bluuebunny/crossref_metadata_2025 huggingface dataset:
Metal Ion-Induced Structural Changes Of Bovine Blood Coagulation Factor X1
Authors: Freyssinet J M
Published: July 2018

Wheat germ lectin (W.G.A.) has been used as a macromolecular probe to investigate metal ion-induced conformational changes of bovine Factor X1 . The classical precipitin reaction between lectin and glycoprotein only occurs when certain metal ions are present. Precipitin reactions were followed by recording turbidity at 330 nm.

It has been verified that W.G.A. lectin activity is insensitive to the presence of either EDTA, Ca(II), Mn(II) or Mg(II). Ca(II) and Mn(II) are able to induce a precipitin reaction between W.G.A. and Factor X1. The reaction is slow at metal ion concentration of 2.5 mM but becomes more rapid when higher divalent cation concentrations are used. Ca(II) and Mn(II) have comparable efficiency, Mg(II) is totally ineffective in promoting such a precipitin reaction. Halftimes of precipitin reactions range from 7 minutes at 2 mM Ca(II) to 0.3 minutes at 100 mM Ca(II). Above 100 mM Ca(II) or Mn(II), increasing metal ion concentration would slow down the reaction ; increasing the ionic strength had a similar effect. Although very slow and weak, the precipitin reaction could still occur at high ionic strength up to 0.8 M. The various conditions that give rise to a precipitin reaction demonstrate that Factor X1 must undergo a cation- catalyzed transit ion which allows binding of lectin to its specific sugar(s). This leads to suggestions on the nature of this structural change. When compared to metal-ion induced protein transition involved in protein-phospholipid binding described in the literature, the difference of divalent cation specificity and active concentration in both of these studies point out that the conformational changes have to be different.
Anticoagulant Effect Isolated From Radix Miltiorrhizae (RSM)
Authors: Stemberger A , Blümel G
Published: July 2018

RSM is a well known Chinese medicinal herb traditionally used as (injectable) decoction to improve circulation+. Decoctions of the root* showed a prolongation of the aPTT and this effect was further characterized.

Standardized RSM decoctions were ultrafiltrated using step by step the following membranes: 300.000 MW, 100.000 MW, 50.000 MW, 5.000 MW, 1.000 MW , 500 MW. The effect of these fractions on blood coagulation was studied in routine tests like thrombin time and aPTT. With chromogenic substrates the inhibitory effect was elucidated on the Xa− and thrombin level after exogen or endogen activation of the coagulation system.

The most potent fraction has a molecular weight of 5.000 daltons with a high anti-Xa and anti-thrombin effect. Using partly purified Xa and thrombin preparations inhibition of these enzymes was independent from antithrombin III. Activation of plasminogen as reported elsewhere was not detectible. In contrast an antiplasmin effect could be demonstrated. The chemical nature of the inhibitor(s) is under investigation.
Influence Of Elastase On Blood Coagulation Factors: Studies In Vitro, In Rats And In Göttinger Miniatur Pigs
Authors: Kasten U , Artmann U , Kaethner T , Burchardi H , Köstering H
Published: July 2018

The influence of blood coagulation factors in pat. with acute respiratory insufficiency of adults, especially of the so called “pancreatitis lungs” is still unknown. In order to find out the effect of elastase, possibly activated by trypsin in pat. with acute pancreatitis, on blood coagulation factors, we performed some studies. In vitro elastase induces in plasma and blood in correlation to the dosages Enhancement of thrombingeneration in the TGT, a shortening of PTT, Thrombin time and of r- and k-time in the TEG, a loss of fibrinogen and an increase of fibrinmono-mercomplexes. In another study, elastase (960 U/ kg b.w.) was injected intravenously in rats. 30 min. later there was found a loss of fibrinogen, number of platelets, Prothrombin and a prolongation of PTT and Thrombin time and an increase of fibrinomonomercomplexes, especially in these rats, which received beside elastase Kalikreininhibitors or antifibrinolytic drugs. After repeated injections (3 times within 30 h) we found histomorpholgically thrombi as well as bleeding complications. In another study we performed (150 min) an infusion of elastase (333 U/kg b.w./h) to 9 pigs. We determined a loss of fibrinogen of platelets, of F. II, F. VII and F. XIII, a prolongation of PTT. F. VIII and F. V remained within the normal range But there was found an enhancement of Thrombin generation in the TGT, too. Compariening the results of blood coagulation tests and of histomorphological findings, elastase induced a DIC. We have to discuss their influence on ARIA and “Pancreatic lungs”.
Viscoelastic Studies Of Coagulation: Cell-Plasma Interactions
Authors: Mitchell J , Zuckerman L , Vagher J P , Caprini J A
Published: July 2018

In order to examine the cell-plasma interactions during coagulation, a thrombelastograph was modified to measure two viscoelastic properties of blood: rigidity (G) and rate of stress relaxation (S). Citrated whole blood was collected from healthy donors and centrifuged to obtain platelet-rich and platelet-poor plasma and washed red cells. The blood was reconstituted at varying hematocrits (HCT) and platelet counts (PLT) and coagulation was initiated by adding CaCl2. Factor XIII (FXIII) activity was inhibited with glycine ethyl ester (GEE). In normal blood, the rigidity increases rapidly after 6-8 min following addition of CaCl2 with a rate constant (kG) of 8.2 min and a saturated rigidity (Gsat ) of 22.3×l03 dyn/cm2 . S decreased in a biphasic minner: the first phase (ΔS1) lasting about 15 min and accounting for a 70% fall in S and with a rate constant (ks1) of 6.7 min; the second phase (ΔS2) from 15 min on with a 15% drop in S and with a rate constant (ks2) of 71.4 min. Increasing PLT resulted in a linearly increasing G up to 31×l03 dyn/cm2 for 3×105 cells/mm3. Increasing HCT had no effect on Gsat , but increased kG linearly for 10<HCT<50%. Inhibition of FXIII activity by GEE had no effect on G, but inhibited ΔS2 while kS2 was linearly dependent on GEE concentration. In cell-free plasma, ΔS1 was about 80% in 10 min with virtually no ΔS2. With the inclusion of even a small number of platelets, ΔS1 and ΔS2 returned to that found in normal blood.

It was concluded that platelets change the configuration of fibrin in the clot and alter the kinetics of orientation and crosslinking. Red cells have little mechanical influence but do seem to have some procoagulant activity.
Isolation And Characterization Of Procoagulant Substances From Human Ascites
Authors: Johnston M F M , Vargo J , Joist J H
Published: July 2018

The treatment of massive, medically intractable ascites by a peritoneovenous shunt (PVS) is associated with variably severe disseminated intravascular coagulation (DIC). Ascitic fluid obtained from cirrhotic patients at the time of placement of a PVS was found to shorten the partial thromboplastin time (PTT) of normal human platelet-poor plasma. This procoagulant activity which was found to reside in the cell-free fraction of ascitic fluid was heat stable and insensitive to pH change over a wide range. Chromatography on Bio-Rad Agarose 1.5 yielded one major high molecular weight component and several smaller fractions of lower molecular weight exhibiting procoagulant activity as determined by the PTT assay. The activity in the major fraction (80% total activity) coprecipitated with human fibrinogen but could be separated from fibrinogen by affinity chromatography using anti-human fibrinogen. This procoagulant did not hydrolyze chromogenic substrates S- 2222 and S-2238 and was not inhibited by diisopropylfluoro-phosphate (DFP). Purification of a minor procoagulant fraction by aluminum hydroxide (Al (OH)3) adsorption and chromatography on Sephadex G-200 yielded a fraction that clotted both citrated normal plasma (without the addition of calcium chloride) and purified human fibrinogen and induced platelet aggregation in citrated human platelet- rich plasma. Chromatography on DEAE cellulose yielded two peaks with procoagulant activity (PTT), one of which hydrolyzed S-2238 and was DFP-sensitive. These studies indicate that at least three distinct, clotting active substances are present in ascitic fluid some or all of which may be responsible for PVS-induced DIC. Further studies on purification and characterization of these ascitic procoagulants are in progress.
Determination Of Reaction Time Changes Per Time Unit In Clot Assays
Authors: Popović Lj
Published: July 2018

Changes in reaction time of clot assays are usually expressed only in time units, which fails to indicate the extent of the increase or decrease of the reaction time of the tested specimens against that of the basic sample. Reaction time increases of, e.g. , 6 seconds in tested samples, compared to basic sample reaction times of 12 and 24 seconds respectively, signify an increase twice as large in the first as in the second instance.

Changes in reaction time of clot assays can be expressed as the increment or decrement of the reaction time per time unit. This amount of increase or decrease (positive or negative alteration of reaction time, T
a
) can be expressed as the quotient of the difference between the reaction times of the tested (T
x
) and basic (To) samples and of the basic sample, e.g. in seconds per second, T
a
=T
x
-To/To. A test sample reaction time 6 seconds longer than basic sample reaction times of 12 and 2k seconds would mean an increase of 0.5 and 0.25 seconds per second, respectively.

Reaction time changes of tested samples against that of the standard sample (T
std
) can be calculated in a similar way, T
a
=T
x
-T
std
/T
std
.

It can be assumed that this parameter reflects the intensity of the increase or decrease of reaction time per time unit. The quotient of the tested and basic samples can be considered as the coefficient of the increase or decrease of the total reaction time (CT=T
x
/To).
Studies Of The Peptide Fractions Derived From Fibrinogen By Digestion With Trypsin Or Plasmin
Authors: Mihalyi E
Published: July 2018

Digestion of fibrinogen with a number of proteolytic enzymes yields a series of large fragments with protein-like structure and a substantial amount of peptidic material. Some of the peptides released possess biological activity, therefore, the study of their formation and properties may be of clinical interest. In the present investigations human or bovine fibrinogen was digested by either plasmin or trypsin. Approximately 35% of the fibrinogen molecule is removed as peptides with either enzyme and these are well divided into two groups: 1. large peptides of 15,000 to 40,000 MW and 2. small peptides of less than 10,000 MW. The distribution of material between these groups is approximately the same with either enzyme and with bovine or human fibrinogen, however, the plasmic peptides in either group are larger than the tryptic ones. All the large peptides are derived from the approximately 40,000 MW C-terminal portion of the α-chain. In this segment, according to the sequence of the human α-chain, there are 33 bonds theoretically susceptible to either plasmin or trypsin. Of these, only 3 are attacked at a fast rate by plasmin in the native molecule, resulting in the appearance of peptide segments of 40,000, 32,000 and 26,000 MW. The first two are short lived, reach only about 25% of the stoichiometric amount, and are converted into the last one, which accumulates as a fairly stable end product. Trypsin digestion of the latter involves again 3 susceptible bonds and through an intermediate of 17,000 MW results in 2 fragments, one of 10,000 and another of 7,000 MW. The restricted cleavage of susceptible bonds in these fragments, their intrinsic viscosity and its increase in denaturing media, suggest that these peptides possess a fairly compact structure, i.e., they are not in the random coil state as hitherto suggested.
Recurrent Venous Thrombosis Related To A Hereditary Dysfibrinogen With Abnormal Crossed Immunoelectrophoretic Pattern
Authors: Papp A C , Snopko R M , Cole E R , Sassetti R J , Wu K K
Published: July 2018

A 24 year old man with a history of multiple recurrent venous thrombosis was found to have a qualitatively abnormal fibrinogen. Plasma fibrinogen concentration by a kinetic thrombin time method was 135mg% as compared to 150mg% by the gravimetric method of Ratnoff and Menzie. The thrombin, reptilase and Russell viper venom times were all approximately 50% longer than the normal values. Neither the thrombin time nor reptilase time was corrected upon addition of CaCl2. Antigenic quantitation by Laurell immunoelectrophoresis gave a value of 320mg%. On Ouchterlony immunodiffusion plate, the patient’s plasma and normal plasma showed a line of identity. The patient’s fibrinogen had a normal rate of migration by one-dimensional immunoelectrophoresis but it exhibited an abnormal pattern upon crossed immunoelectrophoresis. The abnormality is characterized by the presence of a shoulder on the anodal side of the fibrinogen peak. Family studies revealed that the fibrinogen defect was inherited as an autosomal dominant trait. To determine whether recurrent venous thrombosis is related to increased viscosity due to the abnormal fibrinogen, fibrinogen was purified by the method of BlombSck and the relative viscosity of fibrinogen solution was determined. Preliminary data were suggestive of an increased viscosity of the patient’s fibrinogen. These findings indicate that the immunologically abnormal dysfibrinogen may be responsible for recurrent venous thrombosis because of alteration of the physicochemical properties of the fibrinogen molecule.
Clinical And Experimental Studies On The Ionic And Nonionic Contrast Media Interactions With Anticoagulant Drugs
Authors: Parvez Z , Messmore H L , Moncada R , Anderson A C , Fareed J
Published: July 2018

Interactions of ionic and non-ionic radiologic contrast media (CM) with anticoagulant drugs like heparin and Dex- tran-40 have been investigated. Renografin-60 (Squibb and Sons, Princeton, New Jersey), P-297, ioxigalic acid and iothalamic acid (Laboratoire Guerbet, Paris, France) were used. Human, monkey, dog and rabbit plasmas were incubated with CM, CM + heparin, CM + Dextran-40 and saline; prothrombin time (PT), partial thromboplastin time (PTT), and thrombin time (TT) were determined. In human plasma, Renografin-60 (final cone. 30 mg/ml) produced a strong anticoagulant effect and clotting times PT, PTT, and TT were prolonged by 1-1.5X their base value. When Renografin-60 was supplemented with 0.2u/ml heparin, the TT was greatly elevated, >100 secs, (control < 20 secs). No such prolongation of TT was noted when Dextran-40 was mixed into CM + plasma mixture. Iothalamic acid also showed potent anticoagulant action. P-297, which is a .non-ionic CM, showed the least anticoagulant action by itself, but did manifest some synergistic reaction with 0.2 - 0.4u/ml heparin (TT >100 secs, with 10 NIH u/ml thrombin). Similar results were obtained with monkey, dog and rabbit plasmas. In the in vivo experiments, dogs weighing 11-18kg were injected intravenously with 100-150 u sodium heparin/kg body weight, along with 50-100 ml Renografin-60. Blood samples were drawn at 1 hr., 3 hrs., 4 hrs., and 6 hrs., post injection and assayed for clotting times. 5 ml/kg CM injected with 25 u/kg heparin prolonged the activated partial thromboplastin time (APPT) and TT for upto 6 hours, while sodium heparin along produced anticoagulant effects for shorter duration (< 4 hours). Our studies indicate that ionic forms of CM produce relatively stronger antithrombotic response in animals as compared to non-ionic ones and utmost care should be exercised in their usage in patients on anticoagulant drugs.
Synthetic Peptide Assays For Monitoring The Defects Of Extrinsic And Intrinsic Pathways Of Coagulation: Selection Of A Suitable Substrate
Authors: Walenga Jeanine , Fareed Jawed , Messmore Harry L , Kniffin Judith
Published: July 2018

Several assays based on the use of thrombin specific synthetic peptide substrates have been proposed for the amidolytic equivalents of prothrombin time (PT) and partial thromboplastin time (PTT). All of these methods employ the same principle as the clotting assays; the test plasma is activated with either thromboplastin or activator-cephalo-plastin mixture and the generated thrombin activity is measured employing thrombinr specific synthetic substrates such as Bz-Phe-Val-Arg-pNA (S-2160), H-D-Phe-Pip-Arg-pNA (S-2238), Tos-Gly-Pro-Arg-pNA (Chromozym TH), and CH3-Gly- Pro-Arg-pNA. These substrates and their free peptide forms inhibit the amidolytic action of bovine Xa activated human and Xa Russell’s viper venom in the following order: S-2160 > S-2238 > Sarc-Pro-Arg-pNA ≥ Chromozym TH. A new substrate for thrombin Pyro-Glu-Pro-Arg-pNA (S-2366) and a plasminogen activator (tissue) substrate, H-D-Ile-Pro-Arg-pNA (S-2288) have been tested and were found to produce weaker inhibition of bovine and human Xa. Tos-Gly-Pro-Arg-pNA, Sarc-Pro-Arg-pNA and H-D-Ile-Pro-Arg-pNA were found to produce a ≤ 10% inhibition of the activator generated Xa’s amidolytic activity at concentrations which are commonly employed in the PT and PTT assays. Although in the developmental stages the synthetic substrate equivalent assays are more sensitive than the existing PT and PTT assays and provide useful information on the total amount of thrombin generated in each assay, our results suggest that amidolytic equivalent assays for PT and PTT are feasable. However proper selection of a peptide substrate is important as some of the thrombin substrates and their free peptide forms may inhibit Xa and other serine proteases which are generated during the activation step thereby seriously influencing the results.
A Simple Quantitative Methodology For The Analysis Of In Vitro Blood Coagulation Profile Kinetics
Authors: Palmason C , Hewitt L A , Harris C
Published: July 2018

Blood coagulation profilers graphically display the kinetic events which lead to in vitro clot formation. Their potential usefulness in patient monitoring is evident, but at the present time is limited by the lack of a standard data analysis procedure. The present study demonstrates a simple, quantitative, approach to profile analysis.

The profile curve was comprehensively defined in terms of seven easily determined parameters. The average value for each parameter was determined from extensive testing of commercial coagulation control plasmas (n=100), human female volunteer plasmas (n=30), and pooled female plasmas (n=5). Where control and human parameter values were significantly different, correction factors, were calculated to “normalize” the commercial plasma values. The “normalized” commercial plasma parameters could then be used as a highly reproducible reference standard with which a patient’s profiler could be statistically tested for abnormalities.

Profiles of third trimester of pregnant women (n=11) were compared with the reference standard as a positive test of the procedure to identify hypercoagulable states. Similarly profiles of newborn babies (n=10) and factor deficient patients (n=7) were compared as a positive test of the procedure to identify hypocoagulation. The results indicate that the analytical procedure applied to the blood profiles are valid, and may be of use in the clinical setting.
A Laboratory Case Report Of “Hypercoagulation Syndrome” Following Eight Weeks Of Estrogen Replacement Therapy
Authors: Palmason C , Shams S , Harris C
Published: July 2018

A preliminary study was designed to identify if estrogen replacement therapy (E.R.T.) could be correlated with any progressive alterations in blood coagulation function. A total of thirty post menopausal women were assigned to treatment groups and monitored monthly for six months. This presentation relates the laboratory findings of one 55 year old woman who developed significantly altered blood coagulation parameters after two months on conjugated estrogens equine (1.25 mg/day).

Routine biochemistry, F.S.H. levels, haematology, and eight standard coagulation tests were done. However, prothrombin time (P.T.), and partial thromboplastin times (P.T.T.), were done on a Coagulation Profiler (T.M.). Assessment of coagulation function was affected through comparison of the patients present P.T. and P.T.T. profiles with pretreatment and reference standard profiles. All aspects of the patient’s coagulation kinetics were altered significantly, in the direction of hyper-coagulation (p< 0.001) with the exception of the P.T. time alone, which was unchanged. Platelet aggregation was impossible, due to spontaneous aggregation during pre-test standing, fibrinogen and euglobulin lysis levels were also raised. The patient reported breast engorgement, abdominal distension and calf tenderness. Two months following E.R.T. discontinuation, the patient’s blood coagulation parameters had all returned to pretreatment levels.

The case is discussed within the context of an hypercoagulation syndrome resultant upon an idiosyncratic response to estrogen therapy.
Rebound Phenomenon After Discontinuation Of Oral Anticoagulant Therapy?
Authors: Harenberg J , Haas R , Zimmermann R
Published: July 2018

Duration and discontinuation of oral anticoagulant therapy in the prophylaxis of recurrent thrombembolic diseases is often discussed. After stopping of the therapy an elevated incidence of rethromboses are observed. Therefore we determined fibrinopeptide A (FPA) as the most sensitive parameter of thrombin activity in vivo in patients treated with phenprocoumon in order to get information on the effectivity of the oral anticoagulant therapy and on the rebound phenomenon after discontinuation of the treatment.

In 136 outpatients a significant relation between FPA and the effectivity of the anticoagulant therapy was observed: 2.7 ng/ml plasma (mean, thrombotest values < 5%), 3.5 ng/ml (5%-15%) and 4.1 ng/ml (>15%, p < 0.01). Normal FPA were 1.4ng/ml. In 11 patients with low FPA the oral anticoagulants were discontinued. FPA was determined in weekly intervalls. Within four weeks a continous increase of FPA from 1.6 ng/ml (median) to 3.8 ng/ml was observed. After eight weeks FPA was 8.2 ng/ ml (p < 0.05). The in vitro release of FPA after 10 min incubation of blood increased from 3.4ng/ml to 72 ng/ml after eight weeks (p < 0.05, normal release up to 5 ng/ml).

The data indicate the clinical relevance and the different therapeutic implications of the determination of FPA in patients treated with phenprocoumon. They further give the first criterion, which outlines the effectivity of an oral anticoagulation therapy by reducing the thrombin activity in vivo. They give some evidence for a rebound like phenomenon after discontinuation of the oral anticoagulant treatment.
Hypercoagulability In Acute Esophageal Variceal Bleeding
Authors: Hiller E , Hegemann F , Riess H
Published: July 2018

In 15 patients with acute esophageal bleeding selected parameters of hypercoagulability were determined at frequent intervals during intensive care treatment. Estimation of soluble fibrin monomer complexes (SFMC) by gel filtration of plasma samples which were purified by ß-alanine precipitation allowed the determination of the relative amount of SFMC (percentage of SFMC in relation to the total fibrinogen content in plasma). Antithrombin III (AT III) activity was determined photometrically using the chromogenic substance S-2238 and immunologically by one dimensional immunelectrophoresis. Fibrin split products (FSP) were estimated by the staphylococcal clumping test.

Increased levels of SFMC were observed in 10 out of 15 patients on admission. A further increase was noted in most patients in whom bleeding persisted and who needed replacement therapy with blood components. Substitution with prothrombin complex concentrates induced acute DIC in two patients with levels of SFMC up to 24 %. AT III was decreased to levels of 30-50 % in 8 patients during the acute illness. A discrepancy between the functional and immunological AT III value was noted in some instances but more often both values were very low.

High levels of SFMC in addition to levels of AT III of less than 50 % reflect a serious state of hypercoagulability with a very poor prognosis for the patients. Clotting factor concentrates may be especially thrombogenic in these patients with impaired clearing activity. Fresh frozen plasma and AT III concentrates provide an appropriate source of the most important clotting factors.
Clinical Spectrum Of The Lupus Anticoagulant In Pediatrics
Authors: Sternbach M S , Reece E L , Esseltine D , Macdonald E
Published: July 2018

Hemostatic mechanisms were investigated in two patients: one had deep vein thrombosis, the other one bled after a tooth extraction. Standard coagulation methods were used: Coagmate for P.T. and P.T.T., manual water bath methods for thrombin time, factor levels and circulating anticoagulants.

An 11 year old girl presented with a deep vein thrombosis of the left ileo-femoral junction. Her P.T.T. was prolonged 66"/30" with the presence of a circulating anticoagulant with Anti-factor XI activity. All other factors, fibrinogen and thrombin time were normal. A.N.A. was + 1/40; D.N.A. binding was + 1/40; complement (C4) was low. An Anti I was positive in a titer of 1/256. The patient was treated with anticoagulants for 6 months.

A 10 year old boy had prolonged bleeding following a deciduous tooth extraction. A spleen tip was the only pertinent physical finding. P.T. and P.T.T. were markedly prolonged (P.T. 16"/11"; P.T.T. 50"/30"). A circulating anticoagulant with Anti-factor XI and XII activity was detected. The patient’s clot dissolved completely at 4°C for 2 hours (while being stable for 48 hours at 37°C and 20°C). The patient had a + A.N.A., a + anti-D.N.A., and a positive direct Coombs’ with anti-N specificity. Hematuria, cylindruria, and proteinuria were also present. Following prednisone therapy, the circulating anticoagulant and the positive Coombs’ disappeared.

These cases illustrate that the lupus anticoagulant may be present with either thrombotic or hemorrhagic manifestations.
Inhibition Of Prostacyclin Production In Some Women With Lupus Anticoagulant And Repeated Abortion Or Intrauterine Death
Authors: Carreras L O , Van Assche A , Vermylen J
Published: July 2018

The association of the lupus anticoagulant with repeated abortions has recently been recognized. We discovered the lupus anticoagulant in four patients with a history of repeated abortions, intrauterine growth retardation or intrauterine death of unknown origin. The diagnosis of the lupus anticoagulant was based upon prolonged KCT (not corrected in the mixtures with normal plasma) and abnormal dilute tissue thromboplastin assay. Patient 1 had two intrauterine deaths, severe pregnancy induced hypertension and a history of recurrent arterial thrombosis. The patient’s IgG fraction, containing the lupus anticoagulant, decreased the release of prostacyclin (PGI2) from rat aorta rings or human pregnant myometrium and reduced the production of 6-keto prostaglandin F1α by cultured bovine endothelial cells. The plasma level of 6-keto PGF1α was reduced. Patient 2 had one intrauterine death and six spontaneous abortions. During these pregnancies she developed severe hypertension. The production of PGI2-like activity by fresh or exhausted rings of rat aorta was decreased by the patient’s plasma. Patient 3 had three repeated intrauterine growth retardations and patient 4 had four spontaneous abortions. The plasma of these two patients did not prevent the release of PGI2 by vascular tissue. No clinical of laboratory signs of systemic lupus erythematosus were found in these patients. Considering that the lupus anticoagulant acts by interfering with the function of phospholipids in blood coagulation we suggest that the same antibody could interfere with the production of PGI2 by the vessel wall in some of these patients, possibly by interfering with the availability of arachidonic acid. Since there is increasing evidence for a physiological role of PGI2 in pregnancy and fetal life, an inhibition of PGI2 production could be deleterious for fetal outcome.
Antithrombin III Infusion During Fulminant Hepatic Failure
Authors: Braude S , Arias J , Hughes R D , Canalese J , Gimson A E S , Williams R , Scully M F , Kakkar V V
Published: July 2018

The antithrombin III (ATIII) levels in 17 patients with fulminant hepatic failure due to viral hepatitis or paracetamol overdose were found to be 25.8%±SD 12.80 of normal on admission. The levels did not correlate with eventual survival or death and remained essentially unchanged for up to 7 days.

In an attempt to assess the role of the low levels of AT III during the course of hepatic failure and in relation to treatment by charcoal haemoperfusion we have infused patients with commercially purified ATIII. Preliminary measurements of ATIII (chromogenic substrate method) were made and ATIII infused to achieve a plasma concentration of 50 to 70%. Infusion was by an initial bolus of 1500-2000 units followed by up to 500 units every 6 hours. To date 3 patients in Grade IV hepatic coma have been treated, one died 1 day after admission and the other two survived. In the latter the return of the prothrombin time to normal was similar to that in patients without the addition of ATIII. In one of the survivors the platelet count did not fall, suggesting ATIII may have had a protective effect on platelet consumption. There was also an indication, that there was a more uniform and better response to heparin anticoagulation during haemoperfusion than found previously without ATIII infusion.

Further patients will be treated to evaluate whether AT III substitution can reduce the consumptive coagulopathy and platelet destruction which occurs in the course of fulminant hepatic failure.
Antithrombins And Coronary Artery Disease
Authors: Bick R L , Faulstick D
Published: July 2018

The naturally occurring antithrombins, primarily anti- thrombin-III, are key modulators of coagulation and appear to protect against increased fibrin deposition. The role of antithrombin-III with respect to atherogenesis and myocardial infarction remains unclear; however, it has been reported that antithrombin-III levels are decreased in many individuals with coronary artery disease and in those at high risk for coronary artery occlusion. However, the several reports available do not agree and remain inconclusive. To better define the role of antithrombins in coronary artery disease antithrombin-III levels were determined in 86 individuals undergoing coronary artery angiography. Angiography was performed by the brachial or femoral route and evaluation was by cineradiographic technique using standard RAO and LAO projections. Patients were categorized as 0=no coronary artery disease, Class I = 50% stenosis of 1 or 2 vessels, Class II = 75% stenosis of 1 or 2 vessels, Class III = 75% stenosis of 3 vessels, and Class IV = stenosis of
3 vessels and abnormal left ventricular function. Samples for AT-III determinations were drawn at both the initiation and cessation of the procedure. Results of these determinations reveal that the majority of patients with documented coronary artery disease by angiographic studies have normal antithrombin-III levels. Additionally, there was found to be no correlation between antithrombin-III levels and severity of coronary artery disease. The only finding was that during the angiographic procedure antithrombin-III levels tend to decrease, confirming the work of others that angiographic dyes in general may alter hemostasis. The results of this study would suggest that plasma antithrombin-III levels are of no particular significance in determining the presence, absence, or severity of coronary artery disease.
Antithrombin III Treatment In Acute Liver Failure
Authors: Vogel G E , Bottennann P , Clarmann M v , Komm Ch , Oberdorfer A
Published: July 2018

In acute liver failure (alf) there is a defect in synthesis of coagulation factors in addition there is a disseminated intravascular coagulation which is followed by an impairment of the microcirculation. With an early substitution of Antithrombin III (AT III) we tried to stop this situation. In 22 patients (10 female, 12 male, age 10-68) with alf presenting with hepatic coma (grade I-IV) we studied the time course of AT III plasma activity (the study started in December 1978 and is continued until now). AT III was measured with the chromogenic substrate method. When AT III activity fell below the level of 80% of normal, we started to substitute AT III and to give low dose heparin (125-500 U/hrs). In addition in case of bleeding or a decrease of coagulation factors or fibrinogen under the hemostatic active concentration, complexes of prothrombin and fibrinogen were administered. Besides the usual conservative treatment for alf, patients in coma (grade IV) were undergoing baboon liver perfusion. The rapid fall of the hepatic coagulation factors stopped. In patients, who still were able to synthesize coagulation factors a reincrease of these factors after administration of AT III was seen and there was a further fall in fibrinogen. The dosage of AT III in alf required to bring AT III to normal values depended on the degree of intravascular coagulation. The average dose in our study was 250 U/3 hrs. The clinical course of alf was prolonged in all patients and 7 patients with the prognostic deleterious colombindex (sum of factors II + V + VIII) < 75% eventually survived the alf. The coagulation disorders in alf can be treated with an early substitution of AT III; thus, there is more time for liver regeneration. Our results suggest an improved prognosis of the acute liver failure.
Antithrombin III Substitution In Acute Hepatic Failure Due To CCl4 Intoxication
Authors: Egbring R , Klingemann H G , Heimburger N , Karges H E , Beule J , Seitz R , Have-mann K
Published: July 2018

In patients with acute severe hepatic failure the synthesis of clotting factors and inhibitors is considerably diminished. The decrease of clotting factors may be enhanced by liberation of thromboplastic substances from liver cell debris, leading to thrombus formation in the sinusoids and to further cell damage. At the low levels of clotting factors and inhibitors signs of disseminated intravascular coagulation as well as hyperfibrinolysis have been demonstrated. Treatment with heparin to prevent coagulation is insufficient at low levels of antithrombin III (AT III). Therefore, Vogel und Fritsche 1979 suggested the substitution of AT III in these cases.

We now report about 3 patients who were admitted to the clinic together with severe signs of liver damage after oral uptake of CCl4

On the day of admission several clotting factors plasminogen and alpha2-antiplasmin were significantly diminished; AT III levels between 25-45% of the norm were found. (Diss. Eckhardt-Klaßnitz). Therefore we started treatment with AT III concentrate from Behringwerke (1000-2000 I.U. daily for 3 to 14 days) and fresh frozen plasma (total volume of 2 1 within the first 3 days).

AT III was simultaneously determined by clotting test, a chromogenic substrate test, and immunologically. Hemodialysis was necessary in 2 patients. Unter treatment with AT III and fresh frozen plasma no bleeding tendency occured Though two of the patients showed severe intoxication on admission all could be dismissed with only slight histological signs of liver alterations. Treatment with AT III concentrates, therefore, seems of value in patients with acute yellow liver dystrophy.
Changes In Biologically And Immunochemically Measured Antithrombin III (At III) In Total HIP Replacement With Special Regard To Type Of Thromboembolic Prophylaxis
Authors: Fredin H O , Tengborn L
Published: July 2018

The plasma levels of AT III were studied in 71 patients operated with total hip replacement. AT III was determined with amidolytic assay using the chromogenic substrate Coatest S-2238 (KabiDiagnostica, Sweden) as well as electroimmunochemically. Samples were drawn before the operation and 4-5 times the first week postoperatively. The results were adjusted to the hematocrit (Hcr) for each sample.

The patients were randomly allocated to thromboprophylactic prevention with either Macrodex (Pharmacia, Sweden) or heparin with dihydroergotamine (Sandoz, Switzerland). Phlebography of the operated leg and perfusion/ventilation lung scanning was performed on the 10-14th postoperative day.

Results. No difference in the AT III levels were seen in patients who developed postoperatively deep venous thrombosis and/or pulmonary embolism (DVT/PE) as compared to those who did not develop DVT/PE.

No decrease of AT III was found postoperatively, when adjusted to Hcr

The frequency of DVT/PE did not differ significantly between the two types of thromboembolic prophylaxis.

Conclusion. Pre-operative determination of AT III, whether biologically or immunochemically, did not seem to be successful as screening method of patients to develop DVT/PE.
Purification Of Antithrombin Ul Using Chinese Yellow ( Ocher Originated From China ) As A New Adsorbent And Heparin - Sepharose Affinity Chromatography
Authors: Suzuki H , Matsuo S
Published: July 2018

In recent years, we have presented several reports with respect to Chinese Yellow ( ocher originated from China ) as a new adsorbent for blood coagulation factors. Adsorbability of this ocher is strong for many coagulation factors in plasma including fibrinogen, while quite weak for antithrombins. After ocher adsorption treatment, AT HI activity of plasma in comperison with the value before adsorption is 86 - 96 % in clotting assay ( Biggs thrombin time methods ) and 96 - 100 % in immunologic activity (detected by SRID ). In this study, purification of plasma AT HI, we have used this ocher as the adsorbent in pretreatment of plasma together with heparin - sepharose affinity chromatography. As the actual procedure, 5 g of ocher was added to 100 ml of ACD plasma, and the mixture was subjected to adsorption with stirring at room temperature to obtain the ocher adsorbed plasma, crude AT HI fraction, in the supernatant after centrifugal separation. Highly purified AT HI with a molecular weight of approximately 67,000 ( determined by SDS-PAGE ) was obtained from the crude AT HE fraction by means of heparin - sepharose affinity chromatography. The antiserum from rabbits immunized with the purified AT HI showed a single fuse precipitin line against the starting plasma, ocher adsorbed plasma and AT HI fraction. However, it did not exhibit reaction with α2-macroglobulin and α1-antitrypsin. From the above results, it has been found that the ocher is an effective adsorbent in pretreatment of plasma for AT HI purification.
Different Affinities Of Heparin Preparations With Identical Molecualr Weight To Plasma Proteins And Factor Xa
Authors: Harenberg J , Fussi F , Grϋn F , Schlegel A , Mattes K , Zimmermann R , Weber E
Published: July 2018

The study intends to get information on the in vivo comparability of different heparin preparations with the same molecular weight (mean MW 15000). Four heparins (150-154 USP/mg, sodium salt) were applicated s.c. to each of six volunteers randomly at weekly intervalls.

The pharmacodynamic effects were controlled on the factor IIa activity (thrombin clotting time), aPTT and factor Xa activity (chromogenic substrate S2222) over 10 hrs by 12 blood samples. On the factor IIa activity no differences were seen between the heparins. The coagulation times of the aPTT first prolonged and then shortened below the initial value after 4-6 hrs indicating an activating of the intrinsic coagulation system. One of the heparins had a higher anticoagulation effect than the other preparations (p < 0.05). For this heparin a more effective anti-Xa-activity was measured (p < 0.05). In vitro no differences of the heparins were registered on the test systems in a range of 0.01-2.0 USP/ml plasma.

The data indicate, that heparin preparations with the same MW and the same in vitro activities may have different effects in vivo. They let suppose, that these differences may be mediated by different affinites of the heparins to plasma proteins as AT III or lipoproteinlipase, which might have caused the activation of the intrinsic coagulation system by a realease of phospholipids, and by different affinities of the heparin AT III complexes of each heparin to factor Xa.
Analysis Of Subcutaneous Sodium And Calcium Heparin
Authors: Caprini J A , Vagher J P , Zuckerman L , Mitchell J
Published: July 2018

Thirteen normal adults were randomized to receive a 15,000 unit subcutaneous injection of sodium (Na-hep) or calcium (Ca-hep) heparin and seven days later the experiment was repeated with the other heparin. The APTT, Thrombin-Calcium clot time (TCCT), Lee-White clot time (LWCT) and Heparin 10-A (10-A) assays were determined prior to the injection and 1,2,6, and 10 hours post-injection. With the Na-hep injection, the APTT, TCCT, and LWCT showed a more rapid onset and persistently greater anticoagulant activity during the 10 hour period with statistically significant prolongations for 1 and 2 hour samples. The 10-A activity was higher in the Ca-hep group with significance at 10 hours. All the test values showed anticoagulant activity 1.5 to 2.0 times the control at 10 hours for both heparin preparations.

An additional 13 normal adults were randomized to receive 5000 units of Na-hep or Ca-hep subcutaneously every 12 hours for 4 time intervals and after seven days the other heparin preparation was given. The APTT, TCCT, LWCT, and 10-A assays were determined prior to the initial injection and at 2,6,and 10 hours post injections. All APTT values varied between 1 and 1.4 times control with significant elevation at 6 hours in the Ca-hep group. The 10-A assay showed significant differences throughout the study with greater activity in the Na-hep group.

We conclude that subcutaneous injections of 5000 units sodium or calcium heparin are bioequivalent despite minor test differences which are magnified by increasing the subcutaneous dose. Therapeutic levels of heparin are maintained for greater than 10 hours following a single 15,000 unit injection and less than 6 hours for 5000 units of either heparin in normal adults. The lack of correlation of either 10-A with other results and the large standard deviation may indicate lack of sensitivity of this assay, rather than differences between heparin salts.
On The Anticoagulatory Activity Of Different Heparins
Authors: Angelkort B , Petrick A
Published: July 2018

The anticoagulatory effectivity of different heparins is seen as to be equal. The study aims for proving the anticoagulatory effects of two sodium heparins with comparable molecular weights (heparin-Na(ThrR.), Naheparin (LiqR.) made by two different manufacturers and one heparin (CalcR.) in patients undergoing chronic hemodialysis (N=10) in crossover trial. Antithrombin Ill- activity, fibrinogen, hematocrit, platelets, blood lipids were of comparable levels in the three patient groups. 20 and 60 min. after application of heparin-Na (ThrR.) the average plasma levels are constant: 1,46 ± 0,28 and 1,45 ± 0,38 U/ml, after application of Na-heparin (LiqR.) plasma levels and anticoagulatory effect tends to be lower. Ca-heparin has a significant meaner anticoagulatory effect as the two sodium-heparins. Between heparin-Na (ThrR.) and Ca-heparin there is a significant difference, whereas the differences between Na-heparin (LiqR.) and Ca-heparin is not performed to be significant. There endures no dominance for the measurement of heparin in plasma with chromogenic substrate instead of the plasma-thrombin-time in supervising an anticoagulation therapy.
Heparin Kinetics And Coagulation Studies After Subcutaneous And Jet Injection Of Low-Dose Heparin
Authors: Briel R C
Published: July 2018

Administration of low-dose heparin by jet injection is regarded as most suitable for both patient and staff. The present study deals with the comparison of plasma heparin levels and various coagulation parameters after s.c. and jet injection (K3-pistol Sandoz) of 5000 IU calcium heparin in 0.2 ml and 5000 IU sodium heparin + 0.5 mg dihydroergotamin (DHE) in 0.7 ml. The injection sites were upper arm, abdcmen or upper leg. Estimations were performed in 10 healthy volunteers each receiving 12 different injections. Heparin level profiles (chromogenic substrate S-2222), AT III activity (S-2238) and concentration (Mancini technique), antifactor Xa (S-2222), antiheparin activity (S-2222), antiplasmin (S-2251), TT, aPTT and TEG r+k were measured.

There was a positive and significant correlation between postinjection heparin levels and body weight, preinjection “endogenous heparin”, AT III activity and concentration, and a negative correlation to the antiheparin activity found before injection. Heparin levels were slightly higher after heparin/DHE than after heparin alone, but the differences were not significant. 30 min after jet injection heparin levels were slightly higher than after s.c. injection, while after 1 hr they were similar. After 2 to 3 hrs the highest levels were found, being significantly higher after s.c. injection. After 4 and 6 hrs there were no differences between s.c. and jet injection. TT, aPTT and TEG showed similar behavior patterns. The values were independant of the injection site, upper arm or abdomen with both s.c. and jet injection. They were lower, however, if heparin jet injection was done in the upper leg.

Thus, the pretreatment measurement of AT III, endogenous heparin and antiheparin activity with chromogenic substrates may be helpful in the management of an inividualized heparin prophylaxis. After heparin jet injection, altered heparin kinetic has to be considered: a more rapid initial absorption is followed by lower peak heparin levels. The upper leg can not be recommended as injection site for heparin jet injection.
Prophylaxis Against Deep Vein Thrombosis Following Total Hip Replacement Using A Combination Of Heparin And Dihydroergotamine
Authors: Kakkar V V , Fok J , Djazaeri B , Ham R , Fletcher M
Published: July 2018

Venous thromboembolism is a frequent and important complication of total hip replacement. Changes in blood coagulability and stasis in the deep veins of the lower limb are major factors in the pathogenesis of deep vein thrombosis (DVT). Prophylactic efforts aimed at changes in coagulability have met with little success in this group. Dihydroergotamine mesylate (DHE), a potent venoconstrictor acting mainly on the capacitance vessels of the limbs is capable of doubling the velocity of venous flow and should minimise stasis.

Four hundred and six patients undergoing total hip replacement were investigated for the efficacy of a combination of heparin (5000IU) and DHE (0.5mg) given 8 hourly subcutaneously in the prevention of postoperative venous thromboembolism. High risk patients were scanned using the I125-labelled fibrinogen uptake test and all patients under went bilateral ascending phlebography on the 14th postoperative day unless there was clinical or isotopic evidence necessitating earlier investigation.

There were 117 DVT (28.8%), 19 (16.4%) of which were bilateral. Three pulmonary emboli (0.7%) occurred, one of which was fatal. Prophylaxis was discontinued in 6 patients (1.5%) because of postoperative bleeding and wound haematoma developed in 23 patients (5.7%) - 6 of which required evacuation.

Heparin and DHE combination in prophylaxis represents a considerable improvement over the use of heparin alone (52% incidence DVT) and Aspirin (80%).