 4°c/24 hrs.) also adhered to collagen-Sepharose; however, in contrast to disc-platelets (GFP at 37°c/10 min.), the adhering platelets demonstrated a depressed release response (-79%),
Kinetics of the Inhibitory Effect of Factor IXBm in the Activation of Factor X by Factor VII and Bovine Tissue Factor
Authors: Østerud B. , Kasper C.K. , Rapaport S.I.
Published: May 2019

F.IXBm is giving a prolonged prothrombin time with bovine tissue factor (T.F.). In the present study the inhibitory effect of purified F.IXBm was tested in a F.VII assay that measures the ability of F.VII to activate F.X in the presence of bovine T.F. and Ca++, followed by the quantitation of formed F.Xa by the synthetic substrate benzoyl -L-Ile-Glu-Gly-L-Arg-p-nitroanilide. Experiments were carried out with different concentrations of F.IXBm and F.X, keeping the concentrations of F.VII and bovine T.F. constant. Lineweaver-Burk plot of the data showed that all the curves crossed the l/V axis in one point - no inhibition by F.IXBm at infinite concentratlons of F.X. Thus, F.IXBm acts as competitive in inhibitor to F.X in the activation of F.X by F.VII and bovine T.F. By plotting the 1/V against the inhibitor concentrations (F.IXBm conc), a Dixon plot, the Ki for F.IXBm was determined to be 0.017 μM.

Normal F.IX was also found to be a competitive inhibitor to F.X but 4-5 times higher concentrations of F.IX were needed to give the same inhibition as F.IXBm. Utilizing human T.F. instead of bovine T.F. in this assay resulted in no significant inhibition by either F.IX of F.IXBm.
Potentiation of Antithrombin III Activity by Sulphated Polysaccharides
Authors: Kindness G. , Long W.F. , Williamson F.B.
Published: May 2019

Because of current interest in the mechanisms of antithrombin III-mediated anticoagulation by heparin, we examined the anticoagulant effectiveness of a number of other sulphated polysaccharides of relatively well-defined chemical structure using ex vivo assay systems. Our results suggest that heparan sulphate, cellulose sulphate, dextran sulphate, xylan sulphate, and a number of sulphated galactans derived from marine algae are able, like heparin, to potentiate antithrombin III inhibition of thrombin and activated coagulation factor X. Dermatan sulphate exerts anticoagulant activity which appears to be independent of antithrombin III. The relative effectiveness of these polymers as activators of antithrombin III may be related to their molecular rigidity.
Studies of the Anti-Xa Activity of Human Plasma and Purified Antithrombin III
Authors: Barrowcliffe T.W. , Eggleton Anne C.
Published: May 2019

When samples of purified antithrombin (At III) were compared to plasma at the same At III concentration, in the absence of heparin, the anti-Xa activity of plasma was considerably higher. In the presence of heparin the anti-Xa activity of purified At III was much greater than plasma. This was shown to be due to an inhibitory effect on the heparin. At III-Factor Xa interaction in plasma which could be removed by absorption with aluminium hydroxide [Al(OH)3]. This inhibition was dependent on the molecular weight of the heparin; low molecular weight heparin was inhibited less than high molecular weight heparin, and this probably accounts for the apparently high anti-Xa activity of low molecular weight heparin.

A1(OH)3 absorption of plasma also increased its anti-Xa activity in the absence of heparin. Addition of Factor IX concentrate to the absorbed plasma reduced its anti-Xa activity to that of normal plasma, and studies with purified proteins showed that this effect was due to the prothrombin in the concentrate. The addition of Factor IX concentrate or prothrombin to purified At III did not affect its anti-Xa activity.

These results suggest that, in addition to At III, there is another Xa-inhibitor in plasma which competes with prothrombin for binding of Factor Xa.
Drug Treatment of Cerebrovascular Disease
Authors: Hirsh J.
Published: May 2019

Drugs which suppress platelet function and anticoagulants have been evaluated in patients with cerebrovascular disease. There is evidence that aspirin prevents cerebral transient ischemic attacks (TIA), stroke and death in certain patient groups with TIA. Effectiveness is most evident in males, who are non-hypertensive and who have appropriate lesions in their large cerebral arteries. Sulphinpyrazone alone is not effective in the treatment of transient cerebral ischemia but may augment the effects of aspirin. There is some suggestive evidence which requires confirmation that sulphinpyrazone may lower the mortality from cardiovascular causes in elderly patients who have strokes. There is no evidence that dipyridamole used alone is effective in the treatment of cerebrovascular disease. Studies using anticoagulants have been less well designed but the tentative conclusions from these studies are that anticoagulants are ineffective and may be dangerous in the treatment of completed stroke, may be effective in the treatment of stroke in progression (although this is not conclusive) and are effective in reducing recurrent episodes in patients with cerebral embolism. The combined use of anticoagulants with either aspirin or dipyridamole appears to be more effective than oral anticoagulants alone in the treatment of embolic episodes including cerebral embolic episodes in patients with prosthetic heart valves.
Fluid Dynamical Considerations of Platelet-Vessel Wall Interaction
Authors: Turitto V.T.
Published: May 2019

Platelet adhesion to subendothelium is dependent on two distinct processes: (1) diffusive transport (T) of platelets to the surface and (2) platelet-subendothelial reactivity (R). The relative magnitude of T to R will determine which factors control the adhesion rate. Physical factors, specifically, wall shear rate (γ) and platelet diffusivity (D) influence T, whereas chemical alterations modify R. An increase in the magnitude of γ or D or a decrease in R tends toward R controlled adhesion. An increase in low rate or decrease in vessel dimensions increases γ, whereas D increases with both red cell concentration (up to 40 %) and γ.

In flowing blood at shear rates comparable to those found in veins or large arteries (< 650 see-1), T determines platelet adhesion. Moderate alterations in R, such as produced by the addition to blood of 45 mM citrate, 10-100 nM prostacyclin or in von Willebrand factor depleted blood, have little effect on platelet adhesion values under these flow conditions. However, as shear rate is increased to values comparable to those in the microcirculation {1300-2600 sec-1), the same blood samples show values of platelet adhesion which are reduced compared to controls, and the reduction increases with shear rate Thus, measurements of R should be determined under controlled shear conditions which are high enough to be outside the range of predominantly transport controlled adhesion.
The Influence of Purified Factor V, Factor Va, and Activated Platelets on the Factor Xa Catalyzed Activation of Prothrombin
Authors: Nesheim M. E. , Mann K.G.
Published: May 2019

The kinetics of prothrombin activation by factor Xa and Ca++ plus purified factor V and phospholipid or activated bovine platelets were monitored continuously with dansylarginine-4-ethylpiperidine amide which specifically inhibits thrombin and displays enhanced fluorescence upon binding. With purified components and saturating amounts of factor Va a 500,000 fold increase in activity occurred compared to that with factor Xa alone, with phospholipid and Ca++ present, the rate increase with factor Va was 24,000 fold. The reaction rate with factor V was only 0.5 to 1.0% of that with factor Va, although saturating concentrations of either form of the cofactor were approximately equivalent. At fixed levels of factor Xa and Ca++, the process was saturable with platelets; it was also saturable with factor Xa at fixed platelet levels. Saturation occurred at levels consistent with the notion that platelet factor Va (measured by bioassay and radioimmune assay) was limiting in both cases. The maximum velocity with platelets was about 1100 moles thrombin min-1/mole factor Xa. These results suggest that I) factor Va influences prothrombinase activity to a greater degree than previously recognized: 2) unactivated factor V has low but measurable activity; and 3) in terms of maximum catalytic capacity the synthetic prothrombinase utilizing phospholipid and purified factor Va is about equivalent to that utilizing platelets. (Supported by NHLBI Grants HL-07069 and HL-17430D).
X-Ray Investigation of Fragment 1 of Bovine Prothrombin
Authors: Burridge J.M. , Aschaffenburg R. , Blaxe C.C.F. , Esnouf M.P.
Published: May 2019

The conversion of prothrombin to thrombin during the penultimate stage of the blood cloting system is achieved by the cleavage by factor Xa of two specific peptide bonds in the inactive prothrombin molecule; it is thought that prothrombin, factor Xa and factor V are organised into a specific prothrombin-converting complex on a phospholipid surface prothrombin and factor Xa binding via Ca2+ ions which bind to γ carboxyglutamic acid residues in both proteins. Fragment 1 is released by the action of thrombin, and is thought to inhibit excess prothrombin conversion. The fragment consists of the first 1 residues of prothrombin, molecular weight 22,000, it contains all ten γ-carboxyglutamic acid residues, and two of the three carbohydrate chains of the prothrombin molecule, together with one of the so-called “Kringle” units, repeated in fragment 2, and occurring five times in plasminogen. Fragment 1 crystallises from 2M phosphate, pH range 6.0 to 7.5, in one of the tetragonal space groups P43212 or its enantiomorph P41212; cell dimensions a = b = 78.87Å, c = 84.99Å, with one molecule per asymmetric unit. Isomorpho,. derivative survey work has led to the collection of low resolution (6Å) photographic oru jection data and three dimensional difftactometer data for crystals grown at pH 7.5 Experiments are in progress on crystals grown at pH 6.0, and on methods of modifyin protein before crystallisation; trial data collection at 3Å has suggested that st tural analysts at this intermediate resolution should be possible.
Behaviour of Heparin in vivo - Its Metabolism as Followed by a New 125-I Shpp-Conjugate
Authors: Pepper D.S. , Dawes Joan
Published: May 2019

Using the reverse sequence of the Bolton and Hunter reagent (SHPP -succinimidylbydroipbenylpropionic acid) than that normally applied for protein labelling we were able to incorporate 0.08 mole SHFP/mole heparin using the non-iodinated molecule. Following purification of the conjugate on protamine-agarose, it could be labelled in a conventional chloramine-T-iodination to yield 125-I or 125-I heparin with a specific activity of 10-20 uCi/ug, more than 95% of the tracer was atill bound to protamine following iodination. Using other chromatography systems, radioactive labelalways coincided with bioassay or chemical assay of heparin. Alkaline hydrolysis studies showed that the label was bound to amino groups of glucosamine and not to a terminal lylosyl-serine residue. In vivo studies shoved that the half life of disappearance was strongly dose dependent, and that the label re-appeared in the circulation after 2 hours and then persisted for a much longer period. This metabolic product was de-sulphated heparin which was biologically inactive but otherwise an intact polysaccharide which appears to be slowly cleared via the kidneys. Whole body scanning of a 131-I-conjugate heparin at +40 mins (when ca 90% of the label had temporarily disappeared from the circulation) shoved that desulphation took place largly in the liver and spleen.
Inter-Relationships of dysfibrinogenaemia, Fetal Fibrinogen and Liver Cell Growth
Authors: Barr R.D. , Clause M.M. , Johnston M.A.
Published: May 2019

When operationally defined as an excess of immunoassayable fibrinogen (IAF) over thrombin-clottable fibrinogen (TCF) of ≥ 1 g per L, dysfibrinogenaemia (DF) is demonstrable in more than 1/3 of patients with chronic liver diseases (CLD)1. Some features of this protein suggest a similarity to fetal fibrinogen.2

To further pursue these investigations, it was deemed necessary to identify a population of very young patients who were synthesizing fibrinogen rapidly. Twenty (2C Infected neonates, of gestational ages 24-41 weeks, were selected for study on the basic of TCF levels ≥5 g per L, Using antisera raised in sheep and rabbits, there was no demonstrable excess of IAF in the same plasma samples. Similar data were obtained fron assays in an unselected aeries of umbilical cord plasmas. These findings suggest either thet DF in CLD is not due to the renewed synthesis of fetal fibrinogen, or that fetal fibrinogen, like alpha-fetoprotein and fetal pre-albumin,3 is physiologically I evident only during early embryonic development.

1. Barr, R.D. et al. J. Clin. Path. (1978) 31, 89.

2. Barr, R.D. Blomedlcine (1979) in press.

3. Tatarinov, Y, S. et al. Lancet (1978) 2, 1122.
Initial Characterization of a Fast Plasmin Inhibitor from Platelets Different fron the Known Plasma Proteinase Inhibitors
Authors: Hansen Sandbjerg M. , Clemmensen I.
Published: May 2019

An inhibitor (I) of the plasma proteinase plasmin (P)(EC 3.4.21.7) was partially purified from washed and lysed human blood platelets by ammani sulphate fractionation and affinity chromatography. The material contain none of the known plasma proteinase inhibitors when studied by crossed ir munoelectraphoresis (CIE) and elee troimmuno assay, but inhibited a clot lysis time assay and an esterolytic assay using the synthetic substrate 2251 (O-Val-Leu-Lys-pNA), The inhibitory activity was found in the alpha region by preparative agarose gel electrophoresis (AGE). Titration of the I preparation by active-site titrated P showed a linear relationship untj zero activity indicating a strong binding of I to P. The inhibition was complete within one minute. The I changed the mobility in AGE af activator-free P or 125-I-P from the gamma region to the alpha-2 region as demoi strated by CIE against specific immunoglobulins against plasminogen or by autoradiography. An antibody against I has been raised in rabbits. The results strongly suggests the presence in platelets of a plasmin inhibitor different from the known plasma proteinase inhibitors.
A Non-Stasis Rabbit Model for the Detection of Factor IX Concentrate Thrombogenicity.
Authors: Prowse C.V. , Williams A.R.
Published: May 2019

A method has been developed whereby aerial blood samples can be obtained from a rabbit over a period of four hours following infusion of potentially thrombogenic solutions. Infusion of 50 uAg thrombin over JO minutes produced intravascular coagulation for up to three hours after infusion as demonstrated by a decrease in factor VIII, increase in partial thromboplastin time and fibrin(ogen) degradation producta and a positive ethanol gelation teat. No change in fibrinogen, factor DC or platelet count was found. Saline infusion produced no change in any of these parameters.

Infusion of a variety of factor IX concentrates at 100 u/kg shewed that those concentrates active in in vitro thrombogenicity teste produced a similar effect to thrombin in vivo and in addition may result in a drop in platelet count. Infesion of concentrates with low in vitro activity did not induce intravascular coagulation.
Modified Tests for Measuring the Thrombogenic Potential of Factor IX Concentrates.
Authors: Middleton Sarah M. , Douglas Jessie T. , Forbes C.D. , Prentice C.R.M.
Published: May 2019

In vitro tests for screening potential thrombogenicity of factor IX concentrates are unsatisfactory as it has been shown that the non-activated partial thromboplastin time (NAPTT) and the TGt50 (reflecting thrombin generation by the concentrate after recalcification} do not correlate with each other. We have modified the TGt50 by addition of optimum concentrations of factor V and phospholipid, and compared it with the NAPTT and factor Xa generation test using the chromogenic substrate S2222. The modified TGtSO correlates with both the NAPTT and the factor Xa generation test suggesting that these tests are measuring the same entity. Chromatography of concentrates on sepharose 4B indicates that the high MW void volume material has procoagulant activity as measured by the unmodified TGt50 whilst the retained volume he procoagulant activity as measured by the TGt50 and the NAPTT. When, however, phospholipid is added to the unmodified TG150 the activity of the high MW component is lost and only that of the retained volume is still present. The data suggests that the modified TGt50 and the NAPTT measure the same procoagulant activity in these concentrates.
Crude, Clinical Factor IX Concentrates: Comparison of Factor IX Antigen Levels and Demonstration of Factor IXa
Authors: Thompson Arthur R. , Smith Kenneth J.
Published: May 2019

Thrombogenicity complicates the use of crude factor IX concentrates but “activated’ forms may benefit hemophilic patients with inhibitors; an in vitro correlate remains to be defined. We have examined various commercial concentrates (CCs) for Factor IX pro-coagulant (PC) and antigen (Ag) levels and chromatography activated CCs on heparin-agarose CHA). Non-activated CCs contained Ag equal to or in excess of PC. Activated CCs from 2 sources (6 lots) showed PC:Ag ratios of 5.2, 6.5, 7.1 and 5.4 for one and 2.1 and 1.8 for the other. One lot each of the activated CCs (300 Ag units) were chromatographed over HA in 50 mM Tris-1 mM CaCl2 with a linear gradient from 0.1 to 0.7 M NaCl and collected Into tubes containing EDTA. Fractions were analyzed for clotting activities of factors II, X, Xa and IX and IX,, and for factor Xa by chromogenic substrate (S-2222). In the activated CC from the second source, a heparin-llke inhibitor was demonstrated in the breakthrough peak and activated clotting activity trailed factor IX. The latter was characterized as factor IXa by assays in which factor IX deficient plasma was rapidly clotted with or without kaolin and PC was >> Ag; It also shortened the clotting activity of factor VII (but not VIII or X) deficient plasma. Monospecific rabbit anti-Eactor IX, at 1:500 dilution, completely Inhibited these activities.
A Synthetic Thrombin Inhibitor Taking Extremely Active Stereo-Structure
Authors: Okamoto S. , Hijikata A. , Kikumoto R. , Tamao Y.
Published: May 2019

The novel series of synthetic thrombin inhibitors reported by us in 1978 included the extremely potent, highly selective and less toxicinhibitors (No, 407 and No. 700). Extensive studies on the stereo-structure activity relationship around No. 700 were further made and No. 805 was found.

Its I50 for fibrinogen-thrombin system was extremely small (0.03~0.04 μM), approximately 1/10 of that of No. 407. Selectivity to thrombin was satisfactorily high. From the characteristic stereo-structure of No, 805, stereo-geometry of the active site of thrombin, maybe assuming replica structure of No. 805, became more informative. LD50 was 140 mg/kg i. v. in rats and 210 mg/kg i. v. in mice. Per os administration of 20 mg/kg of No. 805 was able to prevent the decrease of platelet number due to thrombin infusion in rabbits. At 0.3 μM of No. 805 in plasma in rabbits, the carotid artery thrombi due to acetic acid application was also well prevented.
The Inhibition of Human Thrombin with 5-Acyloxyoxazoles: High Specificity for Thrombin in Contrast to Plasmin
Authors: Lawson William B. , Valenty Vivian B. , Wos David J.
Published: May 2019

A number of 5-acyloxyoxazoles rapidly inactivate human thrombin by acylating the serine residue at the active site. The inhibition takes place via, an affinity-labeling mechanism dependent upon the binding of an aromatic group at position 2 on the oxazole ring to a hydrophobic site on the enzyme. The compounds do not contain a positively charged group (common to most thrombin inhibitors) that could bind to the active-site pocket specific for arginyl side chains. Evidence to date indicates that the 2-aryl group must be coplanar with the oxazole ring for rapid inhibition. 2-Fhenyl-4-(3-N-nitroguanidinopropyl)-5-pivaloyloxyoxaiole (I) inactivates thrombin 3-4 orders of magnitude faster than human plasmin. The 2-methyl analog of I is about 700 times poorer than I as a thrombin inhibitor, but only about 3 times poorer than I as a plasmin inhibitor. These results can be correlated with the good binding of the hydrophobic dye proflavin to thrombin but not to plasmin.(Supported in part by Grant HL 15BBT from the National Institutes of Health.)
Similar Prolongation of Bleeding-Time After Low and High Doses of Acetylsalicylic Acid (ASA)
Authors: Dybdahl J.H. , Eika C. , Daae L. , Godal H.C. , Frislid K. , Wiik I. , Larsen S.
Published: May 2019

In Lancet, 2, p,780, 1978, O’Grady and Moncada reported that high doses of aspirin did not lead to prolonged bleeding-time. Prompted by this observation, we examined 21 healthy male volunteers, aged 21-56 years. After an overnight fast, the bleeding-time was recorded in duplicate using Trombolette (Mannheim, Solingen, DBR). Thereafter, 0.425 or 3.825 g of ASA (GLOBOID, Nyco, Oslo, Norway) was given in a cross over study with a minimal interval of 14 days. Finally, the bleeding-time was repeated after two more hours of fasting.

As is evident from the table, a significant increase in bleeding-time was observed after low as well as after high doses of ASA (p< 0.001). In addition, the bleeding-times obtained after ingestion of low and high doses of ASA were not significantly different (p = 0.22).
A Simple Method for Estimation of Lipoxygenase and Cyclo-Oxygenase Pathways in Human Platelets - The Use of Thiobarbituric Acid Reaction
Authors: Okuma M. , Takayama H. , Uchino H.
Published: May 2019

As lipid peroxides are produced from arachidonic acid(AA) by platelet lipoxygenase(PLO) and cyclo-oxygenase(PCO), a simple method was developed for estimation of these enzymic pathways by quantitating the thiobarbituric acid-reacting substance(TBARS) produced by the incubation of AA with human platelets. Thin-layer radiochromatographic analysis (J. Biol.Chem. 252:5871, 1977) of lipid products obtained by incubating [1-14C] AA with washed platelets under various conditions revealed that maximal amounts of PCO products were obtained by the incubation at 37°C within 1 min around pH 7.4, while those of PLO products within 10 min around pH 6.5. The incubation at 37°C of AA with aspirin-treated platelets at pH 7.4 for 1 min produced no TBARS, while that at pH 6.5 did TBARS as a function of the incubation time (up to 10 min). Based on these findings, PLO and PCO pathways were estimated by the determination of TBARS (J.Lab.Clin.Med. 75:283, 1970) obtained by incubating AA (0.2 mM, f.c.) either with aspirin-treated or with intact washed platelets (108) at 37°C as shown in the table. Normal values (M ± SD) expresse in terms of nmols of malonaldehyde were: PLO = 1. 76 ± 0 .36 (n=l7), PCO = 1.12 ± 0.42 (n=15). Alterations in these pathways were detecetd by this method in some patients with myeloproliferative disorders.
Phospholipase A2-Induced Platelet Aggregation, Release and Lysis.
Authors: Heinrich D. , Beckmann S.
Published: May 2019

Activation of washed platelets by exogenous phospholipase A2 (PIA2) purified from crotalus terrificus terrificus venom was studied. Platelets were labeled with 14C-serotonin and 51chromium and resuspended in Tyrode/albumin (TA). With 1-5 μg/ml (final conc.) of crotalus PIA2 no direct platelet alterations were observed. These platelets, however, were refractory to collagen - but not to thrombin or HLA-specific antibodies.

10-50 μg/ml crotalus PIA2 rapidly induced platelet aggregation and release 100 μg/ml crotalus PIA2 induced platelet lysis.

PIA2-induced platelet alterations were inhibited by EDTA, PGE1, ASS and apyrase. Crotapotin, an acid peptid isolated from crotalus venom, forms complexes with crotalus PIA2 and specifically inhibits PIA2-induced platelet alterations.

Conclusion: PIA2-induced platelet alterations are due to liberation of arachidonic acid from phospholipids of the platelet membrane inducing prostaglandin and thromboxane synthesis. With high concentrations of PIA2 breakdown of membrane phospholipids will lead to platelet lysis.
Heparin Prophylaxis Combined with DHE or Dextran in HIP Operations
Authors: Schöndorf T.H. , Weber U.
Published: May 2019

162 elective hip operation patients were randomly allocated to: Group I: 5 000 U heparin injected s.c. 10 and 2 hours pre-operatively, then 8 hourly; Group II: heparin as above plus dextran 40 on three days; Group III: heparin as in group I combined with dihydroergotamine (DHE). The 125-I-fibrinogen test was routinely used for diagnosis of deep vein thrombosis (DVT), when positive, phlebography was performed.

DVT occurred in 8/55 patients in group I, in 9/54 in group II and in 2/53 in group III. (Chi-square test: I vs. III p = 0.054; II vs. III p = 0.028) .

Pulmonary embolism (confirmed at autopsy) occurred in one patient in both group I and II.

In the three groups, the intra-operative transfusions, wound blood loss at the pre- and post-operative haematocrit did not differ.

Platelet aggregation was only significantly lowered in group II.

Heparin/DHE were most effective in preventing thromboemboli, dextran infusions did not improve heparin prophylaxis.

Peridural anaesthesia performed in 83 - 87% of patients in all groups led to no complications following pre-operative heparin.
Further Experience in the Prevention of Deep-Vein Thrombosis (DVT) After Elective HIP Surgery Using Lignocaine
Authors: Cooke E.D. , Bowcock Sally A.
Published: May 2019

We have shown that the administration of intravenous (IV) lignocaine 2mg/min commenced prior to surgery is associated with a marked reduction in the total incidence of deepvein thrombosis (DVT), In particular, thigh vein thrombus was absent, after elective hip surgery. (Lancet 2, 797 - 799, 1977). The present investigation was to determine the effect of a lesser dose of IV and of Intramuscular (IM) lignocaine. In 25 patients IV lignocaine 1mg/min was administered by central venous line, starting 4 hours before, and during the first week following, surgery. Thereafter in 11 of these patients lignocaine was administered intramuscularly; 200mg 4 hourly in 7 patients and 300mg 4 hourly in 4 patients. Monitoring for DVT was by thermography, performed pre-operatively and daily postoperatively until discharge. Phlebography was performed in all cases either when thermography became positive or on the 14th postoperative day. During the administration of IV lignocaine the total incidence of DVT was 32 per cent (expected incidence 70 - 80%) wIth an incidence of thigh vein thrombosis of 25 per cent (expected incidence 50%). The incidence of DVT during IM lignocaine using either dosage regime was similar to the expected incidence of DVT in patients receiving no prophylaxis. We conclude that the effect of lignocaine in the prophylaxis of DVT following hip surgery is both route and dosage dependent.
Activation of Purified C1, The First Complement Component, by Purified Plasma Kallikrein and Plasmin
Authors: Cooper N.R. , Miles L.A. , Griffin J.H.
Published: May 2019

Many similarities exist between the proteins, activation mechanisms and control processes of the complement (C) system and the Hageman factor pathways (the coagulation, kinin-forming and fibrinolytic systems). Earlier studies have presented evidence both in favor of and against activation of the classical C pathway by plasmin at the level of the Cls subcomponent of Cl. To study this problem the effects of purified plasma kallikrein and plasmin on isolated Clr, Cls, the calcium-dependent Clrs complex, and macromoleculcr Cl were studied. Kallikrein activated Clr as shown by its increased ability to cleave 125I-Cls. Kallikrein activated Cls as assessed by hydrolysis of synthetic substrates. This activation was associated with cleavage of 125I-Cls into several fragments with the largest being 29,000 daltons; the usual

heavy chain of 59,000 daltons generated by

was not evident. Kallikrein also cleaved 125I-Cls in Clrs and in macromolecular Cl. Plasmin activated Clr and cleaved 125I-Clr into fragments of 35, 000 daltons and smaller; the usual

heavy chain was not evident. Plasmin also activated Cls and cleaved it into fragments similer in size to those generated by kallikrein. Plasmin activated Clrs and Cl, apparently via its action on Clr. Thus, kallikrein and plasmin both directly activate Cl. These studies suggest that activation of prekallikrein or plasminogen in plasma may lead directly to C triggering via activation of Cl.
Efffct of Heparin-CI ina Complex on the Action of Kallikrein
Authors: Ikematsu S. , Tamura H. , Fukutake K.
Published: May 2019

CI INA has been known to show not only the inhibitory action on CI esterase, but also the inhibition of blood clotting factors including factor XI, factor XII, plasma kallikrein and plasmin. In this paper the authors attempt the following studies to elucidate the function of heparin-CI INA complex in the field of contact phase. By the addition of heparin into normal human plasma the pattern of cross immunoelectrophoresls of CI INA becomes to show three peaks indicating a complex formation of heparin. CI INA and other clotting factors, which is reversible to move to an original peak by the addition of Protamin sulfate. And it has been analysed that the first peak consists of a complex of heparin-CI INA complex with factors XI and XII and the second peak is a complex of heparin-CI INA and the third peak shows a complex of heparin-CI INA with kallikrein and plasmin. On the observation of inhibitory action of heparin-CI INA complex by the use of chromogenic substrate (S-2302) method for plasma kallikrein assay, it is conformed that the inhibitory action of CI INA on kallikrein is enhanced in parallel with the increasing amount of heparin added into the tested plasma, and the same tendency is observed on the experiment with AT-III free plasma. In conclusion it is strongly suggested that heparin-CI INA complex might be able to form a complest with kallikrein to inhibit its amidolytic activity more effectively than CI INA without heparin does.
A Solid Phase Radio-Immuno Assay for Platelet-Bound IgG Employing 125-I protein Extraction of 7S IgG and Identification of Its Subclasses
Authors: Hymes K. , Shulman S. , Schur P. , Karpatkin S.
Published: May 2019

Studies on bound anti-platelet Ab have not differentiated Ab from Ag-Ab complexes. A sensitive solid phase radio-immuno assay has been developed for IgG, which detects picogram IgG. Washed platelets are frozen, thawed, sonicated and centrifuged at 20,000 g. The supernatant is applied in serial dilution to a series of plastic wells, capable of adsorbing protein. After incubation, the wells are washed with 1% BSA in PBS, pH 7.2. Commercial rabbit anti-human IgG in 1% BSA-PBS (or specific anti-human IgG subclass) is then applied. Unbound anti-IgG is washed away and the wells treated with 125-I staphprotein A (10,000 cpm/well; 2x107 cpm/μgm). Unbound protein A is washed away and the wells assayed for radioactivity. Twenty-six of 28 thrombocytopenic patients (92%) with autoimmune thrombocytopenic purpura had IgG greater than 2 SD of controls and averaged autoimmune thrombocytopenic purpura had IgG greater than 2 SD of controls and averaged, 172±155 (SD) ng/106 platelets; 11 healthy controls, 12.3±4.9 ng; 5 ‘thrombocytopenic controls, ‘8.2±5.7 ng. The platelet count (X) correlated inversely with IgG/platelet (Y) according to the equation: log Y=-0.61 log X + 4.4; r=0.64. P<0.00l. Sucrose gradient analysis, employing 7S and 19S marker proteins, revealed platelet ‘extract IgG’ activity migrating as 7S protein, not as Ag-Ab complex. Three patients were tested for IgG subclasses: γG1, γG2, γG3 and γG4. These averaged 75±9.7, 13±6.1, 9.7±4.2 and 2.3±1.3% of total bound IgG. We conclude that a 7S immunoglobulin can be extracted from platelets which is IgG and contains all 4 IgG subclasses at a distribution found in normal plasma.