bluuebunny/crossref_metadata_2025_split huggingface dataset:
THE EFFECT OF LOW DOSE HEPARIN ON THE FEEDBACK ACTIVATION OF FACTOR VIII IN VIVO
Authors: Devilée P P , Fiolet H F B M , van Dam-Mieras M C E , HemKer H C 
Published: August 2018

Heparin is a well established drug in the treatment and prophylaxis of thrombosis. However, the in vitro methods for thecontrol of heparin therapy (aPTT, anti-Xa, anti-II) often are not sufficiently sensitive to monitor low-dose heparin therapy. We used an assay that is based upon the in vivo activation of factor VIII at the site of skin punction: the factor VIII activity in blood samples taken from a capillary wound increases with time Hurlet-Birk Jenssen et al. Path. Biol. 24, 6 (1976).Assay: 50 μl blood samples are taken 30, 60 and 90 sec after capillary puncture. The samples are diluted with 950μl, 10 mM sodium citrate, Mich, buffer and centrifugated(3 min, 4 °C). In the diluted samples the factor VIII activity is determined.In healthy volunteers the increase in factor VIII activity in the successive samples is fairly constant (1.4 - 1.8 fold) in spite of distinct interindividual differences in the factor VIII level. In patients receiving continuous intravenous heparin (20000 IE/24 h) and in a hemophiliaA patient there was no increase in factor VIII activity with the time. In individuals receiving 5000 IE heparin subcutaneously the increase in factor VIII activity was dependent upon the time interval between heparin injection and sampling. There was almost no influence of heparin on the 30 s samples but the 90 and 150 sec. samples showed distinctly prolonged clotting times at sampling points from 2 to 4 hours after heparin administration. It was not possible to detect the heparin effectby an automated anti-IIa assay or in a prothrombin time assay because the heparinlevels were below the detection limit of the tests.These results suggest that low-dose heparin therapy has a definite influence on the feedback activation of factor VIII even in concentrations that are not detectable with the usual heparin tests.
ACYL PLASMINOGEN STREPTOKINASE VERSUS HEPARIN IN RECENT ACUTE MYOCARDIAL INFARCTION. PRELIMINARY REPORT OF A FRENCH INTERUNIVERSITY TRIAL
Authors: Bassand J P , Machecourt J , Cassagnes J , Lusson J R , Bourlard P , Anguenot T , Faivre J , Wolff J E , Peycelon J 
Published: August 2018

One hundred and twenty seven patients (pts) suffering from a first acute myocardial infarction (AMI) were included in a multicenter trial involving three University Hospitals and 22 community Hospitals without catheterization facilities. Patientswere randomly allocated within 5 hours following the onset of symptoms either to acyl plasminogen streptokinase (APSAC) 30 IU within 5 min followed 4 hours later by heparin at a dose of 500 IU/kg/day or to conventional Heparin (H) therapy 500 IU/kg/day. 63 pts received APSAC and 64 received H. Both APSAC and H groups were similar in age, location of AMI, Killip class and time of randomization. After initial therapy Pts were referred to the University Hospitals and submitted between Day 1 and Day 7 to a selective coronary angiography and immediate percutaneous transluminal angioplasty (PTCA) of the infarct-related artery if suitable for the procedure.4 APSAC pts and 2 H pts died during the hospital course (NS).The arterial patency rate determined on average 3.4 ± 1.2 days after the onset of symptoms was 88% in APSAC and 42% in H (P<.001). 15 APSAC pts and7 H pts were submitted to PTC A with one failure in each group. LVEF and akinetic score (AK) were determined from left ventricular angioeraDhy :On Day 14, pts were submitted to Ti 201 scan and to radionuclide angiography (RNA). Results are not yet available.Our results showed that early infusion of APSAC in AMI produced a high short term patency-rate of the IRA with a tendancy toward improvement of the left ventricular systolic function which failed to reach statistical significance because of the relatively small size of the cohort.
EFFECTIVENESS OF INTRAVENOUS THROMBOLYTIC TREATMENT IN ACUTE MYOCARDIAL INFARCTION:SHORT AND MEDIUM TERM PROGNOSIS
Authors: Mauri F 
Published: August 2018

An unblinded trial of intravenous stretokinase in early acute myocardial infarction was planned to study wheter the drug produces a clinically relevant benefit in terms of reduction of in-hospital and one year mortality.11806 pts in one hundred and seventy six coronary care units were enrolled over 17 months.Patients admitted within 12 h after o.nSet of symptons and with no contraindications to SK were randomized to receive SK in addition to usual treatment and complete data were obtained in 11712 for what concerns in hospital prognosis.At 21 days overall hospital mortality was 10.7% in SK recipients versus 13%,in controls,an 18% reduction(p=0.0002,relative risk 0.81).The extent of beneficial effect appears to be a function of time from onset of pain to SK infusion(relative risk 0.74,0.80, 0.87 and 1.19 for the 0-3,3-6,6-9 and 9-12 h subgroups).The data of 1-year follow-up concerning 11605 pts(95.3% of the whole population)were available up to December the 31st.1987.4333 pts out of the SK-treated group(74.0%)and 4219 out of the control one (72.1%)were alive,with a significant difference.These results document that the benefit produced by SK in the hospital period remains substantially unchanged.The differences in mortality in favour of SK vs.C remain highly significant specifically for the 0-3 and 3-6 hrs subgroups and is dramatic for patients treated between one hour from onset of symptoms. 503 out of the 637 treated with SK were alive at 1-year follow-up versus 443 out of 641 control group pts:the amplitude of the benefit seems to be further increased in this particular subgroup.The GISSI results document conclusively that an acute thrombolytic treatment with SK in AMI is effective in reducing mortality not only over the short,but also over the medium period.
IDENTIFICATION OF GLYCOPROTEIN lb IN “IN VITRO” TRANSLATES FROM ISOLATED HUMAN PLATELET mRNA
Authors: Wicki A N , Walz A , Clemetson K J 
Published: August 2018

Platelet membrane glycoproteins play a crucial role in platelet adhesion and activation. To understand how they function it is of great importance to know their amino acid sequences and structures. It is rather difficult to purify membrane glycoproteins in amounts that are sufficient to determine their amino acid sequences by protein sequencing techniques. The easier way seems to be molecular cloning of the genes for these proteins.Metabolically stable mRNA derived from nucleated megakaryocytes is known to be present in the anuclear human platelets. We have developed a purification method for the isolation of platelet mRNA. Starting with 100 units of blood platelets we isolated 0.5 mg of mRNA by guanidine chloride/lithium chloride/phenol extraction. Crude mRNA as well as oligo-dT purified, polyadenylated mRNA, was assayed for protein synthesis in a reticulocyte lysate translation system in the presence of different labelled amino acids. SDS-PAGE of nonreduced and reduced samples of the translation products showed molecular masses up to at least 200 kDa. Immunoprecipitation and affinity column chromatography with poly- and monoclonal anti platelet glycoprotein lb antibodies showed that a protein with a molecular mass of 60 kDa crossreacting immunologically with glycocalicin is specifically recognized by these antibodies. The 60 kDa protein seems to be the nonglycosylated form of GPIbケ.
PROTEOGLYCANS AND SULFATED PROTEINS OF PLATELETS: CHARACTERIZATION AND RESPONSE TO THROMBIN AND ADP
Authors: Schick B P , Walsh C J , Jenkins-West T 
Published: August 2018

The proteoglycans (PG) and sulfated proteins of guinea pig platelets were labeled in vivo by intraperitoneal injection of (35S)sulfate. At 3 days after injection, platelets contained 3 distinct populations of chondroitin-6-sulfate proteoglycans which together constitute about 65% of the cellular (35S) label. Most PG elute from a DEAE-Sephacel column with 4M Gdn HC1 (PG-1, 87%), and elute at Kav 0.12 on Sepharose CL-6B. The PG-1 can be resolved by SDS-PAGE into two fractions. The remainder (PG-2, 13%) elutes from the DEAE-Sephacel column with 4M Gdn HCl/2% Triton X-100 or 2% CHAPS, and has a Kav of 0.07 on Sepharose CL-6B. About 20-25% of the cell (35S) label elutes from DEAE-Sephacel in the wash-through or with 0.23M NaCl, and can be resolved by SDS-PAGE into at least 8 distinct bands which we have tentatively characterized as sulfated glycoproteins. The remainder of the (35S) is in low molecular weight (LMW) material which does not adhere to DEAE-Sephacel and has not been further characterized.Platelets were treated with either thrombin or ADP, and the cells were then separated from the supernatant by centrifugation. The radiolabeled molecules in the supernatant and the cells were analyzed by DEAE-Sephacel and Sepharose CL-6B column chromatography. About 65% of the total cell (35S) was released from the cells by thrombin. Most of this radiolabel adhered to the DEAE-Sephacel column, and was found to be PG-1. The remainder of the released (35S) was about half the LMW material. In contrast, only 10-15% of the (35S)labeled material retained by the cells adhered to DEAE-Sephacel and was found to be PG-2. The remainder of the (35S)-labeled material retained by the platelets was the sulfated proteins and the LMW material. ADP caused release of about 15% of the (35S), and this was found to be in part PG-1 and in part the LMW material, but not PG-2. None of the (35S)-labeled molecules appeared to be degraded during platelet activation. We suggest that the PG-1 represent the a-granule and PG-2 the membrane proteoglycans. The sulfated proteins have not been described previously. Their role is not known, but we hypothesize that they may form part of the negative charge of the glycocalix and thus be part of the reactive surface of the platelet.
APPLICATIONS OF PLASMA ASSAYS OF CROSSLINKED FIBRIN DEGRADATION PRODUCTS (XLFDP) IN THE DIAGNOSIS OF THROMOEMBOLIC DISEASE
Authors: Whitaker A N , Masci P , Rowbotham B , Reasbeck P , Guerrini S M , McCaffrey J , Lee K , Elms M J , Bunce I H , Webber A J , Wylie F , Rylatt D B , Bundesen P G 
Published: August 2018

The presence of XLFDP in plasma, a consequence of the balance between fibrin generation and dissolution, is an indirect index of the presence of fibrin and raised levels accompany intravascular thrombosis. The potential of an enzyme immunoassay (EIA) for XLFDP, using the monoclonal antibody DD-3B6/22 was evaluated in 104 patients with suspected DVT receiving lower limb venography. Using a reference level of 400 ng/ml of XLFDP, all patients with a positive venogram had elevated levels (sensitivity 100%), although levels were also elevated due to causes other than DVT in some patients with negative venograms (specificity 52%). Higher elevations of XLFDP occurred in proximal than in calf DVT.A second study of another 48 patients receiving venography for suspected DVT confirmed the sensitivity of EIA while the complementary latex "Dimertest", also based upon DD-3B6/22, was positive in 19 and negative in 3 patients with confirmed DVT. In an ongoing study, 130 patients have been followed after abdominal operations with 125I-fibrinogren leg scanning and venography when appropriate. Levels of XLDFP (EIA) were elevated in all patients developing thrombosis. Latex assays were positive in 42/48 patients with positive leg scans. While elevated levels of XLDFP do not specifically define the location or even the presence of a thrombus they reliably indicate the possibility of thrombosis in patients without antecedent medical or surgical illnesses. In complicated surgical patients,an arbitrary cut-off of 1000 ng/ml is more appropriate for screening for the possibility of DVT. A positive latex test is also suggestive, although it is negative in some patients with calf thrombi. Measurements of XLFDP, with high sensitivity but moderate specificity, are potentially valuable in selecting patients for invasive investigations such as venography, and combinations with other noninvasive screening tests for require evaluation
PLASMA LEVELS OF FRAGMENT D-DIMER OF CROSSLINKED FIBRIN DURING THROMBOLYTIC THERAPY WITH RECOMBINANT TISSUE-TYPE PLASMINOGEN ACTIVATOR
Authors: Declerck P , Mombaerts P , Holvoet P , Collen D 
Published: August 2018

Plasma levels of crosslinked fibrin degradation products (XLDP) were measured before and at the end of the administration of rt-PA (40 to 100 mg over 1.5 to 8 hours) in healthy volunteers (n=5) and patients with deep venous thrombosis (DVT) (n=8), pulmonary embolism (PE) (n=16)and myocardial infarction(MI)(n=10). Determinations were performed using our newly developed ELISA, specific for crosslinked fibrin derivatives, based on two monoclonal antibodies (15C5 and 8D3H2) raised against purified human fragment D-dimer. All plasma samples were collected on citrate and trasylol. Results are expressed as mean and range of D-dimer equivalents (μg/ml).Baseline levels in patients with MI are only slightly elevated. The increased levels inDVT and PE are in agreement with previous studies. After infusion of rt-PA a small increase of XLDP is seen even innormal subjects. A very marked increasof XLDP is detected in patients with PE and DVT but not in patients with MI. This may reflect differences in the amounts of fibrin clot dissolved in these patient groups.No significant correlation was found between the increase of XLDP and success of therapy, although a significant difference in D-dimer levels was formed between the two groups with PE: successful (n=ll): 116 (range 61-192) vs. unsuccessful (n=5): 68 (36-155).Thus, XLDP are already elevated under baseline conditions in patients with DVT and PE and increase very markedly during thrombolytic therapy. The absolute levels after thrombolytic therapy do not strictly correlate with success of therapy. It could be useful to measure D-dimer levels during early stages of therapy, because the rate of increase of XLDP levels might correlate with the efficacy of thrombolytic treatment.
EFFECT OF INTERFERON GAMMA ON PROCOAGULANT ACTIVITY FROM HUMAN PROMYELOCYTIC CELL LINE (HL 60)
Authors: Gamba G , Dezza L , Montani N , Gangale G , Gangale G , Ascari E 
Published: August 2018

Interferon gamma (IFN), a lymphokine acting as biological response modifier, can induce morphological and phenotypic differentiation of some leukemic cell lines, specially along the monocytic pathway.Furthermore, myeloid leukemic cells and normal monocytes have been demonstrated to possess procoagulant activity (PCA).The aim of this study was to investigate the modulation of PCA induced by treatment with IFN on human promyelocytic cells from HL 60 cell line.Cells were cultured in suspension for 5days in RPMI 1640 medium supplemented with 10% fetal calf serum in the absence or in the presence of different concentrations of IFN (100-10.000 u/ml) at37°C and 5% CO2
Differentiation was assessed by morphological and cytochemical methods (MGG, ANAE, CAE, MPO) on cytospin preparations and surface marker analysis with monoclonal antibodies (0KMI, Mo2, MY9, MY7, MY4).PCA was measured as capacity to shortening ricalcification time of normal plasma and of factor VIII, VII and X deficient plasmas by the cells and the conditioned media.Untreated HL 60 cells exhibit high tissue factor-like PCA, related to the cellnumber. IFN treatment (1000 u/ml) induced at the same time, monocytic differentiation and significant increase in PCAboth in the cells and in the conditioned media.The PCA was not further affected by higher concentrations of IFN, unabletodetermine cell maturation.In conclusions the modulation by IFN seems to be dependent onmonocytic differentiation of HL 60 cells.
ANTITHROMBIN ROUEN-I(47 ARG→HIS) AND ROUEN-II (47SER) : TWO NEW VARIANTS WITH DECREASED HEPARIN AFFINITY
Authors: Borg J Y , Owen M C , Soria C , Caen J , Carrell R W 
Published: August 2018

Four families with AT-III variants of decreased heparin affinity have been compared with families with a quantitative deficiency of AT-III. The affinity variants had a lower incidence of thrombosis and, unlike thedefiency variants, had no increase in plasma FDF (defined by anti-D-neo ELISA assay). These results support a major physiological role for the progressive activity of AT-III.The characterisation of the affinity variants provides information onthe heparin-binding site. Variants were isolated from plasma of heterozygotes by NaCl elution gradient from a heparin-Sepharose column. Maps of tryptic and SA V8 peptides were compared on TLC and HPLC with analysis of aberrant peptides. Confirmation was by gas-phase automated sequencing.Two variants were unequivocally identified.1) Rouen-I (47Arg→His) wasfoundin a heterozygote with a history of a single cerebral occlusion.2) Rouen-II (47Arg→Ser) was in a forty years old heterozygote with an acute myocardial infarction but no other history of thrombosis.Three mutations of Arg47 are known: Cys(Toyama), His(Rouen-I) and Se(Rouen-II) all with decreased heparinaffinity.Functional and structural modelling studies are reported indicating apositive heparin-binding site formedby the A and D helices centred on Arg47 (A2). The mutation in Rouen-IIcreates an unutilised oligosaccharide attachment Asn⋆-Arg-Ser suggesting facultative glycosylation. This would, as an incidental observation, support a translational (pre-secretory) origin of the high affinity AT-III beta.
HEPARIN INDEPENDENT PURIFICATION OF ANTITHROMBIN III (AT III) BY IMMUNO-AFFINITY CHROMATOGRAPHY RESULTING IN A FUNCTIONALLY INTACT MOLECULE
Authors: Jørgensen M 
Published: August 2018

Previous methods for purification of AT III are based on its heparin-binding capacity. However, in congenital AT III deficiency abnormal inhibitor molecules with impaired binding of heparin and/or thrombin has been reported. The aim of the present study was to develop a purification method based on immuno-affinity chromatography, and thus independent of the heparin binding capacity.Rabbits were immunized with human AT III purified by a three-step procedure involving dextran sulphate precipitation, affinity chromatography on heparin-Sepharose and ion-exchange chromatography on DEAE-Sephadex A-50. Rabbit immunoglobulins against human AT III were isolated by affinity chromatography using purified human AT III coupled to CNBr-activated Sepharose 4B. Trace amounts of immunoglobulin against human albumin, IgG and IgM were removed by solid phase immunoadsorption. The highly purified immunoglobulins against human AT III were coupled to CNBr-activated Sepharose 4B. This anti-AT III-Sepharose was used for single-step purification of AT III from plasma. Elution was performed by Na-citrate buffer at pH 3.0 and the eluted fractions immediately neutralized. The recovery was more than 60%.The purified AT III appeared as a single protein band in SDS-poly-acrylamide gel electrophoresis with or without reduction. Affinity purified AT III and AT III purified by the three-step procedure were indistinguishable when analyzed by crossed immunoelectrophoresis in the absence and the presence of heparin isoelectrical focusing in polyacrylamid gel at a pH 4-6.5 gradient, and SDS-polyacrylamide gel electrophoresis. AT III antigen concentration was determined by electroimmunoassay and the reactive site concentration determined by titration with purified human thrombin using Phe-Pip-Arg-Nan (S-2238) as substrate. The ratio (active site conc.)/(antigen conc.) was identical in the two AT III preparations. It is concluded that this single-step immuno-affinity chromatography gives a high recovery from plasma of a highly purified functionally intact AT III molecule. The purification method is independent of the heparin binding capacity of AT III. This is of particular importance for the purification and characterization of abnormal AT III molecules with impaired heparin-binding site.
LMW-HEPARIN VERSUS HEPARIN-DHE IN ORTHOPAEDIC SURGERY
Authors: Breyer H G , Rahmanzadeh R , Bacher P , Werner B 
Published: August 2018

The efficiancy and the side effects of a LMW heparin (FRAGMINR, KabiVitrum) and Heparin-DHE (Sandoz) have been compared in a randomized open prospective study of 120 patients (60/60) undergoing elective surgery on the lower limbs (total hip and knee replacement, corrective osteotomies). A radiofibrinogen uptake test (RFUT) was regularly done on all patients. Positive tests were controlled by ascending phlebography. The parameters, clinically obtained, included the intra-and postoperative blood loss, wound closure, and the incidence of haematoma. Hb, Hk, red and white blood cells, thrombocytes, total protein, aPTT, AT III, TT, and anti-Xy-activity were analyzed at the day before operation, the 2nd, 4th, and 6th day after operation.There were three positive RFUT in the group of LMW heparin (5 per cent), and there were six (10 per cent) in the control group. No pulmonary embolism occurred. In no case an operative treatment of deep vein thrombosis was done. There were no statistically significant differences in intra- and postoperative blood loss, and in the laboratory data, except the anti-Xa-activity, which was significantly higher in the LMW heparin group.The comparative study has shown, that a single daily injection of LMW heparin (FRAGMINR) is more effective than the two daily injections of the combination of UF heparin and DHE in order to prevent postoperative thromboembolism
ENOXAPARINE (LOVENOXR), VERSUS STANDARD HEPARIN IN PROPHYLAXIS OF DEEP VEIN THROMBOSIS (DVT) AFTER TOTAL HIP REPLACEMENT (THR)
Authors: LE BALC’H T , LANDAIS a , BUTEL T , WEILL D , PASCARIELLO J C , PLANES A 
Published: August 2018

THR is associated with a high risk of thromboembolic complications. Enoxaparine, LovenoxR, a low-molecular-weight-heparin, and standard heparin were compared in their abilities to prevent DVT in patients undergoing THR. The efficiency and the bleeding risk of each treatment were studied.237 patients, with a non traumatic hip disease, requiringTHR, were included in a multicentric, randomized, double blind trial. Mean age:65.8 years ± 9.2; mean weight :67.3kg ± 1.3.113 patients received standard heparin, 5000 UI/8 hrs, by"subcutaneous (SC) injection. 124 patients received Enoxaparine, 40 mg/24 hrs, by SC injection. Administration of drugs was begun 2 hours before operation for standard heparin, 12 hours before operation for Enoxaparine. Patientswere treated for 10-15 days, until bilateral ascending phlebography (BAP) had been completed.Lower limbs BAP were performed in 228 patients. The incidence of DVT was significantly lower in the Enoxaparine group : a DVT was detected in 15(12.5 %) of 120 patients who received Enoxaparine and in 27(25%) of 108 patients who received standard heparin (p=0.014).A pulmonary embolism occurred in 1 patient of the heparin group, in none of the Enoxaparine group.The frequency of bleeding complications was significantly lower in the Enoxaparine group. A post operative wound hematoma occurred in 1 patient of the Enoxaparine group and in 3 patients of the heparin group. Red cell transfusions requirements were significantly lower in the Enoxaparine group (3.37 U ± 1.81) than in the heparin group (3.84U ± 1.70)(p=0.03). The hemoglobin level was significantly higher, on the 3rd, 4th post operative day, in the Enoxaparine group.Subcutaneous Enoxaparine (40 mg/24 hrs) was significantlymore efficient than subcutaneous heparin (5000 UI/8 hrs) in preventing DVT, in patients undergoing THR. The incidence of bleeding complications was significantly lower in the Enoxaparine group.Enoxaparine (LOVENOXR) - PHARMUKA S.F.