bluuebunny/crossref_metadata_2025 huggingface dataset:
Early Markers of Blood Coagulation and Fibrinolysis Activation in Argentine Hemorrhagic Fever
Authors: Heller M V , Marta R F , Sturk A , Maiztegui J I , Hack C E , ten Cate J W , Molinas F C
Published: July 2018

Junin virus, an arenaviridae, is the etiological agent of Argentine hemorrhagic fever. In addition to thrombocytopenia, patients present several alterations in both the blood coagulation and the fibrinolytic system, but diffuse intravascular coagulation could not be demonstrated. To investigate further the activation status of the two systems, levels of thrombin-antithrombin complexes (TAT), prothrombin fragment 1+2, protein C, total and free protein S, C4bBP, antithrombin III, t-PA, PAI-1 and D-dimer were measured. Fourteen patients with a confirmed diagnosis of Argentine hemorrhagic fever were included in the study, 2 were severe, 3 moderate and 9 mild clinical cases, but hemorrhages were slight throughout. Blood samples were collected for 6 consecutive days on admission and on remission. At admission TAT and F1+2 levels were increased in 13/14 patients, reaching 0.33 nM (0.06-0.87) and 2.16 nM (0.96-6.5), respectively. PC was low in 4 cases, fPS in 6 and tPS in 2, whereas C4bBP and ATIII values were within normal range. t-PA and D-dimer levels were high in 11/14 patients, reaching 20 ng/ml (2.7-106) and 1660 ng/ml (877-3780) respectively, while PAI-1 was considerably increased in the 2 severe cases and normal in the remainder. These results suggest low level though persistent process of blood coagulation and fibrinolysis activation in this viral hemorrhagic disease. We believe these abnormalities may lead to the well described bleeding manifestations in these patients.
Sensitivity to Activated Protein C; Influence of Oral Contraceptives and Sex
Authors: Henkens C M A , Bom V J J , Seinen A J , van der Meer J
Published: July 2018

We investigated sensitivity to activated protein C (APC) in 43 men, 42 women not using oral contraceptives (OC) and 38 women using OC containing 30.0-37.5 μg ethinyl estradiol. A commercially available kit was used. Men were more sensitive to APC as compared with women not using OC (p <0.005), but this difference seems not to be of clinical importance. Women using OC showed to be significantly less sensitive to APC, reflected by a lower APC-ratio, as compared with men (p <0.005) and women not using OC (p <0.05).

So, in normal individuals the APC-sensitivity differs according to sex and estrogen intake. This should be taken into account when interpreting APC-ratios.
Glycoprotein IIb/IIIa Blockade Inhibits Platelet-mediated Force Development and Reduces Gel Elastic Modulus
Authors: Carr Marcus E , Carr Sheryl L , Hantgan Roy R , Braaten Julie
Published: July 2018

The effects of GPIIb/IIIa blockade on clot retraction were studied utilizing an instrument which directly measures force produced by platelets. GPIIb/IIIa disruption by calcium chelation, and GPIIb/IIIa blockade by peptides and anti-GPIIb/IIIa antibodies were investigated. One mM EDTA suppressed ADP-induced platelet aggregation by 72% and reduced force developed at 1200 s by 33%. At 234 μM, the tetrapeptide Arg-Gly-Asp-Ser (RGDS) suppressed platelet aggregation by 74%, reduced force at 1200 s by 45% and reduced gel elastic modulus by 19%. At 10 μM, the peptide D-Arg-Gly-L-Asp-L-Try (D-RGDW) completely suppressed platelet aggregation, reduced force development by 38% and reduced gel elastic modulus by 29%. At 0.133 μM, monoclonal anti-GPIIIa antibody (AP-3) reduced force development by 74% and reduced gel modulus by 60%. Murine antiGPIIb/IIIa antibodies 10E5 and 7E3 markedly suppressed force development. At 0.133 μM, 10E5 reduced force by 89% and reduced gel modulus by 67%. At 0.053 μM, 7E3 completely stopped force development and reduced gel modulus by 46%. Platelet aggregation was blocked by 0.027 μM 7E3. Selective GPIIb blockade by antibodies did not affect force development. None of the agents studied altered fibrin structure as monitored by effects on fibrin mass/length ratios. Suppression of platelet aggregation occurred at inhibitor concentrations substantially lower than those required to suppress force development. Complete suppression of platelet aggregation did not assure inhibition of clot retraction probably due to profound platelet activation by thrombin. These results indicate that inhibition of fibrin(ogen) binding to GPIIb/IIIa, either by disruption of GPIIb/IIIa or by competitive blockade, inhibits platelet mediated force development and results in clot structures which are substantially less resistant to deformation by outside forces.
Anticoagulant-induced Pseudothrombocytopenia and Pseudoleucocytosis
Authors: Schrezenmeier Hubert , Müller Hilke , Gunsilius Eberhard , Heimpel Hermann , Seifried Erhard
Published: July 2018

Pseudothrombocytopenia (PTP) is the phenomenon of falsely low platelet counts due to in vitro platelet clumping in the presence of platelet autoantibodies and anticoagulants. We assessed anticoagulant- dependence, time course of platelet counts and impact of different counter devices on the phenomenon. Blood of 10 persons with previously recognized pronounced EDTA-dependent PTP was collected into 7 different anticoagulants and counted after different intervals in parallel in a Coulter T540 and a Coulter STKS counter and by phase contrast microscopy. With the Coulter T540 model PTP was most pronounced in blood samples anticoagulated with EDTA, Na-oxalate or Na-citrate. In the STKS counter EDTA, heparin and oxalate presented as the worst anticoagulants. The time course of platelet counts was significantly different between the two counters. Our results demonstrate that PTP is not restricted to EDTA, but is also present with other anticoagulants. In contrast, pseudoleucocytosis was observed only in EDTA-anticoagulated blood in the Coulter T540 device.

We investigated the expression of platelet integrins and activation antigens on platelets of persons with anticoagulant-dependent PTP and in healthy controls without PTP. In the presence of EDTA the expression of GpIIb/IIIa was significantly reduced in the PTP subjects compared to control. Activation antigens CD62, CD63 and thrombospondin-antigen were upregulated in the presence of EDTA. These alterations in the expression of platelet antigens could also be induced on platelets of normal donors by incubation with sera of PTP subjects and EDTA. The downregulation of GP IIb/IIIa in EDTA-blood of PTP subjects supports the hypothesis that the GPIIb/IIIa complex might be the binding site for the anti-platelet antibodies present in the sera of PTP subjects. Our results suggest that the binding of the autoantibodies causes platelet activation similar to other platelet-activating stimuli leading to subsequent platelet agglutination.
Arg506GIn Factor V Mutation (Factor V Leiden) in Patients with Ischaemic Cerebrovascular Disease and Survivors of Myocardial Infarction
Authors: Kontula Kimmo , Ylikorkala Antti , Miettinen Helena , Vuorio Alpo , Kauppinen-Mäkelin Ritva , Hämäläinen Liisa , Palomäki Heikki , Kaste Markku
Published: July 2018

The point mutation Arg506->Gln of factor V was recently shown to be an important and relatively common genetic cause of venous thromboembolism. Using a DNA technique based on polymerase chain reaction, we surveyed the blood samples of 236 patients with ischaemic stroke or a transient ischaemic attack, 122 survivors of myocardial infarction and 137 control subjects for the presence of this mutation. Although the frequency of the factor V mutation in patients with arterial disease (4.5%) was not significantly different from that in healthy blood donors (2.9%), a carrier status for this mutant gene was associated with symptoms of migraine and relatively mild angiographic abnormalities among patients with cerebrovascular disease. A more extensive study addressing the occurrence and significance of the mutant factor V mutation in patients with vasospastic cerebrovascular diseases seems to be warranted.
Raised Plasma Fibrinogen Concentrations in Subjects Attending a Metabolic Ward - Relation to Family History and Vascular Risk Factors
Authors: Margaglione Maurizio , Di Minno Giovanni , Grondone Elvira , Vecchione Gennaro , Celentano Egidio , Cappucci Giuseppe , Giordano Mauro , Grilli Massimo , Simone Pasquale , Fusilli Saverio , Panico Salvatore , Mancini Mario
Published: July 2018

We have evaluated plasma fibrinogen levels in 171 subjects attending a metabolic ward. As in the general population, a significant difference in plasma fibrinogen concentrations (p <0.05) was found between subjects with diabetes mellitus or hypertension and those without. However, fibrinogen was also abnormally high (p <0.05) when evaluated according to the presence of a family history of ischemic complications of atherosclerosis (p <0.05). In this setting, fibrinogen correlated with diabetes mellitus or hypertension as well as with familial risk, and the latter interacted with hypertension (p <0.05) in accounting for plasma fibrinogen.

The relationships between certain fibrinogen genotypes and familial risk have then been evaluated. Analysis of a locus (1.3 kb, HAE III digestion) of the promoter region of the BB fibrinogen gene, identified a polymorphic cutting site. The allele with the alternative restriction site (HI) was associated with mean fibrinogen levels which were 0.1-0.3 g/1 lower than those associated with the other allele (H2). This difference was not statistically significant. No obvious association was found between the familial risk and the presence of the H2 allele. We conclude that in a group of subjects from a metabolic ward, a positive family history for ischemic complications of atherosclerosis is consistently associated with high plasma fibrinogen levels. Interaction with hypertension significantly strengthens the association.
Variability of In Vivo Recovery of Factor IX after Infusion of Monoclonal Antibody Purified Factor IX Concentrates in Patients with Hemophilia B
Authors: White Gilbert C , Shapiro Amy D , Kurczynski Elizabeth M , Kim Hugh C , Bergman Garrett E ,   Mononine® Study Group
Published: July 2018

Monoclonal antibody purified factor IX concentrate, Mononine® (Armour Pharmaceutical Company, Kankakee, Illinois, USA), is a recently developed replacement factor concentrate for the treatment of patients with hemophilia B. The pharmacokinetic properties of monoclonal antibody purified factor IX concentrate (MAb Factor IX concentrate) have been evaluated in only small samples of patients, and little is known about those factors that might influence in vivo recovery of factor IX after infusion in a larger patient population. In vivo recovery of factor IX was therefore evaluated for 80 different indications in 72 patients who received MAb Factor IX concentrate for the management of spontaneous or trauma-induced bleeding, or as prophylaxis with surgery. The average recovery after infusions for presurgical pharmacokinetic analysis (mean ± standard deviation) was 1.28 ± 0.56 U/dl rise per U/kg infused (range 0.41-2.80), and the average recovery after all infusions for treatment was 1.23 ± 0.49 U/dl rise per U/kg infused (range -0.35-2.92). Recovery values for multiple MAb Factor IX doses in a given patient were also variable; the average recovery was 1.22 ± 0.53 U/dl rise per U/kg given, and standard deviations ranged from 0.03 to 1.26. Patient age, weight, and MAb Factor IX concentrate dose minimally but significantly influenced factor IX recovery. There was no significant effect of either race, history of previous thrombotic complications during treatment with other replacement factor concentrates, or bleeding state on recovery. All of the patients treated with this preparation experienced excellent hemostasis, and no thrombotic complications were observed.
Cellular Localization of Enzymatically Active Thrombin in Intact Human Tissues by Hirudin Binding
Authors: Zacharski Leo R , Memoli Vincent A , Morain William D , Schlaeppi Jean-Marc , Rousseau Sandra M
Published: July 2018

Cellular sites of coagulation activation within complex, intact tissues have been studied by immunohistochemical techniques. Hirudin, a specific and high affinity inihibitor of the active site of thrombin, together with antibody to hirudin were applied to sections of AMeX-fixed specimens of normal lung, kidney, placenta, freshly incised skin and unperturbed skin obtained at fresh autopsy; to rheumatoid synovial tissue; and to malignant tissue from a variety of tumor types. Staining for thrombin was observed selectively on pulmonary alveolar, rheumatoid synovial, and placental macrophages that express an intact extrinsic coagulation pathway. Staining was also observed restricted to the endothelium of capillaries in freshly incised skin but not in either unperturbed skin or in aged incisions. Staining of tumor cell bodies was observed in small cell carcinoma of the lung, renal cell carcinoma, and malignant melanoma tissues that we found previously to show tumor cell-associated procoagulant activity. This staining occurred commonly on cells within the tumor mass that were distant from stromal fibrinogen/fibrin. By contrast, tumor-associated macrophage but not tumor cell staining was seen in adenocarcinoma and squamous cell carcinoma of the lung, and little or no staining was seen in colon cancer tissue. Negative controls in which either the hirudin probe or its antibody were omitted failed to show staining. These results are in accord with previous findings and suggest that such techniques may be useful for studying the cellular sites of thrombin generation in intact tissues. We postulate that administration of potent and specific thrombin antagonists, such as hirudin, to patients with relevant tumor types might be followed by homing of hirudin to tumor cells in vivo so that effects of local thrombin generation on malignant progression can be determined.
IL-6 Upregulates Protein S Expression in the HepG-2 Hepatoma Cells
Authors: Hooper W Craig , Phillips Donald J , Ribeiro Maria , Benson Jane , Evatt Bruce L
Published: July 2018

Several pro-inflammatory cytokines have been shown to be important in the modulation of the procoagulant response. However, what role these cytokines may have in the regulation of coagulation inhibitors is poorly understood. While the hepatocyte is a primary site of synthesis for the anticoagulant proteins C and S, it is also a major target cell for the proinflammatory cytokine, IL-6. We have found that stimulation of HepG-2 hepatoma cells with IL-6 (5 ng/ml) significantly increased the production of the anticoagulant cofactor, protein S, in both a time and dose dependent fashion. This increase was seen at both the RNA and protein level. A mouse monoclonal neutralizing antibody to human IL-6 suppressed the IL-6 effect in a concentration dependent fashion. IL-6 also increased the release of the C4b-binding protein but had no effect on protein C production. When combined with either dexamethasone or soluble IL-6 receptor, the IL-6 response was significantly enhanced. Oncostatin M, a functionally related cytokine, had a similar effect while other related cytokines, IL-11 and leukemia inhibitory factor, only had a marginal effect. IL-1, TGF-β, and TNF-α had no significant effect on protein S production. These results indicate that IL-6 may play an important regulatory role in the anticoagulant pathway.
Plasminogen Activator Inhibitor Type-1 Determines Plasmin Formation in Patients with Ischaemic Heart Disease
Authors: Pedersen Ole Dyg , Gram Jørgen , Jespersen Jørgen
Published: July 2018

The aim of the present study was to find out whether plasminogen activator inhibitor type-1 (PAI-1) controls the formation of plasmin in patients with ischaemic heart disease.

We examined PAI activity, PAI-1 antigen, tissue type plasminogen activator (t-PA) activity, t-PA antigen, plasmin-α2-antiplasmin complex (PAP-complex) and fibrin degradation products D-dimer in 62 patients before (unstimulated) and after infusion of l-desamino-8- D-arginine vasopressin (DDAVP; stimulated). DDAVP was used in a standardized dose to trigger the release of t-PA from the vascular endothelium.

We observed that under basal conditions (unstimulated) median plasma t-PA activity for the whole group of patients was 86.5 mlU/ml (0-900), and after stimulation 2550 mlU/ml (0-6800), P <0.0001; median plasma concentration of t-PA antigen was 14.7 ng/ml (7.0-115.5) under basal conditions, and after stimulation 34.1 ng/ml (15.8-58.6), P <0.0001; median plasma PAI activity was 16.9 IU/ml (1.5-144.8) under basal conditions, and after stimulation 3.1 IU/ml (0-118.5), P <0.0001; median plasma concentration of PAI-1 antigen was 21.5 ng/ml (8.1-132.2) under basal conditions, and after stimulation 14.9 ng/ml (4.8-149.0), P <0.0001; the median plasma concentration of PAP-complex was 469.5 ng/ml (185.0-1802.0) under basal conditions, and after stimulation 695.5 (243.0-2292.0), P <0.0001; median plasma concentration of D-dimer was 298.0 ng/ml (103.0-948.0) under basal conditions, and after stimulation 296.5 ng/ml (97.0-917.0), P <0.0008.

Under basal conditions plasma PAI activities and plasma concentrations of PAI-1 antigen were both significantly negatively correlated with plasma concentrations of PAP-complex (rs = -0.32; P <0.02 and rs = -0.42; P <0.002, respectively). After stimulation of the fibrinolytic system by infusion of DDAVP, plasma PAI activities and plasma concentrations of PAI-1 antigen were also significantly negatively correlated with plasma concentrations of PAP-complex (rs = -0.41; P <0.002 and rs = - 0.33; P <0.009, respectively).

Our results indicate that PAI-1 regulates formation of plasmin in patients with ischaemic heart disease. These observations support that PAI-1 may play a critical role in the evolution of thrombosis.
Enhanced Vascular Plasminogen Activator (t-PA) Release by Epinephrine in Aged Rats
Authors: lacoviello Licia , D’Adamo Maria Cristina , Curtis Amalia De , Buczko Wlodzimierz , Donati Maria Benedetta
Published: July 2018

Adrenergic stimulation induces an acute fibrinolytic response in the vascular wall and can play an important role in limiting thrombus growth. The incidence of thrombotic disease increases with age. As many adrenergic functions are affected by aging, the vascular response to adrenergic stimulation was studied in young (3 ± 1 months) and aged (18 ± 3 months) rats, using an experimental model of vascular perfusion. Five min epinephrine (EPI) infusion (0.15-25 μM) induced a dose-dependent increase in tissue-type plasminogen activator (t-PA) release and in perfusion pressure, in both young and aged rats. In the latter, however, the basal activity of t-PA was significantly higher than in young animals (0.22 ± 0.02 vs 0.13 ± 0.01 U/ml; means ± SE; p <0.01, n = 40) and EPI infusion induced a significantly higher increase in t-PA activity (0.78 ± 0.06 vs 0.47 ± 0.08 U/ml, p <0.01, for 6.25 μM EPI). The response was characterized by a delayed onset. In contrast, the increase in the perfusion pressure by EPI was significantly lower in aged that in young rats, its basal levels being normal.

To evaluate whether the increased vascular response of t-PA was peculiar for adrenergic stimulation, we infused platelet activating factor (PAF), an inducer of t-PA release, not related to catecholamines. PAF, during five-minute infusion induced a rapid and dose-dependent (100-1250 pM) increase in t-PA release, to the same extent in both young and aged rats.

In conclusion, basal t-PA secretion was increased in aged rats probably as a response of vascular endothelium to chronic stimulation. Moreover, the vascular fibrinolytic response to acute EPI stimulation was greater in aging. This response was specific for adrenoceptor stimulation and could represent an adaptative mechanism of adrenergic receptors, counteracting an increased thrombotic tendency.
The Relationship of Venous Diameter to Reflux, Cephalad Thrombus and Cephalad Reflux Following Deep Venous Thrombosis
Authors: Tullis M J , Meissner M H , Bergelin R O , Caps M T , Manzo R A , Strandness D E
Published: July 2018

Duplex ultrasonography was used to measure the diameters of the common femoral, superficial femoral and popliteal vein segments in 123 patients following DVT. A cross sectional analysis was done based on the most recent visit to determine chronic venous diameter changes following DVT.

Venous diameters in recanalized segments were smaller at all levels compared to those never occluded (p = 0.06 for CFV and p <0.05 for SFV and PV). After accounting for a previous history of occlusion, the diameters of the segments with and without reflux were not significantly different. There was also no evidence of venodilation in segments caudal to cephalad reflux or thrombus.

Recanalized veins are smaller in diameter than those which were never thrombosed. Cephalad thrombus or reflux is not associated with venodilatation of caudal segments. Reflux following DVT is probably secondary to valvular damage rather than hypertension, since there was no diameter difference between refluxing and non-refluxing segments.
Inhibition of Urokinase-type Plasminogen Activator (uPA) Abrogates Myogenesis In Vitro
Authors: Muñoz-Cánoves Pura , Miralles Francesc , Baiget Montserrat , Félez Jordi
Published: July 2018

Urokinase-type plasminogen activator (uPA) is one of the components of blood’s fibrinolytic cascade. uPA acts as a broad spectrum proteolytic enzyme involved in different physio-pathological processes including cellular fibrinolysis, adhesion, migration, invasion and remodeling. Here, we present evidence that uPA participates in myogenesis, a process which requires drastic cell membrane reorganization, leading to the plurinucleated myotube from the progenitor myoblast. We have dissected the expression of uPA throughout the different myogenic compartments and found an increase in uPA enzymatic activity associated with myotube formation in C2C12 myoblast cells, with uPA mRNA increasing prior the onset of fusion and differentiation. When both fusion and differentiation were blocked by specific inhibitors (DMSO, cyto-chalasin B) the levels of uPA were strongly downregulated. This process was reversible and specific: the removal of the inhibitors immediately restored the levels of uPA mRNA while the specific inhibition of uPA enzymatic activity by an anti-uPA antibody resulted in a 50% reduction of the extent of fusion and in the abrogation of muscle-specific gene products, such as a-actin and MyoD. Moreover, the conversion of fibroblasts to muscle-like cells upon acquisition of MyoD resulted in a dramatic increase of uPA mRNA, which was partially due to transcriptional activation of the uPA gene. These results indicate that the increase in uPA expression prior to fusion and differentiation occurs via a MyoD-mediated mechanism whereas the normal MyoD expression requires the plasminogen activation-dependent activity of this protease. Therefore, these studies extend the sphere of influence of myogenic factors to fibrinolysis, an intrinsic component of the hematological system. Taken together, one mechanism used by the myoblast cell to become a differentiated myotube, involving the inductive extracellular proteolysis of urokinase, is proposed.
Ganglioside Content of Human Platelets – Differences in Resting and Activated Platelets
Authors: Ferroni Patrizia , Lenti Luisa , Martini Francesca , Ciatti Filippo , Pontieri Giuseppe M , Gazzaniga Pier Paolo
Published: July 2018

Gangliosides may play functional roles in platelet physiology, therefore this study has been designed to evaluate whether changes in ganglioside composition may occur as a consequence of platelet activation. The results obtained indicate that lactosylceramide and GM3 are the major glycosphingolipids of human platelets. The lipid-bound sialic acid (LBS A) content was 1.27 ± 0.04µεg/mg of protein. Resting platelets did not express GD3; GD3 was synthesized upon platelet activation (24 ± 8 ng/mg of protein). The stimulation of platelets with adenosine diphosphate showed the appearance of GD3 even in the absence of degranulation. Finally, incorporation of pyrene-labeled GM3 into platelet membranes, followed by stimulation with adenosine diphosphate, resulted in the appearance of a fluorescent band comigrating with GD3. The present studies indicate that sialyltransferase activation may occur as an early event following platelet stimulation, leading to GD3 synthesis mainly from the GM3 pool.
The Routine Determination of the Endogenous Thrombin Potential, First Results in Different Forms of Hyper- and Hypocoagulability
Authors: Wielders Simone , Mukherjee Manjari , Michiels Jan , Rijkers Dirk T S , Cambus Jean-Pierre , Knebel Richard W C , Kakkar Vijay , Hemker H Coenraad , Béguin Suzette
Published: July 2018

The area under the thrombin generation curve (the endogenous thrombin potential; ETP) has been proposed as a parameter for plasma-based hypercoagulability and to monitor anticoagulant treatment. We present an ETP assay for the routine laboratory using a centrifugal analyser. Throughput is 30 samples/h, within and between run imprecision is 4-5.6%. Suitable substrates were developed for the ranges of 10-500% and 2-100% of normal.

Independent of tissue factor concentration (if >4 pM), the normal value of the extrinsic ETP is 384.8 ±51.7 nM.min. The intrinsic ETP, triggered by ellagic acid, is 414 ± 41 nM.min.

The ETP is decreased to 15 and 35% of normal by oral anticoagulation (INR 2.5-4.0) and by heparin administration (APTT 1.5-2.5 X control).

The ETP is increased in untreated subjects with congenital antithrombin deficiency and in women using oral contraceptives. In deep vein thrombosis (phlebographically confirmed), it is increased by 29.4% (extrinsic) and 53% (intrinsic). In (angiographically assessed) coronary artery disease the increase is by 10% and 17% respectively.
Comparative Effects of Recombinant Staphylokinase and Streptokinase on Platelet Aggregation
Authors: Abdelouahed Mustapha , Hatmi Mohamed , Helft Gérard , Emadi Sharareh , Elalamy Ismaïl , Samama Meyer Michel
Published: July 2018

Recombinant staphylokinase (RSTA) has been shown to offer promise as a thrombolytic agent. In contrast to streptokinase (SK), few studies have been devoted to possible effects of RSTA on platelets. We have compared the capacity of RSTA and SK to trigger platelet aggregation and to modify ADP (2.5 µM) response in platelet-rich plasma (PRP) of 25 healthy subjects. Thus, exposure of PRP to SK (40 to 50 µg/ml) induced platelet aggregation in 6 out of 25 subjects. However, under the same conditions, RSTA failed to induce platelet aggregation in all cases (25 out of 25 subjects). In contrast to RSTA, SK (0.4 to 50 µg/ml) greatly reduced ADP-induced platelet aggregation in 12 out of 25 subjects. Preincubation of plasma with SK is associated with a decrease in the fibrinogen concentration. Furthermore, there was a good correlation between SK-induced fibrinogenolysis and SK- induced platelet aggregation defect (r2 = 0.9; p = 0.001). No fibrino genolysis was observed when different amounts of RSTA (0.4 to 50 µg/ml) were incubated in plasma for one min. However, there was a marked decrease in fibrinogen level (about 50%) when the plasma was incubated for five min with a very high concentration of RSTA. SK markedly enhanced the platelet response to ADP in 13 out of 25 subjects. In PRP of 6 out of 25 subjects, SK induces platelet aggregation and potentiates platelet response to ADP, however in PRP of 7 out of 25 subjects, SK caused only the increase of platelet response to ADP. The monoclonal antibody anti-FcγRIIal, IV-3 (2 [µ/ml), abolished SK-induced platelet aggregation and SK-enhanced ADP-induced platelet aggregation. In all cases (25 out of 25 subjects), RSTA failed to potentiate platelet response to ADP.

These findings confirm that RSTA has a lesser fibrinogenolytic ability than SK and suggest its negligeable effect on platelet function.
Variation in Plasma Fibrinogen over One Year: Relationships with Genetic Polymorphisms and Non-genetic Factors
Authors: Väisänen Sari , Rauramaa Rainer , Penttilä IIkka , Rankinen Tuomo , Gagnon Jacques , Pérusse Louis , Chagnon Monique , Bouchard Claude
Published: July 2018

We analyzed plasma fibrinogen level in relation to genetic polymorphisms in the a- and β-fibrinogen gene loci. Furthermore, the association of other CVD risk markers with fibrinogen was studied twice, with a time interval of one year in 50 to 60 year old men (n = 183). DNA polymorphisms were detected by PCR and digestion with TaqI (α-fibrinogen), Hind III and Bcl I (β-fibrinogen) restriction enzymes. The correlation coefficient between fibrinogen measurements was 0.48 (p <0.001). Blood leucocytes and waist-to-hip circumference ratio were the strongest correlates of fibrinogen level in both examinations, and the changes in leucocyte count and plasma fibrinogen correlated positively (r = 0.40, p <0.001). In Eastern Finnish men, the Taq I, Hind III or Bcl I restriction fragment length polymorphisms of the a- or β-fibrinogen gene loci did not associate with fibrinogen level, either cross- sectionally or longitudinally.
Monitoring of r-Hirudin Anticoagulation during Cardiopulmonary Bypass – Assessment of the Whole Blood Ecarin Clotting Time
Authors: Pötzsch Bernd , Madlener Katharina , Seelig Christoph , Riess Christian F , Greinacher Andreas , Müller-Berghaus Gert
Published: July 2018

The use of recombinant ® hirudin as an anticoagulant in performing extracorporeal circulation systems including cardiopulmonary bypass (CPB) devices requires a specific and easy to handle monitoring system. The usefulness of the celite-induced activated clotting time (ACT) and the activated partial thromboplastin time (APTT) for r-hirudin monitoring has been tested on ex vivo blood samples obtained from eight patients treated with r-hirudin during open heart surgery. The very poor relationship between the prolongation of the ACT and APTT values and the concentration of r-hirudin as measured using a chromogenic factor Ila assay indicates that both assays are not suitable to monitor r-hirudin anticoagulation. As an alternative approach a whole blood clotting assay based on the prothrombin-activating snake venom ecarin has been tested. In vitro experiments using r-hirudin- spiked whole blood samples showed a linear relationship between the concentration of hirudin added and the prolongation of the clotting times up to a concentration of r-hirudin of 4.0 µg/ml. Interassay coefficients (CV) of variation between 2.1% and 5.4% demonstrate the accuracy of the ecarin clotting time (ECT) assay. Differences in the interindividual responsiveness to r-hirudin were analyzed on r-hirudin- spiked blood samples obtained from 50 healthy blood donors. CV- values between 1.8% and 6% measured at r-hirudin concentrations between 0.5 and 4 µg/ml indicate remarkably slight differences in r-hirudin responsiveness. ECT assay results of the ex vivo blood samples linearily correlate (r = 0.79) to the concentration of r-hirudin. Moreover, assay results were not influenced by treatment with aprotinin or heparin. These findings together with the short measuring time with less than 120 seconds warrant the whole blood ECT to be a suitable assay for monitoring of r-hirudin anticoagulation in cardiac surgery.
Potentially Clinically Important Inaccuracies in Testing for the Lupus Anticoagulant: an Analysis of Results from three Surveys of the UK National External Quality Assessment Scheme (NEQAS) for Blood Coagulation
Authors: Jennings I , Kitchen S , Woods T A.L , Preston F E , Greaves M
Published: July 2018

The identification of the presence of antiphospholipid in plasma is recognised to be of diagnostic and prognostic importance in subjects with thrombotic disease, recurrent miscarriage or collagen vascular disorders. A number of coagulation assays are currently employed for the detection of lupus anticoagulant (LA), many of which are influenced by reagent dependent and methodological variables.

In the present study lyophilised plasma samples from three subjects with “strong”, “weak” and “absent” LA were tested in 220 centres. The most commonly used tests for LA were Activated Partial Thromboplastin Time (APTT), Dilute Russell Viper Venom Time (DRVVT) and Kaolin Clotting Time (KCT). Median DRVVT ratios were 1.75, 1.17 and 1.10 for the three samples. The presence of a strong LA was not detected by 4% of laboratories. The correct diagnosis was made by 94% of users of DRVVT and 85% of users of KCT. A weak LA was not detected by over half of centres. Correction was observed on addition of plasma and also in platelet neutralisation. The correct diagnosis was made by 37% of users of DRVVT and 27% of users of KCT. Lupus Anticoagulant was falsely considered to be present in a Factor IX deficient plasma by approximately one quarter of laboratories. Amongst users of DRVVT and KCT absence of LA in this sample was correctly reported by 73% and 69% of centres respectively.

The accuracy of testing for LA in the present study is suboptimal and this is likely to have important clinical consequences. There is clearly a need for greater conformity in the selection and performance of LA tests to facilitate accurate diagnosis of this important group of disorders.
PCR-RFLP Detection od PAI-2 Gene Variants: Prevelence in Ethnic Groups and Disease Relationship in patients Undergoing corony Angiography
Authors: Foy Carole A , Grant Peter J
Published: July 2018

PAI-2 is a fibrinolytic inhibitor produced predominantly by monocytes. Most PAI-2 is intracellular making study in clinical conditions difficult. Abnormalities in production may be associated with inflammation and fibrinolysis at sites of tissue damage such as the atherosclerotic plaque.

PAI-2 gene variants have been described: variant A consists of Asn120, Asn404 and Ser413 and variant B consists of Asp120, Lys404 and Cys413. We designed a PCR-RFLP assay using primers spanning the region containing Asn/Lys404 and Ser/Cys413. Variant B contains an Mwol restriction site. We analysed 302 Pima Indians and 286 healthy Caucasian volunteers. To investigate relationships between genotype and vascular disease we analysed 333 Caucasian patients undergoing coronary angiography.

Gene variant B was more common in the Pimas than in Caucasians (p <0.0001). There was no significant difference in genotype distribution between the volunteers and patients. In the patients there was no association between genotype and either a history of MI or extent of coronary atheroma.
A Reference Material for Harmonisation of D-Dimer Assays
Authors: Nieuwenhuizen Willem
Published: July 2018

The term “D-dimer assay” suggests, that these assays report the concentrations of the end-stage degradation product of crosslinked fibrin. This hardly occurs in patients. Degradation products of crosslinked fibrin rather occur with a wide range of molecular weights, and comprise various numbers of the D-dimer motif. Moreover, the numerical values obtained with different “D-dimer” assays vary widely. The variations are probably due to differences in reactivity of the various monoclonal antibodies used with the various D-dimer containing degradation products as they occur in patients; and to the various calibrators with a value assigned by the manufacturers. For the reasons indicated above a calibrator in the strict sense (e.g. pure D-dimer) is not feasible. This study shows that: it appears feasible to generate a conversion factor for each of the “D-dimer” assays studied, which will make the widely varying results obtained with these kits comparable; that the conversion factors can be based on a pool of real patient samples; and that the conversion factors are pool-independent.

The next and final step in this study is to prepare and make available an international reference material for use by manufacturers and others facing a comparison problem. This is being carried out, in collaboration with the NIBSC (Dr. P. Gaffney), and the material is expected to be available in early 1997.
Influence of effective channel length in self-aligned coplanar amorphous-indium-gallium-zinc-oxide thin-film transistors with different annealing temperatures
Authors: Kim Hyeong Wook , Kim Eok Su , Park Joon Seok , Lim Jun Hyung , Kim Bo Sung
Published: July 2018

Electrical characteristics of self-aligned coplanar amorphous-indium-gallium-zinc-oxide (a-IGZO) thin-film transistors (TFTs) were investigated for different annealing temperatures. The field-effect mobility of the a-IGZO TFTs increased with the annealing temperature, in particular, for a small channel length. The effective channel length (Leff) of the a-IGZO TFTs was extracted using the transmission line method. The decrease in Leff significantly depended on the annealing temperature, due to the hydrogen diffusion into the a-IGZO channel region. The intrinsic mobility calculated from the channel resistance of the a-IGZO TFTs was in good agreement with the mobility corrected by Leff.
Fabrication of single-electron transistors with electromigrated Ni nanogaps
Authors: Ito Mitsuki , Yagi Mamiko , Shirakashi Jun-ichi
Published: July 2018

We analyze single-electron transistors (SETs) fabricated with electromigrated Ni nanogaps using the Korotkov and Nazarov (KN) model. First, we investigate nanogap-based SETs consisting of multiple Ni islands placed between the source and drain electrodes by a field-emission-induced electromigration technique known as “activation.” After the activation procedure is performed using a preset current Is of 3 μA, the drain current-drain voltage characteristics of SETs with single-island structures are obtained and analyzed by using the KN model and considering the offset charges on the islands. We determine the fitting parameters obtained by the KN model from the electrical properties of the SETs. The parameters can be explained using the geometrical structures of the SETs that are observed in both scanning electron and atomic force microscopy images after the activation procedure. This approach allows the electrical and structural properties of the single-island structures of the SETs fabricated using the activation method to be determined.
Serial integration of Dean-structured sample cores with linear inertial focussing for enhanced particle and cell sorting
Authors: Holloway Paul M. , Butement Jonathan , Hegde Manjunath , West Jonathan
Published: July 2018

In this contribution, a channel aspect ratio of >2 was used to access high velocity regimes to provide confined sample cores by Dean focussing in advance of linear inertial focussing. This produces a singular separation origin with a mirrored transport path for efficient particle and blood cell sorting, while also increasing the spatial resolution for multiscale sorting.
Air-stable, transparent flexible ambipolar organic thin film transistors based on CuPc-F16CuPc bi-channel structure
Authors: Li Yi , Han Xiao , Li Yinghui , Zhou Weixin , Li Pan , Liu Ruiqing , Lin Xiujing , Huang Zhendong , Feng Xiaomiao , Ma Yanwen
Published: July 2018

We report the fabrication of air-stable, transparent flexible ambipolar organic thin film transistors based on CuPc and F16CuPc bi-channel films deposited under 10−1 Pa, by using graphene gate electrode, fluoropolymer Cytop dielectric layer, and Au source/drain electrodes on polyethylene terephthalate substrates. The devices show outstanding air-stability and high optical transparency of above 75% with electron and hole mobilities up to 0.031 and 0.029 cm2/V·s, respectively. The excellent ambipolar performance, along with the outstanding air-stability and high optical transparency, make CuPc-F16CuPc bi-channel transistors as a promising candidate for organic integrated circuits.
Consequences of season of prescribed burning on two spring-flowering terrestrial orchids and their endophytic fungi
Authors: Jasinge N. U. , Huynh T. , Lawrie A. C.
Published: July 2018

Prescribed burning is mandated in seasonally dry regions of the world to reduce fuel load and fire risk. This study investigated the effects of prescribed burning at different seasons on two Australian terrestrial orchids (Glossodia major R.Br. and Thelymitra pauciflora R.Br.) and their orchid mycorrhizal fungi (OMF) to find the least damaging season for a burn. Burns were conducted mid-season in autumn, winter, spring and summer. Orchids were enumerated and measured and their OMF isolated during their flowering seasons. The OMF were grouped and identified using their internal transcribed spacer (ITS) regions and tested for sensitivity to smoke water. Orchid numbers decreased by up to 100% after autumn and winter burns but not spring and summer burns. Plant height, leaf and flower dimensions decreased between the two years. Fire reduced success in the isolation of OMF. The relative frequencies of ITS sequences of Serendipita Roberts from G. major (but not of Tulasnella asymmetrica Warcup and P.H.B. Talbot from Th. pauciflora) changed after burns. OMF from G. major were up to 100% inhibited by smoke water, twice the inhibition of the OMF from Th. pauciflora. Prescribed burning during active orchid growth damaged both the orchids and their OMF. The least damaging practical season for a prescribed burn was in late spring, soon after seed dispersal.
One-Step Synthesis of CdSe Quantum Dots by Using Hydrazine Hydrate Reduction of Selenium Dioxide
Authors: Wang Rongfang , Wei Xingming , Xie Jun , Wang Bomei , He Xiaotong
Published: July 2018

Hydrazine hydrate was used as a novel reducing agent for the synthesis of water soluble and stable cadmium selenide (CdSe) quantum dots (QDs). The as-prepared CdSe quantum dots were investigated by X-ray powder diffraction, high-resolution transmission electron microscopy, photoluminescence, and UV-vis absorption spectroscopy analyses. The results show that the as-prepared CdSe QDs possess a cubic crystal structure and an average size of 2 nm. The effects of the pH of the original solution, thioglycollic acid (TGA)/Cd2+ molar ratio (nTGA/nCd2+), and the Cd/Se molar ratio on the luminescence properties of CdSe QDs were also systematically analysed.
Photo-Luminescence Dynamics of Ionic Liquids Composed of the Dicyanoaurate(I) Anion
Authors: Kimura Yoshifumi , Narita Takamitsu , Tanaka Saki , Taniguchi Mitsuhiro , Fujii Kaori , Endo Takatsugu , Yasaka Yoshiro , Ueno Masakatsu
Published: July 2018

Time-resolved luminescent spectra of ionic liquids of 1-butyl-3-methylimidazolium dicyanoaurate ([C4mim][Au(CN)2]) and N-butyl-N-methylpyrrolidinium dicyanoaurate ([P14][Au(CN)2]) at different excitation wavelengths (310 and 340nm) were measured using a streak camera. Immediately after photoexcitation, an intense luminescence band appeared at ~380nm that rapidly decayed with a time constant of 31 ps for [C4mim][Au(CN)2] and 71 ps for [P14][Au(CN)2]. With the decay of this band, another luminescent band appeared at ~460nm that slowly decayed (88ns for [C4mim][Au(CN)2] and 1.2μs for [P14][Au(CN)2]). The peak position of this second band shifted to a longer wavelength over time for both ionic liquids. The time profile of the peak showed a multi-exponential decay and depended on the excitation wavelength and the cation species. The peak shift is discussed in terms of the aggregation of anions. The excitation wavelength dependence was supposed to reflect the distribution of the larger oligomer in the ground state and the structural heterogeneity of the ionic liquids. The difference as a result of the cation is discussed in relation to the viscosity and the structure of the ionic liquids.
Quantifying wildfire growth rates using smoke plume observations derived from weather radar
Authors: Duff Thomas J. , Chong Derek M. , Penman Trent D.
Published: July 2018

Fast-moving wildfires can result in substantial losses of infrastructure, property and life. During such events, real-time intelligence is critical for managing firefighting activities and public safety. The ability of fixed-site weather radars to detect the plumes from fires has long been recognised; however, quantitative methods to link properties of radar observed plumes to fire behaviour are lacking. We investigated the potential for weather radars to provide real time estimates of the growth of large fires in south-eastern Australia. Specifically, we examined whether the rate of change in fire area could be approximated using the change in volume represented by radar returns. We evaluated a series of linear mixed-effects models predicting fire-area growth using radar data representing a range of dBZ thresholds and search volumes. Models were compared using an information–theoretic approach. Radar return volume was found to be a robust predictor of fire-area change. The best model had a minimum threshold of 10 dBZ and a search radius of 60 km (R2 = 0.64). Fire area and radar relationships did not vary significantly between radar stations, suggesting broad applicability beyond the dataset. Further development of the use of weather radars for wildfire monitoring could yield substantial benefits because of their high frequency of scan and broad coverage over many populated areas.
Cross-cultural literary translation strategies within a Maltese bilingual context
Authors: Grima Kenneth
Published: July 2018

The process of literary translation includes the source culture-specific elements that constitute an integral part of the source text. This paper aims to identify and analyse various translation strategic processes that could be adopted in translating cultural factors within the parameters of a Maltese bilingual, but not necessarily bicultural, context. Each of the suggested strategic procedures is presented in useful flow-chart formats, varying from source language/source culture to target language/target culture bias approach in order to keep cultural losses to a minimum whilst maximising cultural gains and, therefore, to make the transformation of the source text into the target text successful. Such flow-charts are aimed to provide the literary translator with a rapid means of achieving an adequate and satisfying suggested solution for a quality cross-cultural transposition of the cultural elements encountered within a bilingual context. In certain instances, it is also suggested that some strategies are used concurrently with others. To achieve this aim, an extended practical translation exercise by the author himself is used. This paper also helps to strengthen further both the level of research in narrative translation studies in general, and the research done in Maltese narrative literary translation from a cultural point of view.
On large orbits of subgroups of linear groups
Authors: Meng H. , Ballester-Bolinches A. , Esteban-Romero R.
Published: July 2018

The main aim of this paper is to prove an orbit theorem and to apply it to obtain a result that can be regarded as a significant step towards the solution of Gluck’s conjecture on large character degrees of finite solvable groups.
A Rnd3/p190RhoGAP pathway regulates RhoA activity in idiopathic pulmonary fibrosis fibroblasts
Authors: Monaghan-Benson Elizabeth , Wittchen Erika S. , Doerschuk Claire M. , Burridge Keith
Published: July 2018

Idiopathic pulmonary fibrosis (IPF) is an incurable disease of the lung that is characterized by excessive deposition of extracellular matrix (ECM), resulting in disruption of normal lung function. The signals regulating fibrosis include both transforming growth factor beta (TGF-β) and tissue rigidity and a major signaling pathway implicated in fibrosis involves activation of the GTPase RhoA. During studies exploring how elevated RhoA activity is sustained in IPF, we discovered that not only is RhoA activated by profibrotic stimuli but also that the expression of Rnd3, a major antagonist of RhoA activity, and the activity of p190RhoGAP (p190), a Rnd3 effector, are both suppressed in IPF fibroblasts. Restoration of Rnd3 levels in IPF fibroblasts results in an increase in p190 activity, a decrease in RhoA activity and a decrease in the overall fibrotic phenotype. We also find that treatment with IPF drugs nintedanib and pirfenidone decreases the fibrotic phenotype and RhoA activity through up-regulation of Rnd3 expression and p190 activity. These data provide evidence for a pathway in IPF where fibroblasts down-regulate Rnd3 levels and p190 activity to enhance RhoA activity and drive the fibrotic phenotype.