bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Prothrombinase-based Assay for Detection of Resistance to Activated Protein C
Authors: Nicolaes Gerry A F , Christella M , Thomassen L G D , Oerle Rene van , Hamulyak Karly , Hemker H Coenraad , Tans Guido , Rosing Jan 
Published: July 2018

SummaryIn this paper we present a new method for the detection of resistance to activated protein C (APC) that is based on direct measurement of the effect of APC on the cofactor activity of plasma factor Va. The factor V present in a diluted plasma sample was activated with thrombin and its sensitivity towards APC was subsequently determined by incubation with phospholipids and APC. The loss of factor Va cofactor activity was quantified in a prothrombinase system containing purified prothrombin, factor Xa and phospholipid vesicles and using a chromogen-ic assay for quantitation of thrombin formation. The reaction conditions were optimized in order to distinguish normal, heterozygous and homozygous APC-resistant plasmas. Maximal differences in the response of these plasmas towards APC were observed when factor Va was inactivated by APC in the absence of protein S and when the cofactor activity of factor Va was determined at a low factor Xa concentration (0.3 nM).Addition of 0.2 nM APC and 20 μM phospholipid vesicles to a 1000-fold diluted sample of thrombin-activated normal plasma resulted in loss of more than 85% of the cofactor activity factor Va within 6 rnin. Under the same conditions, APC inactivated ∼ 60% and ∼20% of the factor Va present in plasma samples from APC-resistant individuals that were heterozygous or homozygous for the mutation Arg506⟶Gln in factor V, respectively. Discrimination between the plasma samples from normal and heterozygous and homozygous APC-resistant individuals was facilitated by introduction of the so-called APC-sensitivity ratio (APC-sr). The APC-sr was defined as the ratio of the factor Va cofactor activities determined in thrombin-activated plasma samples after 6 min incubation with or without 0.2 nM APC and was multiplied by 100 to obtain integers (APC-sr = {factor Va+APC/factor Va−APC} × 100). Clear differences were observed between the APC-sr of plasmas from normal healthy volunteers (APC-sr: 8-20, n = 33) and from individuals that were heterozygous (APC-sr: 35-50, n = 17) or homozygous APC resistant (APC-sr: 82-88, n = 7). There was no mutual overlap between the APC-sr of normal plasmas and plasmas from heterozygous or homozygous APC resistant individuals (p <0.0001). In all cases our test gave the same result as the DNA-based assay. Since the test is performed on a highly diluted plasma sample there is no interference by conditions that affect APC resistance tests that are based on clotting time determinations (e.g. coagulation factor deficiencies, oral anticoagulation, heparin treatment, the presence of lupus anticoagulants, pregnancy or the use of oral contraceptives). Furthermore, we show that part of the factor Va assay can be performed on an autoanalyzer which increases the number of plasma samples that can be handled simultaneously.
Agonist-induced Actin Polymerization Is Required for the Irreversibility of Platelet Aggregation
Authors: Torti Mauro , Tolnai Festetics Enrico , Bertoni Alessandro , Sinigaglia Fabiola , Balduini Cesare 
Published: July 2018

SummaryCytochalasin D was used to investigate the role of intracellular cyto-skeleton in the stabilization of platelet aggregation induced by strong platelet agonists. Incubation of gel-filtered platelets with increasing concentrations of cytochalasin D resulted in a dose-dependent inhibition of actin polymerization and association of actin-binding proteins with the Triton X-100-insoluble material induced by the thromboxane analogue, U46619, and the thrombin receptor activating peptide, TRAP. The same concentrations of cytochalasin D did not significantly inhibit platelet aggregation promoted by the two agonists. The addition of the chelating agent EDTA to fully aggregated platelets, that had been treated with cytochalasin D, resulted in the rapid and almost complete disaggregation. EDTA did not cause disaggregation of control, solvent-treated, aggregated platelets. The degree of platelet disaggregation induced by EDTA was dependent on the dose of cytochalasin D used, and was correlated with the inhibition of the cytoskeletal reorganization. Aggregation of cytochalasin D-treated platelets stimulated with U46619 or TRAP was also reverted by the addition of the tetrapeptide RGDS or the fibrinogen y-chain dodecapeptide, which competitively interfere with fibrinogen binding to the glycoprotein Ilb-IIIa complex. These results indicate that the intracellular cytoskeleton plays an essential role in the stabilization of the fibrinogen-platelet interaction, and is necessary for the irreversibility of platelet aggregation induced by strong agonists.
Antiplatelet Activity in Onion (Allium cepa) Is Sulfur Dependent
Authors: Goldman Irwin L , Kopelberg Murray , Debaene Jan E P , Schwartz Bradford S 
Published: July 2018

SummaryPlants of the genus Allium such as onion and garlic are often consumed as a source of compounds which inhibit human platelet activity, with the goal of decreasing vascular disease. Antiplatelet activity in these plants is determined in part by native concentrations of organosul-fur compounds. Evaluation of four onion genotypes grown in a field study at four US locations in 1994 demonstrated onions with mild flavor and low sulfur content exhibited significantly lower antiplatelet activity than those containing high levels of sulfur. Antiplatelet activity was significantly positively correlated with genotypically determined bulb sulfur content and dissolved solids, indicating these latter factors are good predictors of antiplatelet strength. These data demonstrate antiplatelet activity is genotype dependent and correlated with bulb sulfur content. Genotype and bulb sulfur content should be taken into account in studies assessing onion antiplatelet effects.
A Multicenter Randomized Double-blind Study of Enoxaparin Compared with Unfractionated Heparin in the Prevention of Venous Thromboembolic Disease in Elderly In-patients Bedridden for an Acute Medical Illness
Authors: Bergmann Jean-Fraçois , Neuhart Eric 
Published: July 2018

SummaryA multicenter, randomized double-blind study compared in two parallel groups the efficacy and safety of a low molecular weight heparin (LMWH) enoxaparin 20 mg once daily, with unfractionated heparin (UFH) 5000 IU twice daily, administered subcutaneously for 10 days, in the prevention of venous thrombosis disease in 442 hospitalized elderly patients bedridden for an acute medical illness. The main efficacy endpoint was defined as the occurrence of venous thrombosis, diagnosed by a daily fibrinogen uptake test, and/or documented clinical pulmonary embolism.Intention-to-treat analysis of efficacy showed that the incidence of venous thromboembolic events was low: 4.8% (10/207) in the LMWH group (9 episodes of isotopic venous thrombosis and one of scintigraphic pulmonary embolism), and 4.6% (10/216) in the UFH group (10 episodes of isotopic venous thrombosis). The two treatments were equivalent, where equivalence was defined as a maximum difference of 7% between the two groups (p = 0.0005).There were no significant differences in terms of safety between the 216 patients in the LMWH group and the 223 patients in the UFH group who received at least one injection of the randomized treatment. During the study period, 15 patients (3.4%) died (7 in the LMWH group and 8 in the UFH group): 2 sudden deaths, one in each group, including one case in which pulmonary embolism could not be excluded since no autopsy was performed, and 13 others deaths unrelated to the study treatments. Six patients (1.4%) presented a bleeding complication: 2 (0.9%) in the enoxaparin group (one major and one minor hemorrhage), and 4 (1.8%) in the UFH group (2 major and 2 minor hemorrhages).These results indicate that subcutaneous enoxaparin 20 mg once daily for 10 days is as effective and well tolerated as subcutaneous UFH 5000 IU twice daily in the prevention of venous thromboembolic disease in bedridden elderly in-patients presenting an acute medical illness.
Serum Thrombopoietin (c-MpI Ligand) Levels in Patients with Liver Cirrhosis
Authors: Shimodaira Shigetaka , Ishida Fumihiro , Ichikawa Naoaki , Tahara Tomoyuki , Kato Takashi , Kodaira Hiroshi , Ito Toshiro , Tanaka Eiji , Sodeyama Takeshi , Kiyosawa Kendo , Kitano Kiyoshi 
Published: July 2018

SummaryTo clarify the role of c-Mpl ligand (thrombopoietin: TPO) in liver cirrhosis (LC), we examined serum TPO levels (sTPO) in patients with LG (N = 44), chronic hepatitis (CH; N = 13) and healthy controls (N = 41) by an enzyme-linked immunosorbent assay. Although platelet counts of all LC patients (89 ± 59 × 109/1; mean ± SD) were lower than those of controls and CH patients, sTPO levels in LC patients (1.23 ± 0.51 fmol/ml) were the same as those in controls (1.22 ± 0.37) and CH patients (1.18 ± 0.36). Platelet counts were significantly higher in splenectomized patients than in unsplenectomized patients, but the sTPO level did not differ between these two groups. In LC patients, the sTPO level was not correlated with the platelet count, but was correlated with prothrombin time, activated partial thromboplastin time, and total bilirubin, indicating that production of TPO in the liver decreases slightly with the development of liver dysfunction. Our findings suggest that production of TPO is maintained in LC patients and their thrombocytopenia is not due to a defect in platelet production.
Measurement of Glycoprotein llb/llla Blockade by Flow Cytometry with Fluorescein Isothiocyanate-conjugated Crotavirin, a Member of Disintegrins
Authors: Liu Chao-Zong , Hur Buo-Tsang , Huang Tur-Fu 
Published: July 2018

SummaryThe blockade of platelet membrane glycoprotein IIb/IIIa by a monoclonal antibody, 7E3, was measured by flow cytometry using a fluorescein isothiocyanate-conjugated disintegrin, FITC-crotavirin, as the probe. After treatment of platelets with 7E3 or 7E3 F(ab’)2, there is a good correlation between the inhibition of platelet aggregation and the blockade of FITC-crotavirin’s binding to platelets. The content of glycoprotein IIb/IIIa for the subsequent binding of FITC-crotavirin to the 7E3-pretreated platelets highly correlated to the extent of glycoprotein Ilb/IIIa, remaining available. It was evidenced by the observation that the sum of glycoprotein Ilb/IIIa occupation by 7E3 and that of FITC-crotavirin approached the total amount of glycoprotein Ilb/IIIa expressed on the platelet membrane. This indicates that the percentage inhibition of FITC-crotavirin’s binding at the saturation dose reflects the extent of glycoprotein Ilb/IIIa blockade by 7E3. At the saturation binding concentration (5 |xg/ml), FITC-crotavirin did not displace platelet bound 7E3. Gating the light-scattering profile for platelets, the binding of FITC-crotavirin to platelet glycoprotein Ilb/IIIa could be easily determined in diluted whole blood by direct stain method. The available unoccupied glycoprotein Ilb/IIIa of platelets in the 7E3 or 7E3 F(ab’)2-pretreated whole blood were measured by flow cytometry at the saturation binding dose of FITC-crotavirin (4 fig/ml) and the data showed that the higher deconcentration of antibody added into whole blood, the lower debinding of FITC-crotavirin to platelets. This technique may provide an alternative rapid method for measuring the blockade of glycoprotein Ilb/IIIa by 7E3, a promising anti-thrombotic agent, thus providing a monitoring method for adjusting the therapeutic dose of 7E3 or its related derivatives.
Human Umbilical Vein Smooth Muscle Cells as a Model to Study Thrombin Generation and Function: Effect of Thrombin Inhibitors
Authors: Catalioto Rose-Marie , Cucchi Paola , Renzetti Anna Rita , Criscuoli Marco , Subissi Alessandro 
Published: July 2018

SummaryThe aim of the present work was to study how human umbilical vein smooth muscle cells (HUVSMC) can initiate the coagulation process and to investigate the responses of these cells to thrombin. Exposure of HUVSMC to recalcified human plasma led to a time-dependent production of thrombin, measured both as amidolytic activity and as release of fibrinopeptide A. Thrombin activity was dose-dependently reduced by an anti-human tissue factor antibody (76 ± 3% at 10 Μg/ml) and by inhibitors like heparin, rec-hirudin, hirulog-1, Napap and hiru-norm, a novel hirudin-like thrombin inhibitor (IC50 = 2 ± 0.4, 8 ± 1, 130 ± 22, 199 ± 29 and 68 ± 8nM, respectively). The release of fibrinopeptide A was similarly prevented (IC50 = 14 ± 1,132 ± 25 and 50 ± 8 nM for rec-hirudin, Napap and hirunorm, respectively). Exogenously added thrombin increased thymidine incorporation into HUVSMC to 240 ± 30% of basal (EC50 = 0.49 ± 0.09 nM) and thrombin inhibitors blocked this effect (IC50 = 10 ± 3, 37 ± 17, 343 ± 165 and 1402 ± 758 nM for rec-hirudin, hirunorm, Napap and hirulog-1, respectively). Also recalcified human plasma was mitogenic for HUVSMC and its effect was mainly due to endogenously generated thrombin, as shown by the use of thrombin inhibitors. In conclusion, HUVSMC are capable of initiating the extrinsic coagulation cascade, leading to the formation of thrombin which promotes clotting and stimulates DNA synthesis. Thrombin inhibitors prevent both coagula-tive and cellular effects of thrombin.
Qualitative Platelet 12-Lipoxygenase Abnormality in a Patient with Essential Thrombocythemia
Authors: Tomo Kenjiro , Takayama Hiroshi , Kaneko Yoshiyuki , Fujita Jun , Nakamura Michihiro , Ueda Natsuo , Yamamoto Shozo , Okuma Minoru 
Published: July 2018

SummaryQuantitative platelet 12-lipoxygenase (12-LOX) deficiency has been reported in some patients with myeloproliferative disorders (MPD). We report here for the first time a novel qualitative abnormality of the 12-LOX enzyme of platelets from a patient with essential thrombocythemia. The anti-12-LOX immunoprecipitates from the patient’s platelet homogenates showed a deficiency of 12-LOX activity, but contained normal amount of 12-LOX protein. There was no difference in subcellular localization of the enzyme between the patient’s platelets and normal ones. This 12-LOX protein lacking its enzyme activity showed slightly larger electrophoretic mobility than normal one, suggesting a molecular abnormality of the enzyme. However, we could not detect any genetic mutation causing such abnormalities in all exons of 12-LOX gene by sequencing the patient’s PCR-amplified DNA. Thus, our results indicate that the deficient activity of this abnormal 12-LOX protein is probably due to a posttranslational modification, and the possibility that platelets of some MPD patients have qualitative abnormality of the 12-LOX enzyme besides quantitative one.
Upper Eyelid Blepharoplasty: A Novel Method to Predict and Improve Outcomes
Authors: Barmettler Anne , Wang Jessie , Heo Moonseong , Gladstone Geoffrey J 
Published: July 2018

Abstract

Background
In some upper eyelid blepharoplasties, maximal skin removal may not result in desired outcomes; raising crease height can therefore be considered. Currently, there is no method to determine the amount of skin to be excised and/or crease elevation required to achieve a specific outcome.

Objective
This study extrapolated an equation to determine amount of skin excision and/or lid crease elevation needed to achieve a specific eyelid margin to fold distance (MFD).

Methods
This institutional review board-approved, HIPAA-compliant study was a prospective, nonrandomized clinical trial. Patients were included if aged 30 to 100 years old and underwent upper eyelid blepharoplasty with one surgeon between 2012 and 2014. Exclusion criteria were thyroid eye disease, myasthenia gravis, myotonic dystrophy, pregnancy, blepharoptosis, prior eyelid surgery or trauma, concurrent brow surgery, and topical alpha-agonists. The following data were collected preoperatively and at postoperative months 1 and 6: age, gender, BMI, brow position, MFD, margin to crease distance (distance between eyelid margin and crease, MCD), and vertical skin distance (distance between eyelid margin and brow, VSD).

Results
A total 322 eyelids of 164 patients underwent 208 skin excisions, 26 crease elevations, and 88 combined skin excision and crease elevation. Age, gender, and BMI category were all nonsignificant and excluded from the final model. This equation was extrapolated with regression analysis: Change in MFD = −0.40 + (−0.28 × Change VSD) + (0.53 × Change MCD) with |R| = 0.28.

Conclusion
To better predict and obtain desired upper eyelid blepharoplasty outcomes, the authors created an equation.

Level of Evidence: 4
Defining the genetic control of human blood plasma N-glycome using genome-wide association study
Authors: Sharapov Sodbo Zh. , Tsepilov Yakov A. , Klaric Lucija , Mangino Massimo , Thareja Gaurav , Simurina Mirna , Dagostino Concetta , Dmitrieva Julia , Vilaj Marija , Vuckovic Frano , Pavic Tamara , Stambuk Jerko , Trbojevic-Akmacic Irena , Kristic Jasminka , Simunovic Jelena , Momcilovic Ana , Campbell Harry , Dunlop Malcolm , Farrington Susan , Pucic-Bakovic Maja , Gieger Christian , Allegri Massimo , Louis Edouard , Georges Michel , Suhre Karsten , Spector Tim , Williams Frances MK , Lauc Gordan , Aulchenko Yurii 
Published: July 2018

AbstractGlycosylation is a common post-translational modification of proteins. It is known, that glycans are directly involved in the pathophysiology of every major disease. Defining genetic factors altering glycosylation may provide a basis for novel approaches to diagnostic and pharmaceutical applications. Here, we report a genome-wide association study of the human blood plasma N-glycome composition in up to 3811 people. We discovered and replicated twelve loci. This allowed us to demonstrate a clear overlap in genetic control between total plasma and IgG glycosylation. Majority of loci contained genes that encode enzymes directly involved in glycosylation (FUT3/FUT6, FUT8, B3GAT1, ST6GAL1, B4GALT1, ST3GAL4, MGAT3, and MGAT5). We, however, also found loci that are likely to reflect other, more complex, aspects of plasma glycosylation process. Functional genomic annotation suggested the role of DERL3, which potentially highlights the role of glycoprotein degradation pathway, and such transcription factor as IKZF1.
Protection of rhesus macaques against vaginal SHIV challenges by VRC01 and an anti-α4β7antibody
Authors: Calenda Giulia , Frank Ines , Arrode-Brusés Géraldine , Pegu Amarendra , Wang Keyun , Arthos James , Cicala Claudia , Grasperge Brooke , Blanchard James L. , Maldonado Stephanie , Roberts Kevin , Gettie Agegnehu , Fauci Anthony S. , Mascola John R. , Martinelli Elena 
Published: July 2018

ABSTRACTVRC01 protects macaques from vaginal SHIV infection after a single high-dose challenge. Infusion of a simianized anti-α4β7mAb (Rh-α4β7) just prior to, and during repeated vaginal exposures to SIVmac251 partially protected macaques from vaginal SIV infection and rescued CD4+T cells. To investigate the impact of combining VRC01 and Rh-α4β7on SHIV infection, 3 groups of macaques were treated with a suboptimal dosing of VRC01 alone or in combination with Rh-α4β7or with control antibodies prior to the initiation of weekly vaginal exposures to a high dose (1000TCID50) of SHIVAD8-EO.The combination Rh-α4β7-VRC01 significantly delayed SHIVAD8-EOvaginal infection. Following infection, VRC01-Rh-α4β7-treated macaques maintained higher CD4+T cell counts and exhibited lower rectal SIV-DNA loads compared to the controls. Interestingly, VRC01-Rh-α4β7-treated macaques had less IL-17 producing cells in the blood and the gut during the acute phase of infection. Moreover, higher T cell responses to the V2-loop of the SHIVAD8-EOenvelope in the VRC01-Rh-α4β7group inversely correlated with set point viremia. The combination of suboptimal amounts of VRC01 and Rh-α4β7delayed infection, altered anti-viral immune responses and minimized CD4+T cell loss. Further exploration of the effect of combining bNAbs with Rh-α4β7on SIV/HIV infection and anti-viral immune responses is warranted and may lead to novel preventive and therapeutic strategies.Short summaryA combination of VRC01 and Rh-α4β7significantly delayed SHIV acquisition, protected CD4 counts, decreased gut viral load and modified the immune response to the virus.
What is a cognitive map? Organising knowledge for flexible behaviour
Authors: Behrens Timothy E.J. , Muller Timothy H. , Whittington James C.R. , Mark Shirley , Baram Alon B. , Stachenfeld Kimberley L. , Kurth-Nelson Zeb 
Published: July 2018

AbstractIt is proposed that a cognitive map encoding the relationships between entities in the world supports flexible behaviour, but the majority of the neural evidence for such a system comes from studies of spatial navigation. Recent work describing neuronal parallels between spatial and non-spatial behaviours has rekindled the notion of a systematic organisation of knowledge across multiple domains. We review experimental evidence and theoretical frameworks that point to principles unifying these apparently disparate functions. These principles describe how to learn and use abstract, generalisable knowledge and suggest map-like representations observed in a spatial context may be an instance of general coding mechanisms capable of organising knowledge of all kinds. We highlight how artificial agents endowed with such principles exhibit flexible behaviour and learn map-like representations observed in the brain. Finally, we speculate on how these principles may offer insight into the extreme generalisations, abstractions and inferences that characterise human cognition.
Deep Semantic Protein Representation for Annotation, Discovery, and Engineering
Authors: Schwartz Ariel S , Hannum Gregory J , Dwiel Zach R , Smoot Michael E , Grant Ana R , Knight Jason M , Becker Scott A , Eads Jonathan R , LaFave Matthew C , Eavani Harini , Liu Yinyin , Bansal Arjun K , Richardson Toby H 
Published: July 2018

AbstractComputational assignment of function to proteins with no known homologs is still an unsolved problem. We have created a novel, function-based approach to protein annotation and discovery called D-SPACE (Deep Semantic Protein Annotation Classification and Exploration), comprised of a multi-task, multi-label deep neural network trained on over 70 million proteins. Distinct from homology and motif-based methods, D-SPACE encodes proteins in high-dimensional representations (embeddings), allowing the accurate assignment of over 180,000 labels for 13 distinct tasks. The embedding representation enables fast searches for functionally related proteins, including homologs undetectable by traditional approaches. D-SPACE annotates all 109 million proteins in UniProt in under 35 hours on a single computer and searches the entirety of these in seconds. D-SPACE further quantifies the relative functional effect of mutations, facilitating rapid in silico mutagenesis for protein engineering applications. D-SPACE incorporates protein annotation, search, and other exploratory efforts into a single cohesive model.