bluuebunny/crossref_metadata_2025_split huggingface dataset:
Various Prothrombin Complex Concentrates and their Effect on Coagulation and Fibrinolysis In Vivo
Authors: Hedner U , Nilsson I.M , Bergentz S.-E 
Published: July 2018

SummaryThromboembolic complications occurring in patients treated with factor IX concentrates have been reported. To study the thrombogenicity various types of factor IX concentrates (50 or 100 units F. IX/kg bodyweight) have been infused in dogs. As control albumin was given. The various components of the coagulation and fibrinolytic system have been assayed before the infusion and at various intervals after the end of infusion (0, 1,4 and 24 hrs). Konyne resulted in marked activation of the coagulation process with decrease of platelets, fibrinogen, F. VIII and appearance of FDP and positive ethanol gelation test. Pro thro mplex and Pre-conativ gave no significant changes. Preconativ is prepared without addition of heparin during the procedure.
Purification of Human Factor IX by Chromatography of a Coagulation Factor Concentrate
Authors: Suomela Hannu 
Published: July 2018

SummaryA purification procedure for, and some properties of, coagulation factor IX are described. The coagulation factor concentrate used for the treatment of hemophilia B patients was employed as the starting material. The isolation procedure consists of chromatography in DEAE-cellulose, two chromatographies in hydroxyapatite gel and two gel filtrations in Sephadex G-200. Only trace amounts of factors II, VII and X were present in the final preparation and the specific activity of factor IX was 159 corresponding 10,300 times purification from plasma. The molecular weight was estimated to be 76,000 in gel filtration and 86,000 in sodium dodecyl sulfate disc gel electrophoresis. Three activity peaks with pis 4.15, 4.25 and 4.40 were obtained by isoelectric focusing.
Oxygen Consumption in Platelets of Newborn Infants before and after Stimulation by Thrombin.
Authors: Principe D. Del , Mancuso G , Menichelli A , Maretto G , Sabetta G 
Published: July 2018

SummaryThe authors compared the oxygen consumption in platelets from the umbilical cord blood of 36 healthy newborn infants with that of 27 adult subjects, before and after thrombin addition (1.67 U/ml). Oxygen consumption at rest was 6 mμmol/109/min in adult control platelets and 5.26 in newborn infants. The burst in oxygen consumption after thrombin addition was 26.30 mμmol/109/min in adults and 24.90 in infants. Dinitrophenol did not inhibit the burst of O2 consumption in platelets in 8 out of 10 newborn infants, while the same concentration caused a decrease in 9 out of 10 adult subjects. Deoxyglucose inhibited the burst in O2 consumption in newborn infant and adult platelets by about 50%. KCN at the concentration of 10−4 M completely inhibited basal oxygen consumption but did not completely inhibit the burst after thrombin. At the concentration of 10−3 M, it inhibited both basal O2 consumption and the burst in infants and adult subjects.
Aggregation of Platelets and Inert Particles Induced by Thrombin
Authors: Chao Frangis C. , Tullis James L. , Conneely Gail S. , Lawler John W. 
Published: July 2018

SummaryThrombin-induced platelet aggregation and release were investigated in washed platelet suspensions and in suspensions of inert particles in order to evaluate the role of fibrinogen-fi-brin transformation in aggregometer tracings. Thrombin (0.25–2.0 U/ml) produced two waves of light transmission increase (LTI) in both platelet and inert particle suspensions containing fibrinogen, and concomittantly aggregates were observed under phase microscopy. Without fibrinogen, thrombin induced rapid release of platelet ADP but failed to cause second wave of LTI. The kinetics of LTI in platelet and inert particle systems were related to both thrombin and fibrinogen concentrations. A rapid second wave of LTI could be produced by direct interaction of thrombin-treated platelets or inert particles with polymerizing fibrin, and was inhibited by sodium sulfite and low pH of 5.1 which prevent fibrin monomer polymerization. No fibrin strands were noted in platelet aggregates fixed at the completion of the second wave of LTI. Apyrase and PGE1 inhibited the rate of first but not that of second wave LTI. The results suggest that the release of platelet ADP induced by thrombin primarily affects the first phase aggregation, and the second phase may result from interaction of thrombin-exposed platelets and polymerizing fibrin. Thus, the blood coagulation mechanism may be directly involved in platelet aggregation.
Lack of Agreement of Chromogenic and Glotting Assays for Factor X and Protein C in Warfarinised Plasma
Authors: Han P , Fung K P , Rahdakrishnan U 
Published: July 2018

SummaryCoagulation serine proteases can be measured with either a chromogenic substrate assay or a clotting assay using deficient plasmas. It is a concern whether both assays give similar quantitative results, in particular in plasma obtained fiom patients on long term warfarin therapy. If these two assay methods were interchangeable, then the chromogenic substrate assay has the advantages of precision as well as laboratory automation. We used the intraclass correlation coefficient (r1) to assess the agreement between the two methods in measuring factor X and protein C levels in warfarinised plasma. The results indicate that the extent and pattern of agreement of the two methods for the measurement of the two variables in warfarinised plasma are poor, despite high Pearson product moment coefficients of correlation.
Plasma Level of IL-1β in Disseminated Intravascular Goagulation
Authors: Wada Hideo , Tamaki Shigehisa , Tanigawa Motoaki , Takagi Mikio , Mori Yoshitaka , Deguchi Akira , Katayama Naoyuki , Yamamoto Toshihiko , Deguchi Katsumi , Shirakawa Shigeru 
Published: July 2018

SummaryThe plasma level of interleukin-1β (IL-1β) was determined in normal individuals, patients with disseminated intravascular coagulation (DIC), patients in the pre-DIC period (within 7 days before the onset of DIC), and non-DIC patients to examine the relationship between DIC and the plasma ILlp level. The plasma IL-1β level was 0-0.085 ng/ml in normal individuals, with little difference being seen according to related age. It was significantly higher in the DIC group (0.19 ± 0.19 ng/ml) than in the pre-DIC group (0.05 ± 0.08 ng/ml) or the non-DIC group (0.09 ± 0.01 ng/ml). The plasma IL-1β level was not markedly elevated in leukemia patients, even in the DIC group, but it was significantly increased in the DIC group of solid cancer patients and was generally elevated in patients with sepsis. It was markedly elevated to 0.39 ± 0.26 ng/ml in patients with organ failure. When mononuclear cells were incubated with lipopolysaccharide, it was found that IL-1β, tumor necrosis factor, and tissue factor (TF) were released into the medium, and there was an increase of TF release from endothelial cells incubated with this medium. These results suggest that the increase in IL-Iβ reflected the activation of monocytes and may be an important factor in DIC and its associated organ failure.
Western Blot Analyses of Prekallikrein and its Activation Products in Human Plasma
Authors: Veloso Dulce , Colman Robert W 
Published: July 2018

SummaryPrekallikrein (PK), a zymogen of the contact system, and its activation products, kallikrein (KAL), KAl-inhibitor complexes and fragments containing KAL epitope(s) have been detected in human plasma by immunoblotting with a monoclonal anti-human plasma PK antibody, MAb 13G1L. Detection of antigen-MAb 13G11 complexes with peroxidase-conjugated anti-IgG showed that the two variants of PK (85- and 88-kDa) are the only major antigen species in normal, non-activated plasma. Upon plasma activation with kaolin, the intensity of the PK bands decreased with formation of complexes of KAL with CL inhibitor (C1 INH) and α2-rrtzcroglobulin (α2M) identical to those formed by the purified proteins. Immunoblots of normal plasma showed good correlation between the PK detected and the amount of plasma assayed. Increasing amounts of KAL incubated with a constant volume of PK-deficient plasma showed increasing amounts of KAL and of KAL-C1 INH and KAL-α2M complexes. Complexes of KALantithrombin III (ATIII) and the ratio of KALα2M/ KAL-CL INH were higher in activated CL INH-deficient plasmas than in activated normal plasmas. Protein resolution by 3-12% gradient SDS-PAGE and epitope detection with [125I]MAb 13G11 showed four KALα2M species and a 45-kDa fragment(s) in both surface-activated normal plasma and complexes formed by purified KAL and α2M. Immunoblots of activated plasma also showed bands at the position of KALCL INH and KALATIII complexes. When α1-antitrypsin Pittsburgh (cα1-AT, Pitts) was added to plasma before activation, KAL-α1-ALPitts was the main complex. The non-activated normal plasma revealed only an overloaded PK band. This is the first report of an antibody that recognizes KAL epitope(s) in KAL-α2M, KALATIII and KALa1-α1Pitts complexes and in the 45-kDa fragment(s). Therefore, MAb 13G11 should be useful for studying the structure of these complexes as well as the mechanism of complex formation. In addition, immunoblotting with MAb 13G11 would allow detection of KAl-inhibitor complexes in patient plasmas as indicators of activation of the contact system.
Effect of Plasminogen Activator Inhibitor-1 on Tissue-Type Plasminogen Activator-Induced Fibrinolysis
Authors: Carr Marcus E , Krishnamurti C , Alving B M 
Published: July 2018

SummaryThe effect of fibrin on the interaction of human recombinant single-chain tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) was studied in normal rabbit plasma and in plasma with high levels of native PAI-1. t-PA was added to diluted plasma containing calcium (10 mM) and 125I-fibrinogen at 37° C. Clotting was initiated with human thrombin, and lysis was monitored both turbidimetrically and by release of 125I-fibrin degradation products (fdp). The activity of t-PA (50 IU/ml) was rapidly reduced to 15% of the initial value in plasma containing PAI-1 (23 AU/ml). When thrombin and t-PA were added simultaneously to the plasma, more than 70% of the activity was retained through incorporation of t-PA into the fibrin clot. t-PA-induced fibrinolysis in PAI-1 enriched plasma was further delayed when the temperature was reduced from 37 to 25° C. Turbidimet-ric and 125I-fdp release data provided complementary information. The former technique traced fiber dissolution, while the latter reflected network integrity. These results indicate that t-PA-induced fibrinolysis in PAI-1 enriched plasma is modulated by the presence of fibrin and by temperature.
No Evidence for Short-Term Regulation of Plasminogen Activator Inhibitor Activity by Insulin in Man
Authors: Vuorinen-Markkola Helena , Puhakainen llpo , Yki-Järvinen Hannele 
Published: July 2018

SummaryIn crossectional studies a positive correlation has been found between circulating insulin, triglycerides and plasminogen activator inhibitor (PAI-1) activity. To directly examine the effect of insulin on PAI-1 activity in vivo, we determined the response of PAI-1 activity in 17 normal subjects to acute hyperinsulinemia (serum free insulin 92 ± 8 mU/l) during maintenance of normoglycemia (plasma glucose 5.1 ± 0.1 mmol/l). In 12 matched control subjects PAI-1 activity was measured during infusion of saline (serum free insulin 3.6 ± 0.3 mU/l, plasma glucose 5.2 ± 0.1 mmol/l). Plasma PAI-1 activity decreased during the insulin infusion from 9.0 α 1.4 to 5.6 α 0.8 U/ml (p <0.01), and during saline infusion from 7.0 ± 1.4 to 4.3 ± 0.6 U/ml (p <0.05). Serum triglyceride concentrations decreased from 1.09 ± 0.20 to 0.76 ± 0.09 mmol/l (p < 0.001) during hyperinsulinemia but remained unchanged during the saline infusion (1.04 ± 0.11 vs. 1.02 ± 0.12 mmol/l, NS). We conclude that insulin does not acutely change plasma PAI-1 activity, and that acute insulin-induced changes in serum triglycerides occur independently from those of PAI-1 activity.
The Venous Antithrombotic Effect of LF 1351 in the Rat following Oral Administration
Authors: Millet Jean , Theveniaux Jocelyne , Brown Neil L 
Published: July 2018

SummaryThe venous antithrombotic effects of a novel chemical entity, LF 1351, were investigated in rats following single oral administration, in comparison with i.v. administered heparin. LF 1351 demonstrated a dose-related antithrombotic effect in three models of venous thrombosis. The compound was approximately equipotent in two models involving complete stasis of the vena cava and administration of factor Xa or porcine serum, giving respective ED50 values of 48.7 mg/kg and 36.7 mg/kg. LF 1351 was less effective in a model involving partial stasis in the presence of an endothelial lesion. In this case, the antithrombotic effect did not exceed 60-65%, the ED50 being 150 mg/kg. Heparin (50-300 μg/kg; 7.5-45.0 U/kg) was effective in all three models. At the approximate ED80 value against factor Xa-induced thrombosis (75 mg/kg) the antithrombotic effect of LF 1351 persisted for 6 h. The antithrombotic effect of LF 1351 (300 mg/kg) occurred without significant changes in APTT or thrombin time.
The Predictive Value of the Hemostasis Parameters in the Development of Preeclampsia
Authors: Ho Chao-Hung , Yang Zwa-Ling 
Published: July 2018

SummaryIn order to find out which hemostasis parameters would have the predictive value for the development of preeclampsia, modified antithrombin III (ATM, representative of the antithrombin Ill-serine esterase complex), tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), beta-throm-boglobulin (BTG), antithrombin III (AT III), fibrinogen, fibrin(ogen) degradation product (FDP), FDP D-dimer and euglobulin lysis time (ELT) were measured in 20 normal nonpregnant women, 21 normal pregnant women, 6 high-risk pregnant women, 14 preeclampsia pregnant women, and 5 patients with disseminated intravascular coagulation (DIC). Only tPA and AT III were found significantly different between the preeclampsia and the normal or high-risk pregnant women: tPA was found progressively and significantly increased from the normal pregnant, to the high-risk pregnant, then to the preeclampsia women (p <0.05). AT III was significantly lower in the preeclampsia than in the normal pregnant (p = 0.0001) or in the high-risk pregnant women (p = 0.002). In the 2nd trimester, tPA, PAI, fibrinogen and FDP were significantly higher, and AT III was significantly lower in the preeclampsia than in the normal pregnant women, whereas in the 3rd trimester, tPA and AT III were significantly higher or lower, respectively, in the preeclampsia than in the normal pregnant women. No significant difference of ATM could be found between the preeclampsia and the normal or high-risk pregnant women. From the present study, we suggest that tPA and AT III would be used as the main predictors, and FDP and D-dimer as the complementary predictors for the development of preeclampsia and should be detected in the normal or high-risk pregnant women.
Biochemical and Biological Properties of a Recombinant Tissue-Type Plasminogen Activator Derived from the Rat JMI-229 Cell Line
Authors: Lijnen H R , Webb P D , Van Hoef B , Cock F De , Stassen J M , Prior S D , Collen D 
Published: July 2018

SummaryRecombinant tissue-type plasminogen activator (rt-PA), produced by expression of the genomic t-PA DNA from the JMI-229 cell line, which is of rat origin, in the host cell line, was purified to homogeneity. JMI-229 rt-PA was obtained essentially as a single chain molecule which was quantitatively converted to a two-chain moiety by treatment with plasmin. The plasminogen activating potential of single chain JMI-229 rt-PA was 5-fold lower than that of commercially available human rt-PA (Actilyse®) in the absence of fibrin, but comparable in the presence of fibrin; it showed a concentration-dependent binding to fibrin, with a significantly more pronounced binding than Actilyse® at low fibrin concentration (85 ± 8% versus 20 ± 7% at 0.025 mg/ml fibrin; p = 0.004). In human plasma in the absence of fibrin, the concentrations of both single chain and two-chain JMI-229 rt-PA required to induce 50% fibrinogen degradation in 2 h, were about 15-fold higher than those of Actilyse®. Both single chain and two-chain forms of JMI-229 rt-PA and of Actilyse® induced a similar time- and concentration-dependent lysis of a 125I-fibrin-labeled plasma clot immersed in human plasma, in the absence of significant systemic fibrinolytic activation. Equally effective concentrations (causing 50% clot lysis in 2 h) were 0.11 or 0.10 pg/ml for single chain or two-chain JMI-229 rt-PA, as compared to 0.11 or 0.15 pg/ml for single chain or two-chain Actilyse®. Continuous infusion over 60 min of single chain JMI-229 rt-PA or Actilyse® in hamsters with a 125I-fibrin-labeled pulmonary embolus, revealed a very similar thrombolytic potency (clot lysis versus dose) and specific thrombolytic activity (clot lysis versus steady state plasma antigen level of t-PA). The initial plasma half-life following intravenous bolus injection of 0.10 mg/kg in hamsters was equally short for JMI-229 rt-PA or Actilyse® (1.2 or 1.4 min respectively).It is concluded that JMI-229 rt-PA has a higher fibrin-affinity and a higher fibrin-specificity in human plasma in the absence of fibrin than Actilyse®, but a comparable thrombolytic potency in a hamster pulmonary embolism model.
Binding of Human Platelet Glycoprotein lb and Actin to Fragments of Actin-Binding Protein
Authors: Aakhus Anne M , Wilkinson J Michael , Solum Nils Olav 
Published: July 2018

SummaryActin-binding protein (ABP) is degraded into fragments of 190 and 90 kDa by calpain. A monoclonal antibody (MAb TI10) against the 90 kDa fragment of ABP coprecipitated with the glycoprotein lb (GP lb) peak observed on crossed immunoelectrophoresis of Triton X-100 extracts of platelets prepared without calpain inhibitors. MAb PM6/317 against the 190 kDa fragment was not coprecipitated with the GP lb peak under such conditions. The 90 kDa fragment was adsorbed on protein A agarose from extracts that had been preincubated with antibodies to GP lb. This supports the idea that the GP Ib-ABP interaction resides in the 90 kDa region of ABP. GP lb was sedimented with the Triton-insoluble actin filaments in trace amounts only, and only after high speed centrifugation (100,000 × g, 3 h). Both the 190 kDa and the 90 kDa fragments of ABP were sedimented with the Triton-insoluble actin filaments.
The Effects of Biguanides on Thrombokinase, Thrombin and Trypsin
Authors: Roberts Phyllis S , Fleming Patricia B 
Published: July 2018

SummaryA purified preparation of bovine thrombokinase (activated Factor X) loses the ability to hydrolyze TAME (p-toluenesulfonyl-L-arginine methyl ester) when it is incubated at 37° in 0.25 M Tris. HCl buffer, pH 7.4 with lauroxypropyl biguanide, N1, N5-dimethyl, N1-lauroxypropyl biguanide, N1-p-chlorophenethyl, N5-phenethyl biguanide, or N1-methyl, N1-p-chlorobenzyl, N5-o,p-dichlorobenzyl biguanide. Activity is lost much more slowly when 0.15 M NaCl is also present. Lauroxypropyl biguanide is the most potent of the compounds tested, 0.22 mM causing thrombokinase to lose almost all of its activity in about 30 minutes at 37° in pH 7.4 buffered saline.Topical bovine thrombin also loses activity when incubated with either of the lauroxypropyl biguanides but not with the diphenethyl or the dibenzyl compound. Instead, the latter biguanides accelerate thrombin’s hydrolysis of TAME. The percent acceleration is not affected or only slightly decreased by the presence of 0.15 M NaCl or KCl, and it is also unaffected by incubating the enzyme with the compounds in buffered saline for 4 to 120 minutes.Purified bovine trypsin is stabilized by both lauroxypropyl and the diphenethyl biguanide when incubated at 37° in pH 7.4 buffered saline for the 60 minute test period but neither compound has any effect on its rate of hydrolysis of TAME.It is postulated that the enzymes first react rapidly and reversibly with all of the test biguanides and, depending upon the enzyme and the substrate, the rate of hydrolysis of the substrate is unaffected, accelerated or inhibited. The lauroxypropyl biguanides also undergo a second, slower reaction with both thrombokinase and thrombin that produces loss of enzymatic activity. The dibenzyl and diphenethyl biguanides also undergo this second slow reaction with thrombokinase but not with thrombin, and none of the biguanides undergo this second reaction with trypsin.
The Influence of Beta-adrenergic Blockade upon Baseline Blood Coagulation and Fibrinolytic Activity and upon the Responses to Venous Occlusion
Authors: Butler M. J , Smith Muriel , Irving M. H , Gordon Y. B , Ratky S. M , Rivers J. W. P , Hawkey C 
Published: July 2018

SummaryTwenty volunteers were assessed for baseline coagulation (Thrombelastography, Factor VIII and platelet count) and fibrinolytic (Euglobulin Lysis Time and Fibrin(ogen) Degradation Products) activity two hours after ingestion of 60 mgm propranolol or an identical placebo administered on a double blind basis. The responses of these parameters to a fifteen minute period of venous occlusion was also assessed.Beta adrenergic blockade reduced baseline FDP titres but did not affect other parameters, nor the local response to venous occlusion. During occlusion no significant change in coagulation activity could be detected in the non-occluded arm, but a small but statistically significant shortening of ELT was observed. Since this effect was prevented by propranolol it is probably mediated through adrenergic mechanisms.Adrenergic stimuli appear to influence episodes of coagulation and fibrinolysis during everyday activity, but do not contribute to baseline levels of coagulation or fibrinolytic function.
Purification and Some Properties of Urokinase
Authors: Ogawa Nobuhisa , Yamamoto Hiroshi , Katamine Tomoaki , Tajima Hideo 
Published: July 2018

SummaryA method for efficient extraction of urokinase from human urine was established by using polyacrylonitrile synthetic fiber as an adsorbent. By a combination of this method and known methods for purification of proteins, such as gel filtration and ion-exchange chromatography, urokinase with a specific activity of 224,000 International Units per mg of protein was obtained.This sample showed homogeneity by ultracentrifugation, moving-boundary electrophoresis at pH 4.8 and 9.0 and Polyacrylamide gel disc electrophoresis at pH 4.0, but was separated into five active fractions by isoelectric focusing and Polyacrylamide gel disc electrophoresis at pH 9.4. This sample showed a single precipitin line in double radial immunodiffusion and Immunoelectrophoresis using rabbit anti-urokinase serum. This precipitin line fused with that of the International Standard preparation of urokinase and its immunological identity was established.The molecular weight of this sample was 33,000, agreeing with that of the International Standard preparation. Its optimal pH as a plasminogen activator was approximately 8.8.