bluuebunny/crossref_metadata_2025_split huggingface dataset:
ACTIVATION AND INACTIVATION OF PURIFIED HUMAN FACTOR VIII BY THROMBIN
Authors: Sewerin K , Lundin A , Hellström E , Larsen K , Sandberg H 
Published: June 2019

Highly purified human factor VIII was activated by human thrombin. The factor VIII material used showed variation in molecular weight and was separated into one heterogeneous form with a molecular weight ranging from 280 to 185 kDa and into one homogeneous 170 kDaform. Incubation of the (280-185) kDa form of factor VIII with thrombin resulted in proteolytic degradation of the heavy chain and formation of a 90 kDa peptide chain. This chain was further split intoone 52 kDa and one 43 kDa peptide chain, as was the 90 kDa chain pesent in the 170 kDa form of factor VIII. The light chain of 80 kDa was split into a 70 kDa peptide chain. The change in peptide composition was followed by scanning and integration of SDS-PAGE gels on samples from the activation of both forms of factor VIII with thrombin. No strict relationship between factor VIII activity and degradation or formation of a new peptide chain was seen.When factor VIII bound to von Willebrand factor matrix was incubated with thrombin, activated factor VIII was released. The released material consisted mainly of a 90 kDa and a 70 kDa peptide chain, but the presence of the 52 and 43 kDa peptide chains increased with the passage of time. The concentration of Ca2+ ions was of great importance for the degree and rate of activation. A high concentration of CaCl2 resulted in a lower degree of activation but also slower inactivation of the factor Villa formed. However, no permanent stabilization of factor Villa was noted at either low or high concentrations of CaCl2.Inhibition or removal of thrombin from the factor VIII system, by the use of protease inhibitors or matrix bound thrombin, did not prevent the loss of factor VIII activity following the activation. Thepeptide composition was stabilized, however, when thrombin was removed. This indicates that the inactivation of factor Villa is due to an intramolecular change in the factor VIII molecule.
ROUTINE MONITORING OF THE β-PROPIOLACTONE/UV STERILIZATION PROCEDURE USING BACTERIOPHAGES
Authors: Dichelmüller H , Stephan W 
Published: June 2019

Cold sterilization of plasma or plasma derivatives such as the serum preserve Biseko® and coagulation factor concentrates by β-Propiolactone (β-PL)/UV has been demonstrated in various studies to be effective in inactivation of Hepatitis viruses and HIV. The methods used in these studies are not applicable in routine monitoring of sterilization processes, as chimpanzees must be involved in the titration of Hepatitis B or NANB viruses.For routine monitoring of the sterilization efficacy we therefore developed a test system using four types of bacteriophages: Øx 174, Øe, Kappa and f2. Using these bacteriophages in 88 single tests, sterilization efficacy was regularly monitored during the period fran 1981 to 1986. For these tests samples were drawn from pooled human plasma, spiked with bacteriophages and subjected to the cold sterilization procedure under identical conditions and apparative equipenent, but strictly separated fran the production process. By treatment with β-Propiolactone and UV all bacteriophages were inactivated by more than 6.7 log10, independent from size or genom structure. This inactivation is in the range of inactivation of Hepatitis B virus by β-PL/UV as a relevant pathogenic virus. During this six year period of monitoring no significant alternations in sterilization efficacy were observed, indicating a constantly safe inactivation rate for numerous production lots and for many years of production.
NORMAL IN VIVO KINETICS OF FACTOR VIII (F VIII) AND FACTOR IX (F IX) TREATED WITH TRI (N-BUIYL) PHOSPHATE (TNBP) AND TWEEN 80 FOR INACTIVATION OF VIRUSES
Authors: Baumgartner C , Perret B A , Meili E , Furlan M , Friedli H , Morgen-thaler J J 
Published: June 2019

Heat treatment has been cxxnmonly used for the sterilisation of coagulation factor concentrates. This causes, however, considerable loss of coagulation factor activity? therefore alternative methods have been developed. Two new virus-inactivated coagulation factor preparations were recently introduced by our institution. Their manufacturing procedure includes a lipid solvent extraction step: The cryoprecipitate (F VIII preparation) or the first DEAE eluate (F IX preparation) is incubated with 0.3 % TNBP and 1 % Tveen 80 at 24°C for at least 12 hours. (Horowitz, Transfusion 25 : 516-522, 1985). Single doses of these preparations (F VIII: median 20.5, range 6-33 U/kg, F IX: median 25, range 9-46 U/kg body weight) were given intravenously to 28 hemophilic patients with minor or no bleeding. F VIII or F IX levels in plasma were determined before and at intervals up to 48 h after injection. The recovery was calculated from the maximum increase of activity and an plasma volume of 41 ml/kg body weight. The plasma half life was calculated according to the procedures described by Morfini (Thrcmb. Res. 42: I-III, 1986). Results are shown in the table below.No side effects were recorded. Hemostasis was satisfactory in all patients with bleedings (n = 13). These results are within the range expected for conventional and heat treated F VIII and F IX preparations. We conclude that the lipid solvent extraction procedure as used here does not influence the in vivo kinetics and the function of F VIII and F IX and does not induce any acute toxicity.
LOSS OF IMMUNE TO RECALL ANTIGENS IN THERE HIV+ HEMOPHILIC CHILDREN
Authors: GOMPERTS E D , WEINBERG K 
Published: June 2019

Three children with severe inherited bleeding disorders have been followed for a number of years at this center. One child (DOB 3/71) initially presented with mild hemophilia A, (Factor VIII 6%). He subsequently developed an inhibitor to Factor VIII (maximum 45 B. U.) and seroconverted to HIV+ Status 12/83. In 12/86 he had virtually lost his antibody response to infused Factor VIII (previously withheld), with a maximum increase in inhibitor titre to1 B. U. on challenge. In addition, his antitetanus antibody titre was very low at 0.01 u/ml earlier in theyear. His absolute T4 cell number at this time was very low at 64 and did not respond to skin antigen testing to PPD, tetanus and Candida.The second patient (severe hemophilia A DOB 7/76) had seroconvertedto HIV+ Status in 9/78. This child has lost his a-HBs seropositive status with an absolute T4 count of 239. His current anti-tetanus titre is 0.01 u/ml.The third patient (von Willebrand disease, Type III, DOB 7/74) seroconverted to HIV+E status by 5/83. His T4 absolute numbers have fallen to 53. His anti-tetanus antibody titre has fallen to extremely low levels (0.01 u/ml), and this failed to respond to re-immunization with tetanus toxoid. These three patients indicate that previously immunized children may lose their immune status and their ability to respond to recall antigens. It is pertinent to note that lymphocytes from all 3 patients failed to respond mitogenically in vitro to tetanus antigen pari passu with the observed very low anti-tetanus antibody titres. These phenomena would indicate that these patients are probably susceptible to previously preventable infectious agents including poliovirus, measles, mumps, rubella, diphtheria, tetanus and hepatitis B virus.
EXPERIMENTAL THROMBUS FORMATION AND HAEMOSTASIS OF DIFFERENT LOW MOLECULAR WEIGHT HEPARINS AND DOSAGES
Authors: Zimmerman R A , Rieger C T , Hübner K , Harenber C W , Kübler W 
Published: June 2019

Low molecular weight heparin induces a higher anti factor Xa (a-Xa) and a lower antithrombin activity in plasma in comparison to conventional heparin. From this constellation a more pronounced antithrombotic effect and a minor incidence of bleeding Complications has been suggested.Therefore the antithrombotic activity of heparins was studied in a standardized experimental thrombosis model in rabbits. Three low molecular weight heparins with a mean molecular weight of 4.200 (heparin I),4.000 (heparin II),4.600 Dalton (heparin III) and standard heparin were tested at different dosages in 120 experiments. In the first series the dose of 60 anti Xa units (a-Xa U) given initially and 60 a-Xa U/kg/h induced a reduction of the thrombus size by 40 % (heparin I),37 % (heparin II) and 53 % (heparin III) and a prolongation of the aPTT to 45 (heparin I),66 (heparin II) and 79 sec (heparin III). The a-Xa activity was minor than 0.1 U/ml. In the second series heparins were given to aim at an a-Xa activity of 0.2-0.3 U/ml. Thereby the thrombus formation could be reduced by 84 % (heparin I), 62 % (heparin II) and 39 % (heparin III). aPTT and a-Xa activity were measured at 65.5 sec and 0.22 a-Xa U/ml (heparin I),67.3 sec and 0.3 a-Xa U/ml (heparin II) and 67.5 and 0.31 a-Xa U/ml (heparin III),respectively. In the third series the increase of the a-Xa activity to more than 0.3 U/ml showed no further reduction of the thrombus formation by heparin I, while heparins II and III already at this level reachedthe antithrombotic activity of heparin I.Our data on three different low molecular weight heparins demonstrate that already a heparin level ranging at a minimal a-Xa activity induces a clear and statistically significant antithrombotic effect. A higher heparin dosage with higher a-Xa activity increases the antithrombitic effect. At a level of 0.2-0.3 a-Xa U/ml an obvious and maximum effect could be reached, but the further elevation of the a-Xa activity produced no further antithrombotic action.
HEPARIN POTENTIATES PLATELET ADHESION AND AGGREGATION ON COLLAGEN SUBSTRATES. ABSENCE OF INHIBITION BY ASPIRIN
Authors: Mazurov A V , Si-nitsin V E , Repin V S 
Published: June 2019

We studied the effects of two commercial heparin (H) preparations, from bovine lungs (HL) and pig intestine (HI), on the interaction of human plateletswith collagen substrates, and tested a possibility of modulating these effects by aspirin. PRP was incubated in the presence or absence of H with human collagens type IV and III (CIV and Cl I I) adsorbed ona plastic surface. Platelet adhesion and spreading on CIV and aggregation on Cl I I was quantitated by scanning electron microscopy. HL and HI were added to platelets in the concentrations approximating to those achieved in vivo at heparin therapy (0.5̲5.0 U/m 1) . HL and H|=I raised the number of adherent platelets, but do not alter the percentage of spread platelets on CIV. Both preparations had approximately equal effect, increasing platelet adhesion by 1.5 to2-fold at 2.5 U/ml . HL and HI potentiate the formation of. surface-bound aggregates on CIII, enlargingthe total substrate surface coated with aggregates. This effect is achieved due to increased size and number of aggregates. At 2.5 U/ml both preparations potentiate aggregation 2- to 3"fold.Aspirin has no effect on platelet adhesion and spreading both in the absence and presence of HL/HI. In the absence of H, aspirin decreased platelet aggregation on CIII, reducing the aggregate-coated area by 40-50%. The inhibition was related mainly to reducednumber, not the size of aggregates. Aspirin failed to prevent potentiation of the Clll-induced aggregation by HL/HI.
IN VITRO DEMONSTRATION OF "HEPARIN REBOUND"
Authors: Taylor C , Baugh R F 
Published: June 2019

"Heparin rebound", the in vivo appearance of measurable heparin anticoagulant activity following theapparent neutralization of heparin by protamine, hasbeen a problem sporadically associated with the use of heparin in cardiovascular surgery. A number of mechanisms have been proposed to explain rebound, and to some extent each may contribute to the phenomena. As yet no reliable, predictable method has been demonstrated for measuring, reproducing or quantifying "heparin rebound".We have demonstrated and measured the appearance of heparin anticoagulant activity following neutralization with protamine in citrated whole blood. The reappearance of heparin anticoagulant activity was associated with a rapid loss of protamine. The loss of protamine followed 1st order enzyme kinetics, and was indicative of the action of an enzyme. The anticoagulant activity which eappeared could be titrated againwith protamine. The loss of protamine neutralizing activity, in whole blood, could be followed by titration with heparin using a recalcified activated clotting time. The rate of loss varied with both individual blood donors and with the type and source of protamine. The rate of loss of protamine was great enough to influence in. vivo heparin/protamine neutralization ratios, i.e. at 4 units of heparin/ml, 1 unit/ml anticoagulant activity was routinely recovered within 30 minutes following initial neutralization. The indications for cardiovascular surgery are:1)the in vivo neutralization ratio should be adjusted to account for loss of protamine activity, 2) the higherthe blood levels of heparin used during surgery, themore significant the potential for heparin rebound, and 3) protamines may be evaluated in an in vitro test which measures the stability of protamine neutralizing activity in whole blood.
SAFETY AND EFFICACY OF SERIAL IMPEDANCE PLETHYSMOGRAPHY IN THE DIAGNOSIS OF DEEP VEIN THROMBOSIS IN AN URBAN HOSPITAL: AN EXTERNAL VALIDATION STUDY
Authors: Heyermans H H S , Huisman M V , Büller H R , Laan J V D , ten Cate J W 
Published: June 2019

The clinical diagnosis of deep vein thrombosis (DVT) is unreliable. Impedance plethysmography (IPG) has become accepted as a highly reliable non-invasive method for DVT detection in symptomatic patients.It has a high sensitivity (95%) and specificity (96%) for proximal vein thrombosis. Studies with impedance plethysmography have however always been carried out in academic hospitals. To evaluate the safety and efficacy of serial impedance plethysmography alone in an urban hospital setting, a prospective study was done in an urban hospital, involving 234 consecutive outpatients with clinically suspected venous thrombosis. IPG was performed on days 1, 2 and 7. If all tesis remained normal the patient was not treated with oral anticoagulants. All patients were followed for 3 months. In 131 of the 234 patients (56%) IPG was repeatedly normal. Of these 131 patients, no patient died from venous thromboembolism during 3 months follow-up, completed in all patients, and no patient returned with signs of pulmonary embolism. One patient (0.8%) returned with objectively documented DVT after two months. In 103 of the 234 patients (44%) the IPG was abnormal. Venography confirmed the diagnosis of deep venous thrombosis in 92%.It is concluded that serial IPG is a safe and effective method to detect DVT in clinically suspected outpatients, referred to an urban hospital.
COMPUTERGRAPHIC ANALYSIS OF OBSTETRIC HEMORRHAGE
Authors: Suzuki S , Ichinoe K 
Published: June 2019

(Purpose) From the viewpoint of maternal and perinatal mortality, it is very important to manage the postpartum hemorrhage (PPH). In order to observe the riskfactors, especially the relationship between the obstetric factors, abnormal bleeding of PPH,. and blood -coagulation-system, we have tried to prognosticate the abnormal bleeding during labor.Next, the relationship of the Hepaplastintest-value of umbilical venous blood and obstetric factors were examined by computer graphic analysis.(Methods) 200 cases of non-DIC and 17 cases of DIC cases, Fibrinogen, FDP, Platelet, Factor XI and Factor XII et were tested in the 1st stage of delivery.(Results)
(1) In the non-DIC cases no relation was proved between bloodcoag-ulation factors excluding Factor XI.(2) The correlation-Index in the DIC cases, we have proved the following order.Platelet (75 %) > Fibrinogen (65 %) > FDP (55 %)(Conclusion)
(1) Even in normal delivery, the Kinin-Kallikrein system plays an important role in the contraction of the uterus, and the duration-of delivery.(2) In the DIC-cases, the platelet count was reversely related and showed a linear relation with Postpartum-Hemorrhage. We also noted that Fibrinogen, FDP and Prothrombin Time are important factors from the viewpoint of DIC.
EFFECT OF ORAL CONTRACEPTIVES ON HEMOSTASIS
Authors: Farag A M , Bottoms S F , Mammen E F , Hosni M , Ali A 
Published: June 2019

Retrospective statistical epidemiological studies have suggested a possible association between the ingestion of oral contraceptives (OC) and thromboembolic disease. Past analyses of the coagulation system have yielded controversial informationWe studied a cross section of 131 women taking different kinds of OC and 36 controls for changes in hemostasis. No significant differences were noted in the levels of fibrino- peptide A (RIA), platelet factor 4, 8 thromboglobulin (RIA), fibrinogen (Multistat III (MCA), clottable), antithrombin III (MCA, S-2238), α2 antiplasmin (MCA, S-2251), pre-kallikrein (MCA, S-2302) and fibronectin (MCA, immune turbi-dometric). However, plasminogen (MCA, S-2251) and protein C antigen (Laurell) levels were significantly elevated (p < 0.001 and p < 0.01), respectively)Canonical correlation analysis was used to examine correlations between hemostasis parameters measured and clinical risk factors, such as age, parity, weight, smoking, family history for thromboembolic diseases and estrogen-progesterone dose. There was a significantly negative correlation between family history for thromboembolisms and antithrombin III levels (p < 0.01). A positive correlation existed between obesity and fibrinogen and fibronectin levels (p < 0.001 for both). The hemostasis data seem to suggest that OC use does not introduce an imbalance in the hemostasis system which fosters "hypercoagulability", and that, if at all, possibly other risk factors determine the incidence of thromboembolisms in OC users. It is suggested that caution be exercised in the use of OCs in patients with a history of thromboembolic diseases and with obesity
EDTA-DEPENDENT MONOCLONAL ANTIBODIES TO HUMAN PROTEIN S
Authors: Marcovina S , Coppola R , Protti M P , Gelfi C , Mannucci P M 
Published: June 2019

Splenocytes from a Balb/c mouse immunized with purified human protein S (PS) were fused with murine hybridoma cell line SP2/0-Agl4 and cultured in Iscove's medium without addition of fetal bovine serum. Hybrid supernatants were screened for the presence of specific antibodies by solid-phase ELISA and cloned by the limiting dilution technique. Pour clones, named S2, S3, S8, and S10, were selected, recloned several times, and injected intraperitoneally into Balb/c mice for the production of antibody-rich ascitic fluid. The monoclonal antibodies (Mabs), all of IgGl subclass with k light chain, were purified from ascitic fluid by Protein-A chromatography. The specificity of Mabs was controlled by the immunoblotting technique: the Mabs appeared to react only with two plasma proteins, one with a MW of about 70.000 dal tons comigrating with purified PS, and the other with a MW >300.000 da that is likely to be the C4b-binding protein-PS complex. No interaction has been observed with PS-depleted plasma. As tested by a fluid phase radio immunoassay, the binding of Mabs to PS was significantly higher in the presence of EDTA while was totally inhibited in the presence of Ca2+. Scatchard plot analysis of the binding between 125I-PS and Mabs showed that the binding affinity of the antibodies ranged from 108 to 109 1/mol. Each EDTA-dependent Mab was then immobilized on Sepharose 4B-CNBr and used to purify PS from barium precipitation of citrated plasma. The fraction eluted with 2 mmol of CaCl2 from the immunoadsorbent appeared to contain only two proteins when stained with Cocmassie Blue. By immuno blotting, all Mabs reacted with both proteins, one comigrating with purified PS and the other with a MW >300.000. Essentially homogeneous PS, free from the high MW component, was obtained when the barium citrate adsorbate was applied to a DEAE-Sephadex column and the protein peack containing the balk of PS was sussequently applied to the immunoadsorbent and eluted with 2 mmol CaCl2.In summary, we have described an unusual set of EDTA-dependent monoclonal antibodies to PS and their use for the purification of homogeneous protein S from human plasma.
SUDAY ON A SUBSTANCE FUNCTIONALLY LIKE PROTEIN C IN PLASMA
Authors: TSUNEIZUMI I , MEGURO T , YAMADA K 
Published: June 2019

The present report first deals with the isolation and characterization of a substance found to demonstrate protein C like activity(PCLA). When it was activated by thrombin, the PCLA substance prolonged the activated partial thromboplastin time(aPTT). The activated PCLA substance also hydrolyzed synthetic substrates such as S�2238, S�2366 and S�2266,while the PCLA substance was not cross reacted with anti-protein C serum(Behring Manheim). The PCLA substance was adsorbed by both aluminium hydroxide gel and barium sulfate. The level of Km, optimum pH and optimum I.S. in the amido-lytic reaction with synthetic substrate is shown in the table.In the chromatography of Sepharose CL-6B gel, the PCLA substance was eluted in the fraction of higher molecular weight, while protein C was eluted with a lower fraction.This prompts an estimate that the molecular weight of the PCLA substance is approximately 200,000. Through DEAE-Sepharose CL-6B gel ion exchange chromatography, the PCLA substance was eluted by a lower ionic strength than was protein C.The levels of a PCLA substance were low in the patients with a vitamin K deficiency and in newborn infants. It is expected that our findings regarding the PCLA substance will be as clinically significant as that which we know about protein C at present.
ASSOCIATION OF HEREDITARY DYSFIBRINOGENEMIA WITH PROTEIN C DEFICIENCY IN TWO PATIENTS WITH THROMBOTIC TENDENCY
Authors: Gandrille S , Priollet P , Capron L , Roncato M , Fiessinger J N , Aiach M 
Published: June 2019

An abnormal fibrinogen was found to be associated with protein C deficiency in two unrelated patients. Both were symptomatic, one having severe complicated atherosclerosis, the other recurrent venous thrombosis. In both cases, the two abnormalities coexisted in several members of the family. The two abnormal fibrinogens (Poitiers and Argenteuil) were purified from the patients' plasma and compared to fibrinogens purified from several normal subjects. Polymerisation was abnormal in both cases in presence of reptilase and thrombin.The kinetic of proteolysis by plasmin was studied during 24 hours using SDS-polyacrylamide gel electrophoresis. In the first case, the appearance of fibrinogen degradation products (FDPs) was slightly delayed. The most sliking abnormalities were observed with fibrinogen Argenteuil which still contains "early" FDPs (X and Y) at 24 h while the normal fibrinogens were all completely degraded.The stimulating effect of fibrin on plasminogen activation by tPA was studied after insolubilization of the purified fibrinogen according to the SOFIA technique described by Angles-Cano (Anal. Biochem. 1985, 153, 201). Only fibrinogen Argenteuil was found to have a decreased catalytic effect on plasmin formation.Thrombin binding by fibrin clots, studied according to Ha-verkate (Thromb. Haemostas. 1986, 55, 131) was found decreased in fibrinogen Poitiers and normal in fibrinogen Argenteuil.Conclusion: In these two cases of hereditary dysfibrinoge-nemia least one of the natural antithrombotic function elicited by fibrinogen-fibrin transformation was found to be strongly abnormal . Both fibrinogens presented abnormal lysis by plasmin, particularly fibrinogen Argenteuil in which Tibnn degradation was also reduced. This functional abnormality have already been described in patients with thrombotic tendency. It is however difficult to conclude, as in both families the dysfibrinogenemia was associated with protein C deficiency.