bluuebunny/crossref_metadata_2025 huggingface dataset:
A rationally assembled graphene nanoribbon/graphene framework for high volumetric energy and power density Li-ion batteries
Authors: Gao Liangliang , Jin Yi , Liu Xiaofang , Xu Ming , Lai Xiaokang , Shui Jianglan
Published: March 2018

A high volumetric performance electrode LFP/GNR/G is assembled by a coupling technique of “spray deposition + vacuum filtration” for Li-ion batteries.
Multifunctional hybrids with versatile types of nanoparticles via self-assembly for complementary tumor therapy
Authors: Duan Shun , Li Jia , Zhao Nana , Xu Fu-Jian
Published: March 2018

Multifunctional nanohybrids with various types of nanoparticles were readily constructed via self-assembly for combined photothermal/gene therapy of cancer.
Sm2O3/Co3O4 catalysts prepared by dealloying for low-temperature CO oxidation
Authors: Duan Dong , Hao Chunxi , Shi Wenyu , Wang Haiyang , Sun Zhanbo
Published: March 2018

A series of Co3O4 catalysts modified by Sm were prepared by a combined dealloying and calcination approach, and the catalytic activities were evaluated using CO catalytic oxidation.
Fe(CN)63− ion-modified MnO2/graphene nanoribbons enabling high energy density asymmetric supercapacitors
Authors: Sheng Lizhi , Jiang Lili , Wei Tong , Zhou Qihang , Jiang Yuting , Jiang Zimu , Liu Zheng , Fan Zhuangjun
Published: March 2018

Fe(CN)63− ion-modified MnO2/graphene ribbons can provide extra pseudocapacitance from the Fe(CN)63−/Fe(CN)64− redox reaction for high energy density supercapacitors.
Enhanced photovoltaic performance of polymer solar cells through design of a fused dithienosilolodithiophene structure with an enlarged π-conjugated system
Authors: Qi Feng , Zhang Yu , Wan Meixiu , Liu Juan , Huo Lijun
Published: March 2018

Polymer solar cells (PSCs) have garnered considerable attention in recent years. However, improving the fill factor (FF) remains a big challenge.
Stable narrowband red emission in fluorotellurate KTeF5:Mn4+viaMn4+noncentral-site occupation
Authors: Deng T. T. , Song E. H. , Su J. , Zhou Y. Y. , Wang L. Y. , Ye S. , Zhang Q. Y.
Published: March 2018

Unlike conventional central-site doping, here, we firstly report a stable Mn4+emission in novel fluorotellurate KTeF5viaMn4+abnormally equivalent and penta-coordinated octahedral vertical-site occupation.
Compact and high isolated triangular split‐ring diplexer
Authors: Xiao Jian‐Kang , Li Yong , Ma Jian‐Guo
Published: March 2018

A compact microstrip diplexer using triangular split‐ring resonator bandpass filters with separated electric and magnetic coupling (SEMC) has been presented. The transmission zeros are attributed to the SEMC without requiring more resonators and can be controlled by the electric and magnetic couplings. The diplexer is fabricated, and the design has been demonstrated by measurement. The diplexer has a compact size of 0.264λ0 × 0.102λ0, and a measured isolation of no less than 38 dB.
Extended cuckoo search‐based kernel correlation filter for abrupt motion tracking
Authors: Zhang Huanlong , Zhang Xiujiao , Wang Yong , Qian Xiaoliang , Wang Yanfeng
Published: March 2018

Kernelised correlation filter (KCF)‐based trackers have recently attracted considerable attention due to their exciting accuracy and efficiency. Numerous improvements have been made later for coping with scales variation or partial occlusion etc. However, when there is an abrupt motion between the consecutive image frames, these trackers would face failure. To alleviate the problem, the authors present an extended cuckoo search (CS)‐based KCF tracker (called ECSKCF). At first, the extended CS algorithm is constructed by the Simplex method (SM). CS has obvious capability in global search while the SM has exceptional advantage in local search. Based on ECS method, motion prediction is transformed to globally search for optimal position intending to enhance the quality of base image. Then, combined ECS with Gaussian distribution, a hybrid motion model is introduced to KCF framework, which has the capability of capturing abrupt motion. Finally, a unified framework is designed to track smooth or abrupt motion simultaneously. Extensive experimental results in both quantitative and qualitative measures demonstrate the effectiveness of the authors’ proposed method for abrupt motion tracking.
Printed quadrifilar square‐helical antennas with folded arms for satellite applications
Authors: Fartookzadeh Mahdi , Mohseni Armaki Seyed Hossein
Published: March 2018

In this study, a new straightforward method for construction of the printed quadrifilar helical antennas is presented. Folded helices with square shapes are used as the arms of the proposed antenna for satellite applications. The proposed structure is consisting of four printed flat walls with similar patterns of arms on each wall. Patterns of the folded arms are printed on four flat substrates with removed ground layers and the antenna structure is obtained by connecting these substrates. Two versions of the proposed antennas are designed for operating at 435 MHz with isolated and connected arms. Both antennas have minor responses at 145 MHz; however the large antenna with connected arms has better results. In addition, the large antenna has wider bandwidth and higher gain at both operating frequencies. Furthermore, the proposed antenna can be used as a wideband antenna with 34% bandwidth.
Wideband circularly polarised antenna with an asymmetric meandered‐shaped monopole and defected ground structure for wireless communication
Authors: Jhajharia Tejpal , Tiwari Vivekanand , Yadav Dinesh , Rawat Sanyog , Bhatnagar Deepak
Published: March 2018

A new geometry of wideband circularly polarised antenna with an asymmetric meandered‐shaped monopole is proposed in this communication. The wideband characteristics of the antenna are accomplished by the two asymmetric meandered‐shaped perpendicular arms of monopole and defected ground structure having slanting edge. The proposed prototype offers good performance in terms of impedance bandwidth (close to 102.04% in the frequency range 2.4–7.4 GHz) and the 3 dB axial‐ratio (AR) bandwidth (close to 37.5% in the frequency range 4.9–6.9 GHz). The realised impedance and AR bandwidths cover the WLAN/Wi‐MAX bands and Wi‐Fi band (5.5 GHz).
De Phlegmasia alba
Authors: Hach W. , Hach-Wunderle V.
Published: March 2018

Heinrich Hoffmann, der Vater des Struwwelpeter, gilt in seiner Heimatstadt Frankfurt am Main als Reformator der Sozialmedizin. Vor allem die Psychiatrie hat unter seiner ärztlichen Leitung den Weg in eine moderne Zukunft angetreten. Aus seiner Klinik für Irre und Epileptische ging die Frankfurter Universitätsklinik für Psychiatrie hervor. Problemstellung: Die Doktorarbeit von Heinrich Hoffmann befasst sich mit der Phlegmasia alba. Sie ist in lateinischer Sprache abgefasst. Bezüglich der Pathogenese gab es zu Beginn des 19. Jahrhunderts noch drei verschiedene Vorstellungen, Metastasen von Milch oder Lochien, die Verletzung oder Erkrankung von Lymphgefäßen und den Verschluss von Venenstämmen im Beckenbereich. Hoffmann griff aus der Literatur eine vierte Variante auf, die Entzündung der Tunica cellulosa, der bindegewebigen Umhüllungen von Muskeln und Sehnen sowie des Neurolemm der Vaginalnerven. Gegen die Theorie der Venenkrankheit hatte er wichtige scheinbare Argumente anzuführen. Die Doktorarbeit endet mit einer ausführlichen Beschreibung des Krankheitsverlaufes einer jungen Soldatenfrau. Konsequenzen: Die Arbeit bringt zwar in medizinhistorischer Hinsicht keine originellen neuen Aspekte. Ihr Studium erinnert an diagnostische Einzelheiten, die unter dem Eindruck der modernen technischen Möglichkeiten verloren zu gehen drohen. Dazu gehören die Charakteristik des Schmerzes vor dem Stadium der Schwellung, eine strenge Seitenbegrenzung auch der Beckenvenenthrombose, die treffende Bezeichnung des Ödems als porzellanartig und nicht zuletzt die Vorausschau des Begriffes des Klinischen Wahrscheinlichkeit in der Art von kritischen Tagen.
Gene Targeting of Components of the Fibrinolytic System
Authors: Ploplis Victoria , Castellino Francis
Published: March 2018

A number of in vitro and in vivo observations have implicated components of the fibrinolytic system in events associated with diverse physiological and pathophysiological processes, ranging from embryo implantation to cancer. Advances in gene targeting technology have led to the generation of mice deficient for components of the fibrinolytic system. Remarkably, these animals survive to adulthood with few spontaneous life threatening events. Thus, these mice are valuable resources for in vivo studies, not only for hemostasis-related research, but also for the relationships of these genes to other disease states associated with cellular growth and mobility, along with angiogenesis, vascular remodeling, inflammation, wound healing, and tumor growth and dissemination.
The North American Immune Tolerance Registry: Practices, Outcomes, Outcome Predictors
Authors: Kroner B.L. , DiMichele D. ,   The North American Immune Tolerance Study Group
Published: March 2018

The North American Immune Tolerance Registry was initiated to study of immune tolerance (ITT) in Canada and the United States with respect to: 1) therapeutic regimens in use for haemophilia A (HA) and B (HB) inhibitor patients; 2) therapeutic outcomes; 3) potential predictors of successful outcome and 4) complications of therapy. Data on 188 ITT courses was collected by questionnaire from 60 haemophilia centers from 1993-99. Among the completed courses, the overall ITT success rate was 70% (115/164) for all HA and 31% (5/16) for all HB. Outcome parameters noted to be predictive of ITT success for all HA were 1) pre-ITT induction (p = 0.003), 2) ITT peak (p = 0.007) and 3) historical pre ITT peak (p = 0.04) inhibitor titres. An inverse correlation between total daily dose (units/kg/day) and success: (80% with under 50; 71% with 50-99; 73% with 100-199; and 41% with ≥ 200, p = 0.01) was found. Outcome predictors were not evaluable for HB, although adverse reactions to therapy, including nephrotic syndrome, and access complications were more common among failed courses. Infection most often complicated the use of access catheters. These results are discussed within the context of the international ITT registry and upcoming prospective ITT study.
Oxidized Low-density Lipoprotein Associates Strongly with Carboxy-terminal Domain of Tissue Factor Pathway Inhibitor and Reduces the Catalytic Activity of the Protein
Authors: Hiraishi Sayuri , Hamuro Tsutomu , Kamikubo Yu-ichi , Matsuda Juzo , Horie Shuichi
Published: March 2018

Tissue factor pathway inhibitor (TFPI) is a physiological protease inhibitor of the extrinsic blood coagulation pathway. Previously we have shown that TFPI associates quite rapidly with oxidized lowdensity lipoprotein (ox-LDL), with a reduction of the inhibitory activity on factor X activation. In the present study, it was found, by means of agarose gel electrophoresis, that the pre-incubation of full-length rTFPI with heparin or the carboxy (C)-terminal part (peptide 240-265) of TFPI prevented the association with ox-LDL in a dose-dependent manner. When rTFPI lacking the C-terminal basic part of the molecule (rTFPI-C) was mixed with ox-LDL, only a small amount of rTFPI-C was shifted to the position of ox-LDL on electrophoresis. Further, ox-LDL did not reduce the activity of rTFPI-C. These results indicate that the C-terminal domain of TFPI molecule plays a predominant role in the binding to ox-LDL and the binding through the C-terminal part is essential for the ox-LDL-dependent reduction of the anticoagulant activity of TFPI.
Tissue-type Plasminogen Activator –7,351C/T Enhancer Polymorphism Is Associated with a First Myocardial Infarction
Authors: Jansson Jan-Håkan , Jern Sverker , Nilsson Torbjörn , Tjärnlund Anna , Boman Kurt , Ladenvall Per , Johansson Lars , Jern Christina
Published: March 2018

We recently identified a polymorphic Sp1 binding site in an enhancer at the tissue-type plasminogen activator (tPA) locus (tPA –7,351C/T), which was associated with vascular tPA release. Subjects homozygous for the –7,351C allele had twice the tPA release rate compared to subjects carrying the –7,351T allele. In this study we tested the hypothesis that the tPA –7,351C/T polymorphism is associated with myocardial infarction (MI). In a population-based prospective nested case-control study within northern Sweden, genotypes were determined among 61 MI cases and 120 controls. In a multivariate model, the tPA –7,351C/T polymorphism (OR 2.68 for T allele carriers; 95% CI 1.31– 5.50), tPA antigen (OR 1.16; 95% CI 1.07–1.25) and apo A-I (OR, 0.997; 95% CI 0.995–0.999) were independently associated with a first MI. These findings suggest that genetic markers of local tPA release and circulating steady-state tPA levels carry independent prognostic information.
The Antimitogenic Action of the Sulphated Polysaccharide Fucoidan Differs from Heparin in Human Vascular Smooth Muscle Cells
Authors: Mulloy Barbara , Gallagher Karen , O’Brien Leisha , Hughes Alun , Patel Mahendra
Published: March 2018

The sulphated polysaccharides fucoidan and heparin both inhibit vascular smooth muscle cell (VSMC) proliferation. In this study we compared their actions on mitogenesis and ERK1/ERK2 activation in human VSMC. Although they displaced cell surface [3H]-heparin binding with similar affinity, they exerted clearly distinguishable actions. Fucoidan potently inhibited DNA synthesis stimulated by foetal calf serum, PDGF-BB and thrombospondin-1. Heparin inhibited the mitogenic action of serum and thrombospondin-1 (though less potently than fucoidan), but failed to inhibit PDGF-BB-induced DNA synthesis. In parallel studies, fucoidan, but not heparin, inhibited ERK1/ERK2 activation by PDGF-BB. Moreover, fucoidan inhibited serum-induced mitogenesis in “heparin resistant” VSMC, which are refractory to heparin’s antimitogenic action. In summary, the structurally different polysaccharides, heparin, fucoidan (and fucans) have distinguishable effects on mitogenesis and ERK1/ERK2 activation, suggesting that different mechanism(s) mediate these actions. The potent antimitogenic action of fucoidan and its efficacy in heparin resistant VSMC emphasise the need to further investigate its mechanism of action in human VSMC and suggest this agent could have therapeutic potential.
Distribution of Th1- and Th2-induced Anti-factor VIII IgG Subclasses in Congenital and Acquired Hemophilia Patients
Authors: Lei Sijin , Lei Howard , Green Daid , Gill Joan , Conti-Fine Bianca , Reding Mark
Published: March 2018

Development of antibodies (Ab) that inhibit the procoagulant function of factor VIII (fVIII) seriously complicates the treatment of hemophilia A patients. It also causes acquired hemophilia, a rare yet serious autoimmune disease. The design of effective fVIII-specific tolerizing procedures will require elucidation of the role of the different CD4+ T cell subsets that drive inhibitor synthesis. To examine the contribution of Th1 and Th2 cells in the anti-fVIII Ab response, we measured the concentration of Th1- and Th2-driven anti-fVIII IgG subclasses in 17 patients with severe hemophilia A and 18 patients with acquired hemophilia. We found that both congenital and acquired hemophilia patients had similar and comparable proportions of Th1- and Th2-induced anti-fVIII Ab, suggesting a more important role of Th1 cells in the immune response to fVIII than previously appreciated. The distribution of anti-fVIII IgG subclasses was stable for periods of up to six months. More intense anti-fVIII Ab responses and higher inhibitor titers correlated with a predominance of Th2-driven subclasses. In contrast, Th1-driven anti-fVIII Ab were predominant in patients who had low anti-fVIII Ab concentrations, even when this was the result of successful immune tolerance or immunosuppressive therapy, which had caused drastic reduction or disappearance of inhibitors.

Thus, synthesis of Th2-driven inhibitors occurs when the anti-fVIII Ab response is intense, while Th1 cells may be involved in the long-term maintenance of anti-fVIII Ab synthesis.
The Thrombogram in Rare Inherited Coagulation Disorders: Its Relation to Clinical Bleeding
Authors: Al Dieri Raed , Peyvandi Flora , Santagostino Elena , Giansily Muriel , Mannucci Pier Mannuccio , Schved Jean , Béguin Suzette , Hemker H.
Published: March 2018

We investigated the relation between clotting factor concentration, the parameters of the thrombin generation curve (the thrombogram) and the severity of clinically observed bleeding in patients with congenital deficiency of prothrombin (n = 21), factor V (n = 22), factor VII (n = 22), factor X (n = 10), factor XI (n = 7) and factor XII (n = 6). The parameters used were: area under the curve (endogenous thrombin potential, ETP), peak concentration of thrombin attained and lag time before manifest formation.

Peak height and ETP varied linearly with the concentration of prothrombin. For the other factors these parameters hyperbolically approached to the 100% limit with increasing clotting factor concentration. Half normal ETP was seen at about the following concentrations: prothrombin (50%), factor V (1%), factor VII (2%), factor X (5%) and factor XI (1%). As a rule, the peak height was somewhat more sensitive to clotting factor decrease than the ETP was.

In all the patients with severe bleeding symptoms the ETP was less than 20% of normal. Bleeding tendency was absent or mild in patients with an ETP of 30% or higher. This value (except for prothrombin) is already obtained at concentrations of clotting factor of 1%-2%, which corroborates the clinical observation that a severe bleeding tendency is only seen in severe clotting factor deficiencies (less than 1%). The one exception was a patient with factor VII deficiency and severe bleeding, who showed a normal ETP value, albeit with a decreased peak height and a prolonged lag-time.
Is APC Resistance a Risk Factor for Venous Thromboembolism in Patients over 70 Years?
Authors: Lacut Karine , Le Gal Grégoire , Van Dreden Patrick , Bressollette Luc , Scarabin Pierre-Yves , Leroyer Christophe , Mottier Dominique , Oger Emmanuel
Published: March 2018

Activated protein C (APC) resistance is the most common risk factor for venous thromboembolism (VTE). Previous studies mostly analysed patients under 70 years and reported a four-to sevenfold increased risk. This case-control study included consecutive patients referred for a clinical suspicion VTE to our medical unit: 621 patients with a well-documented diagnosis (cases) and 406 patients for which the diagnosis was ruled out and who had no personal history of VTE (controls). APC resistance related to factor V Leiden was defined by either a positive DNA analysis or a positive STA® Staclot APC-R assay. Under 70 years, APC resistance was associated with a threefold increased risk of VTE (odds ratio 3.2, 95% CI, 1.7 to 6.0), whereas in patients over 70 years, it appeared to be no longer a strong risk factor (odds ratio 0.8, 95% CI, 0.4 to 1.7). Age appeared as an effectmeasure modifier with a significant interaction (p = 0.005). Our data suggest that APC resistance is not a risk factor for VTE in elderly.
Recombinant Nematode Anticoagulant Protein c2, a Novel Inhibitor of Tissue Factor-Factor VIIa Activity, Abrogates Endotoxin-Induced Coagulation in Chimpanzees
Authors: Moons Arno , Peters Ron , Cate Hugoten , Bauer Kenneth , Vlasuk George , Büller Harry , Levi Marcel
Published: March 2018

Systemic activation of coagulation leading to disseminated intravascular coagulation (DIC) is an important feature in patients with severe sepsis. Tissue factor has been shown to play a primary role in this pathological response, as revealed by the use of specific inhibitors and antagonists of the tissue factor/factor VIIa pathway. This class of agents has been demonstrated to attenuate the coagulation response in human volunteers with induced low-grade endotoxemia and to reduce mortality in primate models of Gram-negative sepsis. The efficacy of these agents in attenuating the activation of coagulation and formation of microvascular thrombosis in sepsis may depend on the mechanism of inhibition. Here we demonstrate the efficacy of recombinant nematode anticoagulant protein c2 (rNAPc2) that specifically inhibits the tissue factor/factor VIIa complex by a novel mechanism, in a model of endotoxin-induced coagulation activation in chimpanzees.

Administration of a low dose of Gram-negative endotoxin induced marked increases of thrombin generation as measured by plasma levels of prothrombin activation fragment F1+2 and thrombin-antithrombin complexes, which were completely blocked by rNAPc2. In chimpanzees receiving rNAPc2 alone, there was a significant reduction in the activation of factor X but not factor IX, compared to animals receiving placebo. In contrast to the effect of rNAPc2 on thrombin generation, there was no effect of this inhibitor on the well known enhanced systemic fibrinolytic response induced by endotoxin.

In conclusion, the recombinant peptide rNAPc2 is an effective inhibitor of tissue factor-driven thrombin generation during low grade endotoxemia. These results suggest that rNAPc2 may be a promising therapeutic option to inhibit coagulation activation in patients with sepsis.
Analysis of the Amino Acid Requirement for a Normal αIIbβ3 Maturation at αIIbGlu324 Commonly Mutated in Glanzmann Thrombasthenia
Authors: Milet-Marsal Sandrine , Breillat Christelle , Peyruchaud Olivier , Nurden Paquita , Combrié Robert , Bourre François , Nurden Alan
Published: March 2018

Glanzmann thrombasthenia is an inherited bleeding disorder arising from quantitative or qualitative defects of the αIIbβ3 integrin of platelets. Here, we report that PCR-SSCP analysis and DNA sequencing revealed a homozygous single base pair substitution in exon 12 of the αIIb gene leading to a Glu324 (E) to Lys (K) substitution in the αIIb subunit in a patient with Type I disease. As this mutation is found on at least 3 continents, the codon for Glu324 may be a mutational hotspot of the disease. To better understand this mutation, we analyzed the effect of substituting E324 with A324, L324, D324, Q324, N324, S324, as well as K324, looking at both αIIbβ3 maturation and cell surface expression in transiently transfected Cos-7 cells. The maturation state of the receptor clearly correlated with the level of cell membrane expression. Maturation efficiency was dependent on the electric charge as well as the size of the side chain of the amino acid present in what is a highly conserved N-terminal position in the third β-strand of blade 5 of the αIIbβ-propeller.
The Mucin-like Macroglycopeptide Region of Glycoprotein Ibα is Required for Cell Adhesion to Immobilized von Willebrand Factor (VWF) Under Flow but not for Static VWF Binding
Authors: Li Chester , Dong Jing-fei , López José
Published: March 2018

A dominant feature of the structure of platelet glycoprotein (GP) Ibα, the von Willebrand factor (VWF)-binding subunit to the GP IbIX-V complex, is the presence of an elongated, heavily glycosylated mucin-like stalk between the plasma membrane and the N-terminal 45-kDa ligand-binding domain. Here, we investigated the function of that region by expressing a mutant lacking residues 318-452 as part of a recombinant GP Ib-IX complex. We studied the VWF-binding function of this mutant under both static conditions and flow. The mutant GP Ibα was expressed normally on the surface of CHO bIX cells (stably expressing GP Ibβ and GP IX) and the proper conformation of the ligand-binding region was verified by the normal binding of 5 conformation-sensitive monoclonal antibodies. Under static conditions, cells expressing mutant GP Ibα bound VWF (binding induced by either botrocetin or ristocetin) in a manner indistinguishable from cells expressing wild-type GP Ibα. We also evaluated the ability of the mutant to mediate cell adhesion to immobilized VWF in the presence of fluid shear stress (at 2 and 10 dyn/cm2). When the mutant-expressing cells were incubated with immobilized VWF for 1 min before being exposed to shear, they rolled on the VWF surface in a manner similar to wild-type cells. However, if the cells were not first allowed to settle on the surface before the application of shear stress, the mutant GP Ibα was unable to capture the cells onto the VWF surface from the fluid stream, an indication that steric hindrance from other cell surface molecules may prevent access of the GP Ibα ligand-binding site to the surfaceimmobilized VWF.
Acquired Activated Protein C Resistance Associated with Anti-Protein S Antibody as a Strong Risk Factor for DVT in Non-SLE Patients
Authors: Kuratsune Hirohiko , Suehisa Etsuji , Kawasaki Tomio , Machii Takashi , Kitani Teruo , Iwatani Yoshinori , Kanakura Yuzuru , Nojima Junzo
Published: March 2018

Anti-phospholipid (aPL) antibodies (Abs) are well known to be associated with thromboembolic events in patients with systemic lupus erythematosus (SLE). However, the clinical relevance of aPL Abs in patients without SLE (non-SLE) who have venous thromboembolism remains unclear. We evaluated 143 non-SLE patients with a first episode of clinically suspected deep vein thrombosis (DVT) by using objective tests for diagnosing DVT and laboratory tests including the activated protein C resistance (APC-R) test, the factor V Leiden test, and various aPL Abs. The prevalence of acquired APC-R, in which case there was no factor V Leiden mutation, was significantly higher in patients with DVT (15/58 cases, 25.9%, p <0.0001) than in those without DVT (3/80 cases, 3.7%), and confirmed that acquired APC-R was a strong risk factor for DVT (odds ratio [OR], 8.95; 95% confidence intervals [CI], 2.45-32.7; p <0.001). Multivariate logistic analysis revealed that the presence of LA, aCL, anti- β2-glycoprotein I, anti-prothrombin and anti-protein C Abs was not reliable as a risk factor for DVT in non-SLE patients, and that the presence of anti-protein S Abs was the most significant risk factor for DVT (OR, 5.88; 95% CI, 1.96-17.7; p <0.002). Furthermore, the presence of anti-protein S Abs was strongly associated with acquired APC-R (OR, 57.8; 95% CI, 8.53-391; p <0.0001). These results suggest that acquired APC-R may reflect functional interference by anti-protein S Abs of the protein C pathway, which action may represent an important mechanism for the development DVT in non-SLE patients.
Interaction Between Hyperhomocysteinemia, Mutated Methylenetetrahydrofolatereductase (MTHFR) and Inherited Thrombophilic Factors in Recurrent Venous Thrombosis
Authors: Keijzer Miranda , Blom Henk , Bos Gerard , Willems Huub , Gerrits Wim , Rosendaal Frits , den Heijer Martin
Published: March 2018

Venous thrombosis is a multicausal disease involving acquired and genetic factors. In this study we investigated a possible interaction between hyperhomocysteinemia (fasting or postload) and factor V Leiden or prothrombin G20210A on the risk of recurrent venous thrombosis. We also looked at the risk due to mutations in the MTHFR-gene (C677T and A1298C).

We performed a case-control study in 171 patients with a history of recurrent venous thrombosis and 461 control subjects from the general population. Hyperhomocysteinemia (fasting or 6 h after an oral methionine load) was defined as a homocysteine concentration above the 90th percentile of the distributions in the control group.

The odds ratio (adjusted for age and sex) for recurrent venous thrombosis was 1.8 (95%CI: 1.1 to 3.0) for fasting hyperhomocysteinemia, 5.1 (95%CI: 3.0 to 8.6) for factor V Leiden and 1.8 (95%CI: 0.7 to 4.2) for prothrombin G20210A. We found 14 patients and 3 controls with both hyperhomocysteinemia and factor V Leiden, which yielded an odds ratio of 11.6 (95%CI: 3.2 to 42.5). We found no interaction between hyperhomocysteinemia and prothrombin G20210A. The relative risk for MTHFR 677CT was 1.6 (95%CI: 1.1 to 2.4) and for MTHFR 677TT was 1.4 (95%CI: 0.7 to 2.8). The combined risk for MTHFR 677TT and factor V Leiden was 18.7 (95%CI: 3.3 to 108).

We conclude that hyperhomocysteinemia and factor V Leiden are risk factors for recurrent venous thrombosis. The risk of thrombosis appeared high for individuals who had both risk factors.
Anti-β2-Glycoprotein I and Anti-Prothrombin Antibodies in Antiphospholipid-Negative Patients with Thrombosis
Authors: Previtali Sara , Barbui Tiziano , Galli Monica
Published: March 2018

We performed a case-control study to assess whether anti-β2-glycoprotein I and anti-prothrombin antibodies are independent risk factors of thrombosis. Cases were 79 patients with arterial and/or venous thrombosis without lupus anticoagulants, anticardiolipin antibodies and systemic lupus erythematosus; controls were 85 normal subjects. The prevalences and titers of IgG and IgM anti-β2-glycoprotein I and antiprothrombin antibodies were similar in both groups. Cases were analyzed with respect to the arterial or venous type of thrombosis and to the presence of congenital or acquired risk factors for thrombosis: no statistically significant relationships with the presence of anti-β2glycoprotein I and anti-prothrombin antibodies were found. Our data indicate that anti- β2-glycoprotein I and anti-prothrombin antibodies are not risk factors for thrombosis independent of lupus anticoagulants and anticardiolipin antibodies. Their measurement, therefore, is not warranted in the laboratory screening of patients with arterial and/or venous thrombosis.
Polymorphic Changes in the 5’ Flanking Region of Factor VII Have a Combined Effect on Promoter Strength
Authors: Kudaravalli Rama , Tidd Theresa , Pinotti Mirko , Ratti Antonia , Santacroce Rosa , Margaglione Maurizio , Dallapiccola Bruno , Bernardi Francesco , Fortina Paolo , Devoto Marcella , Pollak Eleanor
Published: March 2018

Polymorphic differences in the 5’ flanking region of the gene encoding procoagulant protein Factor VII (FVII) are associated with variations in FVII coagulant activity (FVII:C) and FVII antigen (FVII:Ag) levels. A decanucleotide insert polymorphism (CCTATATCCT) at 323 bp upstream of the start site of translation correlates with a decrease of approximately 20% FVII:C levels per allele containing this insert. However, linkage disequilibrium of the decanucleotide polymorphism with two single nucleotide polymorphisms (SNPs) at –122 and –401 have made it difficult to pinpoint the functional role, if any, of these genetic changes in lowering FVII levels. In vitro reporter gene studies in HepG2 cells analyzing the 8 possible combinations of polymorphic sites at –401, –323, and –122 reveal the necessity of the presence of the three concurrent polymorphic changes to maximally decrease promoter strength. In addition, these in vitro results are supported by in vivo studies in 89 individuals of African heritage, 34% of whom display a new haplotype that shows the polymorphic changes at -323 and -401 but lacks the change at -122.
Platelet Aggregation and Activation under Complex Patterns of Shear Stress
Authors: Zhang Jian-ning , Bergeron Angela , Yu Qinghua , Sun Carol , McIntire Larry , López José , Dong Jing-fei
Published: March 2018

Arterial stenosis results in a complex pattern of blood flow containing an extremely fast flow in the throat of stenosis and a post-stenosis low flow. The fast flow generates high shear stress that has been demonstrated in vitro to activate and aggregate platelets. One potential problem of these in vitro studies is that platelets are invariably exposed to a high shear stress for a period that is significantly longer than they would have experienced in vivo. More importantly, the role of the poststenosis low flow in platelet activation and aggregation has not been determined. By exposing platelets to a shear profile that contains both high and low shear segments, we found that platelets aggregate when they are exposed to a high shear stress of 100 dyn/cm2 for as short as 2.5 s, a period that is significantly shorter than those previously reported (30–120 s). Platelet aggregation under this condition requires a low shear exposure immediately after a high shear pulse, suggesting that post-stenosis low flow enhances platelet aggregation. Furthermore, platelet aggregation under this condition is not activation-dependent because the CD62P expression of sheared platelets is significantly less than that of platelets treated with ADP. Based on these findings, we propose that shear-induced platelet aggregation may be a process of mechanical crosslinking of platelets, requiring minimal platelet activation. This process may function as a protective mechanism to prevent in vivo irreversible platelet activation and aggregation under temporary high shear.
Blockade of Platelet GPIIB-IIIA (Integrin αIIbβ3) in Flowing Human Blood Leads to Passivation of Prothrombotic Surfaces
Authors: Riederer Markus , Ginsberg Mark , Steiner Beat
Published: March 2018

We examined the impact of platelet activation on platelet adhesion to collagen in flowing human blood. ADP activation of platelets in ex vivo flowing blood resulted in paradoxical inhibition of platelet deposition on collagen. Blockade of fibrinogen binding to platelets by Lamifiban, a competitive antagonist of GPIIb-IIIa (integrin αIIbβ3), reversed this inhibition, leading to a marked increase in integrin α2β1-dependent platelet adhesion. Analysis of integrin α2β1-dependent platelet adhesion to collagen indicated that ADP-induced suppression of platelet adhesion is the result of trans-dominant inhibition of integrin α2β1 caused by fibrinogen binding to integrin GPIIb-IIIa. Lamifiban blocked fibrinogen binding, reversing the trans-dominant inhibition of α2β1 dependent adhesion to collagen. The GPIIb-IIIa antagonist resulted in the formation of a nonthrombogenic, passivated surface comprised of an adherent platelet monolayer. This unexpected consequence of blocking fibrinogen binding to GPIIb-IIIa may explain the long-term benefits of short-term GPIIb-IIIa antagonist treatment of Acute Coronary Syndrome patients.
Regulation by Nitric Oxide of Endotoxin-Induced Tissue Factor and Plasminogen Activator Inhibitor-1 in Endothelial Cells
Authors: Pérez-Ruiz Ana , Montes Ramón , Velasco Francisco , López-Pedrera Chary , Páramo José , Orbe Josune , Hermida José , Rocha Eduardo
Published: March 2018

The increase in nitric oxide (NO) production in lipopolysaccharide (LPS)-induced sepsis is thought to contribute to the development of shock. However, NO could also play an antithrombotic role. Little is known about the modulating effect of NO on the endothelial overexpression and production of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) occurring in endotoxemia. We analyzed the effect of N(G)-nitro-L-arginine-methyl-ester (L-NAME), an inhibitor of NO synthases, and S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a NO donor, on the expression and synthesis of TF and PAI-1 by LPS-challenged human umbilical vein endothelial cells (HUVEC): L-NAME enhanced the increase in TF mRNA and antigen levels (P <0.05) observed in LPS-treated HUVEC; SNAP down-regulated the LPSinduced TF increment (p <0.05). However, no effects of NO on regulation of the LPS-dependent increase in PAI-1 could be seen. Thus, NO could play an antithrombotic role in sepsis by down-regulating the endothelial overexpression and production of TF.
Thrombinography shows acquired resistance to activated protein C in patients with lupus anticoagulants
Authors: Regnault Véronique , Béguin Suzette , Wahl Denis , de Maistre Emmanuel , Coenraad Hemker H. , Lecompte Thomas
Published: March 2018

In patients with lupus anticoagulants (LA), acquired resistance to activated protein C (APC) is difficult to demonstrate with clot-based assays due to the presence of the anticoagulant. Via the conversion of a fluorogenic substrate (thrombinography), we monitored the complete process of thrombin formation and decay and its delimitation by the protein C system in eight consecutive LA-patients without anticoagulant therapy and non-carriers of the V Leiden polymorphism. Thrombin generation was triggered in platelet-poor and platelet-rich plasma by recalcification in the presence of a low concentration of tissue factor.

In 7 out of 8 patients we observed a long lag-time before the thrombin burst (LA effect) together with a marked inability of APC to diminish the thrombin activity. The lag-phase was however prolonged to some degree by APC. The effects were more outspoken in the presence of phospholipids from patients' platelets than with added phospholipids.

Thrombinography thus demonstrates APC resistance in LA-patients despite the occurrence of long lag-times (clotting times). The amount of thrombin activity generated in the presence of APC could be a better indicator of the thrombotic risk than the moment at which the thrombin burst starts.
Effect of factor VIII on tissue factor-initiated spatial clot growth
Authors: Ovanesov Mikhail , Lopatina Elena , Ananyeva Natalya , Ul’yanova Ljudmila , Plyushch Olga , Butilin Andrey , Ataullakhanov Fazly , Saenko Evgueni
Published: March 2018

Using time-lapse videomicroscopy, we studied the role of coagulation factor VIII (fVIII) in tissue factor-initiated spatial clot growth on fibroblast monolayers in a thin layer of non-stirred recalcified plasma from healthy donors or patients with severe Haemophilia A. Analysis of temporal evolution of light-scattering profiles from a growing clot revealed existence of two phases in the clot growth-initiation phase in a narrow (0.2 mm) zone adjacent to activator surface and elongation phase in plasma volume. While the initiation phase did not differ in normal and haemophilic plasmas, the rate of clot growth in the elongation phase in haemophilic plasma constituted only 30% of that in normal plasma. Supplementation of haemophilic plasma with 0.05 U/ml fVIII restored the normal clot growth rate (44.9 ± 2.5 μm/min) at high but not at low fibroblast density. Our results indicate that the functioning of the intrinsic tenase complex is critical for normal spatial clot growth.
Deficiency of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) impairs nutritionally induced obesity in mice
Authors: Demeulemeester Diego , Van Hoef Berthe , Collen Désiré , Maquoi Erik , Roger Lijnen H.
Published: March 2018

Tissue inhibitor of matrix metalloproteinase-1 deficient (TIMP-1–/–) mice and wild-type (TIMP-1+/+) controls were kept on a standard (SFD) or a high fat diet (HFD) for 15 weeks. At the time of sacrifice, TIMP-1–/– mice on HFD had a significantly lower body weight (29 ± 1.5 versus 41 ± 1.8 g, p <0.005), and significantly less subcutaneous (0.81 ± 0.19 versus 1.78 ± 0.21 g, p <0.05) and gonadal (0.87 ± 0.17 versus 1.85 ± 0.18 g, p <0.005) fat mass. These differences were much less pronounced for mice on SFD. On HFD but not on SFD, adipocyte diameters were significantly lower in the adipose tissue of TIMP-1–/– mice. Plasma leptin levels in TIMP-1–/– mice on HFD were significantly lower as compared to TIMP-1+/– mice, and strongly correlated with adipose tissue mass for both genotypes. Staining with an endothelial cell specific lectin revealed a significantly higher blood vessel density, larger stained area and vessel size in adipose tissue of TIMP-1–/– mice on HFD. This difference disappeared after normalization to the adipocyte number, suggesting that it does not represent a true enhancement of angiogenesis. Thus, in a murine model of nutritionally induced obesity, TIMP-1 promotes adipose tissue development.
In vivo pig models of venous thrombosis mimicking human disease
Authors: Kang Chantal , Bonneau Michel , Brouland Jean-Philippe , dit Sollier Claire Bal , Drouet Ludovic
Published: March 2018

Most animal models of venous thrombosis involve acute thrombosis with hypercoagulability in small rodents. To better replicate human disease, we developed two models in the pig, a species similar to humans in size and in vascular and coagulation reactivity. One model involves de-endothelialisation with 50% or 80% stenosis and the other replacement of a venous segment by a Gore-Tex™ vascular prosthesis. Both models were tested with and without acute induced hypercoagulability (thromboplastin infusion). Thrombi obtained without thromboplastin infusion were composed of a multilayered platelet and a fibrin meshwork structure similar to that usually found in humans. With thromboplastin infusion, the thrombi were homogeneous fibrin structures imprisoning red blood cells. The high incidence of thrombosis obtained with the 80% stenosis model would be useful for studying anticoagulant treatments, whereas the low incidence with 50% stenosis would be useful for evaluating procoagulant effects of conditions or treatments. These new models shed further light on the development of venous thrombi under conditions similar to those seen in humans and may prove useful for investigating anticoagulant and procoagulant effects.
Differential regulation of P-selectin expression by protein kinase A and protein kinase G in thrombin-stimulated human platelets
Authors: Libersan Danielle , Rousseau Guy , Merhi Yahye
Published: March 2018

P-selectin is rapidly translocated from platelet α-granules following activation. Intracellular cyclic AMP (cAMP) is a potent inhibitory pathway that results in global downregulation of platelet activation. While cAMP-dependent protein kinase (PKA) has long been considered as the main mediator of cAMP-dependent effects, no study has yet evaluated its effect on P-selectin expression in human platelets. Pretreatment of thrombin-stimulated platelets with forskolin resulted in a concentration-dependent inhibition of P-selectin expression that correlated with adenylyl cyclase activity. Inhibition of PKA with H-89 reversed cAMP-induced inhibition of P-selectin while cGMP-dependent protein kinase (PKG) inhibition with KT5823 significantly potentiated cAMP-dependent inhibition of P-selectin. Similar results were also observed in a platelet/neutrophil binding assay. In conclusion, cAMP-induced inhibition of P-selectin expression is, in large part, mediated through activation of PKA. PKG appears to be sollicited for P-selectin expression when cAMP levels are elevated which suggest a cAMP/PKG-dependent pathway of platelet activation.
A critical role of lipid rafts in the organization of a key FcγRIIa-mediated signaling pathway in human platelets
Authors: Bodin Stéphane , Viala Cécile , Ragab Ashraf , Payrastre Bernard
Published: March 2018

The involvement of platelet FcγRIIa in heparin-associated thrombocytopenia (HIT) is now well established. However, the precise sequence of molecular events initiated by FcγRIIa cross-linking in platelets remains partly characterized. We investigated here the role of lipid rafts in the spatio-temporal organization of the FcγRIIa-dependent signaling events. Upon cross-linking, FcγRIIa relocated in rafts where the kinase Lyn and the adapter LAT were among the major phosphotyrosyl proteins. Upon stimulation by HIT sera, the second messenger phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) accumulated in rafts in a P2Y12 adenosine diphosphate (ADP) recep- tor-dependent manner. PtdIns(3,4,5)P3 was then essential to specifically recruit phospholipase Cγ2 (PLCγ2) to these membrane microdomains. Controlled disruption of rafts by methyl γ-cyclodextrin reversibly abolished PtdIns(3,4,5)P3 production, PLC activation and platelet responses induced by FcγRIIa cross-linking without affecting the tyrosine phosphorylation events. This work demonstrates that platelet rafts are essential for the integration of a key signaling complex leading to the rapid production of PtdIns(3,4,5)P3 and in turn PLCγ2 activation during HIT.
Activation-dependent surface expression of gC1qR/p33 on human blood platelets
Authors: Murphy Tara , Ghebrehiwet Berhane , Peerschke Ellinor
Published: March 2018

GC1qR/p33 (gC1qR) is expressed by a variety of somatic and cultured cells, including blood platelets. It interacts with several cellular, viral, bacterial, and plasma proteins, suggesting a potential role in thrombosis, inflammation, and infection. Considerable controversy has surrounded the surface membrane localization of gC1qR, however, since its cDNA sequence does not predict a traditional membrane-anchoring domain, and bears a typical mitochondrial targeting sequence. The present study examined gC1qR expression on resting and activated human blood platelets using flow cytometry and confocal microscopy with two monoclonal antibodies, 74.5.2 and 60.11, directed against gC1qR C-terminal amino acids 204-218, and N-terminal amino acids 76-93, respectively. Unstimulated platelets reacted minimally with either antibody. In contrast, platelet activation with TRAP, epinephrine, or ADP produced markedly increased gC1qR expression as reflected by 74.5.2 binding but not 60.11 binding. Platelet activation was verified using PAC-1 and anti CD 62 antibodies. Whereas PAC-1 binding to activated platelets could be reversed following platelet incubation with PGE1, 74.5.2 binding remained unchanged, suggesting the sustained expression of gC1qR following platelet stimulation. The data further demonstrate that detection of cell surface gC1qR may be dependent on antibody specificity. The ability of gC1qR to bind proteins involved in complement, coagulation, and kinin systems, as well as viral and bacterial pathogens including S. aureus protein A, supports the hypothesis that gC1qR expressed on activated platelets may contribute directly to thrombosis, inflammation, and endovascular infections.
Aging of stromal-derived human breast fibroblasts might contribute to breast cancer progression
Authors: Sieuwerts Anieta , Bolt-de Vries Joan , Bosma Peter , Swiggers Susan , Klijn Jan , Foekens John , Martens John
Published: March 2018

Age is an important factor in the development and spread of breast cancer. Stromal cells also contribute to breast cancer growth and metastasis through the production of extracellular matrix (ECM) modifiers such as urokinase type plasminogen activator (uPA), its receptor (uPAR), its inhibitors (PAI-1 and PAI-2), matrix metalloproteinases (MMPs), and growth factors, including the fibroblast and insulin-like growth factors (FGF’s and IGF’s). In the present study we have investigated whether breast fibroblasts aged in vitro through passage in culture display altered levels of the plasminogen activator system and growth factors that are known to modulate that system.

With real-time RT-PCR we found that during passage human breast fibroblasts, whether derived from the tumour burden or from matched adjacent normal breast tissue, exhibited a consistent increase in PAI-1 and FGF-1 and a decrease in MMP-2 mRNA expression. In addition, in 5 out of 7 fibroblast strains we observed an induction of uPA expression in combination with a reduced IGF-1 expression. Interestingly, while during aging MMP-2 protein increased in all tumour-derived fibroblast strains, these protein levels were reduced in all normal-tissue-derived fibroblasts. No other clear-cut age-dependent alterations were found in the all-together 25 factors investigated. We furthermore demonstrate in one tumour-derived fibroblast strain that the increases in uPA and PAI-1 mRNA and MMP-2 protein production are inversely related to the telomere length. Artificially increasing telomere length in this fibroblast strain by expressing human telomerase reverse transcriptase (hTERT) prevented senescence and resulted in late passage cultures with early passage uPA, PAI-1 and MMP-2 levels.

Our results show that aging accompanied by telomere loss induces PAI-1 and FGF-1 mRNA expression in all breast fibroblast strains, increases uPA and decreases IGF-1 mRNA expression in a subset, and increases MMP-2 protein expression only in tumour-derived breast fibroblasts. These age-induced levels of PAI-1, FGF-1, uPA and MMP-2 in stromal breast fibroblast could contribute to breast cancer progression.