bluuebunny/crossref_metadata_2025_split huggingface dataset:
DOSE-FINDING STUDY OF THROMBOLYSIS IN ACUTE MYOCARDIAL INFARCTION BY SINGLE-CHAIN PROUROKINASE
Authors: Diefenbach C , Erbel R , Meyer J , Hamm C W , Schofer J , Mathey D G 
Published: August 2018

In 24 patients (pts)(mean age 54.2 ± 12.1 years) with acute myocardial infarction (onset of symptoms <6 hrs; mean duration 181.8±54.4 minutes) single-chain prourokinase (PUK) was applied intravenously. PUK is an inactive proenzyme of human urokinase isolated from recombinant Escherichia coli. Efficiency and safety of low and high dose application were analysed. Complete coronary artery occlusion was proved before start of thrombolysis therapy. After 5000 IU heparine intravenously the first 12 pts were given 10 mg as a bolus followed by 30 mg within 1 hour (group A), the next 12 pts received the double dose: 20 mg as a bolus and 60 mg within 1 hour (group B). After PUK infusion 20 IU/ kg heparine were administered continously in all pts for 72 hours. There were no significant differences in age, bodyweight and duration of symptoms in both groups In group A in 6 of 12 pts (50%) reperfusion of the occluded coronary artery was achieved within 58.3±17.2 minutes; in group B 9 of 12 (75%) vessels reopened within 44.0±29.9 minutes. The remaining stenosis of coronary artery was 91% in both groups. In group A no drop of fibrinogen or plasminogen was observed; only 2 pts of group B showed significant drops of fibrinogen (below 100 mg%) and plasminogen (below 40%) plasma level. In no patient major bleeding or any other acute adverse effect occurred.Thus, intravenous application of 80 mg PUK is more effective than 40 mg PUK with low incidence of systemic side effects.
CEREBRAL THROMBOLYSIS WITH INTRAVENOUSLY ADMINISTERED RECOMBINANT LOW- MOLECULAR-WEIGHT-UROKINASE AND RECOMBINANT PRO-UROKINASE IN A DOG MODEL
Authors: Hirschberg M , Manoutchei A , Klemens B , Hofferberth B 
Published: August 2018

There is increasing evidence that recombinant prourokinase (rec-pro-UK) is a proenzyme which in vivo systems may induce activation of the fibrinolytic system with a better thrombus selectivity than that obtained with active urokinase.In order to study the effects of rec-pro-UK and low-molecular-weight-urokinase (LMW-UK) on acute stroke, a thrombus was induced in the middle cerebral artery (MCA) of anesthetized mongrel dogs (n=12). Occlusion of the vessel was confirmed by angiography. Following a 1 hour period of MCA occlusion, in six animals LMW-UK was administered intravenously at a dose of 4000 lU/kg/min. Angiographically confirmed thrombolysis occurred after 30 minutes. Thrombolysis by LMW-UK was accompanied by bleeding from all surgical wounds and consumption of plasminogen, alpha-2-antiplasmin and fibrinogen. Rec-pro-UK was administered to six other dogs in a LMW-UK-equivalent dosis. Thrombolysis was achieved after 30 minutes in all six cases without inducing a systemic lytic state. Neither in the LMW-UK-group nor in the group treated with rec-pro-UK intracerebral bleeding complications were observed on post mortem examination.Our findings indicate that intravenous administration of rec-pro-UK - because of the lack of systemic side-effects - may be a safe way of rapid thrombolysis of occluded cerebral arteries in acute stroke.
INTRAVENOUS THROMBOLYTIC THERAPY WITH SINGLE-CHAIN UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN PATIENTS WITH ACUTE MYOCARDIAL INFARCTION
Authors: Bode C , Schwarz F , Schuler G , Zimmermann R , Kubler W 
Published: August 2018

Seventeen patients with acute myocardial infarction were treated with heparin combined with intravenous single-chain urokinase-type plasminogen activator (scu-PA),obtained from transformed human kidney cells. 4 patients in group I and 13 patients in group II received a bolus of 4 mg and 7-5 mg followed by an infusion of 11 mg and 40.5 mg over 60 minutes, respectively. Thrombolysis was achieved in no patient of group I during intravenous infusion of scu-PA, however, upon subsequent intracoronary infusion of 250000 IE streptokinase, reperfusion could be established in two out of three patients. In group II seven patients were successfully treated with intravenous infusion of scu-PA. In 2 of the 6 patients unsuccessfully treated with intravenous scu-PA,intracoronary streptokinase was subsequently administered. In both cases recanalization could not be achieved. The effects on the hemostatic system are summarized below. In all patients a severe residual stenosis persisted after thrombolytic treatment and 13 patients underwent PTCA.Group Reperfusion Fibrinogen Plasminogen AntiplasminThere was no difference between sucessfully and unsuccessfully treated patients in group II with respect to the effect of treatment on serum parameters.It is concluded that intravenous infusion of scu-PA at a dose of 15 mg over 60 minutes is ineffective treatment for patients with acute myocardial infarction. However,at a dose of 48 mg over 60 minutes this form therapy is effective, save and specific.
MAY LMW (CY 216) BE ADMINISTERED DURING PREGNANCY ?
Authors: Forestier F , Daffos F , Rainaut M , Cornu P , Deschamps A , Toulemonde F 
Published: August 2018

One of the main problems related to the use of fractionated heparin during pregnancy concerns its transplacental passage.Previous studies showed LMW heparin fraction CY 216 has no teratogenic effects, and when labelled, does not cross the placental barrier in animal, and does not appear into the milk.We studied the transplacental passage following subcutaneous administration of large dosage (17.500 AXa IC u) in 7 women who where going to have an abortion during the third trimester of gestation because of severe fetal malformation, after informed consent.Blood samples were taken before and 3 h after injection from che mother , from the fetuses 3 h after mother injection -using ultrasound guidance of the needle and aspiration of blood in the umbilical vein.Biological assays showed that the effects are clearly observable in mother, whereas no change was observed from the fetus.Thus, it was justifiable to treat, for several reasons, 22 patients using CY 216 during a period of 2 to 5 weeks before delivery. Treatments were successful and no complication has been observed. The cord blood samples at birth never showed any biological activity.These data seem to clearly indicate that there is no passage through the placental barrier of CY 216 which offers a new possibility of treatment during pregnancy.
IMMUNOASSAYS FOR SINGLE CHAIN URINARY-TYPE PLASMINOGEN ACTIVATOR (SCUPA) IN PLASMA AND IN CELL CULTURE SUPERNATANTS
Authors: Mahmoud M , Hammerschmidt F , Scheuerlein H , Gaffney P J 
Published: August 2018

Monoclonal antibodies (mabs) to SCUPA have been generated in balb/C mice by conventional means and have been demonstrated to have no crossreactivity with two chain urinary-type plasminogen activator (TCUPA). These mabs have been used to develop two types of specific assay for SCUPA. Mabs coated on polyvinyl plates in conjunction with polyclonal antibodies (pabs) to TCUPA have allowed the development of a catcher-tag ELISA using alkaline phosphatase-label led goat anti-rabbit IgG as a final step. The sensitivity range of the assay was 0.5 - 10.0 iu/ml. A second bioimmunoassay (BIA) using SCUPA mabs as catcher and inplate development with glu-plasminogen and S-2251 has yielded an assay with a sensitivity range of 0.5 - 10.0 iu/ml. The international unitage ascribed in these assays was derived by comparing the hydrolysis of S-2444 by the I.S. for TCUPA with the purified SCUPA following full activation with plasmin.Using these assays it was found that normal pooled plasmas contained about 1.0 iu of SCUPA antigen which was fully inhibited such that no activity was evident by the BIA assay for SCUPA. This suggests that urokinase in plasma is present in two forms: SCTJPA bound to inhibitor and TCUPA which is biologically active when assayed using a BIA based on immobilised pabs to TCUPA. Cell supernatants from cultured human lung fibroblasts yield a SCTJPA/TCUPA ratio of 70/30 using S-2444 chromogenic assay following a plasmin-mediated SCTJPA-TCUPA conversion step. It was also shown that, in these cell supernatants, SCTJPA was secreted with no inhibitor bound to it, since the BIA and ELISA data were quite similar. A curious feature of these assays, which is as yet unexplained, is the observation that urokinase (both plasmin activated SCTJPA and TCUPA), when immunologically adsorbed on to the PVC-immobilised specific mabs or pabs used in this study, readily activated plasminogen but showed no hydrolytic activity on the chromogenic substrate, S-2444.
ANTITHROMBOTIC PROPERTIES OF HEPARIN IN A NEONATAL MODEL OF THROMBIN INDUCED VENOUS STASIS THROMBOSIS
Authors: Schmidt B , Buchanan M R , Ofosu F , Brooker L A , Andrew M , Univ McMaster 
Published: August 2018

Anecdotal clinical experience suggests that more heparin is required in newborn infants that in adult patients to effectively treat thrombotic disease. We compared the ability of heparin to inhibit thrombus formation induced by a pathological bolus of thrombin and stasis in newborn piglets and 3 week old pigs. The coagulation system of the newborn piglet closely resembles that of the human neonate Including low antithrombin III (AT-III) activity (0.5U/ml). By 3 weeks, adult porcine values for coagulation factors and inhibitors are reached, while blood volume/kg body weight still approximates that of the newborn piglet. Piglets and pigs were pretreated with saline, 10 or 25U/kg heparin (n ≥16/group/dose. Following an injection of 100U/kg thrombin, systemic 125I-fibrinogen consumption and local 125I-fibrinogen incorporation into jugular venous stasis thrombi were measured. Peak heparin levels were identical In both age groups (Anti-factor Xa assay and protamine sulphate titration). Heparin was less effective in preventing thrombus formation in piglets than in pigs (Table). Heparin was also less effective in preventing systemic neonatal 125I-fibrinogen consumption (p<0.0001 at both heparin doses).Raising AT-III levels to adult values significantly improved the antithrombotic properties of heparin in neonatal piglets. Thrombus formation was completely abolished in 19 or 22 piglets who received a combination of human or porcine AT-III concentrate and 25U/kg heparin. Raising the heparin dose to 50U/kg had the same effect. We conclude that the efficacy of heparin in neutralizing thrombin is decreased in newborn piglets. Treatment with AT-III concentrate overcomes this relative heparin resistance, and may help reduce high heparin doses otherwise required in neonatal thrombotic disease.
FREQUENCY AND IMPLICATIONS OF SEVERE NEONATAL PROTEIN C DEFICIENCY
Authors: Manco-Johnson M J , Abshire T C , Jacobson L J 
Published: August 2018

The newborn infant has a physiologically low level of protein C which rises very slowly in postnatal life. The frequency and significance of severe neonatal protein C deficiency has not been reported. In this study, protein C levels were measured in 110 newborn infants at the time of birth using functional (amidolytic, Cact) and immunologic (Laurell rocket, Cag) assays. The protein C levels were compared with a marker of thrombin activation (D-dimer fragment of fibrin, +D-D) and infants were subsequently followed for signs and symptoms of thrombosis. Results are summarized below (protein C levels are expressed as U/ml).Thirteen infants had protein C levels compatible with the homozygous deficiency state. Extremely low levels of protein C (<0.20 U/ml) were not found in well term infants and were rarely noted in stable preterm infants. D-D were infrequently present and no thrombosis occurred. Near term infants born with fetal distress frequently showed +D-D but rarely demonstrated extremely low levels of protein C. None of these infants required indwelling arterial catheters and no thromboses occurred. Preterm infants with severe respiratory distress showed lower protein C levels at birth (p <0.01). Although 71% had +D-D, thromboses in these infants were all related to invasive catheterizations. In contrast, the study population of twins demonstrated a high frequency of severe protein C deficiency with negative D-D and frequent thromboses, three of which occurred in the absence of instrumentation. In summary, severe protein C deficiency and thrombin activation are common in sick preterm infants with the risk of thrombosis increased by intravascular catheterization. In contrast, twins with severe protein C deficiency may manifest a thrombotic risk which is independent of thrombin activation or catheterization.
THE PHOSPHOLIPID-BINDING SITE OF FACTOR VIII IS LOCATED ON THE 80 kD LIGHT CHAIN
Authors: Kemball-Cook G , Edwards S J A , Sewerin K , Andersson L-O , Barrowcliffe T W 
Published: August 2018

The binding of Factoi. VIII (F.VIII) peptides to phospholipid (PL) vesicles has been studied by two different methods involving the use of fractionated anti-F.VIII:C I-Fab123’pre
viously reported, i-Fab123’ was fractionated by immunoadsorptionwith F.VIII-PL complexes into two pools:one binding only to PL-binding sites on F.VIIIsAg (PL-site antibody), the other directed against other antigenic sites (non-PL-site antibody).The first technique used was a modification of the method of Weinstein et al. (Proc.Natl.Acad.Sci.USA, 78, 5137-5141, 1981), and involved incubation of the two anti-F.VIII pool swith F.VIII-containing samples, followed by electrophoretic separation of the complexes on the basis of size in non-denaturing SDS gels: this technique allows qualitative analysis of antibody reactive peptides in highly impure samples. Non-PL-site pool reacted with a range of peptides with MrMapparent Mr 90 kD up to 280 kD, a similar pattern to that of ’heavy chain’(HC) peptides of F.VIII seen on SDS-PAGE under reducing conditions; the PL-site antibody, however, reacted only with peptides at apparent Mrs of 80 kD and sometimes150 kD, but not with bands of higher Mr a pattern more consistent with binding to light chain (LC) peptides. Thesame patterns with the two labels were seen in both plasma and F.VIII concentrateThe second approach employed the two labels described above in direct immunoradiometric assays (IFMA’s) on purified human F.VIII peptides prepared by immunoaffinity chromatography and ion exchange on Mono Q gel. Both PL-site and non-PL-site labels measured similar amounts of F.VIII m a sample containing both HC and LC peptides; however, on assaying a sample containing purified HC peptides alone, PL-site antibody measured only 2% of F.VIII:Ag found by non-PL-site label, indicating that PL-binding sites present in samples containing both HC and LC are absent in HC alone.Results from both these immunological methods indicate that the 80 kD LC peptide of F.VIII carries the PL-binding site.
APSAC versus placebo: A Multicenter Study of Safty and Early Morality in Acute Myocardial Infarction
Authors: Julian D G , Borthwick L S , Reid D , Jennings K P , Wainwright R J , Rodger J C , Wood D , Flax J S , Phillips W S 
Published: August 2018

A prelimiry study was carried out of obtain data on mortality,benefit and the safety of anisoylated plasminogen streptokinase activator complec (APSAC).90 patients from 7 centers,with symptons of acute myocardial infarction of not more than 4 hours duration,were randomised to receiveeithere 30U of APSAC as an intravenous injection over 4 to 5 minutes or placebo given by the same route.45 patients recevied APSAC and placedo,mean time to treatment was 3 hours 20 minutes and 3 hours, respectively.Mortality at 30 days was 7 deaths in theplacebo group and 1 death in the APSAC group (p=0.058),these were all related either directly or indirectly to the infarct:cardiogenic shock (2),ventricular asystole(2) further acute myocardial infraction(2),ventricular fibrillation (1) and pulmonry embolism (1).The patient who died after APSAC therany had presented with an anterior infraction; 5 patients who died inthe placebo group presented with anterior infarcts and 2 with inferior infarcts. Most daeths occured in the group randomised 2 hours 30 minutes to 4 hours post-infarcation.Both systolic and diastolic blood pressure were generally similar in each groupfrom 1 hour to 1 month after treatment. 35 percent of petients in the placebo group experienced cardiovasular events (2 of whom died from cardiogenic shock) compared with 20 percent of APSAC of treated patients (1 of whom died from pulmonary embolism).Minor haematuria was apparent in a grater number of patients in the APSAC group after 12 hours, but there was no differnce between groups after 1 or 2 weeks.The result of this study ,although encouraging,should be interpreted with caution,mainly due to the small numbers of patients enrolled.There was however a trend towards incresed survival in the APSAC treated group (97.8 per cent) compared with84.4 cent survival in the placenbo groupat 30 days post-infarction.
PROTEIN C, PROTEIN S AND THE FIBRINOLYTIC SYSTEM IN PATIENTS WITH A HISTORY OF THROMBOSIS
Authors: Malm J , Laurell M , Nilsson I M , Dahlbäck B 
Published: August 2018

Consecutive patients with a history of thrombo-embolic disease (n = 241, 109 males, 132 females, mean age 46 y), referred to the Coagulation Laboratory during an 18 month period, were analysed for defects in their coagulation and fibrinolytic systems. The diagnosis of thrombosis had been verified with phlebography and that of pulmonary embolus with scintigraphy or angiography. Retinal venous thrombosis was found in 15 of the patients. In 15 cases the thrombotic episodes occurred postoperatively, in 15 during pregnancy, in 12 during the postpartum period and in 20 during use of oral contraceptives. In the remaining cases no clinical riskfactors were identified.The concentration of protein C zymogen was measured with an immunoradiometric assay. Functional protein C was determined with a clotting inhibition assay. Protein C deficiency was found in 8 cases. Two of these had a functional protein C deficiency with normal zymogen levels. The concentration of total, as well as free (not in complex with C4b-binding protein), protein S was determined with a radioimmunoassay. Two cases of protein S deficiency were detected. Three patients with antithrombin III deficiency and two with plasminogen deficiency were found.The fibrinolytic activity after venous occlusion was analysed in 216 patients. Decreased levels were found in 32 %. The concentration of tissue plasminogen activator inhibitor (PAI) was measured in 110 patients and found to be increased in 65 % of the cases. In 99 patients both the fibrinolytic activity and the PAI concentration were measured. A combination of decreased fibrinolytic activity and increased levels of PAI was found in 44 cases. The concentration of tissue plasminogen activator antigen was decreased in 22 % of 105 cases analysed.Thus, in this material of patients with thrombo-embolic disease, abnormalities were found in 47 %. Defects in the fibrinolytic system were the most common findings. Protein C or protein S deficiency was diagnosed in less than 5 % of the cases.
MONOCLONAL ANTIBODIES THAT RECOGNIZE Ca2+-INDUCED CONFORMER OF PROTEIN C, INDEPENDENT OF GLA RESIDUES
Authors: Sugo T , Tanabe S , Shinoda K , Matsuda M 
Published: August 2018

Monoclonal antibodies (MCA’s) were prepared against human protein C (PC) according to Köhler & Milstein, and those that recognize the Ca2+-dependent PC conformers were screened by direct ELISA in the presence of 2 mM either CaCl2 or EDTA. Out of nine MCAߣs thus screened, five MCA's designated as HPC-1˜5, respectively, were found to react with PC in the presence of Ca2+ but not EDTA. By SDS-PAGE coupled with Western Blotting performed in the presence of 2 mM CaCl2, we found that two MCA’s HPC-1 and 2, recognized the light chain, and two others, HPC-3 and 4, recognized the heavy chain of PC. But another MCA, HPC-5 was found to react with only non-reduced antigens. Further study showed that HPC-1 and 5 failed to react with the Gla-domainless PC, i.e. PC from which the N-terminal Gla-domain of the light chain had been cleaved off by α-chymotrypsin. However, all the other three MCA's retained the reactivity with the antigen in the presence of Ca2+ even after the Gla-domain had been removed. The binding of these MCA’s to PC in the presence of Ca2+ was found to be saturable with respect to the Ca2+ concentration and the half maximal binding for each MCA was calculated to be about 0.5+mM. Moreover, many other divalent cations such as Mg2+, Mn2+ , Ba2+, Zn2+, Co2+, Sr2+, were found to substitute for Ca2+ in inducing the metal ion-dependent but Gla-domain-independent conformer of PC.Cross-reactivity to other vitamin K-aependent plasma proteins was examined by direct ELISA; HPC-2 and 3 reacted solely to PC, but HPC-1 and 4 also reacted with prothrombin and HPC-5 with both prothrombin and factor X.These findings indicated that there are two or more metal binding sites besides the Gla-domain, possibly one in the light chain and the other(s) in the heavy chain. The presence of these metal binding sites may contribute to the unique conformer of vitamin K-dependent plasma proteins including protein C.
HETEROGENEOUS REACTIVITY OF HUMAN MONOCLONAL ANTIPHOSPHOLIPID ANTIBODIES (HMA) WITH ENDOTHELIAL CELLS (EC), PLATELETS AND DNA
Authors: IZAGUIRRE C A , DUFF S , HASHMENT S , SMITH C D , MENG Q H , RAUGH J 
Published: August 2018

Antibodies that react with cardiolipin (ACL) and/or have lupus anticoagulant (LA) properties are implicated in the thrombotic diathesis of patients with the antiphospholipid antibody syndromes: recurrent venousand arterial thrombosis, fetal wastage, thrombocytopenia and CNS disease, often associated with systemic lupus erythematosus. We studied a panel of 16 HMA with LA and/or ACL properties (J Biol Chem 261:9672 1986) for their reactivity with platelets (ELISA),DNA (ELISA), with surface antigensonEC (EC-ELISA) and with whole cell EC extracts as detected by western blot and immunoperoxidase (EC-WB). Three control HMA werenonreactive in all assays.Five HMA with LA activity alone did not react with EC. Five of 6 HMA-ACL reacted with EC both on ELISA and WB. All of the HMA-ACL also reacted with DNA and platelets but 4 did not react as LA. Our findings indicate that HMA antiphospholipid antibodies(LA and ACL) appear to be heterogeneous in terms of their spectrum of reactivities, thus, the ACL and LA tests may in some cases recognize antibodies with different properties and perhaps different pathogeneticmechanisms.xs