bluuebunny/crossref_metadata_2025 huggingface dataset:
Thrombosis: A Function of Surface Charge and/or Chemistry of Collagen
Authors: Sawyer P. , Stanczewskl B. , Stillman R. , Kirschenbaum D. , Sivakoff S. , Lucas T.
Published: May 2019

Examination of porcine prosthesis and bovine heterografts using histologic, SEM and physical examination revealed that ; (1) they have a measurable half life in terms of function; (2) the areas of failure are predictable; (3) the structure of the observed thrombus is unusual.

A number of collagen and human umbilical cord prostheses were prepared by bonding a variety of aliphatic acids to the helical polypetide collagen side chains to produce a repeating surface chemistry. The control (fresh or ficin digested collagen) was compared to (i) glutaraldehyde tanned (GT)(-) charged, (ii) GT plus (+) charged and (iii) GT neutral surface. These resulting tubular grafts were implanted in dog carotid and femoral arteries.

The results indicate the most important factor in long term patency and function is a coval-ently bonded, GT (-) surface.

Histologic examination reveals that manipulation of the surface charge of collagen results in bizarre thrombus formation with aggregation of a singie cell type or protein element.

The (-) charged grafts were free from thrombus. Control collagen grafts contained classical fibrinogen and cellular thrombus. The (+) charged grafts had a pseudo-thrombus i.e. increased fibrinogen fragments (& B) monomers plus trapped cell deposition. The neutral grafts had a less dense, branched fibrinogen deposit with some celluiar elements.

In summary, the structural, chemical and surface charge configuration of collagen can be manipulated to produce a biological cardiovascular prosthesis with improved performance.
The Use of Thermography for Screening of Postoperative Deep Venous Thrombosis (DVT); A Comparison With 125I-Fibrinogen Test
Authors: Bergqvist D. , Hallböök T.
Published: May 2019

At the last congress on Thrombosis and Haemostasis we showed a high degree of diagnostic agreement between thermography and phlebography in patients with clinical suspicion of DVT (Thromb. Diath. Haemorrh., 34:905, 1975). These findings have been verified in a large number of patients. On the other hand the value of thermography as a screening method for postoperative DVT is not known. The aim of this study was to evaluate the screening possibilities of thermography postoperatively with 125I-fibrinogen test as the reference method. 272 patients (544 legs) were investigated with the two methods simultaneously for one week to ten days postoperatively. The overall agreement between the methods was 79%, the sensitivity being 64% and the specificity 90%. One explanation for the relatively poor precision is that 125I-fibrinogen test is extremely sensitive, detecting also very minute thrombi in the calves. The 125I-fibrinogen positive thrombi were therefore analysed in relation to their localization. In cases with thrombi extending above the knee level the agreement was 72%. It gan be concluded from this study that thermography gives a lower frequency of DVT than 125I-fibrinogen test in postoperative patients. It must be further evaluated if any dangerous thrombi are missed or if thermography can be used as a non-invasive screening method in the postoperative course.
Actin and Myosin from Platelets and Muscle are Substrates for Fibrinoligase
Authors: Cohen Isaac , Bruner-Lorand Joyce , Blankenberg Tikoes A. , Lorand Laszlo
Published: May 2019

Fibrinoligase (thrombin and calcium activated factor XIII) from human plasma catalyzes the incorporation of dansylcadaverine and 14C-putrescine into myosin and actin prepared from either human platelets or rabbit skeletal muscle. An average of 3 to 4 moles of amine are incorporated per mole of myosin of either type when the enzyme is used under saturating conditions. The rate of incorporation is 1.4 to 2 fold faster for filamentous than for soluble myosin.

Electrophoretic analysis on Polyacrylamide and agarose gels in the presence of sodium dodecyl sulfate and 2-mercaptoethanol was used for locating the acceptor sites and for the detection of possible cross-links. Both heavy and light chains of platelet as well as muscle myosin incorporate dansylcadaverine and 14C-putrescine. However, in quantitative terms, the incorporation into the light chains of either type is only a small fraction of that incorporated into the heavy chains. Fibrinoligase also catalyzes the formation of highly cross-linked polymers in both myosin and actin (platelet or muscle) indicating the presence of acceptor and donor sites in the two contractile proteins.
Ultrastructural Studies on the Surface Coat of Human Platelets Aggregated by Polylysine and Dextran
Authors: Taketomi Y. , Kuramoto A.
Published: May 2019

The surface coat of human platelets was stained with positive charged colloidal Thorotrast particles and ruthenium red to elucidate the mechanism of platelet aggregation by macromolecules.

Positive charged macromolecule, polylysine (Mw.l5,000; 23,000; 180,000) could induce the aggregation in low concentration but high concentration was needed in the case of neutral macromolecule, dextran (Mw.40,000; 250,000; 2000,000). The larger molecules of polylysine and dextran were more effective in inducing platelet aggregation. In the dextran induced aggregation, Thorotrast particles on the cell surface did not decrease significantly. On the other hand, the surface membranes of platelets treated with neuraminidase and aggregated by polylysine were essentially devoid of bound particles. These findings suggested that polylysine induced aggregation more effectively than dextran by reducing the negative surface charge and giving stronger adsorption force on cell surface.

In washed platelet suspension, polylysine induced aggregation without release reaction of 14C-serotonin, but average distance between plasma membranes of aggregated platelets did not vary with the degrees of polymerization. In this respect, it seems probable that after forming macromolecular bridging of cell surface, another interaction may follow between cell surfaces.
Production of a High-Titred Sheep Antiserum Against Human Factor IX and Immunological Demonstration of Factor IX Aggregates
Authors: Ørstavik K.H. , Vennerød A.M.
Published: May 2019

Plasma factor IX was purified from a factor IX concentrate by a five step procedure including absorption onto aluminium hydroxide, affinity chromatography on heparin-coupled Sepharose 4B, preparative disc gel electrophoresis, affinity chromatography on an immunosorbent column with rabbit antiserum against factor X and chromatography on DE-52 cellulose. The pooled fractions had a specific activity of approximately 250 U/mg protein. A sheep was immunized with pooled and concentrated fractions. An antiserum was produced which gave one main precipitin band and occasionally an additional weak band against normal plasma in double immunodiffusion. At a dilution of 1/100-1/200 the antiserum neutralized 90% of the factor IX activity in an equal volume of normal plasma.

Polyacrylamide disc gel electrophoresis of the fractions from DE-52 cellulose revealed one major and three minor bands with lower electrophoretic mobility and intensity. The three minor bands disappeared on disc gel electrophoresis in the presence of 10 M urea. When the disc electrophoresis gel was submitted to electrophoresis into anagarose gel containing the sheep antiserum or a previously characterized rabbit antiserum against factor IX, four precipitin arcs corresponding to the four bands were seen. A reaction of identity was seen between the four arcs. This study demonstrates that a highly potent antiserum may be produced against factor IX in sheep.
Turnover of Plasma Kininogens in Rhesus Monkeys
Authors: Yamada T. , Wing D.A. , Pierce J.V.
Published: May 2019

Radiolabelled purified plasma kininogens were injected into healthy rhesus monkeys for the purpose of studying their normal metabolic turnover. Human kininogen was purified using affinity chromatography and further fractionated into high molecular weight (HMW) and low molecular weight (LMW) proteins as described by Pierce and Guimaraes (Fogarty International Center Proceedings No. 27, pp. 113-119, 1976). The kininogens were labelled with 125I using either the choramine-T or microdiffusion techniques. Labelled mixed kininogen, LMW kininogen, and HMW kininogen preparations were injected into 6, 4, and 6 rhesus monkeys respectively. Blood samples were drawn at various time intervals, and assayed in a gamma counter. The disappearance of injected labelled kininogen from the intravascular compartment could be expressed as a double exponential equation. The difference between the half-life of HMW kininogen (26.0 ± 1.6 h) (± SE) and that of LMW kininogen (20.2 ± 2.1 h) was only marginal (P >0.05). In contrast there was a highly significant (P < 0.001) difference in their fractional catabolic rates (1.12 ± 0.09 day-1 for HMW kininogen and 2.05 ± 0.12 day-1 for LMW kininogen). These observations indicate that although there is a minimal difference in the metabolic rates of intravascular HMW and LMW kininogens, a considerable difference exists in their rates of flux between the intravascular and extravascular spaces.
Kinetic Studies of Soluble Fibrin Complexes in Vivo
Authors: Schönbach F. , Mahn I. , Müller-Berghaus G.
Published: May 2019

Soluble fibrin complexes may occur in vivo in a variety of coagulation disorders. The aim of this investigation was to elucidate the in vivo behaviour of fibrin-fibrinogen complexes prepared in vitro in a ratio of 1 part fibrin to 20 parts fibrinogen. 12 rabbits (group A) were injected with soluble fibrin complexes containing homologeous I-131-fibrin and I-125-fibrinogen. Another group of 12 rabbits served as control (group B) and received I-131-fibrin solubilized in urea and I-125-fibrinogen separately from each other. Studies were performed over a period of 6 days.

The mean distribution volume of fibrin as well as of fibrinogen did not significantly differ between both groups. The elimination characteristics of I-131-fibrin of the soluble fibrin complexes (group A) as well as of the solubilized fibrin (group B) were similiar. The fibrinogen elimination did not differ significantly between the groups: a mean T 1/2 of 47.8 h in group A and a T 1/2 of 46.7 h in group B was calculated.

The experiments demonstrate that non-crosslinked soluble fibrin complexes distribute homogeneously in the circulation and dissociate into its subunits. Fibrin is eliminated from the circulating blood without influencing the normal catabolism of fibrinogen.
Thrombocytopenia in Septicemia: ? The Role of Disseminated Intravascular Coagulation
Authors: Neame Peter B. , Hirsh Jack , Nossel Hymie L.
Published: May 2019

Thrombocytopenia is frequently seen in septicemia but its mechanism is uncertain. We have carried out detailed studies in 18 septicemic patients who had associated thrombocytopenia to determine whether this was associated with coagulation abnormalities indicative of disseminated intravascular coagulation (DIC) or whether it was more frequently an isolated event.

Tests performed as indicators of DIC included fibrinogen degradation products (FDP), protamine sulfate test (PS), T½ I125 fibrinogen survival and in some patients fibrinopeptide A assay (FPA). Two distinct patterns of results emerged. In one, made up of patients with mild to moderate thrombocytopenia (platelet count 48,000-135,000/μl), there was no evidence of DIC on the basis of FDP, PS and FPA assays. In the second group, in which there was more severe thrombocytopenia (platelet count 2,000-39,000,/μl), there was definite evidence of DIC on the basis of FDP and protamine sulfate assays. The levels of fibrinogen, Factor VIII and Factor V were significantly lower in group 2 than in group 1.

It is concluded that thrombocytopenia can occur in septicemia without evidence of associated DIC, but in those cases with severe thrombocytopenia the fall in platelet count is usually accompanied by consumption of fibrinogen.
Fibrin Degradation Products and Vascular Permeability
Authors: Gerdin Bengt , Högstorp Herman , Lindquist Olle , Saldeen Tom , Svensjö Erik
Published: May 2019

Increased vascular permeability plays an important role in the pathogenesis of the delayed microembolism syndrome. Fibrin degradation products (FDP) may play a role for this permeability disturbance. Fractions of lymph from the cannulated right lymphatic duct in dogs with induced microembolism syndrome and lysate from fibrin clots obtained by gel chromatography were used. The effect on vascular permeability was determined in the hamster cheek pouch and in the dorsal skin of the rat. Increased permeability was determined by leakage of fluorescein labelled dextran in the first model and by use of isotope labelled albumin in the second model. Lymph from the lymphatic duct and fractions of lysate from fibrin clots caused an increased vascular permeability of the same character in both models, the effect being partly due to high molecular weight products and partly due to low molecular weight products. The effect of high molecular weight products may possibly be due to their continous cleavage releasing low molecular weight vasoactive FDP. The effect of FDP on vascular permeability was enhanced by pretreatment with the β-adrenergic inhibitor propranolol and inhibited by the β2-adrenergie stimulator terbutaline. Bredykinin and PGE1 both increased macromolecular leakage in the hamster cheek pouch. This increase was also counteracted by terbutaline. The FDP effect on permeability might be due to contraction of the endothelial cells.
Platelet Prostaglandin Endoperoxides Formation in Hyperlipidemias
Authors: Bizios R. , Wong L.K. , Vaillancourt R. , Lees R.S. , Carvalho A.C.
Published: May 2019

To evaluate the role of prostaglandin endoperoxide formation in the mechanism(s) of platelet hypersensitivity in hyperlipoproteinemias, a study was undertaken in 8 type II patients, 3 type IV, and 8 normal volunteers. Platelet aggregation and release were studied in all subjects. Simultaneously, EDTA-platelets were washed by centrifugation and suspended in modified Ringer’s solution. One ml suspensions (0.8-1x106/μl) were added to 4.5μg of 14C-Arachidonic Acid (AA; sp. act. 50 mCi/mmol) and incubated for 30 sec and 5 min at 37°C. The methylated reaction products were separated by thin layer chromatography. AA, PGE2, PGF2α, and thromboxane B2 (PHD) (Upjohn) were used to identify and locate the respective bands. The products were analyzed by mass spectrometry. TLC-plates were both scanned and scraped for radioactive counting. The chromatograms displayed a pattern similar to that reported by Samuelsson et al. Compared with normal, hypersensitive platelets of type II patients generated 25% more thromboxane A2 (TXA2) after 30 sec incubation with AA, while at 5 min PHD was increased by 30% and TXA2 was still elevated by a factor of 2. The behavior of type IV platelets was similar to normal.

Aspirin-treated platelets produced one product (HETE). Among the AA derivatives studied, only eluted TXA2 induced platelet release.

Our data suggest that platelet hyperreactivity in type II patients may be mediated by the increased production and long survival of thromboxane A2.
Human Endothelial Cell Injury Mechanisms in Vitro
Authors: Wall R.T. , Harker L.A. , Striker G. , Quadracci L.
Published: May 2019

Since endothelial cell desquamation constitutes the initial event leading to acute and chronic vascular diseases, we have developed an in vitro system for studying injury mechanisms using cultured cells. The system involves the incubation of test material with confluent, 51Cr-labeled, cultured human umbilical vein endothelial cells in 100% pooled human serum; injury was measured as% endothelial cell 51Cr release (ECR) into the supernatant media. Baseline spontaneous ECR was 6.0% ± 1.5 while specific rabbit antihuman endothelial cell antibody (1:64 dilution) in the presence of fresh complement released 90% ± 3 of the total releasable 51Cr activity. Consistent with in vivo predictions, homocysteine in concentrations of 0.5-40 mM iduced dose response ECR up to 20%. Neither methionine nor homocystine increased ECR over controls. Also as expected from experimental work endotoxin (S. enteritidis) caused dose response ECR, i.e., 24% ± 2 at 10 ug/ml. Control studies with cultured human smooth muscle cells (SMC) demonstrated no ECR with homocysteine but significant release occurred with endotoxin.

Specific complement dependent cytotoxic anti-endothelial cell antibody was demonstrated in the secum of two thrombotic thrombocytopenic purpura (TTP) patients at a 1:1 dilution, inducing ECR of 23.1% ± 1.4 and 21.0% ± 0.25. The antibody was also demonstrated by immunoflourescent techniques and was absorbed from the serum using human endothelial cells. One disease-free, six month survivor showed no cytotoxic activity. Serum from a patient with adult hemolytic-uremic syndrome demonstrated antibody dependent cell mediated cytotoxicity with release of 22% ± 1.1 when normal non-immune lymphocytes were added to heat inactivated seriun. Control studies with SMC showed no ECR with TTP sera. We conclude that assays of endothelial cell 51Cr release are stable, reproducible and useful in the characterization of injury mechanisms.
The Functional Significance of Platelet Macromolecules
Authors: Lüscher E.F.
Published: May 2019

Besides the equipment of enzymes, substrates and structural proteins common to all living cells, the platelet contains a variety of macromolecules which are directly involved in its specific biological functions. Although any selection may be arbitrary, the following functions primarily involve macro-molecules:

- contractility based on cytoplasmic and membrane linked actomyosin (AM), responsible not only for active contraction of pseudopodes and clot retraction, but also for shape change. Disputed is the participation of AM in the release reaction.

- upkeep of distinct shape, mediated by the microtubular (MT) system. The activation of AM leading to shape change is incompatible with intact MT, suggesting a subtle interplay between both systems.

- influence on extracellular activities by means of released macromolecules.

- membrane function, involving adhesivity, aggregability, as well as the presence of receptors for external stimulators of platelet activity and the capacity to react upon stimulation in such a way that a signal for the induction of sequential reactions is created. Examples of such secondary reactions of membrane constituents will be discussed and compared with other cells.
The Biochemistry of Fibrin Cross-Linking
Authors: Doolittle Russell F.
Published: May 2019

The final steps in the formation of a fibrin clot involve the factor XIII-catalyzed introduction of covalent peptide bonds. These bonds are the result of the condensation of specific lysine and glutamine sidechains in γ-chains on the one hand and α-chains on the other. Function apart, these bonds have provided great insight into the way the individual molecular units are arranged in the fibrin polymer. Thus, the reciprocal pairing of the carboxy-terminal segments of γ-chains to yield γγ dimers indicates that all molecules in the polymer have the same orientation, and, because of the dimeric nature of the fibrinogen molecule, the abbutting chains of neighboring molecules are therefore antiparallel. Until now the three-dimensional involvement of α-chains—which in contrast to γ-chains become multimerically cross-linked--has been completely mysterious. Recently, however, we have isolated those portions of α-chains involved in multimeric cross-linking by fragmenting fully cross-linked fibrin with cyanogen bromide. Thus, we were able to identify the linked fragments as two segments which, when not cross-linked, have mol. wts. of 29,000 and 6,000 respectively. The latter fragment has aminoterminal leucine and is thought to be the carboxy-terminal penultimate CNBr fragment in the α-chain. It is linked in equimolar quantities to the large mol. wt. glutamic acid-ending fragment. The total mol. wt. of the multimerically linked units is greater than 500,000. The nature of the fragment network is such that the orientation of the α-chains in fibrin could be either parallel or antiparallel. In either case the architecture is well suited to lateral cross-linking between fibrin polymers.
Aspirin Prophylaxis Against Thromboembolic Disease
Authors: Harris W.H.
Published: May 2019

The efficacy and safety of aspirin prophylaxis in preventing deep vein thrombosis is now scientifically established. This was done in a prospective, randomized, simultaneously controlled, double-blind, placebo study using objective diagnostic methods in 98 patients over age 40 undergoing total hip replacement surgery. All patients had fibrinogen uptake tests, cuff impedance phlebography, and radiographic phlebography. The one patient with postoperative pulmonary signs or symptoms had Q/V pulmonary scans and selective pulmonary angiography. Twenty-three of fifty-one patients in the control group developed thromboembolic disease, compared to eleven of forty-seven in the aspirin group (P < 0.02, Fisher Exact Test). The embolus occurred in the control group. There were no deaths. Aspirin was given as 1.2 grams daily starting preoperatively.

To study the efficacy of this prophylaxis without the use of early surveillance diagnostic methods, four hundred ninety-eight consecutive patients over 40 years of age undergoing total hip replacement had aspirin prophylaxis but only clinical diagnosis was used. No fatal pulmonary emboli occurred. The only fatality was shown at postmortum not to be caused by an embolus. All patients with pulmonary signs or symptoms had perfusion pulmonary scans. Only seven patients had objectively demonstrated, nonfatal, pulmonary emboli (1.4%).
Prevention of Fatal Pulmonary Thromboembolism by Heparin Prophylaxis after Surgery for Hip Fractures
Authors: Sharnoff J.G.
Published: May 2019

From 1960 through 1975, 337 patients with surgically treated acute fracture of the hip received subcutaneously administered aqueous heparin sodium to prevent thromboembolic episodes. Four hundred and three patients received no heparin. Their incidence of fatal pulmonary embolism was 3.5 percent. Ninety-five patients receiving the original “small dose” heparin regimen from August 1960 to November 1967 had a 4.2 percent incidence of fatal thromboembolism. These had been administered heparin 8-10 hours or less before surgery. Beginning November 1967 the “small dose” heparin schedule was altered in hip fracture patients to start heparin prophylaxis immediately following hospital admission. One hundred and forty-seven patients treated with the latter schedule had 0.0 percent fatal thromboembolism with the dose modified according to a coagulation time test. The patients received 2,500 units on admission and every 6 hours until the day before operation. Then they were given 5,000 to 10,000 units. 8 to 10 hours before surgery and 2,500 units every 6 hours after surgery until they were fully mobilized. The altered “small dose” heparin regimen adequately monitored by the blood coagulation test, the Dale and Laidlaw Coagulometer, proved highly effective as measured by fatality rates.
Apparent Viscosity of Artificial Red-White and White Thrombi During Anaesthesia and Surgery
Authors: Dintenfass L. , Aronson B.
Published: May 2019

This is a pilot study intended to explore some aspects of thrombus formation which might be of value in understanding the dynamics of anaesthesia, on the one hand, and the effects of anaesthesia and surgery on tissue perfusion, on the other. Patients studied included six subjects undergoing operations for retinal detachement (5), upper abdominal surgery (1), and a major orthopaedic procedure (1). Artificial thrombi, of morphology of red/ /white and white arterial thrombi, were formed in vitro by means of VFTV, variable frequency thrombo viscometer, at temperature 37C, on freshly shed blood, at mean shear rates of 26.8 and 80 sec-1. Blood samples were drawn immediately prior to commencement of anaesthesia, and then at half-hourly intervals. Anaesthesia was induced with thiopentone sodium, and halothane or meperidine and droperidol. In general, the apparent viscosity of artificial thrombi increased during surgery.
Hematologic Changes Following Burns
Authors: Caprini J.A. , Lipp V. , Zuckerman L. , Vagher J.P. , Winchester D.P.
Published: May 2019

Forty-five burn victims were studied at 48 hours and weekly with hematocrit, platelet count (PC), fibrinogen level, prothrombin, and activated partial thromboplastin time, fibrin split products (FSP), plasminogen (PL), complement C'3, C'4, and C'H 50, baseline, and 1 and 5 minute Kallikrein activity (K). Prognostic index was used for grouping blood data. Group I (GI) had a prognostic index under 20% survival and 8 of 12 died. Group II (GII) had a prognostic index of 20-80% survival and 4 of 11 died. Group III (GIII) prognostic index was over 80% survival and all 22 survived. At 48 hours GI had significantly elevated FSP compared to GII (32±6 vs 12±4*) and 5'K of 22±7 vs 50±5*µMoles/hr/ml. At one week PC and C'3 between GI and GII were 157,000±49,000/mm3 vs 326,000±59,000/mm3* and 61±5 vs 113±11 mg/ml, respectively. Also in the first week, PL and 5'K in GI were 6.9±1.0 mg/ml and 5.2±3.2μM/hr/ml compared to GUI 13.5±1.8 mg/ml** and 37.1±4.0μM/hr/ml*, respectively. C'3 in GI was low compared to GII or GIII for two weeks**. Summarizing, severe burns had statistically significant depression of PL, 5'K, C'3, PC, and elevated FSP. These results show activation of coagulation, fibrinolytic, complement, and kinin pathways after thermal injury and at least one measurement from each of these systems shows statistically significant alterations that correlate with the prognostic index. The results strongly support the concept of intravascular contamination with multiple systems activation following thermal injury.

* P<.05

** P<.01
A Study of Antithrombin III Levels in Healthy Male and Female Subjects Tested Monthly for Eleven Months by Four Methods
Authors: Sirridge M.S. , Shannon R.
Published: May 2019

Because of the increased interest in the role of Antithrombin III as a physiologic coagulation inhibitor and because of some previous problems we encountered in sequential measurement of this protein, the present study was designed to answer 3 questions: 1) Which of four methods (2 functional activity methods and 2 immunologic methods) is the most practical, accurate and reproducible? 2) Do normal subjects have relatively constant levels when tested sequentially by these methods? 3) What is the range of Antithrombin III levels in young healthy male and female subjects? A serum pool was tested by each of the 3 serum methods at least 20 different times throughout the study. The von Kaulla functional activity method gave the lowest standard deviation and coefficient of variation. In the group of 29 subjects this method proved to be the most practical and also gave very low coefficients of variation for individual subjects when they were tested from 11 to 16 times (range .023-.054). The other methods did not give such low coefficients of variation for individual subjects. The range of levels was 77 to 110% on 344 samples tested by the von Kaulla method With a mean of 92.8% and a standard deviation of 6.4%. The standard deviations for the pool and for individual subjects were greater but quite satisfactory for all methods.
Purification and Characterization of Human Platelet Factor 3.
Authors: Sandberg H. , Andersson L-O. , Hoglund S.
Published: May 2019

In the purification of platelet factor 3 fresh human platelets prepared by centrifugation were used as starting material. Release material having platelet factor 3 activity, as measured by the Stypven time, was obtained by treating the platelet suspension with collagen or thrombin followed by removal of the platelets by centrifugation. Residual platelets left in supernatant were removed by membrane filtration. Purification of platelet factor 3 from the other components in the release material was accomplished by affinity chromathography followed by gel filtration on Sepharose 2B. The gel filtration fraction having platelet factor 3 activity was studied with respect to chemical composition and appearance in electron mincroscopy. Phospholipids and cholesterol were the main components but small amounts of protein could also be detected. The electron microscopy revealed the presence of numerous membranous particles having diameters between 50-150 Å. The purified fraction was used for immunization of rabbits and an antiserum was obtained which inhibited the platelet factor 3 activity.
The Effects of Different Routes of Administration and Injection Schedules of Thrombopoietin on Platelet Production in Assay Mice
Authors: McDonald T.P.
Published: May 2019

Mice have been used in several studies for the assay of a thrombocytopoiesis-stimulating factor (TSF or thrombopoietin). Little information however, is available on the optimum assay time or injection schedule. Therefore, the present work compares the % 35S incorporation into platelets of mice in rebound-thrombocytosis after subcutaneous (sc) and intraperitoneal (ip) injections of TSF-rich preparations with different injection schedules. For production of thrombocythernia, mice were given a single ip injection of rabbit anti-mouse platelet serum 5 days before injection of TSF-rich test materials. TSF administered sc or ip resulted in almost identical % 35S incorporation values. Various doses of TSF injected sc 4 times over a 2 day period to thrombocytotic mice gave a linear dose-response. Also, this injection schedule gave the greatest response to the same total dose of TSF when compared to other injection schedules. Single injections of TSF did not give as great a response as did multiple injections using the same total dose. Results of this work indicate that for maximum % 35S incorporation values, TSF-rich materials should be administered in multiple injections to mice, either sc or ip, over a period of approximately 2 days.
Specificity of Factor VIII Inhibitor for Procoagulant Activity
Authors: Yang H. , Kuzur M.
Published: May 2019

Nine hemophilia A patients with an inhibitor to factor VIII procoagulant and eight without an inhibitor were studied for the presence of an inhibitor to von Willebrand factor (vWf). After a 2 hour incubation at 37°C, test plasma, normal pooled plasma and test plasma mixed with normal pooled plasma were assayed for vWf in the quantitative ristocetin-induced platelet aggregation system. The mean vWf was 1.31±.59 unit/ml in the non-inhibitor group and 1.46±.52 in the inhibitor group. In the non-inhibitor group the predicted vWf (summation of the test sample and normal plasma) was 1.38±.37 unit/ml, whereas the actual vWf of the mixture was 1.23±.35. In the inhibitor group the predicted vWf was 1.46±.33 unit/ml, whereas the actual vWf of the mixture was 1.44±.40. The results in the two groups were not significantly different. Neither group had inhibitory activity for vWf. The factor VIII inhibitor is highly specific for the procoagulant part of the factor VIII complex.
The Relationship Between Regulation of Prostaglandin Metabolism and Platelet Aggregation
Authors: Tohjima T. , Takahashi Y. , Funayama H. , Shiokawa Y.
Published: May 2019

Platelet aggregation is mediated by prostaglandin (PG) endoperoxide and cAMP. But exogenous PGs might modify platelet function and might play a major role on the PG metabolism in platelet as well as endogenous PGs do. We studied the relationship between quantitative and qualitative regulation of PGs and cAMP and platelet aggregation, by incubation method adding exogenous PGs (PGE1, etc) and antiaggregating agents such as heparin, dextran sulfate, carbocromen, α-tocopherol, persein human platelet suspension. Platelet aggregation was studied photometrically adding thrombin. PGE1, E2, F1α and F2α were assayed by radioimmunoassay (RIA). cAMP was assayed by RIA. PG formation after incubation with the agents in platelet suspension was as follows; 1) PGF system was stimulated by PGE1 and PGE2. 2) PGE system was depressed by PGF2α. 3) PGF system was depressed by PGA1. 4) PGE1, stimulated cAMP and PGF system. 5)Aspirin, heparin, dextran sulfate and carbocromen had dose dependent inhibitory effect on thrombin induced platelet aggregation and had reducing effect on formation before adding thrombin. 6) Plenylamine and CDP-choline had inhibitory effect in vitro on platelet aggregation at higher concentrations than that of clinical use. CDP-choline reduced PGE levels but did not influence PGF levels. Prenylamine reduced PGF levels and stimulated cAMP formation at higher concentrations. 7)Dipyridamole and α-topcopherol did not reduce PGs levels. This approach provided the results that platelet function was regulated by PG metabolism which is modified by exogenous PGs or some drugs. Antiaggregating agents can be classified by the quantitative and qualitative difference of PG formation.
Formation of Prostaglandin Cyclo-Peroxides and Prostaglandins by Phospholipase A2 in Platelets of Normal and Essential Fatty Acid Deficient Rats
Authors: Vincent J.E. , Zijlstra F.J.
Published: May 2019

Phospholipase A2 has a biphasic action upon rat platelet aggregation, enhancing after shorter and inhibiting after longer incubation periods. The first effect is inhibited by indomethacin.

Cycloperoxides and thromboxane-like substances and prostaglandins are formed on incubation with the enzyme in the same amounts whether aggregation is induced or not. Formation of cycloperoxides and thromboxanes is not leading to aggregation. Phospholipase A2 enhances the aggregation of the platelets of essential fatty acid deficient rats. Only very small amounts of cycloperoxides and thromboxane-like substances are formed. It is concluded that cycloperoxides and thromboxanes can enhance, but not induce aggregation in rat platelets.
Aortic Valve Replacement in a Patient with Severe Hemophilia B.
Authors: Ehrman M.L. , O’Donnell M. , Cooper B.A.
Published: May 2019

Thrombotic effects of prothrombin complexes are well known; successful valvular surgery with its attendant dangers of clot formation on synthetic surfaces have not been reported in patients with severe hemophilia B. The appearance of Connaught Factor IX Complex, prepared by ion exchange resin, with non-detectable levels of Factor IXa and Xa seemed to minimize this hazard. A 30 year old male with severe Factor IX deficiency (.5 to 2%) had proaressive cardiac failure, secondary to aortic insufficiency, which necessitated valve replacement. A trial infusion of Factor IX Complex did not alter platelets, fibrinogen, thrombin time, or Factor V & VIII levels, but did raise Factors II & VII to 640% and 215% respectively. The T½ for Factor IX was calculated to be 13.5 hours in this patient. Aortic valve replacement was complicated by significant clot formation on the afferent loop of the pump oxygenator, despite adequate heparin levels. 72 hours post-operatively, disseminated intravascular coagulation developed in a setting devoid of classical stimuli; this was easily reversed by increasing heparin infusion to full therapeutic levels. Heparin was admixed with all further infusions and no further thrombotic episodes occurred in his hospital course. 3 months later the patient has returned to full activity on no replacement therapy. This case demonstrates that while ion exchange preparations like those used in Britain have substantially less risk of coagulopathy than salt eluates, they do create a potentially hypercoaaulable state.
Fibrinolytic Response in Retinal vein Occlusion and Diabetic Retinopathy
Authors: De Rosa V. , Pe-duzzi M. , Montanari F. , Coccheri S.
Published: May 2019

Coagulation and fibrinolysis studies were performed in 40 patients during the second week after occurrence of Retinal Vein Occlusion (RVO), in 35 patients with Diabetic Retinopathy of various degree and no thrombosis (DR), and in an age-sex matched control group.

Euglobulin Lysis Time was markedly prolonged in 30% of RVO and in 44% of DR patients. Poor responders to venous stasis ranked around 35% in both groups. Urokinase inhibition was increased in one third of RVO and in about two thirds of DR patients. These and other findings like increase of Factor VIII or platelet aggregation were only seldom associated in the same subjects.

Decreased plasma or tissue fibrinolysis is a frequent finding both in RVO and in DR and trials with drugs inducing a sustained increase of fibrinolytic activity are therefore justified in these conditions.
Influence of Low Dose Heparin and of Acetylsalicylic Acid on Alterations of the Hemostatic Mechanism by Venous Occlusion
Authors: Vinazzer H. , Loew D.
Published: May 2019

Venous occlusion was used as a model to examine the action of antithrombotic substances. In 15 healthy volunteers, a blood pressure cuff attached to an arm was inflated to 90 mm Hg for 15 minutes. Blood was collected from an antecubital vein prior to and at the end of occlusion. Parameters of coagulation, of fibrinolysis, and of platelet function were examined.

At the end of occlusion, factors XIa and Xa were demonstrable in intact plasma. There was also a diminution of plasminogen and a shortening of the euglobulin lysis time. Collagen induced platelet aggregation was increased. Platelet factor 4 (PF 4) was diminished in platelets, and was demonstrable in plasma after occlusion.

Three groups were formed from these volunteers. Group 1 received 1500 mg acetylsalicylic acid (ASA) per day, group 2 was injected 5000 units heparin s.c. at 12 hr interval s, and group 3 received both drugs. After one week of therapy, blood samples were again collected before and after occlusion. In the ASA group, platelet aggregation was diminished. This was not altered by occlusion. PF 4 was neither diminished in platelets nor was it demonstrable in plasma. There was no influence on plasmatic coagulation factors nor on fibrinolysis. In the heparin group, factors XIa and Xa were not demonstrable in plasma after occlusion. There was no PF 4 in plasma though PF 4 in platelets was diminished. Platelet aggregation was not influenced. In the heparin plus ASA group, factors XIa and Xa as well as plasma PF 4 were missing after occlusion. PF 4 in platelets was not diminished whilst platelet aggregation was inhibited.

These experiments demonstrate a mutual support of ASA and heparin in the prevention of alterations of the hemostatic mechanism which are likely to provoke an increased thrombotic tendency.
Laboratory Control of Fibrinolytic Therapy
Authors: Theiss W. , Wirtzfeld A. , Sauer E. , Lutilsky L. , Kriessmann A.
Published: May 2019

Under fibrinolytic therapy, the levels of fibrinogen and of fibrin(ogen)degradation products (FDP) grossly influence most of the laboratory tests usually performed for clinical control of fibrinolytic treatment. Although these interferences are basically known, there is a lack of more detailed analysis under true clinical conditions. Therefore, fibrinogen determination according to Clauss (1957), thrombin time, and thromboplastin time were used for routine monitoring of 29 patients undergoing therapy with streptokinase for up to 6 days, and the results of these tests were correlated with true clottable fibrinogen as determined according to Ratnoff and Menzie (1961) and with FDP as determined according to Merskey et al. (1966).

Fibrinogen determination by the Clauss method yielded poor results; for immediate clinical control, however, they were sufficient, when values above 80 mg/dl were obtained (r=0.60 when compared to actual fibrinogen concentration). Values below 80 mg/dl were completely unreliable and more closely reflected FDP (r=0.63) than actual fibrinogen levels (r<0.20). The Clauss assay is entirely insufficient, when exact fibrinogen determinations are required as for rheological or pharmacological considerations. Quite in contrast, fibrinogen levels measured according to Ratnoff and Menzie were independent of FDP levels.

After bilogarithmic transformation, the thrombin time closely correlated to the FDP level (r=0.86); it thus is a valuable test for quickly estimating FDP. Since FDP also greatly influence thromboplastin time determinations (r=0.79), they must duely be considered, when overlapping oral anticoagulation is planned towards the end of fibrinolytic therapy.
Abnormal Platelets in Fulminant Hepatic Failure
Authors: Weston M.J. , Rubin M.H. , Bullock G. , Roberts J. , Langley P.G. , White Y. , Williams R.
Published: May 2019

Because bleeding occurs in fulminant hepatic failure, platelet function was investigated. In 12 patients, platelet counts were slightly lower than in controls (197 and 255 × 10 9/1, p<0.05), platelet volumes were smaller (5.89 and 6.68 μ3, p<0.001) and bleeding times were double (15.9 and 7.6 minutes, p<0.001). Patients’ platelets required ten times more ADP than controls to reach second phase aggregation (21.4 and 2.2 × 10-6m, p<0.001). Abnormalities of aggregation correlated with the prolonged bleeding times. Patient platelet poor plasma (PPP) did not prevent aggregation reaching second phase in controls and PPP from controls did not facilitate second phase aggregation in patients platelets. These defects of platelet function in fulminant hepatic failure are related to the severity of hepatic necrosis as platelet function in 16 jaundiced but non comatose patients did not differ from controls. Abnormalities of platelet ultrastructure are presented in an accompanying abstract.
Effects of a Splenic Extract on Coagulation
Authors: Spillert C. R. , Parmer L.P. , Hastings O.M. , Lazaro E.J.
Published: May 2019

We have previously described the ability of a lipoidal splenic extract (LSE) to decrease the coagulability of blood in laboratory animals. (Spillert, C. R. et al. : Hematological effects of a splenic extract. In Microcirculation. Vol 1, Grayson, J and Zingg, W. (eds) New York and London, Plenum Press 1976, p207). We report here the effects of LSE on platelet aggregation (PA) in blood of myocardial infarction prone and advanced malignant disease patients, (2) thromboelastographic values on the blood of patients with a variety of clinical states, and (3) survival of elderly mice with E. Coli endotoxemia (ECE). LSE was added to platelet rich plasma prior to determination of PA, and to citrated whole blood before recalcification in the thromboelastographic studies. One-year-old Swiss mice received LSE (10 mg/kg), or saline (controls), 2 hours prior to ECE (5 mg/kg) challenge. The decrease in PA (LSE versus controls) is indicated by the 16.0% (p<0.001) and 20.2% (p<0.05) inhibition of ADP induced PA in the blood of myocardial infarction prone (n = 27) and advanced malignant disease (n = 6) patients respectively. Thromboelastographic parameters R, R+K, and T50mm increased 17.5%, 17.7% and 21.2% respectively. (p<0.005). Forty-eight hour survivals in mice with ECE were 44.1% (saline, n = 34) and 79.5% (LSE, n = 39) p < 0.01). These results demonstrate the inhibitory effects of LSE on certain parameters of the coagulation process. It appears likely that the protective property of LSE in ECE is related to these anti-coagulation effects.
Alteration of Platelet Cyclic AMP (cAMP) by Ethanol
Authors: Cowan D.H. , Kikta M. , Baunach D.
Published: May 2019

Studies of cAMP in human platelets exposed to ethanol were done to assess one possible mechanism for ethanol-related platelet dysfunction. Ingestion of ethanol by 3 subjects produced blood ethanol levels from 65-76 mM. Thrombocytopenia occurred in 1 subject and impaired platelet function occurred in all. Platelet cAMP decreased 36,51, and 59% below control levels. Infusion of ethanol to 2 normals produced blood ethanol levels of 43 mM and decreased platelet cAMP by 15% and 22%. Incubation of normal platelets with 86 mM ethanol in vitro decreased cAMP from 13.8 ± 2.9 (1 SD) to 9.4 ± 3.5 (p<0.02). By contrast, ethanol did not impair the increase in cAMP that occurred with 1.3 μM PGE1. Further, ethanol enhanced the increase in cAMP produced by 2.0 mM papaverine (Pap) by 160-220% and that produced by Pap + PGE1 by 58%. Dopamine, 0.1 mM, caused a 23% decrease in the basal level of cAMP, a 31% decrease below the subnormal level of cAMP seen with ethanol alone, and a 41% reduction in the increased level of cAMP produced by Pap + ethanol. The effect of ethanol on platelet cAMP metabolism is complex. Ethanol reduces basal levels of cAMP, does not decrease elevated levels that result from PGE1 stimulation of adenylate cyclase, and augments the inhibitory effect of Pap on platelet phosphodiesterase (PDE). Despite causing a decrease in basal cAMP levels, ethanol may impair platelet function by potentiating the effect of agents or other conditions which increase cAMP. The effect of ethanol on Pap-stimulated PDE activity may be blocked by dopamine, a neuropharmacologic agent that is actively accumulated by platelets.
Some Aspects of Biliary Fibrinolysis in Man
Authors: Oshiba S. , Ariga T. , Sawai H. , Imai H.
Published: May 2019

The co-existence of a biliary plasminogen activator, bilokinase, and a streptokinase responsive factor, so-called proactivator, has recently been demonstrated in the bile of several mammalian species including man by Oshiba et al. (The 16th International Congress of Hematology, Kyoto, 1976). Our further investigations have revealed certain characteristic features of the biliary fibrinolysis in man which differ from those in other species. Human bile was obtained from patients with cholelithiasis who had received a surgical operation. The bilokinase activity and proactivator activity in the bile were measured in either the original form or in a quantitatively extracted form.

The main results were as follows: (1) The bilokinase activity of human bile was weak in comparison with that of certain other species. (2) Proactivator in human bile showed the highest activity, and the levels in the other species were generally less than the bilokinase activity. Thus, the activity ratio of proactivator to bilokinase in human bile far exceeded that in the other species. (3) Human bile proactivator and plasma proactivator demonstrated different affinity chromatographic behavior on lysine-sepharose. (4) There was apparently no significant relationship between biliary and plasma fibrinolysis.
Problems with Antiplatelet Therapy in Children with Cyanotic Heart Disease
Authors: Sutor A.H. , Schmidt B. , Lorenz H.P. , Vogt J.
Published: May 2019

Children with cyanotic heart disease are endangered by thromboembolic complications. However, in our 17 patients we could not find laboratory evidence for consumption coagulopathy but found evidence for isolated platelet consumption with increased platelet turnover. We therefore applied antiplatelet drugs (acetylsalicylic acid and dipyridamol) in order to prevent thromboembolic complications. With antiplatelet therapy the platelet count increased in average, however, 12 of 17 children with cyanotic heart disease did not show prolongation of the bleeding time as did controls. There is strong evidence that in these children an adequate blood-level of antiplatelet drugs was not obtained either by non-absorption (due to cardiac decompensation) or because of relative underdosage (due to increased platelet turnover) of the antiplatelet drugs. 2 of these patients who had a normal bleeding time, inspite of antiplatelet drugs, suffered from thromboembolic complications requiring fibrinolytic therapy. As a concequence we increase the dosage of antiplatelet drugs when the bleeding time does not prolong.