bluuebunny/crossref_metadata_2025 huggingface dataset:
Purification and Immunological Studies of Factors XI and XI a
Authors: Connellan J. M. , Castaldi P. A.
Published: June 2019

The inert form of factor XI has been purified from the supernatant of cryoprecipitated human plasma, and activated factor XI was prepared from the eluate of a celite extraction of human plasma. The purification steps involved were chromatography on both sepharose-heparin and hydroxylapatite columns, followed by preparative electrophoresis using 3.75% polyacrylamide in 8 M urea.

Antibodies were raised in rabbits by injecting 50 μg quantities of the antigen in multiple subcutaneous sites using Freuds adjuvant. Neutralizing antibodies were detected after the third injection and the nature of the antibody was studied by immunodiffusion and cross-over electrophoresis.

The antisera to factor XI a were monospecific and showed a biphasic precipitin line when human plasma was run in the first dimension. The antisera produced to factor XI contained an additional precipitin line which could be removed by absorption with plasma, which was depleted of factor XI by celite. Such antisera produced a monophasic precipitation reaction. These results suggest the possibility of distinguishing the inert and activated states of factor XI applicable to the investigation of thrombotic states.
The Change of Fibrin Split Products and Plasmin-Inhibitor in the Newborn Infant
Authors: Suzuki Shigenori
Published: June 2019

It is very important for an newborn to keep a certain balance of its bloodcoagulation and fibrinolytic system in order to maintain extrauterine life.

We have tried a follow up of coagulation abnormalities, fibrin split products and plasmininhibitors in severe asphyxia and respiratory distress syndrome. An other factor which is very dangerous for the fetus is acidosis.

The following results were obtained from 102 newborns:

1. A great number of FDP positive cases occurs in newborn who died from IRDS (80 μg/ml— 160μg/ml).

2. The plasminogen levels of cases who died from IRDS and severe asphyxia are lower than that of normal newborn infants.

3. α1-antitrypsin and antithrombin levels are relatively lower in the newborn period while the value of α1-macroglobulin maintains a normal level.

4. Correlation was seen between the coagulation abnormalities and acid base status (below actual pH 7.1).

The results shows the possibility that DIC is of great importants to RDS and FDP and are associated with hemorrhage and thrombosis.
Differences in Platelet Storage Pool Deficiency (SPD) of Hermansky Pudlak Syndrome (HPS) and Non-Albinos (NA)
Authors: White J. G. , Gerrard J. M. , Rao G. H. R. , Edson J. R. , Witkop G. J.
Published: June 2019

Recent reports of SDP platelets from patients with HPS (Albinism, accumulation of ceroid-like pigment, and mild hemorrhagic disease) and NA have suggested that the platelet defect in both conditions is identical. However, investigation of six HPS patients and three NA siblings with platelet SPD have indicated significant differences. The three NA patients were mildly thrombocytopenic while HPS patients had normal platelet counts. Levels of platelet serotonin (5 HT) were consistently lower in HPS than in NA with SPD, and approached normal concentrations in 2 of 3 NA patients. Dense bodies (DB), the organelles which contain the storage pool of serotonin and adenine nucleotides, were virtually absent in HPS platelets. SPD platelets from NA patients had significant numbers of DB which were markedly abnormal in their ultrastructure compared to normal platelet DB. Analysis of lipid peroxides and other substances reacting with thiobarbituric acid (TBA) revealed levels 5 to 10 times normal in HPS platelets while concentrations in SPD platelets from NA patients were similar to control values. Although the bleeding problems, aggregation defects, and abnormal platelet adenine nucleotide profiles were similar in HPS and NA patients, results of this study suggest that basic differences exist. The bizarre appearance of dense bodies in NA platelets is compatible with a structural abnormality interfering with storage organelle formation while accumulation of TBA reactive substances by HPS platelets may be the critical factor preventing development of the storage pool.
Heparin Neutralizing Effect of Lipoproteins
Authors: Bleyl H. , Addo O. , Róka L.
Published: June 2019

The effect of heparin on coagulation depends on its activation of Antithrombin III (ATB III). Substances which bind or neutralize heparin may compete with ATB III for heparin and inhibit the anticoagulatory effect of heparin-activated ATB III. Addition of isolated lipoprotein to a mixture of heparin and serum prevents the activation of ATB III. Lipoprotein isolated from 1 ml normal human serum binds about 40 mUnits (USP) of heparin. The antiheparin effect of lipoprotein is heat stable. As ATB III activity increases about 60% when serum is delipidated, the presence of lipoproteins in normal human blood may represent a natural inhibitor of antithrombin activity by complexing with heparin.
Binding of (14C) Dipyradamole (RA 8) to Platelets and Inhibition of ADP Induced Platelet Aggregation
Authors: Subbarao K. , Rucinski B. , Niewiarowski S.
Published: June 2019

RA 8 (cone. 10-5-10-6 M) strongly inhibits ADP induced aggregation of washed platelets but has no similar effect if platelets are suspended in the plasma medium, α, Acid glyco protein (GP) of human plasma has been found to complex PA 8 and to block its antiplatelet aggregation activity (Mewiarowski, S. et al. J. Lab. Clin. Med. 1975, in press). Incubation of washed platelets of human or rabbit with (14C) RA 8 (cone. 10-6 M) resulted in a binding of 20-30% radioactivity to platelets reaching a maximum within 1 minute incubation at 37° C and remaining constant for the length of time. Addition of human or rabbit plasma and oc, GP inhibited binding of RA 8 whereas other fractions such as fibrinogen, Cohn fraction II, IV and albumin did not show any effect. The binding of RA 8 to platelets correlated with the inhibition of ADP induced aggregation. The binding was independent of temperature at the range of 4-50° C and of pH at the range of 6.0-8.0. Intravenous injection of (14C) dipyradamole to rabbits at a dose of 25 mg/kg and 10 mg/kg weight resulted in a 32.6% and 6.0% (respectively) inhibition of ADP induced aggregation in platelet rich plasma (PRP). In both instances 3.0% of the total radioactivity recovered in PRP was associated with platelets. It can be suggested that α, GP present in blood may also inhibit binding of drug to platelets in vivo and interfere with its antiplatelet aggregation activity. Large doses of RA 8 are needed to overcome this effect.
Platelet Secretion Induced by Calcium Ionophores: Effect of Anti-Inflammatory Drugs
Authors: Mürer E. H. , Davenport K. , Day H. J.
Published: June 2019

Washed human platelets prelabeled with 3 H-serotonin and 14 C-adenine were incubated at 37° C with ionophores A23187 and X537A. 0.1 μM A23187 released the serotonin store of preincubated platelets after 1 min at 37° C, but was less effective when added in the cold. An increase in incubation time at 37° C did not result in increased release. Platelets preincubated with indomethacin showed reduction of up to 85% in released serotonin, while the metabolic parameters 14 C-ATP and 14 C-IMP were not significantly altered. The platelets from some donors did not show reduced release after treatment with indomethacin. This may indicate a variation in sensitivity to the release inducer similar to that described for Sr++-induced release (Biochim. Biophys. Acta 53-59, 362, 1974), or to the effect of indomethacin. 1 μM X537A caused a time-dependent serotonin release which increased from 2% at 1 min to 58% at 10 min incubation at 37° C. There was little change in 14 C-ATP following release and none in intra- or extracellular 14 C-IMP. 10 μM X537A caused release of 75-80% of the platelet serotonin after 1 min incubation. Longer incubation resulted in 14 C-IMP accumulation and leakage of 14 C-IMP to the surrounding medium. The results do not support the view that X537A and A23187 cause release from platelets by different mechanisms.

(Supported by grant HL 14217 from NHLI. A23187 was a gift from Dr. R. L. Hamill, Eli Lilly and Co., Indianapolis, Ind., and X537A from Drs. Zane Gaut and W. E. Scott, Hoffmann-LaRoche, Inc., Nutley, N. J.)
Complete Primary Structure of Prothrombin. Partial Primary Structures oí Plasminogen and Hirudin
Authors: Magnusson S. , Sottrup-Jensen L. , Glaeys H. , Zajdel M. , Petersen T. E.
Published: June 2019

The complete amino acid sequence (582 residues), positions of the twelve disulfide bridges and three carbohydrate substituents, position and identity of the ten vitamin Independent γ-carboxyglutamic acid (Gla) residues has been determined. A synthetic substrate specific for Factor Xa has been developed on the basis of the prothrombin structure. The primary structure of hirudin has been nearly completed and suggests a mechanism for its thrombin specificity.

The pro-part of prothrombin contains two 83 residue regions with identical disulfide-bridge pattern and 31 sequence identities. These “kringle” structures show sequence similarities with the pancreatic secretory trypsin inhibitor and hirudin. A chymotryptic digest of CM-plasminogen was investigated to find the heavy chain-light chain overlap region. This was isolated and sequenced (28 from h.c., 10 from l.c.) to study the specificity of plasminogen activators. From the same digest sequence evidence has been obtained which indicates that the heavy chain part of plasmin consists mainly of four kringle structures like the two in prothrombin, suggesting a common general structure and evolutionary origin of these two large zymogen structures.
The Relationship between 125I-Fibrinogen Detected Thrombosis and Pulmonary Embolism
Authors: Ruckley G. V. , Jones D. R. B. , Macintyre I. M. C. , Vasilescu C.
Published: June 2019

A total of 846 surgical and medical patients have been screened by the 125I-fibrinogen test (I.F.T.). Deep vein thrombosis (D.V.T.) was detected in 174 (21%). In 49% of positives the condition was bilateral and in 33% the D.V.T. involved popliteal veins or above. In 14 patients pulmonary embolism was diagnosed by lung scan or at autopsy. In one massive embolism was the cause of death. All 14 had positive I.F.T’s. ; 10 bilateral and 10 with both calf and thigh vein thrombus. The relationship between positive I.F.T. and iliofemoral D.V.T. was studied with bilateral phlebography in 60 patients. In no case was isolated iliofemoral D.V.T found. We confirm the value of the I.F.T. as a screening test for potential embolism and observe that embolism from isolated iliofemoral D.V.T. is extremely rare.
Localization of Microfilaments and Microtubules During Functional Shape Changes of Human Platelets (Adhesion, Aggregation, Retraction)
Authors: Lindner E. , Inczédy-Marcsek M. , Dierichs R.
Published: June 2019

Light and electron microscopic observations on spreading platelets show that microfilaments appear in relation to shape changes. Cell shape and in particular the types of process formation were correlated to the ability of platelets to adhere and aggregate. After activation or inhibition of spreading by Mg+2 and metabolic inhibitors (NaF and Malonic acid) respectively microfilaments and microtubules were localized in agreement with protoplasmic streaming associated with shape change. Contractility of intracellular microfilaments and motile activity of processes is demonstrated in the retracting blood clot. The significance of protoplasmic streaming for platelet function is discussed.
A New Case of Dysfibrinogenemia, Fibrinogen Marburg
Authors: Fuchs G. , Egbring R. , Havemann K.
Published: June 2019

A new case of congenital dysfibrinogenemia has been discovered in a 20 years old female, who suffered severe postpartal hemorrhage after delivery of her first child, followed by repeated episodes of thrombosis. Coagulation studies revealed prolongation of thrombin time. Reptilase time and Arvin time was markly prolonged. Very low fibrinogen levels were obtained by techniques which depend on the rate of fibrin formation following addition of thrombin, while heating and immunological procedures gave normal values of fibrinogen.

The patient’s fibrinogen had an inhibitory effect on the fibrin formation in normal plasma. Factor XIII activity was found to be normal using an isotopic method, whereas clot lysis time is shortened. The presence of an inhibitor and increased fibrinolysis were excluded. The coagulation studies indicate that delayed fibrin formation is due to a disturbance of fibrin monomer aggregation. In the turbidimetric assay patient’s monomers aggregated more slowly and incompletely compared to normal monomers.

Preliminary studies suggest that the release of fibrinopeptides A and B is normal.
A New Congenital Dysfibrinogenemia with Hemorrhagic Diathesis (Fibrinogen Montreal II)
Authors: Angelo G. , Lacombe M. , Lemay J. , Lavallée R. , Bonny Y. , Boileau J.
Published: June 2019

This report comprises of a new congenital dysfibrinogenemia in a Caucasian family with hemorrhagic diathesis.

Based on a study of three generations it shows to be an autosomal-dominant hereditary transmission.

The thrombin and the Reptilase time are prolonged, the prothrombin time and the partial thromboplastin slightly prolonged.

The fibrinogen concentration is normal when tested immunologically spectrophoto-metrically or by the heat denaturation method.

Abnormal aggregation of fibrin monomers, and low optical density polymerisation curve with a correcting effect whenever the ionic strength is decreased below 0.15 M.

The patient’s plasma has no inhibitory effect on the coagulation of normal plasma or fibrinogen.

Satisfactory results of treatment with fibrinogen during dental extraction will be presented.
Fibrin/Fibrinogen Degradation Products in Amniotic Fluid
Authors: Purdie D. W. , Howie P. W. , Edgar W. , Prentice G. R. M.
Published: June 2019

Fibrin/fibrinogen degradation products (F. D. P.) have been studied in amniotic fluid from normal pregnancies and in pregnancies complicated by hypertension, anencephaly, and hydramnios without foetal anomaly.

Using the haemagglutination inhibition immunoassay, F. D. P. were identified in 51 of 53 (mean 3.75 mcg/ml, S.D.±2.80) samples of amniotic fluid from healthy patients with normal pregnancies between 14 and 40 weeks gestation. In the hypertensive group of 16 patients the mean F. D. P. level of 5.00 mcg/ml (S.D.±3.25), was not significantly different from the normal group. In 7 patients with anencephalic foetuses the amniotic fluid F. D. P. level was significantly elevated (mean 17.5 mcg/ml, S.D.±5.75), while in 6 patients with hydramnios without foetal anomaly the F. D. P. level (mean 6.00 mcg/ml ±3.25) was not significantly different from the normal group.

Characterization of the amniotic fluid F. D. P. was carried out using double immunodiffusion polyacrylamide gel electrophoresis, and gel filtration on Sephadex G-200. The F. D. P. were present in two distinct components. The major fraction was fragment X (M. W. 240,000) and in a lower concentration, fragment D was identified.

The detection of raised amniotic fluid F. D. P. levels in early pregnancy may provide a diagnostic test for open central nervous system foetal abnormalities.
Determination of Factor Xa Activity by Means of Chromogenic Substrates Based on the Primary Structure of Prothrombin
Authors: Aurell L. , Olausson A. , Claeson G.
Published: June 2019

Through the work of Magnusson and co-workers leading to the elucidation of the primary structure of prothrombin including the amino acid sequences around the two bonds split by factor Xa it has been possible to design a synthetic chromogenic peptide substrate. Bz-Ile-Glu-Gly-Arg-pNA, specifically intended for the determination of factor Xa. Furthermore, additional substrates have been synthezised with various alterations in the amino acid sequence. The activity of factor Xa and other serine proteases within the coagulation and fibrinolytic systems towards these substrates will be discussed with special regard to their possible use in coagulation studies.
Studies on Platelet Factor 4 Like Activity in Atherosclerosis Susceptible and Resistant Pigeons
Authors: Fuster V. , Kottke B. A. , Ruiz G. E. , Lewis J. C. , Bowie E. J. W. , Owen C. A.
Published: June 2019

Recent studies suggest that a platelet factor(s) may be implicated in the stimulation of the smooth muscle cell proliferation occurring at the initial stages of atherosclerosis. Platelet factor 4 like activity in serum (PF4 = units of heparin neutralized by serum heated to 60° C) was measured at sequential ages in 9 White Carneau pigeons which develop spontaneous atherosclerosis first grossly apparent a 1 year of age (susceptible pigeons) and in 9 Show Racer pigeons which do not develop atherosclerosis (resistant pigeons). The susceptible pigeons developed progressive elevation of serum PF4, the maximal rate of increase being between 6 months (2.05u.) and 1 year (4.05 u.) (p < 0.001) and persisting at 3 years (3.40 u.). By contrast, in the resistant pigeons serum PF4 fell progressively from age 6 months (2.05 u.) to 1 year (1.85 u.) and 3 years (1.45 u.) (p < 0.001). Preliminary studies on thrombocyte count, size and ultrastructure transmission and scanning electronmicroscopy show no obvious differences between the two species of pigeons except for a more irregular and less pseudopodic surface in the thrombocyte of the susceptible pigeons.

The study suggests that platelet factor 4 may be correlated with the initiation of atherosclerosis in the pigeon.
Multiple Molecular Forms of Factor VIII
Authors: Seghatchian M. J. , Gaffney P. J.
Published: June 2019

The notion that factor VIII is present naturally in plasma as a large molecular weight aggregate (MW about 2 × 106) has recently come under scrutiny. This preliminary report of molecular size chromatographic separations (Sepharose 2B, 4B and 6B) of factor VIII in plasma and concentrate preparations suggests that factor VIII activity can exist in at least four different molecular sizes (a) over 2 × 106 (b) about 350,000 (c) about 180,000 and (d) about 60,000.

Chromatographic eluates of factor VIII concentrate were examined by polyacrylamide gel electrophoresis (with and without sodium dodecyl sulphate) and by immunoelectro-phoresis/immunoabsorption techniques against antisera to albumin, fibrinogen and factor VIII (void volume antigen). Varying conditions of chromatography suggested that the different size forms of factor VIII were molecularly interchangable, suggesting that a basic subunit (possibly inactive or nearly so) can associate with itself and/or with other plasma proteins of different sizes. Support for this proposition was found in the consistent presence of factor VIII activity and a low molecular weight (about 60,000) protein in all the chromatographic fractions of factor VIII concentrate from molecular sizes of over 2 × 106 to about 60,000. Another interpretation of the heterogeneity is that factor VIII in vivo is in a state of dynamic equilibrium which can be disrupted in vitro by proteolytic enzymes and/or by processing procedures.
Effect of Ethyloestrenol on Fibrinolysis in the Vessel Wall
Authors: Hedner U. , Nilsson I. M. , Isacson S.
Published: June 2019

It has been shown that phenformin plus ethyloestrenol causes a significant increase in the fibrinolytic activity of the vessel wall in most patients with recurrent idiopathic venous thrombosis and with an initially low fibrinolytic activity in the vein walls. At the same time a marked decrease in the frequency of thrombotic episodes occurred (Nilsson et al. 1974). Due to side effects of phenformin attempts have been made to administrate ethyloestrenol alone to patients with an abnormally low plasminogen activator content of the vessel wall and/or a decreased capacity to release plasminogen activator from the vein walls on venous occlusion of the arms. Ethyloestrenol in a dose of 8 mg/d has been given to 34 patients for 3 to 6 months and to 6 patients for 12 months. Ethyloestrenol caused a significant increase in the fibrinolytic activity in 30 of the 34 patients treated for 3-6 months and in 5 of those 6 treated for 12 months. Ethyloestrenol in a dose of 4 mg/d had no effect.
Intravascular Thrombosis and Platelet function in Diabetic Neuropathy
Authors: Preston F. E. , Timperley W. R. , O’Malley B. C. , Ward J. D.
Published: June 2019

Peripheral neuropathy is a serious complication of Diabetes Mellitus, The precise pathogenesis is unknown. Histological examination of biopsy material obtained from patients with diabetic neuropathy has revealed intravascular fibrin in vessels supplying the sural nerve in 10 out of 25 cases. This finding prompted us to study platelet function in this same group of patients.

Blood samples were obtained from fifteen patients. After preparation of platelet-rich plasma, platelet aggregation was measured following the addition of various concentrations of ADP, collagen and adrenaline. The responses were assessed photoelectrically in a temperature-controlled, constantly stirred, non-glass system. In addition the samples were tested for spontaneous platelet aggregation.

Spontaneous platelet aggregation was detected in one-third of samples studied, but in none of the controls. Enhanced ADP-induced aggregation was observed in 53% of samples and enhanced adrenaline-induced aggregation in 50%. Similar results were obtained with collagen.

The data supports the view that enhanced platelet aggregation and intravascular thrombosis is of considerable importance in the pathogenesis of diabetic neuropathy.
Coagulation and Fibrinolysis During and after Hepatic Lobectomy in 17 Patients
Authors: Gonard J. , Morisot P. , Huguet C. , Samanta M.
Published: June 2019

Hepatic lobectomy was performed in 17 patients for tumor (15 cases) or for hepatic cyst (2 cases). With regard to the hepatic lobes resected, there are 2 left lobectomies, 3 right, 9 right extended to the medial segment of the left lobe, 1 atypical resection of the right lobe and 1 of the left lobe, and 1 total hepatectomy followed by hepatic homotransplantation. In 9 patients, laparotomy showed that resection was not possible and they are used as controls.

Blood samples were drawn every 30 minutes during operation, every day or every other day during the two post-operative weeks.

Increased fibrinolytic activity and positive ethanol gelation tests are frequent but transient during surgery. However hemorragic problems are not encountered if hepatectomy is regulated. Routine administration of PPSB, antifibrinolytics or heparin does not seem to be required.

Variations of clotting factors and antithrombin III are observed during the days following surgery.
Influence of Volume Expanders on Endotoxin Induced Glomerular Fibrin Deposition
Authors: Schmidt E. H. , Fleitmann L. , Graubner F. , Beller F. K.
Published: June 2019

The continous infusion of endotoxin in virgin rats is followed by glomerular fibrin deposition. The same effect could be reached by a single injection of endotoxin followed by an infusion of normal saline. (Theiss, W. and Beller, F. K., J. Lab. Clin. Med. 3, 431-442, 1973). This experimental model was used to compare a variety of fluid solutions. Low and high molecular dextrane did not result in glomerular fibrin deposition but depressed the plasma fibrinogen level strongly. The infusion of gelatine produced glomerular fibrin deposition, which was significantly less pronounced than after infusion of normal saline. It is suggested that dextrane prevents glomerular deposition by interference with plasma fibrinogen.
Effects of Dietary α-Tocopherol and Polyunsaturated Fats on the Fatty Acid Composition of Platelet Phospholipids and on Blood Coagulation
Authors: Korsan-Bengtsen K. , Hallgren B. , Teger-Nilsson A.-C
Published: June 2019

The study group was 40 male post myocardial infarction patients 47-57 years old. All the participants were investigated two times with two weeks interval after which they were randomly divided into four groups with 10 subjects in each. Group 1 was given alpha-tocopherol 300 mg/day, group 2 was given alpha tocopherol 300 mg/day and a diet containing extra polyunsaturated fats, group 3 was given extra polyunsaturated fats but no extra alpha-tocopherol and group 4 served as a control group – thus continued their ordinary diet. After three months all participants were again investigated twice with two weeks interval.

On the values from all 40 subjects before the start of the dietary regimens linear regression analyses showed that there was a significant correlation between the content of the fatty acid 18 : 0 in the serin cephalin fraction and re calcification time in platelet rich plasma (RPRP), and a negative correlation between 20: 4 and RPRP. There was also a correlation between the ratio 18: 0/20: 4 and RPRP and a negative correlation between 18: 0/20: 4 and platelet factor 3 activity in plasma.

In group 2 there was a significant decrease in 18:0 and an increase in 20: 4 in the serin cephalin fraction from platelets after the diet period compared to preexperimental values. Russel’s viper venom clotting time (RVV) decreased significantly in group 1. There was a significant correlation between the decrease in RVV and the increase in plasma alphatocopherol.
Effect of Lipid Oxidation Products on Blood Coagulation
Authors: Barrowcliffe T. W. , Gutteridge J. M. C. , Stocks J. , Dormandy T. L.
Published: June 2019

Pure fatty acids in buffered aqueous suspension were allowed to autoxidise for up to 4 days. At intervals, lipid-free extracts were tested for effect on coagulation. 1-day extracts accelerated recalcification and RVV times, but products after 2-4 days oxidation showed progressive inhibition in these tests, and prolonged the P. T. and P. T. T. Addition of extracts to phospholipid altered its activity, and preparations of phospholipid extracted with an without antioxidant had different activity. In the absence of phospholipid, oxidation products had a threefold effect on thrombin generation in plasma: I), delay in appearance of thrombin; 2). increase in amount of thrombin formed; 3). less rapid neutralisation of thrombin. After fractionation of extracts by T. L. C. all these activities appeared in the most polar fration, which had the highest content of lipid peroxides. Thromboplastin generation tests showed that extracts delayed the formation of Xa, but increased the amount formed. Pre-formed Xa was strongly inhibited by oxidation products, but inhibition of thrombin was slight and could be abolished by glutathione. The presence of oxidation products in phospholipids could affect their activity in in vitro tests, and such products could also be involved in pathological coagulation in vivo.
Fibrinogen-Fibrin Interaction Studied by Affinity Chromatography
Authors: Starnberger A. , Hörmann H.
Published: June 2019

From human plasma, fibrinogen was adsorbed at 4° C on thrombin activated immobilized fibrinogen. Additionally retained plasma proteins were eluted at 37° C with 1.0 M Trisphosphate buffer, pH 7.6. Fibrinogen was subsequently eluted with 1.0 M KBr. It was free of contaminating proteins and showed a high clottability after removing KBr. Fibrin covalently linked to Sepharose 6B was reduced with dithioerythritol in 8 M urea. Washing with 8 M urea removed only औ-chains and γ-chains, while α-chains remained fixed to the insoluble matrix. Following carboxymethylation, the affinity of the immobilized α-chains to plasma fibrinogen was tested. Fibrinogen was adsorbed as described above. This may indicate that the α-chains are predominantly responsible for fibrinogen-fibrin interaction.

Supported by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 51.
Molecular Alterations of the Factor VIII/von Willebrand Factor (f. VIII/VWF) in von Willebrand’s Disease (vWd)
Authors: Gralnick H. R. , Coller B. S. , Sultan Y.
Published: June 2019

vWd is thought to be due to a deficiency of the f. VIII/VWf protein. Nine patients with vWd were investigated and their f. VIII/VWF protein characterized. In patients with < 20% procoagulant activity chromatography of cryoprecipitate revealed only small amounts of antigen and procoagulant activity. The f. VIII /VWF protein was not detectable by Polyacrylamide gel electrophoresis (PAGE). In patients with 20-50% f. VIII, both procoagulant and antigenic activity had delayed elution with a plateau of protein in the void volume (Vo). The f. VIII/VWF protein was present in the Vo fractions by PAGE; however, the protein was deficient in carbohydrate (C) as judged by PAS staining. Three patients had > 50% f. VIII, Two patients had normal elution of antigen, protein and procoagulant activity, the other had delayed elution with a unique pattern. In all 3 patients the f. VHI/VWF was seen on PAGE when stained for protein but not C. Plasma f. VIII/VWF arcs in crossed antigen-antibody electrophoresis were anomalous in all 3 patients. vWd may result from gene defects which lead to (1) decreased or absent production of the protein, (2) proteins of altered size and/or (3) proteins with altered C content.
The Effect of Platelet Basic Platelet Proteins on Cell Membranes and Small Vessels
Authors: Molander H. , Jørgensen L. , Seljelid R.
Published: June 2019

Basic proteins were extracted from washed human platelets and partially purified by DEAE-cellulose chromatography. The platelet basic proteins caused immediate release of histamine from rat mast colls taken from peritoneal cavity and isolated on Ficoll.

Platelet basic proteins added to cultures of mouse peritoneal macrophages, lead to loss of unit membrane function at 1–6 hours, as evidenced by slCr-leakage and loss of trypan-blue exclution function. This shows that high dosages of the platelet basic protein may have a delayed toxic effect on cell membranes. In rabbits, the basic proteins have both an immediate permeability increasing effect in small vessels inhibited by antihistamine, and a delayed permeability increasing effect on the vessel wall, not inhibited by antihistamine.
Brinase Treatment of Advanced Obliterative Arterial Disease
Authors: Ekeström S. , Frisch E. P. , Lund F. , Magaard F.
Published: June 2019

Thirtyfive patients with chronic peripheral arterial disease (10 Stage II, 23 Stage III and IV, 2 acute occlusions) were treated with brinase. Arteriosclerosis was the cause of disease in 27 patients, 8 had non-arteriosclerotic obstructions (5 thrombangiitis obliterans, 2 necrotizing vasculitis and 1 systemic lupus erythematodes). Twentyfive patients received full series of brinase treatment, in 10 patients treatment was discontinued (4/10 of reasons related to treatment).

Twenty patients were treated by i. v. brinase infusions. Dosage was based on determination of the brinase inhibitor capacity in plasma. In 5 cases brinase was instilled intra-arterially during vascular surgery, treatment was continued by i. v. brinase infusions. All patients except two were anticoagulated (dicoumarol and/or heparin). After initial arteriography the patients were followed by peripheral blood pressure measurement and in part by dynamic fluorescein angiography. Patients, treated peroperatively with brinase, were examined by electromagnetic flowmetry.

At termination of brinase treatment 20/25 patients showed clinical improvement. Increase of flow in the superficial femoral artery was recorded after peroperative intra-arterial instillation of brinase. The blood pressure gradient (arm-leg) before treatment (25 patients) was 112±24 mmHg. In patients with arteriosclerotic obstructions showing clinical improvement (14/19) the gradient was lowered to 49±20 mmHg, as compared to 112±27 mmHg in clinical failures. In patients with non-arteriosclerotic occlusions (6/6 improved) the gradient was lowered to 32±21 mmHg. Fluorescein angiography indicated improved microcirculation. Bleeding complications were not observed.
Surface Location of the Soluble Glycoprotein of Human Platelets
Authors: Okumura T. , Jamieson G. A.
Published: June 2019

A soluble glycoprotein has been isolated in homogenous form from the soluble fraction of human platelet homogenates (FEBS Letters 41, 30, 1974). Chicken antiglycoprotein antisera caused the agglutination of intact platelets and showed specificities directed against the glyeopeptide and peptide portions of the molecule. Treatment of intact platelets with neuraminidase prior to homogenization results in quantitative removal of sialic acid from the isolated glycoprotein. Similarly, radioactivity from 14C-glycine ethyl ester is incorporated into the glycoprotein by extracellular labeling with transglutaminase and it is cleaved to the macroglycopeptide by treatment of intact platelets with chymotrypsin, which does not cause the release reaction. These data indicate a surface location of the glycoprotein in intact platelets.

(Supported, in part, by USPHS grants.)
Factor XIII Deficiency in Congenital and Acquired Coagulation Disorders
Authors: Egbring R. , Schmidt W. , Havemann K. , Fuchs G.
Published: June 2019

According to Bohn the factor XIII molecule consist of 3 subunits (2 subunits A, MW 80,000 and subunit S, MW 180,000 named FSF-binding globulin).

In 3 patients with congenital defect (homozygotes) a total deficiency of enzyme activity and subunit A could be demonstrated, whereas subunit S was found to be 50% of normal. After substitution with human factor XIII concentrate an increase of subunit A can be observed by immunochemical methods. During a long term treatment of one patients with factor XIII concentrate since 1970 no antibodies against factor XIII could be detected.

In patients with acute leucemia (AML) acquired factor XIII deficiency occured. In this cases both subunits A and S were found to be reduced. This penomenon is not consistent with the concept of DIC.

However granulocytic proteases digest both subunits A and S of factor XIII in vitro. Moreover an elastase-like granulocytic protease could be found in plasma of these patients. These results demonstrate that factor XIII deficiency in AML may be due to direct proteolysis by enzymes released from granulocytes.

Further investigations suggest that this mechanism may bo operative in patients with septicemia too.
Venographic Studies on the Role of Valve Pockets in Thrombogenesis with Clinical Comments
Authors: Diener L. , Ericsson J.L.E. , Lund F.
Published: June 2019

In postmortem exploration of the veins of the lower limbs for studies of the frequency and significance of thromboembolism intraosseous venography with injection of contrast medium into the calcaneus was performed followed by complete dissection of the leg veins. A good agreement was found between venographic results and dissection findings. Out of the 596 legs examined, deep venous thrombosis was found in 229 (38%). In 32 of the lower limbs (5.4%) a definite localization and adherence of thrombi to valve pockets was noted. Altogether, 37 such thrombi were encountered. In addition 9 small thrombi of early type were found floating freely in the venous lumen just above a pocket. Microscopically the thrombi showed the greatest degree of organization at the bottom of the valve pocket speaking for this localization as the origin of formation. This is in accordance with the fact that stasis of blood would be most pronounced at the bottom of valve pockets as shown by retention of contrast medium in pockets in clinical venography. In a clinical series of venographies performed after hip surgery several thrombi located to valve pockets were found. Some of these thrombi had proximally free floating tails forming a potential menace of fatal pulmonary embolism.
Study of the Distribution of Platelet Volumes in Thrombocytopenia
Authors: Tohic F. Le , Prost-Dvojakovic R. J. , Samama M.
Published: June 2019

The distribution of platelet volumes was studied by Coulter F Counter and Coulter Z Bic Channelyser Model C 1000. Platelet rich plasma from severe thrombocytopenia often gives a curve with an abnormal pattern.

A first peak due to particles smaller than 3.5 μ3 is often present and due to elements of discussed origin. A second distribution, of elements larger than 3.5 μ3, corresponds with platelets, and, for sizes over 25 μ3, the shape of the curve can be altered by the presence of red cells or of cell stroma particularity frequent in thrombocytopenic plasma.

These abnormal results are however not found when mixtures of platelet rich plasma and platelet poor plasma are used.

For a better analysis of the distribution of platelet volumes in severe thrombocytopenia the albumin-gradient separation according to Levy-Toledano was found to be the method of choice in most cases.
Surface Structures of Human Platelets after Adhesion and Aggregation
Authors: Dierichs R. , Marcsek M. , Lindner E.
Published: June 2019

Human platelets that have adhered, spread out, and aggregated on the artificial surface - of an epoxy resin show an intact cellular membrane. After normal fixation and staining a filamentous material is seen in the extraneous coat which forms intercellular bridges. Incubation with colloidal iron demonstrates the existence of proteoglycans and unsaturated lipid acids. A distinction of proteinous and lipoid components can be achieved by ruthenium red which binds to proteoglycans and to fibrinogen and iodoplatinate which reacts with quarternary ammonium compounds. The latter method delineates the outer lamella of the unit membrane and liberated phospholipids whose relationship to platelet factor 3 is discussed.
Platelet Survival Time in Angina Pectoris Treated with Clofibrate or Placebo
Authors: Jonker J. J. C. , Schopman W. , Riel L. H. M. v. , Steur J. C. v. d. , Stoets W. , Ottolander G.J.H. den
Published: June 2019

With platelet survival time (PST) determinations we observed in a group of 20 normal volunteers a platelet half life time (T½) of 99.05 hours. In a group of 35 patients with angina pectoris (A. P.) we obtained a T½ of 84.6 hours (p < 0.05).

In a second trial on the effect of Clofibrate in patients with E. C. G. proven A. P., we obtained in 46 patients a T½ of 84.9 hours.

The difference with the “normal” group was again significant (p < 0.02). When looking at patients with A. P. + hyperliporoteinaemie (HLP), we found in comparison with the “normal” group in A. P. + HLP type IIA: acc. to Frederickson: T½ = 96,7 (ns); in A. P. + HLP type IIB: T½ = 88.3 (p < 0.2), in A. P. + HLP type IV: T½ = 82.2 (p < 0,02).

After 6 months treatment with Clofibrate or Placebo there was in the 27 A. P. treated with Clofebrate a significant increase in T½ of 106.8 hours (p < 0.01). In the group A. P. + HLP type IIB T½ = 90 (ns) and in the group A. P. + HLP type IV T½ = 94,3 (ns).

In the 27 Clofibrate treated patients cholesterol level decreased from 257 mg% to 217 mg% (p < 0.001), the fibrinogen level from 397 mg% to 247 mg% (p < 0.001) and the ESR from 22 mm to 14,7 mm (p < 0.01). Results and possible explanations will be discussed.
Acquired von Willebrand’s Syndrome and Acquired Thrombopathy in a Patient with a Chronic Lymphocytic Leukemia
Authors: Wautier J. L. , Levy-Toledano S. , Sultan Y. , Bodevin E.
Published: June 2019

The patient was 65 years old when she developped spontaneous bleeding. She did not have a family history of abnormal bleeding and profered clear evidence of normal hemostasis before.

Bleeding time, platelet aggregation induced by collagen were abnormal. VIII related antigen, ristocetin cofactor were 10 per cent the normal range while VIII procoagulant activity was 50%.

Interaction with rabbit sub endothelium was studied: contact and adhesion were 50% of the normal range and thrombi were absent.

Infusion of cryoprecipitate corrected partially von Willebrand abnormalities while platelet aggregation induced by collagen remained unchanged. Velocity of platelet aggregation induced by collagen, VIII related antigen and ristocetin cofactor increased moderately after corticotherapy.

The association of an acquired thrombopathy and von Willebrand’s disease in a patient with chronic lymphocytic leukemia lead one to wonder about possible relationship between these diseases in this case.
Immunological Separation and Study of a F. VIII Inhibitor
Authors: Casillas G. , Sinionetti G. , Pavlovsky A.
Published: June 2019

A potent F. VIII inhibitor developed by a patient with hemophilia A, poly transfused with homologous material, was studied. The ability of the inhibitor to inactivate human, bovine, and others mammals F. VIII activity showed its low species specificity. A crude gamma globulin preparation of the plasma with the inhibitor was mixed with higly purified bovine F. VIII, When the complex “bovine F. VIII-inhibitor” formed in the mixture was purified by gel filtration it did not show F. VIII activity or inhibitory action. By the use of specific antisera to bovine F. VIII and to human gamma globulins two antigens were found in the complex: a) bovine F. VIII and b) human IgG gamma globulin. Thus the nature of the inhibitor was seen in its purified form. For further evidence of the results, rabbit antiserum was prepared with the “bovine F. VIII-inhibitor” complex. This antiserum gave precipitation bands with: a) bovine F. VIII, b) human plasma (gamma globulins) and c) “Bovine F. VIII-inhibitor” complex. Even though this antiserum has a good precipitating capacity for bovine F. VIII, its neutralizing capacity against F. VIII biological activity is very low. Any way when the mixture antiserum-bovine F. VIII is ultracentrifuged, F. VIII activity disappears from the supernatant.
Heterogeneity of Platelets: Origin of Light Platelet Fraction from 32N Megakaryocytes
Authors: Penington D. G. , Lee N. Y. T. , Moore A. , Streatfield K.
Published: June 2019

Heterogeneity of circulating platelets has been ascribed to ageing but demonstration of differing cytoplasmic features in 8N, 16N and 32N megakaryocytes (Ser. Haemat. 8, 21, 1975) suggests that heterogeneity arises during platelet production.

Rat platelets have been separated on continuous density gradients (whole blood on Ludox: PVP) and platelets sized using a Coulter Counter with 24 channel pulse: height analyser. Approx. 80% of rat platelets settle between S. G. 1,058 and 1.067 with marked, consistent size variation (modal channel, 6). Approx. 12% settle between 1,042 and 1,058 and are small in size (modal channel, 3) with few large platelets. Platelets between 1,007 and 1,074 are a mixed population, both large and small.

In vivo 75Se selenomethionine injection gives synchronous labelling of the three density fractions.

IN vitro incubation of platelets with colloidal gold 188Au (200–400 Å) gives more avid labelling of light than of median and heavy platelets. Mature megakaryocytes in vitro also take up colloid and 32N cells, with abundant demarcation tubular system, are particularly heavily labelled. Origin of small light platelets from 32N megakaryocytes is suggested.
Image theory for a sphere with negative permittivity
Authors: Majić Matt
Published: June 2019

An image system for a point charge outside a dielectric sphere is presented for all complex values of relative permittivity ϵ = ϵ′ + iϵ″. The standard image integral solution of a point charge outside a dielectric sphere involving an image point charge plus a line source is shown to diverge for ϵ′ < −1, and a correction is proposed for this case, involving image multipoles of infinite magnitude which regularize the divergent line integral. The number of these multipoles depends on the position of ϵ relative to the resonant values ϵ = −1 − 1/n for positive integer n. The internal potential and dipole sources are also considered.
Enhanced thermoelectric properties in vacuum-annealed Bi0.5Sb1.5Te3 thin films fabricated using pulsed laser deposition
Authors: Symeou E. , Nicolaou Ch. , Kyratsi Th. , Giapintzakis J.
Published: June 2019

Localized cooling in microelectronics and nanoelectronics as well as energy autonomy in applications such as wireless sensor networks and wearable electronics could be well served by thin-film thermoelectric devices fabricated on rigid and/or flexible substrates. Bi0.5Sb1.5Te3 is considered to be a state-of-the-art p-type thermoelectric material at the desired temperature range, i.e., near room temperature (RT). Fabrication of Bi0.5Sb1.5Te3 thin films with bulklike thermoelectric properties (∼3900 μW/mK2 at 380 K) remains, however, a great challenge. In this study, we have successfully fabricated Bi0.5Sb1.5Te3 thin films on fused silica and Kapton substrates using a two-step process. The films were deposited at RT using pulsed laser deposition and then subjected to a postdeposition ex situ vacuum annealing process. The as-grown films were nearly amorphous. However, the annealing process enhanced both their crystallinity and texture, resulting in thin films with bulklike thermoelectric power factor values. Bi0.5Sb1.5Te3 thin films grown on fused silica and annealed at 350 °C for 16 h exhibit a power factor of 3750 μW/mK2 at 380 K. In addition, Bi0.5Sb1.5Te3 films grown on Kapton and annealed at 250 °C for 5 h and also grown on Kapton substrates at 250 °C exhibit a power factor of 2600 μW/mK2 at 390 K. Both of these power factor values are among the highest reported in the literature to date for Bi0.5Sb1.5Te3 thin films grown on fused silica and Kapton substrates, respectively.
Interfacial coupling modulation to the electrocaloric effect of Ba(Zr, Ti)O3 multilayered thick films
Authors: Hou Ying , Huang Xiaohua , Zhao Xiaobo , Yang Lu , Liang Ruihong
Published: June 2019

Negative environmental consequences of traditional cooling technologies have spurred the development of newly emerged green cooling devices rooting in electrocaloric effect (ECE). The relaxor ferroelectrics represented by BaZrTiO3 ceramics are being actively explored as a promising candidate for constructing ECE cooling devices. However, the reported electrocaloric (EC) response of BaZrTiO3 is still limited, particularly under low and moderate electric fields, which greatly inhibits its practical application. In this work, we propose an interfacial coupling modulation approach to enhance the EC response of BaZrTiO3 by introducing composition homogeneous and composition gradient multilayered structures. A systematic work was performed to study the dielectric, ferroelectric, and EC performances of BaZrTiO3 multilayered ceramics consisting of BaZr0.20Ti0.80O3 and/or BaZr0.17Ti0.83O3. It was found that the optimized BaZr0.17Ti0.83O3/BaZr0.20Ti0.80O3 bilayered films displayed the maximum EC response of 4.1 K under 10 MV m−1 at ambient conditions, corresponding to a high level of the coefficient of performance up to 18.33. The results demonstrate the efficiency of interfacial coupling modulation in boosting the EC response of relaxor ferroelectric ceramics, opening up an avenue to explore advanced electrocaloric systems with a giant electrocaloric effect.
GW quasiparticle energies of atoms in strong magnetic fields
Authors: Holzer Christof , Teale Andrew M. , Hampe Florian , Stopkowicz Stella , Helgaker Trygve , Klopper Wim
Published: June 2019

Quasiparticle energies of the atoms H–Ne have been computed in the GW approximation in the presence of strong magnetic fields with field strengths varying from 0 to 0.25 atomic units (0.25 B0=0.25 ℏe−1a0−2≈58 763 T). The GW quasiparticle energies are compared with equation-of-motion ionization-potential (EOM-IP) coupled-cluster singles-and-doubles (CCSD) calculations of the first ionization energies. The best results are obtained with the evGW@PBE0 method, which agrees with the EOM-IP-CCSD model to within about 0.20 eV. Ionization potentials have been calculated for all atoms in the series, representing the first systematic study of ionization potentials for the first-row atoms at field strengths characteristic of magnetic white dwarf stars. Under these conditions, the ionization potentials increase in a near-linear fashion with the field strength, reflecting the linear field dependence of the Landau energy of the ionized electron. The calculated ionization potentials agree well with the best available literature data for He, Li, and Be.