bluuebunny/crossref_metadata_2025_split huggingface dataset:
DDE COMPLEX DETERMINATION AS A SPECIFIC MARKER FOR FIBRINOLYSIS
Authors: Soria J , Soria C , Mirshahi Me , Mirshahi S , Mirshahi M , Pujade E , Boucheix C , Samama M , Bernadou A , Caen J P 
Published: August 2018

Monoclonal antibodies (McAb) reacting with fibrin degradation products (FbDP), but not with fibrinogen have been produced in order to determine specifically FbDP directly in plasma. Most of the McAb available however do also react with fragment D. Our anti D neo McAb reacts about 10 times less with fragment D than with FbDP but does not react with fibrinogen, fragment X or Y.In clinical investigation, even in pathological conditions in which there is a great release of tissue-type plasminogen activator (tpA), we have shown that fragment D is not generated in patients plasma. Therefore, the reactivity of our McAb with fragment D did not alter the specificity of FbDP assay.On the contrary, using polyacrylamide gel electrophoresis in the presence of SDS followed by immunorevelation, we have evidenced that fragment D is generated in patients undergoing thrombolytic therapy even with rtpA. Therefore, using conventional Elisa procedure (capture of FbDP on polystyrene-immobilized anti D neo antibody and detection by peroxidase-labelled anti fragment D immunoglobulins), the presence of fragment D in patients plasma leads to an overevaluation of FbDP. To avoid this overestimation we have modified the Elisa procedure. The structure of FbDP was taken into acount in order to render the technique specific of FbDP. In FbDP fragment D coming from one fibrin monomer is always associated with fragment E from another fibrin monomer, as DDE complex for example. Therefore after capture of fragment D by the polystyrene-bound anti D neo McAb, FbDP were specifically revealed by peroxidase-labelled anti E antibody (polyclonal or monoclonal anti E may be used). For this reason, this test was named DDE determination and DDE determination can be used in any circumstances to evaluate fibrin degradation.
ORAL ANTICOAGULANTS IN BREAST CANCER PATIENTS
Authors: Sagripant A , Carpi A , Baicchi U , Nicolini A , Ferdeghint M 
Published: August 2018

As the fibrin clot may play a role in the intravascular metastatic spread of breast cancer, we have treated women with advanced breast cancer with oral anticoagulants. After mastectomy, 11 consentient women with postsurgical stage Nl (6 patients), N2 (3 patients) and N3 (2 patients) and without evidence of distantmetastasis have been treated with acenocoumarol (INR 2-4,5) for 4-22 months (mean 12 months). All the patients received adjuvant therapy (radiotherapy, polichemiotherapy and/or tamoxifen) in conformity with theclassic indications. FibrinopeptideA plasmatic level, checked in everyone on stable anticoagulation, was always lower than 2 ng/ml (mean 0,78 ng/ml). No major bleeding occurred, except a copious hematuria caused by overdosage. Until now all the 11 women are alive. Two of them stopped anticoagulant, one because of hemorrhagic cystitis, the other because of awareness of visceral metastases; all the other 9 women arebeing treated now. Three patients(2N2 and 1 N3) have evidence of visceral metastases; no sign of relapsehas been observed by serial instrumental and laboratory examinations inthe other 8 women. A control group of 13 patients (9 N2 and 4 N3)was compared with the group of anticoagulated patients in more advanced stages (2 N2 and 3 N3):The follow-up of the anticoagulatedNl patients is too brief as yet forcomparative evaluation. Our preliminary data seem to indicate an useful role of oral anticoagulants inbreast cancer and oblige us to prolong investigation.
ANTITHROMBIN III (AT III) REPLACEMENT FOR THE PREVENTION OF REOCCLUSION AFTER INTRACORONARY THROMBOLYSIS
Authors: Egbring R , Bethge C , Behling A , Seitz R A 
Published: August 2018

Within 4 hours of acute myocardial infarction thrombolysis of an intracoronary thrombus leads to reperfusion in 40-707. of all cases, when streptokinase (SK), urokinase (UK) andtissue plasminogen activato(TPA) areused as thrombolytic agent.Although reperfusion could be demonstrated after thrombolysis, reocclusion occurred to different extents. There are several reasons for reocclusion: 1. Reocclusion occurs at the site of theprevious thrombus, as the vessel wall is deprived of endothelium and thethrombogenic surface is exposed; 2. The post-thrombolytic anticoagulant treatment (ACT) is not sufficient oris not performed correctly. When ACT was carried out with high doses of heparin, bleeding complications could be observed. Therefore we have tried in an open pilot study to increase the anticoagulant effect by additional replacement of AT III concentrates for 5 days as follows. The first group of patients (30) was given 1000 U AT III concentrate daily with an average dose of 24000 U heparin daily for 5 days. The second group (9)received more than 30000 U heparin daily. In the first group of 30 patients neither hemorrhage nor reocclusion of the infarction vessel was observed, the AT III levels rose gradually. In 3 of 9 patients of the secondgroup, hemorrhage occurred, necessitating reduction of the heparin dose.Subsequent reocclusion of the infarction arteries occurred in all 3 cases. The mean AT III level decreased in these patients to the lower limit.Conclusion: After reperfusion of coronary arteries following thrombolytic therapy reocclusion may occur for several reasons. The present studies show that reocclusion is rare ifanticoagulant treatment is performedwith AT III concentrate replacement in combination with ACT with heparin.Although the initial AT III levelsare not decreased beneath the critical level of 707o, the substitution leading to a further gradual increase seems to prevent reocclusive thrombosis. According to Reeve (1982) the amount of circulating thrombin is dependent on the equation 1:(AT III)2.
EXPRESSION OF BIOLOGICALLY ACTIVE HUMAN ANTITHROMBIN III IN CHINESE HAMSTER OVARY CELLS
Authors: Zettlmeiβl G , Ragg H , Karges H 
Published: August 2018

Expression of human antithrombin III (AT III) at high levels has been achieved in Chinese hamster ovary (CH0) cells by cotransfection and subsequent coamplification of the transfected sequences. Expression vectors containing the AT III cDNA gene and a dihydrofolate reductase (DHFR) cDNA gene were transfected into CH0 DHFR-deficient cells. About 20% of the DHFR+ transformants secreted recombinant human AT III into the medium. Stepwise selection of the AT III producing DHFR+ -transformants in increasing concentrations of methotrexate generated cells which had amplified the AT III géne. We determined the copy number of the AT III cDNA and the relative amounts of AT III specific mRNA at different stages of the amplification process. Transfected CH0 cell lines expressed elevated immunreactive levels of human AT III.AT III secreted from these cell lines had the same molecular weight (60 kDa), immunological properties and biological activities as AT III obtained from human plasma. In vivo data concerning the inhibition of glycosylation by different drugs suggest that recombinant AT III from CHO cells is glycosylated according to a complex type pattern.
PURIFICATION AND CHARACTERIZATION OF GENTECHNOLOGICALLY PREPARED ANTITHROMBIN III
Authors: Karges H E , Zettlemeiβl G , Naumann H , Eberhard U , Bröker M 
Published: August 2018

Isolation and purification of antithrombin III (AT III) by affinity chromatography on immobilized heparin is a standard method for the large scale preparation of this protein from human or animal plasma. Hence, after AT III became available by gentechnological methods, we tried to adapt this procedure for the isolation of AT III from supernatants of mammalian- and yeast-cells. Indeed, it was possible to use this method also for the isolation of the recombinant gene products. Since, however, the cell growth media contain heterologous protein or peptide mixtures like fetal calf serum, the method had to be improved to avoid the adsorption of non human proteins or peptides. We are now able to purify AT III from CHO-cell-superna-tants to more than 95 % purity. The characterization of this AT III-product by double immuno diffusion revealed that it is immunologically totally identical with the authentic material from plasma. AT III antigen content, progressive inhibitor activity and heparin cofactor activity compare very well in the final product; hence, it is totally active compared to AT III from plasma.In polyacrylamidegel electrophoresis most of the material migrated differently to the authentic material showing 9 bands in equal distance to each other, instead four in the At III from plasma. After degradation with sialinidase from both AT III preparations identical cleavage products were obtained migrating predominantly as a single band. Hence, the electrophoretic heterogeneity seems to be due to a different degree of sialinyla-tion of the products.
CHARACTERIZATION OF THE FIBRINOGEN BINDING SITES USING MONOCLONAL ANTIBODIES TO HOMAN PLATELET MEMBRANE GLYCOPROTEINS IIb/IIIa*
Authors: Ruan C , Du X , Wan H , Hu X , Xi X , Li P 
Published: August 2018

Two new murine monoclonal antibodies, SZ-21 and SZ-22, (both IgG1 subclass), were produced by the hybridoma technique using washed human platelets as the immunogen. Both SZ-21 and SZ-22 reanted specifically with normal platelets and megakaryocytes but not with Glanzmann's thrombasthenic platelets which lack the membrane glycoprotein(GP)IIb/IIIa complex. Platelets from 10 normal donors bound 64,500±20,300(x+SD) SZ-21 molecules/platelet with KD4.4±1.5nM and 61,000±19,900 SZ-22 molecules/platelet with KD18.8±6.7nM respectively.Affinity chromatography confirmedthat SZ-21 and SZ-22 reacted with theGPIIb/IIIa complex. SZ-21 inhibited the platelet aggregation and secretioninduced by collagen, arachidonic acidand thrombin,and the second wave of aggregation in response to ADP, adrenaline and ristocetin. The fibrinogen binding to human platelets induced by ADP, arachidonic acid and PAF was also inhibited by SZ-21. But SZ-22 hado effects on platelet aggregation andfibrinogen binding. Western blot analyses indicated that SZ-21reanted with GPIIIa and that SZ-22 bound to GPIIb. When the protein was reduced by 2-mercaptoethand, SZ-22 reacted with the enchain of GPIIb, but SZ-21 lost its ability of binding to GPIIIa, suggested that the antigenic determinants of SZ-21 depended on the integrity ofthe intra αchain disulfide bond of GPIIIa. On chymotrypsin treated platelets,the epitope for SZ-21wasidentified on a 66 KD membrane-bound fragment of GPIIIa. The appearanceofthe 66 KD fragment was related withthechymotrypsin-mediated fibrinogen binding and aggregation. However the prolonged treatment with chymotrypsin reduced the platelet aggregation, which coincided with the appearance of the 66KD fragment, a product of thefurther hydrolysis of the 66KD fragment.These results suggested that the domainbetween the 66KD and 60KD fragments of GPIIIa might be essential for the maintenance of the fibrinogen binding sites.* Supported by the Science fund of the Chinese Academy of Sciences ( project No.82-172 ) and the National Science fund of China ( project No.3860713 ).
EFFECT OF TISSUE-TYPE PLASMINOGEN ACTIVATOR (t-PA) ON BACTERIAL ENDOCARDITIS
Authors: Buiting A G M , Thomson J , Emeis J J , Mattie H , Brommer E J P , Van Furth R 
Published: August 2018

In bacterial endocarditis the causing microorganisms are located in a fibrin-platelet matrix, making them less accessible to host-defence mechanisms and antibiotic therapy. Trombolytic treatment could break down the fibrin of these endocardial vegetations and thus eliminate the focus of infection. This approach was studied in vitroand in vivo using recombinant t-PA (rt-PA,Wellcome Biotech) having a fibrinolytic activity comparable with melanoma t-PA. The following results were obtained.1.Incubation of Streptococcus sanguis infected plasma clots in the presence of t-PA resulted in lysis of the clots as evidenced by a significant decrease in the weight of the clots and by an increase in the number of streptococci in the medium.No effect of t-PA was found on the antimicrobial action of penicillin G (PenG)onthe streptococci in the non-lysed part ofthe clots.2. Vegetations isolated from the heart of rabbits with a S. sanguis endocarditis,incubated in medium with t-PA were also lysed. This resulted in a rise in the mediumof both the number of bacteria and the concentration of fibrin degradation products.3.Treatment of rabbits with a S. sanguis endocarditis using a combination of rt-PA, given as bolus injections at one hour intervals (4x0.5 or 4x1 mg per kg/day), andPenG decreased the weight of the endocardial vegetations significantly compared toa control group treated with PenGonly (43.3 and 81.8 mg respectively). No additional decrease in weight was obtained with repeated administration of t-PA at daily intervals up to 2 days. The significant decrease of the number of streptococci per gram of vegetation as a result of the treatmentwith PenG was not influenced by t-PA. Treatment with 4x1 mg t-PA per kg caused a small decrease (about 30%) in the plasma concentration of plasminogen, fibrinogen and aα2~antiplasmin.However, no bleeding complications were observed.In conclusion rt-PA can influence the treatment of bacterial endocarditis in rabbits by decreasing the size of the vegetations but not by influencing antimicrobial action on the bacteria in the non-lysed part of the vegetation.
EFFECT OF RECOMBINANT TISSUE TYPE PLASMINOGEN ACTIVATOR(rt-Pa)ON THE PREVENTION OF INTRAABDOMINAL ADHESION FORMATION
Authors: Mohler M , HollenBach S , Nguyen T , Reger V , Hothkiss A 
Published: August 2018

The formation of intraperitoneal adhesions, the pathological adherence of organs and tissue surfaces, is the leading cause of postoperative intestinal obstruction following abdominal surgery, as well as the major contributing factor to infertility after reconstructive tubal surgery. At present, there is no generally accepted therapy for the prevention of adhesion formation. Since fibrin has been shown to be the physiological basis of adhesion formation, the current study was undertaken to determine if an application of exogenous rt-PA to the site of peritoneal injury would prevent adhesion formation. New Zealand white rabbits (3 kg) were anesthetized and midline laparotomies performed. A nine cm area of the peritoneal wall was surgically removed and resutured in place, creating an ischemic patch of peritoneal tissue. A proximal area of the cecum (75cm2 ) was abraided with dry gauze until punctate bleeding occurred. Approximately 2.5 grams of an ointment containing 0, 0.16, 0.31, or 0.63 mg rt-PA was applied to the ischemic peritoneal tissue and the abraided cecum. After seven days the rabbits were euthanized and the adhesions scored. In the rabbits which received placebo ointment (n=6) the cecum was adhered to the entire area of the ischemic patch. In the rabbits which received the ointment containing 0.63 mg of rt-PA, five had no adhesions and one rabbit had a very minor adhesion. The high dose of rt-PA was equally effective in preventing reformation of adhesions after surgical lysis. No evidence of systemic fibrinogen degradation or abnormal wound healing were evident in this model.In summary, these studies demonstrate that intraperitoneal administration of rt-PA is effective in preventing initial adhesion formation as well as the reformation of adhesions after surgical trauma or injury.
ON THE MECHANISM OF THE IN VIVO SYNERGISM BETWEEN TISSUE-TYPE PLASMINOGEN ACTIVATOR (t-PA) AND SINGLE CHAIN UROKINASE-TYPE PLASMINOGEN ACTIVATOR (scu-PA)
Authors: Stassen J M , Collen D 
Published: August 2018

t-PA and scu-PA, in molar ratios between 1:4 and 4:1 do not act synergically in vitro (Thromb. Haemost. 56,35,1986) but display marked synergism in a rabbit model (Circulation 74, 838, 1986) and in man (Am. Heart J. 112, 1083, 1986). To investigate the mechanism of in vivo synergism in the rabbit model (J. Clin. Invest. 71, 368, 1983), t-PA and scu-PA were infused 1) simultaneously over 4 hrs, 2) t-PA over 1 hr, then 15 min later scu-PA over 2 hrs and 3) scu-PA over 1 hr, then 15 min later t-PA over 2 hrs.Significant synergism on thrombolysis is observed when t-PA and scu-PA are infused simultaneously or when t-PA is followed by scu-PA but not when scu-PA is followed by t-PA. These results suggest that low dose t-PA induces some plasminogen activation, sufficient to partially degrade fibrin, exposing COOH-terminal lysines with high affinity for plasminogen (Eur. J. Biochem. 140, 513, 1984). scu-PA might then activate surface-bound Glu-pla-minogen more efficiently.Sequential therapy with t-PA (or any other agent which "predigests" the thrombus), followed by scu-PA might constitute an alternative to simultaneous infusion of synergistic thrombolytic agents.
THE EFFECT OF TWO CHOLESTEROL-LOWERING AGENTS ON PLATELETS IN PATIENTS WITH HYPERCHOLESTEROLEMIA
Authors: Norϕ A , Smimonsen T , Ytre-Arene K , Lyngmo B , Svensson B 
Published: August 2018

Twenty-one subjects with type Ila hyperlipoproteinemia, receiving dietary treatment were given Synvinolin (MK-733), a HMG-CoA reductase inhibitor, 40 mg or Cholestyramin (Questran) 24 g daily for a period of 12 weeks.Serum lipids, platelet cholesterol, phospholipids and fatty acid composition and platelet function were measured before and after the intake of the two drugs.Both drugs reduced serum total cholesterol with approximately 50%. No significant changes were observed in platelet lipid concentrations or in the primary bleeding time. Collagen induced aggregation and thromboxane (TXA2) production were reduced, whereas thrombin induced aggregation and TXA2 production were unaffected. This study shows that both a cholesterol synthetase inhibitor and Cholestyramin reduce the total serum cholesterol concentration and also reduce platelet aggregation and thromboxane synthesis without changing the platelet cholesterol content or the cholesterol/phospholipid ratio. The effect on serum lipids and platelet function may indicate a beneficial effect of both drugs on arterial disease in patients with hypercholesterolemia.
EFFECT OF ASPIRIN AND ASPIRIN COMBINED WITH DIPYRIDAMOLE IN EARLY DIABETIC RETINOPATHY
Authors: Samama M , Baudoin C E 
Published: August 2018

In a double blind randomised controlled clinical trial the effect of antiplatelet agents (aspirin 330 mg × 3 × day) or in combination with dipyridamole (75 mg 3 × day) versus placebo, was tested in 475 patients with early diabetic retinopathy. Patients were follewed fourmonthly for 3 years. Ophtalmological examinations were carried out initially and at yearly intervals. The assessment of retinopathy was based on changes in the number of microaneurysms (MA) present in the macular field as seen on fluorescein angiograms over a period of three years. Forty one patients did not complete the study. Among the others at least three readable initial and yearly angiograms were available on 420 patients who had a 3 year follow up (266 insulin treated and 154 non insulin treated). The results are based on these patients.It is concluded that either Aspirin alone or in conjunction with dipyridamole significantly slows down the progression of MA evolution in early diabetic retinopathy. Because of the very small, although significant changes, the clinical relevance of these drugs has not been established.
Effects of intermittent dosage of ASA on the increased in vivo formation of thromboxane during acute myocardial infarction
Authors: Vesterqvist O , Rasmanis G , Henriksson P , Edhag O , Gréen K 
Published: August 2018

We have earlier demonstrated considerable increase of the in vivo formation of thromboxane (Tx) in connection with myocardial infarction (AMI). The in vivo formation of prostacyklin (PGI) was also increased, in some cases very much so, with a maximum after the CK peak. In normals ASA reduces the in vivo Tx synthesis to about 15% for several days while the PGI synthesis is only inhibited for 2-3 hrs. The in vivo synthesis of those prostanoids was measured by gas chromatographic mass spectrometric quantitation of the major urinary metabolites. In a serie of twenty patients with AMI we measured the in vivo synthesis of Tx and PGI during seven consecutive days. Ten patients received 0.5 g ASA every third day. In the nontreated group the Tx synthesis slowly decreased to about 70%, in the treated group to about 25% on day 3 as compared to the synthesis at admission. There was no difference in the in vivo synthesis of PGI between the two groups.These data demonstrate that in most individuals with AMI it is possible to inhibit Tx synthesis considerably with intermittent dosages of 0.5g ASA while the PGI synthesis is essentially maintained intact. Therefore this regimen should be more beneficial to the patient than daily doses of any magnitude.
A NEW BLEEDING TENDENCY DUE TO HERITABLE HYPER α2-MACROGLOBULIN-EMIA (α2-MACROGLOBULIN-AKITA)
Authors: Endo Y , Iwamoto K , Mamiya S , Niitsu H , Itoh T , Miura A 
Published: August 2018

A heritable elevation in α2-macroglobulin (α2M) was identified in a 9-year-old girl with bleeding tendency and prolonged APTT to 49.1 sec (normal 27-38) and recalcification time to 438 sec (<180). Plasma α2M level in her family tree i.e. sister, mother, maternal grand mother, father and paternal grand mother was 406, 380, 352, 339, 166 and 236 mg/dl (140-285), respectively. Thus it is thought to be an autosomal dominant disease, but other relatives show no apparent clinical syndrome. The significance of a possible causal association between elevated α2M and orolonged APTT was showed.The activity of α2M of the patient determined as trypsin-protein esterase was 351 mg/dl (197%), also. The α2M of the patient showed normal mobility to anti-α2M plasma with acceralated anodal mobility by crossed immunoelectrophoresis and normal mobility by immuno-fixation electrophoresis. In addition, by the analysis with SDS-polyacrylamidegel electrophoresis we demonstrated no qualitative abnormality in α2M of the patient. The new familial abnormality is tentatively designated "α2-Macroglobulin -Akita".
INACTIVATION OF HEPARIN COFACTOR II (HC II) BY POLYMORPHONUCLEAR LEUKOCYTES (PMNL)
Authors: Sié P , Dupouy D , Dol F , Boneu B 
Published: August 2018

Several authors have shown that antithrombin III (AT III) was catalytically inactivated by neutrophil elastase, an observation relevant to pathophysiological processes in the vicinity of inflammatory sites. The aim of this study was to investigate whether HC II, another natural thrombin inhibitor, is also inactivated by PMNL.A rapid loss of HC II activity occured upon incubation with fresh human PMNL stimulated by phorbol myristate acetate (Tl/2 1 uM HC II, 0.35 108 cells/mm : -2 min) or with PMNL extracts prepared by nitrogen cavitation. Antithrombin (dermatan sulfate cofactor) and antichymotrypsin activities of HC II were lost at the same rate. Resting PMNL were ineffective. Inactivation was prevented by several serine-protease inhibitors but was Ca++ /Mg++ independent. Inactivation coincided with the formation of a 54 KD peptide after a first non-inactivating degradation into a 62 KD peptide (native HC II : 76 KD). These reaction products are reminiscent of those described upon incubation with proteinase I from Echis carinatus venom. HC II was inactivated more rapidly than AT III (T 1/2 of AT III in the same conditions -15 min). However, heparin (1-10 ug/ml) strongly accelerated the rate of AT III inactivation and slightly protected HC II, thus reversing the order of inactivation. Dermatan sulfate had no effect on this process.In conclusion, this study shows ; l)both AT III and HC II are rapidly inactivated by PMNL enzymes, thus favoring locally thrombin-mediated processes ; 2) heparin increases AT III degradation by PMNL, a possible route of catabolism operating in patients with low AT III levels during heparin treatment.