bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitative Abnormalities of Aα Chain Molecular Weight in the Fibrinogen of Cirrhotic Patients
Authors: Weinstein Mark , Deykin Daniel
Published: May 2019

A novel two-stage SDS gel electrophoretic procedure was devised to examine the molecular weight heterogeneity of fibrinogen in small samples of whole plasma from 12 normal and 7 cirrhotic individuals. Fibrinogen was first separated from other plasma proteins on a large pore gel, cut out of the gel, reduced, and separated into its component Aα, Bβ, and γ chains on a second gel. Two major mol wt species—fibrinogen I and II—were observed on the first gel. The ~ 25000 mol wt difference between these two forms reflected a decrease primarily in the size of one of the fibrinogen II Aα chains. In both normals and cirrhotic patients fibrinogen II comprised 30% of the total (range 20–35%). Fibrinogen I and II each contained two major high mol wt Aα chains—Aα/1 and a smaller Aα/2—that differ by 3000 mol wt. In normal fibrinogen I, Aα/2 comprised 33% of the total Aα chains (range 27–41%). In contrast the fibrinogen I of 6 out of the 7 patients had a lower per cent of Aα/2 (range 10–25%). Similar quantitative differences were seen in the decreased fraction of Aα/2 in the fibrinogen II of cirrhotic patients compared to normals. No correlation was found between per cent fibrinogen II and per cent Aα/2 in either normal subjects or cirrhotics. These results suggest that at least two independent processes are responsible for the observed levels of Aα chain heterogeneity in normals and cirrhotics and that one of these processes yields a lower than normal fraction of Aa/2 chains in the fibrinogen of cirrhotic individuals.
The Structural Characteristics of Chicken Fibrinogen
Authors: Pindyck J. , Mosesson M. W. , Bannerjee D. , Galanakis D.
Published: May 2019

The structure and subunit composition of chicken fibrinogen(ϕ) have been investigated. Dodecyl sulfate gel electrophoresis of unreduced specimens revealed a single ϕ band with a molecular weight of approximately 320,000. ϕ and fibrin specimens were also electrophoresed after reduction with dithiothreitol, and after crosslinking of unreduced specimens in the presence of Factor Xllla. Chromatographically separated S-sulfo chains were also studied after reptilase or thrombin treatment,and certain samples were stained with periodic acid Schiff reagent(PAS). Chicken Aα chains weresmaller than human Aα chains (54,500 vs.70,900, respectively) but, like mammalian Aα chains, they possessed a reptilase and thrombin sensitive site, were PAS negative,and undergo Factor XIIIa catalyzed α-polymer formation. The sizes of chicken Bβ and γ chains were nearly thesame as their mammalian counterparts, (i. e. 60,000 and 49,000 respectively) ; both types of chains were PAS positive. Chicken Bβ chains possessed a slowly reactive thrombin sensitive site apparently corresponding to the one in human ϕ; the chicken β chains, like mammalian β chains, did not undergo Factor XIIIa catalyzed cross-linking. Like mammalian γ chains, chicken γ chains could undergo Factor XIIIa catalyzed γ-γ dimerization and did not possess thrombin or reptilase sensitive sites. These findings indicate that the chicken fibrinogen molecule is composed of three pairs of disulfide-bridged chains corresponding in most respects to mammalian fibrinogen chains.
Size and Shape of Human Fibrinogen in Solution
Authors: Hofmann V. , Straub P.W. , Binkert T. , Serrallach E. , Känzig W. , Zulauf M.
Published: May 2019

In order to obtain information on size and shape of the fibrinogen molecule in solution the translational diffusion coefficient (DT), the rotational diffusion coefficients (DR⊥ and DR//) and the sedimentation coefficients (S) have been measured on human fibrinogen with a clottaoility above 95%. The methods used were dynamic light scattering, nanosecond fluorescence depolarization and analytical ultracentrifugation. Dynamic light scattering yields DT = 2.0 ± 3% x 10-7 cm2sec–1 at a concentration of 7 mg/ml in 0.15 M Tris-NaCl, pH 7.4. DT is strongly dependent on concentration, being 3.4 ± 10% × 10-7 cm2 sec-1 at 0.1 mg/ml. The rotation along the minor axis as measured with the same method is DR = // 1.5 × 106 sec-1 at 0.1 mg/ml. The rotation along the major axis as measured on fibrinogen labeled with dansylchloride is DR// = 1.5 x106 sec–1. S is also strongly dependent on concentration, being 7.9 S at 0.1 mg/ml, 8.1 S at 1 mg/ml and 6.6 S at 10 mg/ml.

These results fit with an elongated molecule having an axial ratio of 7. They are compatible with a MW of 340’000 only for concentrations above 2 mg/ml, while at lower concentrations (0.1 mg/ml) they agree with a MW of approximately half the accepted value.
Prospective Study on the Value of a Single Infusion of Dextran 70 During Surgical Operation in the Prevention of Fatal Postoperative Pulmonary Embolism (PE)
Authors: Ljungström Karl-Gösta
Published: May 2019

At Danderyd’s Hospital a controlled clinical trial of dextran 70 using phlebography in all patients demonstrated a significant reduction in postoperative deep venous thrombosis (DVT) from 52% in the untreated control group to 4% in the dextran group. Johnson et al. 1968. As a consequence, routine prophylaxis of postoperative DVT with dextran 70 was instituted for all patients over 50 years of age undergoing abdominal, orthopedic or urological surgery. A retrospective investigation of. the result of this two-year period of routine prevention with dextran 70, which was preceded and superceded by 2 two-year periods when no dextran was given, revealed the following sequence of fatal PE verified at autopsy over the six-year period:15 (no prophylaxis), 6 (dextran) and 15 (no prophylaxis), Ljungström, 1975.

At this stage a prospective study for a new two-year period was started. Infusion of 100 ml of dextran 70 was started during operation in all patients filling one of the following criteria: 1. All patients over 50 years of age undergoing abdominal, orthopedic or urological surgery. 2. All patients undergoing re-operation regardless of age. 3. All patients undergoing major surgery with a history of previous thromboembolic disease regardless of age.

During this last two-year period only 3 patients died of fatal postoperative PE. The difference between 15 fatal PE in the non-dextran and 3 in the dextran group was significant, P 0.05. A comparable number of surgical operations were performed during each year of the eight-year period.
Concentrated and Diluted Heparin-Prophylaxis of Postoperative Deep Vein Thrombosis
Authors: Törngren S. , Forsberg K.
Published: May 2019

Two calcium heparin solutions (Vitrum AB) have been evaluated in a controlled blind study with standard dosage of 5000 IE subcutaneously. The injections were given 2 hours before operation and 12-hourly for 6-8 days after operation. 175 patients undergoing abdominal surgery were included and randomized into 3 groups. 75% of the patients had malignant disease. 66 patients recieved heparin 500 IE/ml (1 ml), 47 patients recieved heparin 25000 IE/ml (0.2 ml) and 62 patients recieved 5% glueos. Deep vein thrombosis (DVT) was diagnosed with the 125I-fibrinogen method. Finally the heparin solutions with different viscocity were compared in an in vitro experiment. The nurses who injected the patients gave injections into weighed bottles with the usual technique.

DVT was diagnosed in 32% of the control patients, 15% of those on heparin 5000 IE/ml and 21% of those on heparin 25000 IE/ml. The difference between controls and heparin 5000 IE/ml is statistically significant (p<0.05). In the in vitro experiment the mean dosage of heparin 5000 IE/ml was 5100 IE and the mean dosage of heparin 25000 IE/ml was 4415 IE. The dosage of heparin 25000 IE/ml was significantly less than 5000 IE (p<0.0l).

We conclude that the frequency of DVT is reduced with calcium heparin 5000 IE/ml (1 ml) 12-hourly for 6-8 days after operation. The concentrated heparin 25000 IE/ml (0.2 ml) does not significantly reduce postoperative DVT, which probably can be explained from our results in the in vitro study.
Coagulation Studies on Synthetic Benzyl Glutamate-Leucine Copolymers
Authors: Parks P.J. , Malhotra O.P. , Gibbons D.F.
Published: May 2019

To obtain a polymer(s) which is passive to blood proteins so that it can be used as a prosthetic material, a series of uncharged amino-acid copolymers containing:a) 20% benzyl ester of glutamic acid and 80% leucine (GL-14); b) equal amounts of glutamic acid and leucine (GL-11); c) 80% glutamic acid and 20% leucine (GL-41); d) 100% benzyl glutamate (G-1); and e) 80% benzyl ester of glutamic acid and 20% phenylalanine (GP-41) were tested for initiation of clot formation. The effect of solvent was also taken into consideration because films of a particular polymer cast from dioxane, dichloroethane and benzene exhibited different morphology and different polar and total surface energies. We investigated the activation of the Intrinsic and extrinsic pathways of blood coagulation including the adhesion of platelets and release of platelet Factor 3. In all these studies, glass and siliconized glassware were included to serve as negative and positive controls, respectively. For the extrinsic system, glass and GP-41 had the greatest effect in the activation of Factor VII and GL-14 and G-1 the least; siliconized glass, GL-11, and GL-41 were between these two extremes (p < .001). For the intrinsic pathway, when the solvents used for casting purposes were benzene or dichloroethane, glass and GL-11 had the greatest effect, GL-14 and siliconized glass the least, and G-1, GL-41 and GP-41 an intermediate effect (p < .001). However, when the solvent used for casting purposes was dioxane, GL-14 was even more passive than siliconized glass in the activation of the intrinsic system; G-1 was as effective as siliconized glass. Again, the effects of the polymer (p < .001) and of solvent (p < .0025) were significant. For adhesion of platelets, the polymer containing 100% benzyl glutamate was most compatible; the number of platelets attached to this surface was minimal.
Interactions of the ADP Analogs, 2′-0-Methyl ADP and Adenine 9-β-D-Arabinofuranoside 5′-Diphosphate and Catecholamines with Human Platelets
Authors: Ragatz B.H. , Iatridis P.G. , Iatridis S.G. , Markidou S.G. , Asterita M.F. , Gadarowski J.
Published: May 2019

The two analogs 2'0-methyl ADP (Me'ADP) and adenine 9-β-D-arabinofuranoside 5'-diphosphate (A-Ara-DP) are compounds with stereochemical modification in the pentose moiety of ADP. These compounds have been added individually to citrated platelet rich plasma (PRP) and the concentration dependent interaction of each with platelets has been studied turbidimetrically using a dual channel Payton Aggregometer under the following experimental conditions:variable amounts of compound only; variable amounts of compound challenged at 30" or 5' by an aggregation-stimulating ADP concentration; or variable amounts of compound plus aggregation-stimulating concentrations of epinephrine (EPI) or norepinephrine (NOR) added after 30".

Data obtained indicate that both compounds induce reversible aggregation at high concentrations, and that neither compound competes effectively with ADP added after 30". Both compounds delay the secondary wave of aggregation when ADP is added after 5'. Both compounds also exhibit mild dose dependent cooperative effects when epinephrine or norepinephrine are added 30" later.

The ADP analogs, Me'ADP and A-Ara-DP, exhibit weak interactions with the platelet ADP receptor, indicating the importance of the ribose conformation in this case. The mild cooperative effects for either of these analogs plus EPI or NOR also suggest that the platelet ADP and catecholamine receptors are independent but interacting. Supported by USPHS HL-15425.
Platelet Factor XI
Authors: Castaldi P.A. , Connellan J. , Smith I. , Michel H.
Published: May 2019

Contact factors are known to be associated with platelets. There is evidence to suggest that F XI may mediate an enhancement of coagulant activity in response to collagen stimulation of platelets. The aim of this study was to further explore this relationship using fresh platelets and extracts prepared from normals and a F XI deficient subject. Antisera against plasma F XII and F XI were used in tests of collagen induced coagulant activity and the latter antibody in immunoprecipitation studies with platelet extracts. Comparisons were made with antisera to isolated platelet membranes.

Collagen induced coagulant activity was inhibited by anti F XI and to a slight degree by anti F XII. F XI-like coagulant activity could be detected in extracts of washed platelets and this activity adsorbed to heparin-sepharose as does plasma F XI. Similar activity could be detected in platelet extracts from the F XI deficient patient. Normal platelets contained an antigen that precipitated with anti F XI.

F XI activity and antigen are closely associated with platelets. This may provide a mechanism for enhancement of coagulation.
Significance of Fibrinogen/Fibrin Degradation Products (FDP) after Cardiovascular Surgery
Authors: Altman R. , Zielinsky A. , Rouvier J. , Favaloro R.G.
Published: May 2019

The frequent appearance of FDP during the postoperative of cardiovascular surgery using extracorporeal circulation, induced us to determine its diagnostic value. 17 patients were studied. Fibrinolytic activity was determinated using the staphylococcal clumping test (SCT) and the latex plasmmin-antiplasmin immunoassay (LPA). Normal value of LPA in plasma previously studied in 15 normal patients was negative to positive up to a dilution of 1/4. Normal value of SCT in our laboratory (in 200 normal patients studied) was less than lug/ml. All postoperative periods were normal and from 1 to 6 tests using both methods were performed. From 36 SCT readings, 27 were positive (in 13 patients) with a mean of 10.2ug/ml (range 1.2-96.8). Positive LPA was found in 5 readings in 4 patients. In only 2 cases the SCT detected high concentration of FDP together with strong positive LPA (dilutions 1/10 and 1/20). It is concluded that the frequent appearance of FDP during early postoperative period of cardiovascular surgery, because of the low incidence of positive LPA, does not necessarily indicate plasmatic fibrinolytic activity, as it is possible that this should appear as a result of absortion of extravascular resolution of fibrin related material.
Characterisation of Serum FDP Produced During Enhanced Fibrinolysis
Authors: Lane D.A. , Kakkar V.V.
Published: May 2019

The serum FDP produced in patients in response to defibrination with ancrod and to thrombolytic therapy with intermittent streptokinase/plasminogen infusion have been characterised using a method of solid phase immunoprecipitation followed by Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (1). Using this method it is possible to distinguish between the plasmin degradation products of fibrinogen and Factor IIIa induced crosslinked fibrin. During ancrod administration, fragments of similar mobility to fibrinogen fragments X, Y, D and E are present in serum samples taken 24 hours after the initiation of treatment, while subsequent sera contain mainly fragments with the same mobility as fibrinogen fragment D. The intermittent nature of the streptokinase/plasminogen infusions produces fibrinogen fragments X, Y, D and E in the sera 1 hour after each daily streptokinase/plasminogen administration. There was an early clearance of fragment E from the circulation of patients receiving either ancrod or streptokinase/plasminogen infusions. The presence of the D dimer fragment, which is produced only by plasmin lysis of fibrin crosslinked with Factor XIIIa, could not be conclusively confirmed in either group of patients. 1. Lane, D.A., Scully, M.F. and Kakkar, V.V. Thrombosis Research, 9, 191, 1976.
Microembolism, Diffuse Intravascular Coagulation and Respiratory Insufficiency
Authors: Edmunds Henry L.
Published: May 2019

Clinically significant pulmonary microembolism and/or diffuse intravascular coagulation (DIC) may occur in critically ill patients with hemorrhagic shock, septicemia, multiple injuries, transfusions, open heart surgery and certain other diseases. Platelet aggregates, fibrin, fibrin-platelet aggregates and fat emboli may form in stored and extracorporeally circulated blood or during DIC. While evidence indicates that many microemboli break up in lung capillaries, others block capillaries or induce a local reaction. In DIC emboli and thrombosis are thought to occlude pulmonary capillaries. The result is a mismatch in ventilation and perfusion, arterial hypoxemia, decreased compliance, increased pulmonary vascular resistance and respiratory insufficiency which may progress to acute respiratory failure. In severe cases lung biopsies and autopsies show hemorrhagic atelectasis with blood found in peribronchial and perivascular spaces and in alveoli. Few emboli or thrombi are seen. Since platelet aggregates may disaggregate and fibrin may lyse, the cause and effect relationship between pulmonary microembolism and respiratory insufficiency in critically ill patients is implied rather than proven. The pathogenesis of the pulmonary injury may be related to circulating substances or substances released from emboli that cause leakage of blood through capillaries. Experimental bland microemboli do not reproduce respiratory insufficiency as seen clinically.
Anticoagulation in Cardiopulmonary Bypass
Authors: Thomas Kisker C. , Young John A. , Doty Donald B. , Taylor Barbara J.
Published: May 2019

Prolonging the activated clotting time (ACT) 2 to 3 times normal is said to provide a “safe” level of anticoagulation during cardiopulmonary bypass. To test this level of anticoagulation 9 monkeys were anticoagulated with heparin at the start of cardiopulmonary bypass so that ACT’s ranged from 201 sec to > 1000 sec (normal 91 sec). ACT, platelet count (P), fibrinogen (F), and fibrin monomer (FM) were measured at 10, 30, 60, 90, and 120 minutes during bypass. Antithrombin III (AT3) was measured before and after bypass. Six monkeys developed increased FM indicating active coagulation beginning from 10 to 60 minutes on bypass. ACT’s were > 200 sec in all animals at the time of FM detection. P fell below 100,000/mm3 in the 6 animals with elevated FM, but remained above 100,000/mm3 in the other 3 animals. The mean value of AT3 (69%) decreased to 24.4% after bypass in the 6 animals with elevated FM, but was 61% after bypass in the others. Scanning electron microscopy of the oxygenator membranes showed significant amounts of fibrin on the membranes used in monkeys who developed increased FM levels, but not on those with normal FM concentrations. F decreased from 167 mgm/dl to 80.5 mgm/dl in monkeys with elevated FM and to 117 mgm/dl in those with normal FM concentrations. Excessive bleeding did not occur in the animals without increased FM although ACT’s were in excess of 1000 sec. Subsequently three human subjects on cardiopulmonary bypass whose ACT’s were maintained above 400 sec have not shown increased FM levels. The results suggest that prolonging the ACT more than 2 - 3 times normal is required to prevent activation of clotting during cardiopulmonary bypass.
Aes-, Esca-, Fibrinogen Adsorption and Bioluminescence Studies of Glutardialdehyde-Stabilized Heparin Surfaces
Authors: Larsson Rolf , Eriksson Jan Christer , Lagergren Hans , Olsson Per , Rosengren Åke
Published: May 2019

The chemical constitution of glutardialdehyde-stabilized cetylamine hydrochloride heparin (CAHG) surfaces was investigated with AES (Auger Electron Spectroscopy) and ESCA (Electron Spectroscopy for Chemical Analysis) before and after plasma exposure.

In comparison with a pure heparin film, the CAHG-surface contains a substantial amount of water-insoluble Schiff’s bases and its heparin surface concentration is about a factor of ten lower.

On exposure to plasma or blood, proteins are heavily adsorbed onto the CAHG-surface. In contrast to the protein adsorbate on a polyethylene surface, which contains 200 nanograms of fibrinogen/cm2 surface area, the protein adsorbate on CAHG includes no functional fibrinogen. The surface-bound fibrinogen has been measured by radio-immunoassay of fibrino-peptide A split off when the surface is treated with thrombin or Defibrase.

The adhesion of ATP-containing cellular material on the surfaces after exposure to blood has been explored with the aid of bioluminescence. These studies show that platelets and some other blood cells adhere to polyethylene surfaces, whereas adsorption of cells does not occur on CAGH-surfaces.

It is concluded that the adsence of fibrinogen and cellular material on the CAHG-surface is associated with its non-thrombogenic properties.
Systemic Streptokinase Treatment of Chronic Arterial Occlusions - Clinical Results
Authors: Martin M. , Auel H.
Published: May 2019

435 arterial occlusions were treated with streptokinase over a period of 3 days. A continuous streptokinase regimen with a maintenance dose of 100 000 u SK/h was administered. The successful removal of femoral obstructions was possible in 8.9%. A complete dissolution of the iliac thrombus masses had been achieved in 19%. 41 aortic occlusions were treated. The treatment was capable of removing the aortic obstructions in 24%. It became evident that there was a close relationship between the average occlusion time and the respective clearance rate. Por example, in patients undergoing lysis treatment during the very first two weeks after femoral occlusion, clearance was established in 75% and best lytic results were achieved if the history of iliac occlusion was well below 3 months duration. The relationship of vessel clearance rate and walking distance showed, that on the average there was no lengthening in walking distance where treatment had failed (no vessel clearance obtained). On the other hand, a significant average rise in walking distance from initially 325 up to 800 meters (p < 0.05) was seen when vessels clearance had been recorded.
Natural Plasma Inhibitors in Individuals with High Risk for Thrombotic Disease
Authors: Engel Araceli M.
Published: May 2019

Our modification of the assay for the inhibitor of factor Stuart activated (Anti-Xa) was evaluated together with the antiactivator of plasminogen (Antiplasm.) and the nature of the fibrinogen degradation products (FDP) as a test of the risk for thrombotic disorders. Thirty patients with acute myocardial infarction (AMI), twenty eight in the chronic phase (CMI) and thirty healthy controls (C) were studied. The age range for the three groups was 45-85 with a mean age of 57-62. The activity of Anti-Xa in C was 46.7 U/ml (± SD 11.5); 31.5 U/ml (± SD 16.5) with p<0.001 for the AMI group and 42.1 u/ml (± SD 17.9) for the GMI group. Very high activity for Antiplasm. in the AMI group was found (11 min ± SD 7.1) with p < 0.001 for a C value of 3 min 36 sec (± SD 1.2). For the CMI group the value was 6 min 11 sec (± SD 3.3) and p < 0.001. The FDP were elevated in the AMI group with a mean value of 15.5 polimerization inhibition units (PIU/ml) being O PIU/ml the C value. The CMI group had a mean value of 4.3 PIU/ml. The ratio between PIU/ml and µg/ml of the FDP indicated products of “early” nature in AMI and “late” type in CMI, The findings mentioned above were similar when the patients received anticoagulants except for the nature of FDP that was changed to the “late” type in both groups. The simultaneous determination of Anti-Xa, Antiplasm. and the nature of the FDP can be applied to other pathologies. The comparison of these parameters may indicate the oncoming of an acute thrombotic episode and can be useful in evaluating the status of the cardiovascular system during cardiac rehabilitation.
Skin Necrosis Complicating Oral Anticoagulant Therapy
Authors: Krzywanek H.J.
Published: May 2019

Since the first report in 1943 far more than 100 cases of skin necrosis as a complication of oral anticoagulant therapy have been reported, mainly in European countries. Breast, thigh, legs and abdomen are the predominant sites of the lesions, obese women are most frequently affected.

10 cases which we observed over the last ten years are presented and the etiology is discussed. The common feature of our patients is a history of fever, probably caused by a bacterial or viral infection, treated with a variety of antibiotics. A complicating or preexisting thrombosis or thrombophlebitis of the leg leads to the initiation of oral anticoagulant therapy. Regularly on the 4th (3.-5.) day of anticoagulant treatment the skin necrosis develops within hours. We suggest that the administration of antibiotics, and perhaps also the infectious process are predisposing factors. The high loading dose at the beginning of coumarin treatment and an abnormal sensitivity of the patient seem to be other important etiologic factors for coumarin necrosis. These points are substantiated by the facts that skin necrosis occurs only in the first days after the start of oral anticoagulant therapy, and by one of our patients who developed another skin necrosis when coumarin therapy was repeated several years later. Oral anticoagulant therapy should not be initiated when antibiotics must be given at the same time. In case anticoagulant treatment is necessary during antibiotic therapy we propose the administration of heparin instead of using coumarin derivatives.
Some Platelet Function Tests Normalized by ICI 55,897 and Clofibrate
Authors: O’Brien J.R. , Etherington M.D. , Jamieson Sandra , Shuttleworth R.
Published: May 2019

We have shown (Lancet, 2, 995, 1976) that in normal men a diet substituting linoleic acid for most of the normal saturated fat intake is followed by marked changes in some of those platelet function tests which are abnormal in the prothrombotic state. Are these changes due to alterations in the plasma lipids?

Clofibrate (Atromid-S) and an analogue, ICI 55,897, were given in double blind cross over studies to respectively 10 and 19 patients with severe peripheral arterial disease and/or a past history of MI. Preliminary analysis indicates that in 3 weeks on 55,897 the mean heparin thrombin clotting time was normalized from 25 seconds pre-treatment to 30 seconds. The fibrinogen decreased from a mean of 371 to 332 mg/%, but this treatment had no effect on the anti-thrombic activity and no effect on the plasma lipids. In the Clofibrate study the lipids and the anti-thrombic activity decreased; the HTCT was probably ameliorated but a marked placebo effect confounded the statistical analysis. However, clinical records of patients to be presented strongly support these conclusions. Thus no simple single mechanism, e.g. a change in lipids, can account for the changes achieved. These observations, however, establish that these tests regularly found to be abnormal in groups of patients at risk of further thrombosis can be changed towards normal in patients by two ICI compounds. It is now vital to find out if these changes are beneficial to the patient.
Comparison of Two Tests for Prediction and/or Diagnosis of Thrombosis
Authors: Ridgway H. , Weirich C. , Broom V. , Speer R. J. , Hill J. M.
Published: May 2019

Two laboratory procedures: the heparin thrombin clotting time (HTCT) of O’Brien et al and the platelet catalytic activity (PCA) of King and Joffe are being investigated for their usefulness in prediction and/or diagnosis of a thrombotic or hypercoagulable state in patients with a variety of diseases. Included are patients with current thrombosis, patients with a past history of thrombotic disease, as well as those with a negative history. The HTCT has been reported to correlate with release of platelet factor 4 from platelets destroyed in thrombotic episodes. The PCA was designed and used by King and Joffe to predict successfully postoperative venous thrombosis. In this study, both the PCA and HTCT showed a useful correlation with other laboratory and clinical findings in patients with previous and current thrombosis. In addition, clinical situations in which these tests appear to give misleading results will be presented and analyzed. While this study represents relatively small numbers of patients, the degree of correlation seems to indicate that both the HTCT and PCA have value in assessment of hypercoagulability and thrombosis.
Activation of the Coagulation System by Endotoxin in Frog (Rana Esculenta L.)
Authors: Csåko G. , Gazdy E. , Csernyånszky H. , Toth S. , Szilågyi T.
Published: May 2019

The frogs, unlike mammals, are extremely resistant to the lethal effect of gram-negative endotoxins. Even injecting enormous doses we were unable to determine the DL50 value at their usual ambient temperature. The finding by Levin and Bang that endotoxin activates the coagulation in the invertebrate Limulus suggested, however, a similar effect in frogs as well. Frogs were, therefore, injected with a high dose of E. coli Olll endotoxin (Boivin). Four hours later the fibrinogen level was 30 %, the thrombocyte count 50 %, and the neutrophil count 70% lower than the initial value. By 24 hours a further drop of fibrinogen level appeared, whereas the thrombocyte and neutrophil counts started to normalize. At that time, repeating the endotoxin injection (generalized Shwartzman reaction) a decrease of all 3 parameters studied was again observable in 4 hours. When frogs were treated i.p. with high endotoxin doses 72: 72 and 48; or 72, 48 and 24 hours before blood sampling, progressively prolonged prothrombin (Quick) and thrombin times were obtained, indicating a consumption coagulopathy. (Unfortunately, reptilase times are hardly measurable in frog plasma- Blut 30: 233, 1975) Also, abnormal clotting times were found when according to the previous schedule protamine HCl, known to exert multiple effects on the coagulation system, was given frogs (50 mg/100 g). The results show that endotoxin is able to trigger, like in mammals and Limulus, the clotting mechanism in frog.
Effect of Cryoprecipitate Infusion on Factor VIII Response to Venous Congestion in Patients with von Willebrand’s Disease
Authors: Morfini M. , Longo G. , Ferrini Rossi P. , Seri A.
Published: May 2019

In patients with Von Willebrand’s disease(VWD) venous congestion(v.c.) is un able to increase the VIIIAHF and VIIIAGN level as in normal subjects. To evaluate the effect of rise in VIIIAHF and VIIIAGN levels, 5 patients with VWD received single infusion of lyophilised cryoprecipitate(15 U./Kg) for treatment of bleeding episodes. Two v.c, at 15min and at 300min after the completion of infusion, one on each arm, was performed. VIIIAHF plasma level, raised by infusion, did not show further increase after v.c. at 15min. This finding may suggest that increased levels after v.c. in normal subjects is not achieved by activation of plasma VIIIAHF. In contrast at 300min a significant increase of VIIIAHF was obtained(F test p<0.05). On the other hand VIIIAGN high plasma level immediately after infusion showed a significant rise(F test p<0.05) after v.c. at 15min but much less at 300min when VIIIAGN is reduced. The divergence on response of VIIIAGN and VIIIAHF to v.c. at different times after infusion may support the mechanism of VIIIAGN conversion to VIIIAHF in VWD. As the plasma VIIIAHF activation appears to be unlikely, the endothelial cells may be responsable for release of AGN and AHF i.e. for conversion of AGN to AHF.
Platelet-Like Bodies in Urine in Some Blood Disorders
Authors: Kōno N. , Chen T.P. , Suzawa N. , Mizokami T.
Published: May 2019

We had reported that it is always observed a small body resembling a platelet in urine of normal and various sick persons and the small body is a platelet itself or a ruin of platelet (Lancet, 1, 1339, 1975., N. Engl. J. Med., 293, 44, 1975., Nihon Univ. J. Med., 17, 117, 1975., Japan. J. Clin. Hemat., 17, 632, 1976., Jap. J. Urol., 67, 903, 1976.). We had studied the Platelet-like bodies in urine of 3 cases suffered from ITP, 3 cases suffered from AML, 2 cases suffered from CML, and one case suffered from MOL, SLE and reticulosarcoma, respectively. The small bodies were also observed in all urine of these patients. By transmission electron microscopy, these small bodies were a platelet itself or a ruin of platelet. And the TEM findings showed differencein each blood disorder and treatment.
The Effect of Enhanced Fibrinolytic Activity on the Red Cell Membrane Transport
Authors: Ulutin S.B. , Emekli N. B. , Yardimici T. U.
Published: May 2019

In dogs and in human subjects, using hepatic vein catheterization before and after the activation of the fibrinolytic system, the blood samples were obtained and the red cell amino acid transport was investigated. The time course accumulation of radioactive histidine in isolated red cells was followed together with the measurements of the fibrinolytic activity.

A decrease in the active transport of histidine was observed in the red cells after the stimulation of the fibrinolytic system. Also a correlation between the decrease of active transport and the increase of fibrin-fibrinogen degradation products was seen.
The Sonoclot: A New Thrombokinetic Coagulation Instrument
Authors: Hamstra R.D. , Ens G.E. , Simons S.
Published: May 2019

A new coagulation instrument (Sonoclot) has been evaluated in the laboratory, at the bedside, and in surgery. It was compared to the prothrombin time, activated partial thromboplastin time, activated clotting time, Lee-White clotting time, fibrinogen and thrombelastograph. Most coagulation tests are reported as time to a clot end point while the Sonoclot records a continuous thrombokinetic pattern by measuring the energy required to maintain axial vibration of a hollow plastic probe in 0.4 ml of whole blood while it clots. Disposable plastic probes and non-siliconized glass cuvettes are rapidly changed after testing. Results are a line pattern which has been analysed for Clot Onset (C0) and Clot Formation Rate (CFR). Native whole blood from normal adults (54 studied) has a C0 of 2-4.5 minutes and CFR of 4-8 chart units/minute. The only similar clotting instrument is the thrombelastograph (TEG). Sonoclot and TEG patterns correlate, the Sonoclot sensing change before the TEG. Whole blood clotting measured with the Sonoclot is a new method which distinguishes hypercoagulation, various levels of heparin, and severe bleeding problems from normal. The device is durable, stable, easy to operate, and portable, providing a permanent record in a few minutes.
Testing of the Aggregating Activity from Various Batches of a Platelet Specific Collagen Preparation
Authors: Schulte F.
Published: May 2019

Producing of a collagen preparation for determining platelet functions requires the testing of the activity of each single batch to make possible a standardization. For this purpose methods using PRP (platelet rich plasma) to measure the platelet aggregation are not very appropriate. Since four years a collagen centrifugation test proves to be suitable for determining the activity of various batches of Collagenreagent “Horm”. In this test the platelets of citrated whole blood are mixed with graduated doses of collagen directly by the withdrawal of blood. The turbidimetrical evaluation of the PRP obtained from the collagen treated whole blood samples shows 15% (SD ± 3. 5 %) of the O.D. (optical density) of the PFP (platelet free plasma) after a dose of 1 meg and 80% (SD ± 2 %) after a dose of 2 meg collagen per ml whole blood. The aggregation of platelets in citrated whole blood simultaneously with graduated doses of collagen allows to standardize different batches of platelet specific collagen preparations.
Studies on Human Serum Inhibitors of Plasmin, Urokinase and Plasma Kallikrein Using Chromogenic Substrates
Authors: Gallimore M.J. , Fareid E.
Published: May 2019

Human serum inhibitors of plasmin, urokinase and kallikrein were studied using chromogenic substrate assays,(Plasmin substrate, S-22 51, Kabi, Sweden, Urokinase substrate, Chromozyme -UK and Plasma Kallikrein substrate, Chromozyme -PK, Pentapharm, Basle, Switzerland). In whole serum both “immediate” and “time dependent” inhibition of plasmin and kallikrein was observed, whilst only very weak inhibition of urokinase was detected. When serum samples were fractionated by Sephadex G-200 gel-filtration the “immediate” plasmin inhibitors were identified as α2-macroglobulin and low molecular weight antiplasmin whilst α2-macroglobulin and C1-esterase inhibitorwere immediate inhibitors of kallikrein.“Time-dependent” inhibition of both enzymes was observed in the ai-antitrypsin containing fractions.
Haemostatic Factors in Umbilical Artery and Vein Blood
Authors: Foley M.E. , Clayton J.K. , McNicol G.P.
Published: May 2019

Increased clotting factor activity has been demonstrated by other workers in uterine vein blood at the time of placental separation. To examine some effects of placental separation on the fetus, haemostatic factors in umbilical artery and vein blood were studied immediately following delivery in 80 normal babies. Clotting activity was increased in umbilical vein blood as compared with umbilical artery blood; the most obvious difference was a significant shortening of the activated partial thromboplastin time. Cord blood showed a high level of fibrinolytic activity; activity measured in plasma by the fibrin plate method and with the euglobulin lysis test was significantly greater in the umbilical vein than in the artery. Normal levels of fibrinogen degradation products indicated no active fibrinogenolysis or fibrinolysis. The mean umbilical vein blood platelet count (233 × 109/litre) was significantly higher than the mean umbilical artery blood platelet count (205 × 109/litre). Platelet factor 3 availability showed no arteriovenous difference. Platelet aggregation to 2 and 4 micromolar adenosinediphosphate was significantly greater in the umbilical vein than in umbilical artery blood. The data indicate that at the time of placental separation, there is increased potential haemostatic activity in the fetus.
Interaction of Human Platelets with Subndotrelium: Implications for Measurement of Defects in Platelet Function
Authors: Turitto V.T. , Weiss H.J. , Cohen D.S.
Published: May 2019

An in Vitro perfusion technique introduced by Baumgartner has been used to investigate the interaction of human blood platelets with subendothelium of rabbit aortas. Platelet contact, adhesion (spreading) and the formation of ndcxothrombi were measured directly by use of a morphometric technique for a variety of exposure times (1 to 40 min) and a physiological range of flow rates (shear rates varying from 50 to 830 sec-1). A theory accounting for platelet transport through the blood and the platelet reactivity at the vessel surface Indicated platelet transport to be the controlling influence on platelet attachment. Under low shear conditions platelet attachment is unaltered by moderate changes in platelet reactivity. The results suggest that high shear rates (greater than 800 sec-1) are necessary for a sensitive measurement of defects in platelet attachment and that experimental devices which employ low shear conditions are limited in measuring such defects.
FVIII: A Homologous Series of Covalently Linked Oligomers
Authors: Cockburn C.G. , Reay Diana M.L.
Published: May 2019

Purified human FVIII was electrophoresed through SDS agarose gels of different concentrations and through SDS Polyacrylamide (PA) gels of exceptionally large pore size (3.75% monomer, 1% cross linking). The gels were calibrated (a) with the FVIII subunit (b) with dimethyl-suberimidate cross-linked human fibrinogen. After Coomassie-blue staining the PA gels were scanned densito-metrically.

The results show that freshly purified FVIII at pH 6.5 exists as a homologous series of oligomers and that a linear relationship obtains between the mobility of oligomers 1-6 and log molecular weight. Another linear relationship exists between the higher oligomers due to their significantly raised mobility. This suggests that the higher oligomers have a compact structure which is still partially ordered after SDS denaturation. A plot of log oligomer molarity vs. log subunit number has a gradient of approximately -1, suddenly decreasing to -5.5 after the hexamer, strongly indicating a decrease in the rate of polymerisation after this point. Oligomers 1-6 comprised 88% of the total FVIII, and oligomers 7-9 a further 6%.FVIII left at room temperature for 24 hours associated into extremely large oligomers.
Comparative Study on the Effect of Carbenicillin, Inhibitors of Platelet Function and Heparin on Experimental Thrombus Formation
Authors: Zimmermann R. , Andrassy K. , Zeltsch C. , Lange D. , Hof F.
Published: May 2019

Previous studies documented an impairment of haemostasis by synthetic penicillins (Thromb. Haem. 34: 115, 1976) and penicillin (Lancet II : 1039, 1976). Therefore the antithrombotic activity of synthetic penicillin. (carbenicillin)(C) was compared with that of aspirin (ASA), dipyridamole (DIPY) and heparin in 150 rabbits. Thrombus formation was induced by standardized endothelial lesions. The dose of C was adjusted to a 4.2 fold prolongation of bleeding time, similar to that seen in clinical patients. Analysis and composition of thrombi was done by measurement of incorporation of labeled blood elements (51cr labeled platelets, 125J-fibrinogen and 59Fe labeled red cells). The ‘specific thrombus/blood ratio’ with values of 19.1 and 50.9 (51cr) in venous and arterial thrombi evidenced the significance of platelets in this model. In the venous system C reduced formation of thrombi by 43%, ASA by 34%, ASA and DIPY by 55% and heparin by 90%. In the arterial system C inhibited thrombus formation by 89%, ASA by 15%, ASA and DIPY by 46% and heparin by 60%. It is concluded, that C effectively prevents thrombus formation in the arterial system and to lower extent in the venous system. The results prove the importance of platelets in arterial thrombogenesis and the efficacy of platelet aggregation inhibitors in preventing thrombi in the arterial system. In comparison with other known antiplatelet drugs it seems, that C is the most effective platelet aggregation inhibitor to date.
Hemostatis Disorder Under Penicillin G Therapy
Authors: Andrassy K. , Scherz M. , Ritz E. , Walter E. , Zimmermann R. , Vömel W. , Storch H. , Hasper B.
Published: May 2019

Systemic administration of penicillin G (P) has been shown to cause a coagulation disorder (Andrassy et al., Lancet II, 1039, 1976). In this study time course, dose effect relationship and effect on viscosity were investigated. - 20 pat. with normal GFR received P either intermittently (i.i.) (2x20 mio U/d. as lh short inf.) or by constant drip inf.(c.i.) (40 mio/d.); Analysis:Bleeding time, (B.T.); platelet aggreg. test; coagulation factors (AT III, fXa); rotation/capil. viscosimetry of blood; serum P cone.(diff. test). An increase in B.T. and dimin. collagen rsp. ristocetin ind. platelet aggreg. was usually found 6 h and consistently 24 h after P infusion. After stopping P, the abnormalities were demonstrable for 4 days (i.i.) and for 8 days (c.i.). Serum P levels (preinfusion) were 0.5-8 μg/ml (i.i.) and 40-80 μg/ml (c.i.) 8 days after stopping P, reactive thrombocytosis was noted in the c.i. group. Blood viscosity, if anything, increased; therefore the prolongation of B.T. is not accounted for by abnormalities of blood viscosity. In additional pat., 2x5 mio P were without noticeable effect on hemostasis, whereas definite abnormalities were demonstrable with 20 mio P/day. In the plasmatic system, increased AT III and diminished f. Xa were found irresp. of the mode of administration. Minor bleeding (epistaxis, macrohematuria, wound bleeding) was observed in 5/20 patients.
Fibrinolytic Activity and Effects on the Coagulation System of a Micro-Enzyme of Bacillus Cereus
Authors: Ekenstam B.T. af , Friberger P.
Published: May 2019

A microenzyme of Bacillus Cereus isolated by J. Björksten et al., Björksten Research Foundation, Madison, Wise, USA, showed a caseinolytic activity of 2600 CU/mg. This enzyme has a potent fibrinolytic activity tested on a plasminogen free fibrin plate. The effect on the coagulation system has been compared with that of plasmin and brinase. The microenzyme shows an inhibiting activity on the coagulation of plasma of the same magnitude as that of plasmin and brinase. This inhibition is also correlated to the results obtained from coagulation studies on whole blood using a modified Chandler loop technique. The chromogenic substrate S-2238 has been used for the indirect assay of the activity of the microenzym over a standardized thrombin system.
Plasma Concentrate Consumption - A Five Year Study
Authors: Boone D.C. , Dietrich S.L. , Kasper C.K.
Published: May 2019

Outpatient consumption of plasma concentrates by 225 hemophilic patients of Orthopaedic Hospital was reviewed for the five year period, 1969–1973. Patient data were grouped by age, by hemophilia type and deficiency level. Further subdivisions were made based upon the locality of transfusion; only at home, only in the outpatient facility of Orthopaedic Hospital, or a combination of both. The subdivided data were compared for 1969 and 1973 for transfusions/patient factor units/transfusion, units/patient, and percentage of joint, nonjoint and prophylactic transfusions. Significant differences in consumption were found between the defined age, hemophilia type and deficiency groupings. An important finding was the variation in numbers of transfusions between patients with the same hemophilia type/deficiency and age. Markedly different patterns of consumption were found depending upon the locality of transfusion. Patients with severe hemophilia on home transfusion showed significant increases in transfusions/patient in 1973 contrasted with 1969. Units/transfusion remained essentially unchanged. These results have broad implications for the planning of blood resources since the more significant changes in concentrate consumption occurred among those patients on home transfusion who tend to have more frequent hemorrhages and who possess the freedom to decide to transfuse.
An Improved Method for Preparation of Antihemophilic Factor (AHF)
Authors: Wickerhauser M. , Mercer J.E. , Eckenrode J.W.
Published: May 2019

Intermediate purity AHF prepared by the American Red Cross method (James, H. L. and Wicker-hauser, M., Vox Sang. 23:402, 1972) contains about 10 Factor VIII units/ml at 20-25 fold purification over plasma. A more concentrated and purified product would facilitate administration of AHF in home treatment of hemophiliacs. Our original method was improved by the following modifications: (1) A cold extraction step was incorporated to remove cold-soluble impurities. The cryoprecipitate was extracted by stirring with 0°02 M Tris buffer, pH 7°0 (4 ml/g cryo) for 30 minutes at 0°CoFactor VIII loss in this step was negligible,,(2) AHF was then recovered from the cold-insoluble portion of the cryoprecipitate by extraction at 21°C with the same buffer. To increase the AHF concentration, this second extraction step was carried out with a smaller buffer volume (2 ml instead of 3-4 ml/g cryo). The subseguent steps, deprothrombinazation, filtration, and lyophi1ization were essentially unchanged,, To further increase Factor VIII concentration, the dried AHF concentrate was reconstituted to 40% rather than 50% of the volume prior to lyophilization. AHF concentrate prepared on a large scale by this method was 20-30 fold concentrated and 40-50 fold purified over plasma at a recovery of about 250 Factor VIII units per liter of plasma. The final product was readily soluble, clear and almost colorless upon reconstitution.