bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protein Tyrosine Phosphatase SHP-1 Fails to Associate with Cytoskeleton but is Normally Phosphorylated upon Thrombin Stimulation of Thrombasthenic Platelets
Authors: Li Ruo Ya , Gaits Frédérique , Ragab-Thomas Jeannie M F , Maclouf Jacques , Caen Jacques P , Lévy-Toledano Sylviane , Chap Hugues 
Published: July 2018

SummarySHP-1 is a cytoplasmic protein tyrosine phosphatase predominantly expressed in hematopoietic cells. Upon thrombin stimulation of human platelets, SHP-1 is rapidly phosphorylated on both serine and tyrosine residues, and becomes associated with the cytoskeleton, where it could participate in the formation of multiprotein signalling complexes. In order to discriminate between signalling events occurring downstream of G-protein-coupled thrombin receptor and those subsequent to integrin αIIbβ3engagement, SHP-1 behaviour was examined in platelets from two patients lacking integrin αIIbβ3
(Glanzmann’s thrombasthenia). Upon thrombin stimulation, phosphorylation of SHP-1 occurred normally in thrombasthenic platelets, whereas association with the cytoskeleton was abolished. Moreover, inhibition of normal platelet aggregation with the tetrapeptide arg-gly-asp-ser (RGDS), which impairs fibrinogen binding to integrin aIIb(33, did not alter significantly SHP-1 phosphorylation. It is concluded that SHP-1 phosphorylation is not a consequence of integrin signalling but might rather occur downstream of thrombin receptor and heterotrimeric G-proteins.
Abnormal Proteolytic Processing of von Willebrand Factor Arg611 Cys and Arg 611 His
Authors: Nishikubo Toshiya , Christophe Olivier , Lavergne Jean-Maurice , Obert Bernadette , Nonami Kyoko , Takahashi Yukihiro , Yoshioka Akira , Meyer Dominique , Girma Jean-Pierre 
Published: July 2018

SummaryThe structural and functional properties of plasma and platelet vWF were studied in 8 patients (5 unrelated families) with vWD demonstrating a mutation at position 611 (R611C or R611H). Following reduction, electrophoresis and immunoblotting with a polyclonal anti-reduced vWF antibody, abnormal proteolysis of vWF was demonstrated in plasma and to a lesser extent in platelets from all patients, leading to the formation of a unique 209 kDa fragment undetectable in control as well as in type 2A, 2B or 2N vWF. Immunoblotting with MoAbs to reduced vWF showed that the C-terminal end of the 209 kDa fragment was located beyond residue 1744 of the subunit and that its N-terminus was between residues 523 and 1114. Multimeric analysis of patients vWF showed an abnormal pattern in both plasma and platelets, with a moderate decrease of the HMW multimers together with a significant increase of the lowest MW forms. The specific sensitivity of vWF R611C and vWF R611H to proteolysis was further evidenced using V-8 protease. In all patient’s samples the enzyme produced a unique monomeric 80 kDa fragment, absent in V-8 digested normal vWF, which overlapped the N-terminal part of the subunit. The functional analysis of vWF showed a markedly decreased affinity of mutated plasma vWF for platelet GPIb in the presence of ristocetin. Infusion of DDAVP in two of these patients did not lead to significant platelet count change. It induced a limited increase of the HMW multimers in plasma together with a poor correction of the vWF binding to platelet GPIb. In conclusion, our data demonstrate that in addition to a normal proteolysis, vWF mutated at position 611 undergoes a specific cleavage in plasma and platelets. In contrast to the increased proteolysis observed in type 2A and 2B patients’ plasma, this additional cleavage produced a unique 209 kDa species but maintained a HMW multimer-like structure of vWF R611C and R611H.
Two New Closely Related Rat Models with Relevance to Arterial Thrombosis – Efficacies of Different Antithrombotic Drugs
Authors: Vogel Gerard M T , van Amsterdam Ronald G M , Zandberg Pieter , van Houwelingen Piet , Kop Wim J , van Mensvoort Frans W J , Meuleman Dirk G 
Published: July 2018

SummaryTwo thrombosis models in rats are described in which mixed type thrombi are formed at arterial and venous flow rates. The models, containing a silk thread in the aorta and vena cava, respectively, were characterised for the activity of three platelet inhibitors, three thrombin active site inhibitors and five glycosaminoglycans (GAGs).In the two models a similar highly platelet-dependent thrombus developed both in size and composition during the first 10 min after insertion of the silk thread. The thrombotic processes were self-limiting, thus maintaining blood flow, but persisted twice as long in the vena cava model. In both models the thrombus consisted for more than 65% of platelets. Thrombus development under arterial as well as under venous flow conditions was inhibited dose dependently by all tested compounds including aspirin and the synthetic α-methyl glycoside copy of the ATIII binding pentasaccharide within heparin, Org31540/SR90107A. Simultaneous fibrin deposition and platelet activation, which represents an essential element of arterial thrombosis, initially dominated in both models. The gradual thrombus outgrowth, in the cava model, was more sensitive to factor Xa selective anticoagulants, as is venous thrombosis.
Haemostatic Properties of Human Pulmonary and Cerebral Microvascular Endothelial Cells
Authors: Grau Georges E , de Moerloose Philippe , Bulla Oana , Lou Jinning , Lei Zheng , Reber Guido , Mili Nabil , Ricou Bara , Morel Denis R , Suter Peter M 
Published: July 2018

SummaryLittle is known on the haemostatic profiles of human microvascular endothelial cells (MVEC) from different tissues. In addition it is not known whether MVEC from patients display the same haemostatic pattern as MVEC coming from healthy controls. To address these questions MVEC from human lung and brain were isolated and stimulated with tumour necrosis factor α (TNF) and E. coli lipopolysaccharide (LPS) for 24 h. The level and the kinetics of procoagulant activity (PCA) and thrombomodulin (TM) expression were found to be different depending on the tissue of origin and on the agonist used. In particular, the inducible PCA was higher in lung than in brain MVEC, an observation that may be related to the frequency of lung involvement in septic shock. Differences were also observed for tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) with MVEC supernatants or cell lysates. These variables were then measured in lung MVEC purified from patients with acute respiratory distress syndrome (ARDS) and compared to controls. Cells from ARDS patients constitutively expressed more PCA and PAI-1 than controls. The fibrinolytic potential, expressed as t-PA/PAI-1 ratio, was lower in ARDS than in lung MVEC. It is concluded that MVEC display different haemostatic features depending on the tissue they come from and that lung MVEC from ARDS patients present a procoagulant profile when compared with those from controls.
A Multicenter Study of Recombinant Factor VIII (RecombinateTM) in Previously Treated Patients with Hemophilia A
Authors: White G C , Courter S , Bray G L , Lee M , Gomperts E D ,   The Recombinate Previously Treated Patient Study Group
Published: July 2018

SummaryA prospective, open-label multicenter investigation has been conducted to compare pharmacokinetic parameters of recombinant DNA-derived FVIII (rFVIII) and plasma-derived FVIII concentrate (pdFVIII) and to assess safety and efficacy of long-term home-treat- ment with rFVIII for subjects with hemophilia A. Following comparative in vivo pharmacokinetic studies, 69 patients with severe (n = 67) or moderate (n = 2) hemophilia A commenced a program of home treatment using rFVIII exclusively for prophylaxis and treatment of all bleeding episodes for a period of 1.0 to 5.7 years (median 3.7 years). The mean in vivo half-lives of rFVIII and pdFVIII were both 14.7 h. In vivo incremental recoveries at baseline were 2.40%/IU/kg and 2.47%/IU/kg, respectively (p = 0.59). The response to home treatment with rFVIII was categorized as good or excellent in 3,195 (91.2%) of 3,481 evaluated bleeding episodes. Thirteen patients received rFVIII for prophylaxis for twenty-four surgical procedures. In all cases, hemostasis was excellent. Adverse reactions were observed in only 13 of 13,591 (0.096%) infusions of rFVIII; none was serious. No patient developed an inhibitor to r FVIII.
Pretreatment of Human Platelets with Plasmin Inhibits Responses toThrombin, but Potentiates Responses to Low Concentrations of Aggregating Agents, Including the Thrombin Receptor Activating Peptide, SFLLRN
Authors: Kinlough-Rathbone R L , Perry D W , Rand M L , Packham M A 
Published: July 2018

SummaryEffects of plasmin on platelets, that influence subsequent responses to aggregating agents, are relevant to attempts to prevent rethrombosis following administration of fibrinolytic agents. We describe plasmin-induced inhibition of platelet responses to thrombin, but potentiation of responses to other aggregating agents. Washed human platelets were labeled with 14C-serotonin, treated for 30 min at 37° C with 0, 0.1 or 0.2 CU/ml of plasmin, followed by aprotinin, washed and resuspended in a Tyrode-albumin solution with apyrase. Incubation with 0.2 CU/ml of plasmin almost completely inhibited thrombin-induced (0.1 U/ml) aggregation, release of 14C-serotonin, and increase in cytosolic [Ca2+]. In contrast, with plasmin-pretreated platelets, aggregation and release of 14C-serotonin were strongly potentiated in response to low concentrations of the thrombin receptor-activating peptide SFLLRN, ADP, platelet-activating factor, collagen, arachidonic acid, the thromboxane mimetic U46619, and the calcium ionophores A23187 and ionomycin. Aspirin or RGDS partially inhibited potentiation. Plasmin-pretreated platelets resuspended in plasma anticoagulated with FPRCH2C1 (PPACK) also showed enhanced responses to aggregating agents other than thrombin. The contrasting effects on responses to thrombin and SFLLRN are noteworthy. Plasmin cleaves GPIIb/IIIa so that it becomes a competent fibrinogen receptor, and binding of 125I-fibrinogen during ADP-induced aggregation was greatly potentiated within 10 s. Potentiation of aggregation by other agonists may be due to increased binding of released fibrinogen. Thus, platelets freed from a thrombus may have increased responsiveness to low concentrations of aggregating agents other than thrombin. These results provide further support for the use of inhibitors of platelet reactions in conjunction with administration of fibrinolytic agents.
Construction and Expression of Mouse-human Chimeric Antibody SZ-51 Specific for Activated Platelet P-selectin
Authors: Gu Jianming , Liu Yue , Xia Lijun , Wan Haiying , Li Peixia , Zhang Xiaomin , Ruan Changgeng 
Published: July 2018

SummaryA murine monoclonal (mAb) SZ-51 specific for human P-selectin may be used for in vivo thrombus imaging and for the targeting of fibrinolytic agents to thrombi. In order to reduce the immunogenicity of the murine mAb SZ-51 in humans, we cloned and sequenced the cDNAs encoding the variable region of mAb SZ-51 in order to develop mouse/human chimeric reagents. The E. coli expression vector. pHENl-SZ51 Fab/Hu was constructed by fusing the variable regions of mAb SZ-51 with human IgG γICHI and Cκ genes. The constructs were introduced into E. coli HB2151 for expression of soluble chimeric Fab fragment. We also constructed two fusion products by joining the variable regions of mouse antibody to the appropriate constant regions of human Igγl and κ. These chimeras were cloned into two eukaryotic selectable expression vectors separately, which were then cotransfected into a non-Ig secreting murine myeloma line SP2/0 with lipofectin reagent. Six cell lines remained positive for Ig secretion. The highest producing cell line, which showed stable integration and expression at 5 mg/1 of culture, was selected for the large scale production of chimeric antibody. Immunoblotting analysis demonstrated that both of the chimeric antibodies (SZ51Fab/Hu, SZ51/Hu) in the culture supernatants, like the native mAb SZ-51, bind P-selectin. In addition, the whole chimeric antibody can compete for binding to activated platelets with murine SZ-51. Therefore, the SZ-51 chimeric antibody may be a potential agent for diagnosis and treatment of thrombotic diseases in the future.
Prothrombin Fragment F1+2 Is not Predictive for Recurrent Venous Thromboembolism
Authors: Kyrle P A , Eichinger S , Pabinger I , Stümpflen A , Hirschl M , Bialonczyk C , Schneider B , Mannhalter C , Melichart M , Traxler G , Weltermann A , Speiser W , Lechner K 
Published: July 2018

SummaryIt would be important to estimate in advance the risk of recurrent thrombosis. Deficiencies of antithrombin, protein C or protein S, or resistance to activated protein C are associated with a biochemically detectable prethrombotic state. It is thus far unknown whether in patients with a history of thromboembolism but without a defined clotting abnormality a heightened coagulation activation is detectable.We investigated the value of prothrombin fragment Fl+2 (FI+2) as a predictor of recurrent venous thromboembolism. Furthermore, we compared the Fl+2 levels of thrombosis patients without a defined clotting defect to those of Factor V Leiden patients with a history of venous thrombosis and to those of healthy controls. 180 patients without a defined clotting abnormality and 73 patients with Factor V Leiden were prospectively followed after discontinuation of oral anticoagulants for venous thrombosis and Fl+2 was measured at regular intervals.Recurrent venous thromboembolism occurred in 23 (9%) of the 253 patients. Before or at several time points after oral anticoagulants, no significant difference in Fl+2 levels was found in patients with and without recurrent thrombosis. Fl+2 levels at 3 weeks and prior to recurrence were not significantly different in both patient groups. Over a one-year observation period, Fl+2 levels of both patients with and without Factor V Leiden were higher than those of the controls. No difference in Fl+2 was seen between patients with and without Factor V Leiden.We conclude that monitoring of Fl+2 is not suitable for identification of individuals at risk of recurrent venous thrombosis. Permanent hemostatic system activation is detectable both in patients with a defined abnormality of the clotting system and in patients in whom a particular defect has not (yet) been identified.
Hepatitis G Viral RNA in Serum and in Peripheral Blood Mononuclear Cells and Its Relation to HCV-RNA in Patients with Clotting Disorders
Authors: Sheng Li , Soumillion Ann , Peerlinck Kathelijne , Verslype Chris , Lin Lan , Pelt Jos van , Hess Georg , Vermylen Jozef , Yap Sing Hiem 
Published: July 2018

SummaryThe hepatitis G virus (HGV) has recently been identified as a new member of the Flaviviridae family. Infection by this virus is thought to be associated with blood borne hepatitis. In this study, the presence of HCV- and HGV-RNAs in serum or plasma (175 patients) and in peripheral blood mononuclear cells (PBMC) (133 patients) was investigated in patients with clotting disorders using a sensitive reverse transcriptase polymerase chain reaction (RT-PCR). HGV-RNA was detected in serum of 26 patients (14.8%). In apparently healthy blood donors, serum HGV-RNA was detected in 4 of 358 individuals investigated (1.12%). Ninety two percent of the 26 serum HGV-RNA positive patients had coinfection with the hepatitis C virus (HGV), especially with HCV genotype lb, the most common genotype in Belgium. Of these coinfected patients, 15 (62.5%) showed elevated serum ALT levels. Two patients who were solely infected with HGV had normal serum ALT. HGV-RNA in PBMC was found in 18 patients, of whom 3 were negative for serum HGV-RNA. As in case of HCV, HGV-RNA in PBMC is preferentially sensitive to interferon treatment. Nevertheless, rapid reappearance of HGV-RNA in PBMC was observed after cessation of treatment. In one patient, persistent serum ALT elevation seems to be associated with continued HGV viremia, despite the disappearance of serum HCV-RNA.
Platelet Hyporeactivity in Very Low Birth Weight Neonates
Authors: Rajasekhar Damodara , Barnard Marc R , Bednarek Francis J , Michelson Alan D 
Published: July 2018

SummaryVery few studies have examined platelet function in very low birth weight (VLBW) preterm neonates, because of the relatively large volumes of blood required. In this study, platelet function in clinically stable VLBW neonates was examined by whole blood flow cytometry, which requires only 5 |jl1 of whole blood per assay. The following monoclonal antibodies were used: S12 (P-selectin-specific, reflecting a granule secretion), PAC1 (directed against the fibrinogen binding site exposed on the GPIIb-IIIa complex of activated platelets), F26 (directed against a conformational change in fibrinogen bound to the GPIIb-IIIa complex), and 6D1 (directed against the von Willebrand factor binding site on the GPIb-IX-V complex). VLBW neonates, like normal adults, did not have circulating activated platelets, as determined by the lack of binding of SI2, PAC1, and F26 in the absence of an added agonist. VLBW neonatal platelets were markedly less reactive than adult platelets to thrombin, ADP/epinephrine, and U46619 (a stable thromboxane A2 analogue), as determined by the extent of increase in the platelet binding of SI2, PAC1, and F26, and the extent of decrease in the platelet binding of 6D1. In summary, compared to adults, the platelets of VLBW neonates are markedly hyporeactive to thrombin, ADP/epinephrine and a thromboxane A2 analogue in the physiologic milieu of whole blood, as determined by: 1) the increase in platelet surface P-selectin; 2) the exposure of the fibrinogen binding site on the GPIIb-IIIa complex; 3) fibrinogen binding; and 4) the decrease in platelet surface GPIb. This platelet hyporeactivity may be a factor in the propensity of VLBW neonates to intraventricular hemorrhage. In addition to its previously defined use as a test of platelet hyperreactivity, the present study suggests that whole blood flow cytometry may be useful in the clinical assessment of platelet hyporeactivity.
Performance of Conventional Cytology and Colposcopy for the Diagnosis of Cervical Squamous and Glandular Neoplasias
Authors: Fachetti-Machado Giselle , Figueiredo-Alves Rosane Ribeiro , Moreira Marise Amaral Rebouças 
Published: July 2018

Objective To estimate the cytological and colposcopic performances for the diagnosis of cervical neoplasias.
Methods Cross-sectional retrospective study with data from patients' charts. The participants underwent colposcopy, guided biopsies, and excision when needed. The cytological and colposcopic categorization followed the Bethesda System and the international colposcopic terminologies. The cytology and colposcopy performances were evaluated by sensitivity (SE), specificity (SP), positive predictive value (PPV), and negative predictive value (NPV) analyses with 95% confidence interval (95% CI).
Results From 1,571 participants, a total of 1,154 (73.4%) were diagnosed with cervical squamous intraepithelial neoplasia grade 2 or worse (CIN 2+), 114 (7.2%) with adenocarcinoma in situ or worse (AIS+), 615 (39.2%) presented atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion or worse (ASC-H+) cytology, and 934 (59.4%) presented major or suspicious for invasion colposcopic abnormalities. The SE, SP, PPV, and NPV of ASC-H+ for diagnoses of CIN 2+ and AIS+ were, respectively: 44% (95% CI: 41–47) and 72% (95% CI: 67–76), 79% (95% CI: 77–81) and 79% (95% CI: 75–83), 88% (95% CI: 87–90) and 55% (95% CI: 50–60), and 28% (95% CI: 26–31) and 88% (95% CI: 85–91). The SE, SP, PPV, and NPV of major or suspicious for invasion colposcopic abnormalities for diagnoses of CIN 2+ and AIS+ were, respectively: 62% (95% CI: 60–65) and 86% (95% CI: 83–89), 59% (95% CI: 57–62) and 59% (95% CI: 55–64), 85% (95% CI: 83–87) and 44% (95% CI: 40–49), and 29% (95% CI: 27–32) and 92% (95% CI: 89–94).
Conclusion The SE analyses results of ASC-H+ and major or suspicious for invasion colposcopic abnormalities were higher for diagnoses of glandular neoplasias. These results confirm the role of cytology in identifying women at risk who will have their final diagnoses settled by colposcopy and histology.
Plasticity based on compensatory effector use in the association but not primary sensorimotor cortex of people born without hands
Authors: Striem-Amit Ella , Vannuscorps Gilles , Caramazza Alfonso 
Published: July 2018

SignificanceWhat determines the role of brain regions and their plasticity when typical inputs or experience is not provided? To what extent can extreme compensatory use affect brain organization? We tested the reorganization of primary and association sensorimotor cortex hand-selective areas in people born without hands, who use their feet for everyday tasks. We found that their primary sensorimotor hand area is preferentially activated for nearby body parts that cannot serve as effectors. In contrast, foot-selective compensatory plasticity was found in the association cortex, in an area typically involved in manual tool use. This shows limitations of compensatory plasticity and experience in modifying brain organization of early topographical cortex, as compared with association cortices where function-based organization is the driving factor.
Polymerization pathway of mammalian nonmuscle myosin 2s
Authors: Liu Xiong , Shu Shi , Korn Edward D. 
Published: July 2018

Significance
Nonmuscle myosin 2 (NM2) filaments are highly dynamic, polymerizing and depolymerizing in situ at different cellular sites where they are essential for multiple functions including cell division, cell motility, endocytosis, and exocytosis. Therefore, the mechanism of myosin filament assembly and disassembly has high importance. NM2 monomers can be either folded or unfolded. It has been thought that unfolded monomers and antiparallel bipolar dimers are the fundamental polymerization units. We now show that antiparallel, bipolar filaments are formed in vitro by the conversion of folded monomers to folded antiparallel dimers and then antiparallel tetramers that unfold, forming bipolar tetramers. Mature filaments consist of multiple unfolded tetramers with entwined bare zones. These results should assist our understanding of myosin filament dynamics in vivo.
Narrative and persistence
Authors: Olson Eric T. , Witt Karsten 
Published: July 2018

AbstractMany philosophers say that the nature of personal identity has to do with narratives: the stories we tell about ourselves. While different narrativists address different questions of personal identity, some propose narrativist accounts of personal identity over time. The paper argues that such accounts have troubling consequences about the beginning and end of our lives, lead to inconsistencies, and involve backwards causation. The problems can be solved, but only by modifying the accounts in ways that deprive them of their appeal.