bluuebunny/crossref_metadata_2025_split huggingface dataset:
EVIDENCE FOR BIIIARY EXCRETION OF PHENPROCOUMON AND ITS METABOLITES IN HUMANS; IDENTIFICATION BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY AND GAS CHROMATOGRAPHY-MASS SPECTROMETRY
Authors: de Vries J X , Raedsch R , Stiehl A , Voelker U , Walter-Sack I , Weber E 
Published: August 2018

Recently it has been shown that in man the oral couma-rin anticoagulant phenprocoumon is eliminated up to 60-70 % in urine and 30-40 % in faeces; in urine phenprocoumon (PH) and its metabolites 7-hydroxy-(7-OH),6-hydroxy-(6-OH) and 4'-hydroxy-(4'-OH) phenprocoumon are present mainly as conjugates. No data so far were available on the biliary excretion of these compounds.We examined bile obtained from four in-patients during PH treatment; bile samples were aspirated in the duodenum at the papilla during routine diagnostic endoscopy and immediately deep frozen before analysis. Samples were extracted both untreated as well as after hydrolysis with 6-glucuronidase/aryl sulfatase and separated by reversed phase gradient elution high performance liquid chromatography (HPLC) with fluorescence detection; for confirmation, the same extracts were methylated and analysed by gas chromatography-mass spectrometry (CG-MS) (J.X.de Vries et al J Chromatogr., 338 (1985) 325). PH, 7-OH, 6-OH and 4'-OH were identified by comparison with synthetic authentic samples'''''''
A NEW DISORDER CHARACTERIZED BY FACTOR V DEFICIENCY AND MOLECULAR ABNORMALITY OF VON willEBRAND FACTOR ANTIGEN
Authors: Akutsu Y , Mori K , Suzuki S , Ishikawa M , Sakai H , Hiwatashi K , Fujimoto H , Endo E , Yasuda H 
Published: August 2018

Two patients with a new type of combined factor V and factor Vm deficiency is described. The propositus was 41 year-old woman who had never experienced any bleeding episode. We considered that this case was to be one of the combined factor V/Vin deficiency from the results of prolonged prothrombin time, 23.5 sec. (control 12.6 sec.) and kaolin partial thromboplastin time, 58.9 sec. (control 39.2 sec.) before surgery because of the right postoperative maxillary cyst. Coagulation studies at the time showed that the mean F. V and F. Vm levels were 11.5 % and 27 %, respectively. Von willebrand factor (vWF) which is the biological activity of F. VIII related platelet adhesion and vWF:Ag were also decreased (vWF:Ag 34 %, vwF 36.5 %). Bleeding time, platelet aggregation induced by ADP, collagen and ristocetin were normal. Another coagulation factors were normal and pathological inhibitors were not detected. The antigen levels of protein C (PC) and protein S (PS) were found to be normal. Her younger brother, who had no*past history of spontaneous bleeding, was found to be same abnormality (F. V 13 %, F. VIE 38 %, vwF:Ag 64 %, vWF 58 %). Bleeding time, platelet aggregation, another coagulation factors, PC and PS were normal. Furthermore, decreased intermediate multimers of vWF:Ag in these cases were observed on two-dimensional crossed immunoelectrophoresis.Since first case described by Oeri et al in 1954, approximately 40 combined factor V/Vm deficiency have been reported. In most such cases, vWF:Ag, vwF levels, and the electrophoretic mobility were within the normal range. In our cases, vwF:Ag and vwF were apparently decreased, and appeared to be the same abnormality as the von willebrand disease type-I. From the above mentioned findings, these patients were thought to be a new disorder of combined F. V deficiency and molecular abnormality of F. VIE ( vWF:Ag).
DETERMINATION OF ACTIVATED FACTOR VII BY BOVINE AND RABBIT THROMBOPLASTINS.
Authors: Kitchen S , Malia R G , Greaves M , Preston F E 
Published: August 2018

Thromboplastins from different animal species vary in their relative sensitivity to non-activated VII and activated VII (Vila) in plasma and this has formed the basis of Vila determinations using bovine and human thromboplastins. Since the use of human thromboplastin is now being discontinued we have re-evaluated the assessment of Vila using bovine and rabbit thromboplastins (BT and RT respectively). Citrated plasma was prepared from four normal individuals and maintained at 4°C in glass containers for 24 hours to promote the formation of Vila. Each plasma was subsampled at 0, 2, 4, 6 and 24 hours and factor VII assays performed using BT and two different rabbit preparations viz, (a) Manchester Reagent (RTmr, ISI = 1.16) and (b) Diagen phenolised (RT diagenlSI = 1.4). Results were expressed as the ratio BT/RT. With all thromboplastins factor VII activity increased with time. The changes were most marked using BT with much smaller increments occurring using the rabbit preparatrions. RTmr was less sensitive to Vila than RT diagen. The ratio BT/RTmr thus provides an index of Vila.Using this method we determined Vila in plasma samples from 29 normal subjects. Results obtained were x = 1 ,00;s.d. = 0.126; normal range 0.72 - 1.25 The method was applied to 29 plasma samples from 19 patients with disseminated intravascular coagulation giving BT/RTmr = 1.71 (range 0.92 - 3.38). We conclude that the ratio of Factor VII measured by bovine and rabbit thromboplastins respectively provides an index of Factor VII activation in plasma.
TISSUE FACTOR-FACTOR VII INHIBITION REQUIRES FACTOR Xa AND PLASMA LIPOPROTEINS.
Authors: Hubbard A R , Jennings C A 
Published: August 2018

Tissue factor rapidly loses procoagulant activity when incubated with defibrmated normal plasma and calcium ions. Inhibition is apparently directed against the tissue factor-Factor VII complex (TF-EVII) and requires Factor Xa and a component(s) found in A1 (OH)3-adsorbed plasma (AP). We have developed a two stage assay for the inhibitor which involves first, the incubation of a TF-FVII complex with test material in the presence of Factor Xa, followed by the amidolytic assay of residual TF-EVII activity.Our studies have indicated that the component of AP responsible for this effect is lipoprotein. Incubation of AP with antiserum to apo-lipoprotein B (apo B) reduced the inhibitory activity by 73%, whereas antisera to antithrombin III and a2-macroglobulin had no effect. Inhibition by AP does not appear to be caused by an artefact of adsorption, since the inhibitory-capacity of AP was 59% of normal, defibrinated plasma. This correlated well with the apo B antigen in AP, which was 64% of normal. Moreover, the dose/response lines of AP and normal plasma were parallel, suggesting that the inhibitor assay is not affected by the presence of normal levels of coagulation factors.Purified lipoprotein-rich fractions prepared from AP using density gradient ultracentrifugation all contained inhibitory activity. Incubation of these fractions with anti-apo B greatly reduced the inhibition by the very low density and low density lipoprotein-rich fractions (VLDL and LDL) but had essentially no effect on the high density lipoprotein-rich fractions (HDL). Incubation of LDL with Factor Xa produced an inhibitory component which eluted together with the apo B antigen during gel filtration. Inhibition appears to require the interaction of Factor Xa with plasma lipoproteins, particularly LDL. The product of this interaction is then able to bind and inhibit the TF EVII complex. The requirement of Factor Xa in order for inhibition to be expressed is indicative of a feedback anticoagulant response which may have physiological significance.
INVOLVEMENT OF FACTOR XII (F XII) AND PREKALLIKREIN (PKK) IN THE ACTIVATION OF UROKINASE (UK)-RELATED PROTEINS IN HUMAN PLASMA.
Authors: Binnema D J , Dooijewaard G 
Published: August 2018

Recently it has been shown that in human plasma two types of UK-related proteins occur: Type I, plasma UK, with UK-related antigenic determinants directly accessible to anti-UK antibodies and Type II with UK-related antigenic determinants which become accessible only after SDS treatment and separation of polypeptides on PAGE. In this study we compared the molecular and enzymic properties of the two types in: 1. plasma activated by dextran sulphate (DXS) euglobulin precipitation, 2. plasma that was not activated and 3. plasma deficient in F XII, depleted in PKK and subsequently activated by DXS. ACA 34 gel chromatography, SDS PAGE, fibrin underlay zymography and immunoblotting were used. Results:Conclusions: 1. The UK-related subunits of T1 and TII are active when cleaved, but relatively inactive in the single-chain form. 2. The presence of F XII and PKK is indispensable for activation of TII, but not for that of TI; TII contributes to the F Xll-de-pendent plasminogen activator activity reported earlier, TI to the F Xll-independent part. 3. Activation of TI by DXS with no F XII and PKK present impairs the formation of the 150,000 form. 4. The specific activity of TII is rather low, but its concentration in plasma (not shown) is at least ten times that of TI.
NORMAL APTT IN A PATIENT WITH A PERSISTENT ACQUIRED FACTOR XI INHIBITOR.
Authors: McKenna R , Cole E R , Yuk A , Whisler W 
Published: August 2018

A 55 year-old white male (MR) with documented normal hemostatic tests at our institution for 14 years, was admitted for a laminectomy. His pre-operative APTT and prothrombin time activity (Q) were again normal. A prolonged APTT of 58 seconds (21-31 N) and Q of 18% (> . 70% N) was noted on the 14th post-operative day; patient had received I unit PRBC during surgery.Mixtures of pooled normal plasma (NPP) incubated with MR’ s plasma (301, 37° C) yielded normal APTTs with as little as 20% NPP in these mixtures, suggesting a plasma factor(s) deficiency. Factor XI level was undetectable with normal values of factors Vlll-C, IX and XII. A factor XI level of 7% was found in incubated mixtures (60’ , 37° C) of equal parts of NPP + MR’ s plasma as compared to 70% in NPP + buffer and < 1% in MR’ s native plasma + buffer, indicating the presence of an inhibitor in MR’ s plasma. Infusion of 1540 ml of FFP (22 ml/kg) to the patient resulted in an APTT of 28 seconds and a Q of 36% with a persistent undetectable XI (expected ∿ 45%). The in-vivo lack of response to FFP was also suggestive of an inhibitor.Additional in-vitro tests showed that the response of MR’ s plasma was no different from that of NPP with the addition of increasing dilutions of human brain thromboplastin, differentiating this inhibitor from the Lupus type anticoagulant. The severely reduced Q was due to severe temporary reductions in factors II (20%), V (34%), Vll/X (2%) and X (20%) without any evidence of hepatocellular dysfunction or vitamin K deficiency. Fibrinogen, thrombin time, FSP, platelet counts and bleeding times remained normal throughout.There was no external evidence of bleeding after the first operation at any site, despite 1M injections and venepunctures. MR was reoperated on day 15 for a cauda equina hematoma; adequate intra and 72 hour post-operative management was with 1 U FFP Q 6 hourly and Amicar; this was associated with a normal PTT, Q %35% and undetectable XI, but without generalized bleeding. Discontinuation of this regimen resulted in onset of bleeding from drainage tubes only, without bleeding from the rest of the incision. Reinstitution of I U FFP Q 12 hourly and Amicar resulted in a prompt cessation of bleeding. Patient had no evidence for an autoimmune disease, dyspro-teinemia or any other illness. The potential role of the 1 unit of PRBC transfused cannot be discounted. Further data on this inhibitor will be presented.
COAGULATION FINDINGS IN ULCERATIVE COLITIS AND THE BENEFICIAL EFFECT OF FACTOR XIII CONCENTRATE SUBSTITUTION.
Authors: Suzuki R , Takamura Y 
Published: August 2018

Reports suggest that there is a tendency towards hypercoagulability in ulcerative colitis (UC). However, few reports deal with factor XIII which plays an important role in the wound healing process.In the present study, 8 coagulation parameters including factor XIII activity were determined in 5 patients with UC at the active stage and in 15 patients at remission. A comparison of the two groups shows that, in the active-stage patients, the levels of factor XIII activity were significantly lower, platelets and fibrinogen higher, and the PT prolonged.Furthermore, Factor XIII concentrate (Fibrogammin P ) was administered to 4 patients with active-stage UC and abdominal symptoms. Here, the symptoms (i.e. abdominal pain, melaena, diarrhea, etc.) disappeared in accordance with the increase in factor XIII activity. Endoscopyprevealed that treatment with factor XIII concentrate (Fibrogammin P ) had a beneficial effect on mucosal edema, redness and hemorrhage as well as on healing of erosions and ulcars.The results suggest that the level of factor XIII activity is a remarkably good measure of the severity of UC and that administration of factor XIII concentrate may be useful for treatment.
ON THE DETERMINANTS OF FIBRE THICKNESS AND FIBRIN NETWORK CHARACTERISTICS.
Authors: Nair C H , Shah G A , Dhall D P 
Published: August 2018

Spectacular differences are found between characteristics of networks developed in plasma and those developed in pure fibrinogen solution. Networks in plasma have thicker fibres, are more permeable and have lower tensile strength. In this investigation determinants of network structure under "hycio-logical conditions of clotting have been examined in an attempt to account for differences in network structure in plasma and in fibrinogen solution.Two independent variations of mass-length ratio ( µp and µr-.) from permeability and turbidity respectively were used. Effect of varying fibrinogen and thrombin concentrations and the effect of physiological concentrations of Antithrombin III, fibronectin, albumin, γ-globulin and platelet extract on fibrin network structure was examined.Fibrinogen and thrombin alter network characteristics through the modification of kinetics of network development.In these and several other studies it has been found that the kinetics of fibrin formation ultimately determine the final network structure through events preceding the appearance of visible fibrin. In separate experiments it was found that spectacular differences in network structure developed in plasma and fibrinogen were not entirely accounted for by alterations induced in network properties by albumin, y-globulin, fibronectin, ATIII and platelet extract.It is concluded that the final network structure is determined by kinetics of fibrin fibre growth and is highly responsive to the presence of plasma proteins and platelets.The findings may have fundamental applications to haemostasis and thrombosis.
SHEAR AND FIBRIN CLOT FORMATION; ROLE OF FPB RELEASE
Authors: Berk H R , Clark H 
Published: August 2018

It is well known that flow conditions affect many aspects of hemostasis. These effects range from gross morphological differences of thrombi formed at different flow rates to platelet activation to lysis. In addition, fibrin degradation products can influence blood flow. This investigation concerns itself with the effect of shear on the later stages of fibrin polymerization.Fibrinogen (human, Kabi) is reacted with thrombin (human, Sigma) under Couette flow conditions (volume-averaged shear 0-250 sec-1) in a pH 6.8, 4mM CaC12 HEPES buffered solution (I.S.-.15). Reaction times are chosen to achieve 30% FPA removal for various IT], Reptilase and Crotalase. Low Reynolds number hydrodynamic theory is used to investigate theoretical effects of shear on fibrils.The effect of shear on final clot formation is measured in terms of clot mass (%Pt). The effects of shear on orientation, diffusion and coagulation on the protofibril-fibril stage account for the initial decrease in %Pt at low shear. It is determined that the shear at which %Pt reaches a minimum (Gcrit) is thrombin concentration dependent. Lateral aggregation of fibrils is identified as the critical variable, with fibril breakage accounting for the increase in %Pt with increasing shear. The theory that fibrin formation proceeds by oligomer-oligomer addition is supported.Using hydrodynamic theory, a relationship betv/een axial ratio and Gcrit is obtained (assuming tensional or buckling failure). Using a hypothetical relationship between thrombin concentration and fiber axial ratio, a theoretical Gcrit versus IT] is obtained. This theoretical relationship is similar to that obtained experimentally, supporting the notion of shear induced breakage.Calcium is shown to increase Gcrit for a given [T]Experiments with Crotalase and Reptilase implicate FPB removal as an important factor in the minimum position. This provides an important insight into the significance of FPB removal in in vivo clot formation and shear stability.
DIFFERENCES IN PEPTIDE MAPS OF a POLYMERS FROM FIBRIN PRODUCED IN THE PRESENCE AND ABSENCE OF ERYTHROCYTES
Authors: Hoser M , Savidge G F 
Published: August 2018

α chain polymerisation during clot formation is accelerated in the presence of erythrocytes. This effect is abrogated if the erythrocytes are obtained from patients with various haemoglo-binopathies. Enzyme digests of the α polymers produced in the presence or absence of erythrocytes were prepared to further define any differences between them.Clots were produced from citrate/EACA plasma samples or plasma/erythrocyte mixtures by the addition of thrombin and calcium. After five hours, clots were washed iii 8 M urea until traces of haemoglobin were removed. After reduction and alkylation clots were dissolved in 0.5% SDS. a polymers were purified on sephacryl S-300 and protein concentrations were adjusted to 0.5 mg/ml. These were digested with S. Aureus V8 protease (150 mg/ml), papain (50 mg/ml) or chymotrypsin (100 mg/ml) at 37°C at sequential time intervals.After the addition of 2% SDS samples were analysed on 15% SDS polyacrylamide gels.In all cases digestion of a polymers from clots formed in the absence of erythrocytes took place more rapidly and contained peptide bands not apparent in other digests.The observations suggest that α polymers formed in the presence or absence of erythrocytes exhibit differing kinetics of response to proteolytic cleavage and indicate that erythrocytes may influence the primary and/or quartenary structure of the polymers studied.
IMPORTANCE OF CONFORMATIONAL CHANGE OF FIBRINOGEN INDUCED BY SURFACE ADSORPTION
Authors: He Lu , Mirshahi Mc , Soria J , Soria C , Mirshahi M , Thomaidis A , Boucheix C , Caen J P 
Published: August 2018

Concerning the problem of blood - foreign surface interaction, it is a great importance to know whether fibrinogen adsorbed on a solid surface is conformationally modified.Therefore, we tested the reactivities of soluble and immobilized fibrinogen against monoclonal antibodies obtained by mouse immunisation with fibrin derivatives, by an immunoenzymological assay using a monoclonal antibody which recognized an epitope available in the D domain of the molecule, but which is poorly exposed on soluble undegraded fibrinogen, we found that the epitope became accessible to this monoclonal antibody after the binding of fibrinogen to polystyrene. We concluded that fibrinogen undergoes conformational change when absorbed to a solide phase.Conformational change of fibrinogen induced by immobilization on polystyrene may modify the biological properties of fibrinogen molecule. We have shown that after its adsorption, fibrinogen allows plasminogen activation by tissue type plasminogen activator (tpA) in a similar manner as fragment D and fibrin. Therefore, this modification of fibrinogen structure may allow the exposure of the epitope which is masked on native fibrinogen and demasked on fragment D and fibrin and which is essential for tpA-induced plasminogen activation.These results might be very useful in the screening of synthetic material used for grafts and extracorporeal circulation.A similar conformational change occurs when fibrinogen binds to ADP-treated platelets. Further work however will be required to determine whether conformational change is essential for ADP-induced platelet aggregation.
IMMUNO-VISUALIZATION OF FIBRINOGEN Aa-CHAIN HETEROGENEITY IN NORMAL PLASMA COMPARED TO PLASMA FROM PATIENTS WITH DIC
Authors: Grøn B , Brosstad F 
Published: August 2018

Citrated plasma from healthy individuals or fran patients with disseminated intravascular coagulation(DIC) was diluted, and elec-trophoresed on 11% SDS-PAG slabs after reduction.Subsequently, the electrophoretic pattern was Western-blotted onto nitrocellulose and,-after blocking with gelatine-, reacted with either:a)polyclonal antibodies to human fibrinogen,b)polyclonal antibodies to FPA, or c)monoclonal antibody to non-released FPA (gift from Dr.Nieuwenhuizen, Leyden, Holland).Visualization of fibrinogen-related material was then done with peroxidase-conjugated secondary antibodies.In normal plasma a heterogeneity of the Aa-chain which was substantially more pronounced than hitherto described was noted. This pattern of fibrinogen Aa-chain heterogeneity was even more noticeable in plasma from patients with DIC. The presence/absense of all appropriate fibrinolytic inhibitors during sampling of blood had no effect upon the Aa-chain heterogenity patterns described above. This picture of heterogeneity was also seen when unreduced plasma was electrophoresed on SDS-PAG(2.5%)/AGAROSE(0.5%) slabs.When comparing the patterns produced by the three different antibodies used for identification/visualization, it could be deduced that: l)The pattern observed was mainly due to Aa-chain heterogeneity, and that the N-terminus of almost all species of Aa-chains were intact, confirming earlier observations that cell- or plasmin -mediated proteolysis of this chain is mainly C-terminal; 2) The present technique allows extranely sensitive characterization of both reduced and non-reduced plasma fibrinogen in the presence of all other plasma proteins.
ON THE IDENTIFICATION OF POLYMORPHIC SITES IN HUMAN FIBRINOGEN PEPTIDE CHAINS
Authors: Wilhelm S , Henschen A 
Published: August 2018

Human fibrinogen has repeatedly been shown to occur in a great number of different molecular forms. Some types of heterogeneity are evident already from variations in solubility properties and in ion-exchange-chromatographic as well as gel-electrophoretic behaviour of the total molecule and of its peptide chain components. The reason for these variations are partly known.Thus, degradation of the C-terminal parts and phosphorylation of two serine residues gives rise to heterogeneity in the Aα-chain. Differences in the sialylation of the carbohydrate side chain causes heterogeneity both in the Bβ- and the γ-chain. Differences in chain length at the C-terminus of the γ-chain are responsible for additional variation. All these variants are expected to exist in each human being. An other category of human fibrinogen variants may be due to genetic polymorphism within the population, i.e. the presence of inherited, common, normal variants. Seven sites of microheterogeneity have so far been tentatively identified, mainly by disagreements between protein and DNA sequence analyses. Three of the sites are located in the Aα-chain (positions 47, 296 and 312), three in the Bβ-chain (positions 162, 296 and 44-8) and one in the γ-chain (position 88). The aim of the present study was to identify these sites on the proteinchemical level in pooled plasma as well as in plasma from single individuals, especially various members of the same family. For this purpose suitable fibrinogen fragments containing the tentatively microheterogeneous sites were isolated after cleavage of fibrinogen with cyanogen bromide, trypsin and/or chymotrypsin by repeated fractionations by means of conventional and reversed-phase high-performance liquid chromatography and counter-current distribution. The components were characterized by N-terminal sequence and amino acid composition. Polymorphism in human fibrinogen has previously only once been identified by restriction fragment length analysis in a non-transcribed region of the Aα-chain locus but never in the transcribed regions, i.e. the peptide chains.The present investigation will allow the estimation of the number of peptide chain haplotypes and their possible correlation to other genetic variants of fibrinogen.