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  1. Article ; Online: Detection of tissue factor in platelets: why is it so troublesome?

    Østerud, Bjarne / Bouchard, Beth A

    Platelets

    2019  Volume 30, Issue 8, Page(s) 957–961

    Abstract: Tissue factor (TF) is the most important trigger for the extrinsic coagulation pathway. TF, earlier denoted as thromboplastin, has always been a mystery since its discovery due to its abundant presence in most human tissues but not blood. The latter has ... ...

    Abstract Tissue factor (TF) is the most important trigger for the extrinsic coagulation pathway. TF, earlier denoted as thromboplastin, has always been a mystery since its discovery due to its abundant presence in most human tissues but not blood. The latter has been extensively studied in a vast quest for possible sources of blood-borne TF yielding many conflicting findings and confusing conclusions regarding the presence of TF mRNA, protein or functional procoagulant activity in virtually all blood cells. Platelets, in particular, have been heavily scrutinized by investigators eager to demonstrate expression of TF. However, some investigators including our own groups have not found evidence for TF in platelets. This article discusses notable reports and possible reasons for erroneous detection of platelet TF antigen and activity including artificially hyper-stimulated platelets, suboptimal purity of cell preparations, flaws in study design and/or choice of reagents.
    MeSH term(s) Blood Platelets/metabolism ; Humans ; Megakaryocytes/metabolism ; Thromboplastin/metabolism
    Chemical Substances Thromboplastin (9035-58-9)
    Language English
    Publishing date 2019-05-30
    Publishing country England
    Document type Journal Article
    ZDB-ID 1034283-7
    ISSN 1369-1635 ; 0953-7104
    ISSN (online) 1369-1635
    ISSN 0953-7104
    DOI 10.1080/09537104.2019.1624708
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  2. Article ; Online: Venous thromboembolism, factor VIII and chronic kidney disease.

    Cheung, Katharine L / Bouchard, Beth A / Cushman, Mary

    Thrombosis research

    2018  Volume 170, Page(s) 10–19

    Abstract: Chronic kidney disease (CKD) affects 30 million Americans and is associated with approximately a two-fold increased risk of venous thromboembolism (VTE). There is a graded increased risk of VTE across declining kidney function, as measured by estimated ... ...

    Abstract Chronic kidney disease (CKD) affects 30 million Americans and is associated with approximately a two-fold increased risk of venous thromboembolism (VTE). There is a graded increased risk of VTE across declining kidney function, as measured by estimated glomerular filtration rate (eGFR) and albuminuria. When patients with end-stage kidney disease (ESKD) experience VTE they are more likely than the general population to be hospitalized and they have a higher mortality. The incidence and consequences of VTE may also differ depending on the cause of kidney disease. In addition, kidney transplant patients with VTE are at a greater risk for death and graft loss than transplant patients without VTE. The reasons that patients with CKD are at increased risk of VTE are not well understood, but recent data suggest that factor VIII is a mediator. Factor VIII is an essential cofactor in the coagulation cascade and a strong risk factor for VTE in general. It is inversely correlated with eGFR and prospective studies demonstrate that factor VIII activity predicts incident CKD and rapid eGFR decline. The etiology of CKD may also influence factor VIII levels. This review summarizes the epidemiology VTE in CKD and reviews the biochemistry of factor VIII and determinants of its levels, including von Willebrand factor and ABO blood group. We explore mechanisms by which the complications of CKD might give rise to higher factor VIII and suggests future research directions to understand how factor VIII and CKD are linked.
    MeSH term(s) Factor VIII/metabolism ; Female ; Humans ; Male ; Renal Insufficiency, Chronic/complications ; Renal Insufficiency, Chronic/pathology ; Risk Factors ; Venous Thromboembolism/complications ; Venous Thromboembolism/pathology
    Chemical Substances Factor VIII (9001-27-8)
    Language English
    Publishing date 2018-07-31
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 121852-9
    ISSN 1879-2472 ; 0049-3848
    ISSN (online) 1879-2472
    ISSN 0049-3848
    DOI 10.1016/j.thromres.2018.07.029
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  3. Article ; Online: Mechanisms Regulating Acquisition of Platelet-Derived Factor V/Va by Megakaryocytes.

    Gertz, Jacqueline M / Bouchard, Beth A

    Journal of cellular biochemistry

    2015  Volume 116, Issue 10, Page(s) 2121–2126

    Abstract: Factor Va serves as the nonenzymatic protein cofactor for the prothrombinase complex, which converts prothrombin to thrombin in the events leading to formation of a hemostatic plug. Several observations support the concept that platelet-derived factor V/ ... ...

    Abstract Factor Va serves as the nonenzymatic protein cofactor for the prothrombinase complex, which converts prothrombin to thrombin in the events leading to formation of a hemostatic plug. Several observations support the concept that platelet-derived factor V/Va is physically and functionally distinct and plays a more important role in thrombin generation at sites of vascular injury as compared to its plasma counterpart. Platelet-derived factor V/Va is generated following endocytosis of the plasma-derived molecule by the platelet precursor cells, megakaryocytes, via a two receptor system consisting of low density lipoprotein (LDL) receptor-related protein-1 (LRP-1) and an unidentified specific "binding site". More recently, it was suggested that a cell surface-expressed β-galactoside binding protein, galectin-8, was involved in factor V endocytosis. Endocytosed factor V is trafficked through the cell and retailored prior to its storage in α-granules. Given the essential role of platelet-derived factor Va in clot formation, understanding the cellular and molecular mechanisms that regulate how platelets acquire this molecule will be important for the treatment of excessive bleeding or clotting.
    MeSH term(s) Blood Coagulation/genetics ; Blood Coagulation Factors/genetics ; Blood Coagulation Factors/metabolism ; Blood Platelets/metabolism ; Blood Platelets/pathology ; Endocytosis/genetics ; Galectins/metabolism ; Hemorrhage/genetics ; Hemorrhage/metabolism ; Hemorrhage/pathology ; Humans ; Low Density Lipoprotein Receptor-Related Protein-1/metabolism ; Megakaryocytes/metabolism ; Thrombin/metabolism ; Vascular System Injuries/metabolism ; Vascular System Injuries/pathology
    Chemical Substances Blood Coagulation Factors ; Galectins ; LDLR-related protein 1A, human ; LGALS8 protein, human ; Low Density Lipoprotein Receptor-Related Protein-1 ; platelet factor V ; Thrombin (EC 3.4.21.5)
    Language English
    Publishing date 2015-10
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 392402-6
    ISSN 1097-4644 ; 0730-2312
    ISSN (online) 1097-4644
    ISSN 0730-2312
    DOI 10.1002/jcb.25163
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  4. Article ; Online: Simultaneous flow cytometric analysis of megakaryocyte polyploidy and a labile intracellular protein using zinc-based fixation.

    Gertz, Jacqueline M / Meuser, Megan / Bouchard, Beth A

    Cytometry. Part A : the journal of the International Society for Analytical Cytology

    2017  Volume 91, Issue 7, Page(s) 713–720

    Abstract: Differentiating megakaryocytes undergo a unique endomitotic cell cycle leading to large polyploidal cells, which fragment to generate platelets, blood cells important for normal hemostasis. Simultaneous assessment of DNA content and cellular proteins by ... ...

    Abstract Differentiating megakaryocytes undergo a unique endomitotic cell cycle leading to large polyploidal cells, which fragment to generate platelets, blood cells important for normal hemostasis. Simultaneous assessment of DNA content and cellular proteins by flow cytometry is a useful tool to study megakaryocyte differentiation and to define expression of proteins important for megakaryocyte development and platelet formation. The usefulness of zinc salt-based fixation (ZBF), a non-crosslinking method of cell fixation that permits downstream analysis of nucleic acids (Jensen et al., Cytometry A 2010;77A:798-804), in flow cytometric analysis of megakaryocyte ploidy in conjunction with extracellular and intracellular proteins was assessed. ZBF of a megakaryocyte-like cell line resulted in preservation of proteins similar to paraformaldehyde fixation, and preservation of DNA content in a manner similar to methanol fixation. This is highlighted by experiments in which polyploidal megakaryocytes were analyzed simultaneously for endocytosis of a fluorescently-labeled, endocytosed labile protein or expression of a cell surface integrin and DNA content. These studies demonstrate that ZBF will be a valuable tool to study the molecular events leading to platelet formation. © 2017 International Society for Advancement of Cytometry.
    Language English
    Publishing date 2017-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2099868-5
    ISSN 1552-4930 ; 0196-4763 ; 1552-4922
    ISSN (online) 1552-4930
    ISSN 0196-4763 ; 1552-4922
    DOI 10.1002/cyto.a.23161
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  5. Article ; Online: Endocytosed factor V is trafficked to CD42b

    Gertz, Jacqueline M / McLean, Kelley C / Bouchard, Beth A

    Journal of cellular physiology

    2018  Volume 233, Issue 11, Page(s) 8691–8700

    Abstract: Plasma- and platelet-derived factor Va are essential for thrombin generation catalyzed by the prothrombinase complex; however, several observations demonstrate that the platelet-derived cofactor, which is formed following megakaryocyte endocytosis and ... ...

    Abstract Plasma- and platelet-derived factor Va are essential for thrombin generation catalyzed by the prothrombinase complex; however, several observations demonstrate that the platelet-derived cofactor, which is formed following megakaryocyte endocytosis and modification of the plasma procofactor, factor V, is more hemostatically relevant. Factor V endocytosis, as a function of megakaryocyte differentiation and proplatelet formation, was assessed by flow cytometry and microscopy in CD34
    MeSH term(s) Antigens, CD34/genetics ; Blood Platelets/metabolism ; Cell Differentiation/genetics ; Cell Movement/genetics ; Endocytosis/genetics ; Factor V/genetics ; Fetal Blood/cytology ; Fetal Blood/metabolism ; Gene Expression Regulation/genetics ; Hematopoiesis/genetics ; Hematopoietic Stem Cells/metabolism ; Humans ; Megakaryocytes/cytology ; Megakaryocytes/metabolism ; Platelet Glycoprotein GPIb-IX Complex/genetics ; Thrombopoietin/genetics
    Chemical Substances Antigens, CD34 ; Platelet Glycoprotein GPIb-IX Complex ; Factor V (9001-24-5) ; Thrombopoietin (9014-42-0)
    Language English
    Publishing date 2018-05-15
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 3116-1
    ISSN 1097-4652 ; 0021-9541
    ISSN (online) 1097-4652
    ISSN 0021-9541
    DOI 10.1002/jcp.26749
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  6. Article ; Online: The Polyanionic Drug Suramin Neutralizes Histones and Prevents Endotheliopathy.

    Villalba, Nuria / Sackheim, Adrian M / Lawson, Michael A / Haines, Laurel / Chen, Yen-Lin / Sonkusare, Swapnil K / Ma, Yong-Tao / Li, Jianing / Majumdar, Devdoot / Bouchard, Beth A / Boyson, Jonathan E / Poynter, Matthew E / Nelson, Mark T / Freeman, Kalev

    Journal of immunology (Baltimore, Md. : 1950)

    2023  Volume 211, Issue 4, Page(s) 648–657

    Abstract: Drugs are needed to protect against the neutrophil-derived histones responsible for endothelial injury in acute inflammatory conditions such as trauma and sepsis. Heparin and other polyanions can neutralize histones but challenges with dosing or side ... ...

    Abstract Drugs are needed to protect against the neutrophil-derived histones responsible for endothelial injury in acute inflammatory conditions such as trauma and sepsis. Heparin and other polyanions can neutralize histones but challenges with dosing or side effects such as bleeding limit clinical application. In this study, we demonstrate that suramin, a widely available polyanionic drug, completely neutralizes the toxic effects of individual histones, but not citrullinated histones from neutrophil extracellular traps. The sulfate groups on suramin form stable electrostatic interactions with hydrogen bonds in the histone octamer with a dissociation constant of 250 nM. In cultured endothelial cells (Ea.Hy926), histone-induced thrombin generation was significantly decreased by suramin. In isolated murine blood vessels, suramin abolished aberrant endothelial cell calcium signals and rescued impaired endothelial-dependent vasodilation caused by histones. Suramin significantly decreased pulmonary endothelial cell ICAM-1 expression and neutrophil recruitment caused by infusion of sublethal doses of histones in vivo. Suramin also prevented histone-induced lung endothelial cell cytotoxicity in vitro and lung edema, intra-alveolar hemorrhage, and mortality in mice receiving a lethal dose of histones. Protection of vascular endothelial function from histone-induced damage is a novel mechanism of action for suramin with therapeutic implications for conditions characterized by elevated histone levels.
    MeSH term(s) Mice ; Animals ; Histones/metabolism ; Suramin/pharmacology ; Endothelial Cells/metabolism ; Endothelium/metabolism ; Hemorrhage
    Chemical Substances Histones ; Suramin (6032D45BEM) ; polyanions
    Language English
    Publishing date 2023-07-03
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
    ZDB-ID 3056-9
    ISSN 1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
    ISSN (online) 1550-6606
    ISSN 0022-1767 ; 1048-3233 ; 1047-7381
    DOI 10.4049/jimmunol.2200703
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  7. Article ; Online: Plasma-based assays distinguish hyperfibrinolysis and shutdown subgroups in trauma-induced coagulopathy.

    Lawson, Michael A / Holle, Lori A / Dow, Nathan E / Hennig, Grant / de Laat, Bas / Moore, Hunter B / Moore, Ernest E / Cohen, Mitchell J / Bouchard, Beth A / Freeman, Kalev / Wolberg, Alisa S

    The journal of trauma and acute care surgery

    2022  Volume 93, Issue 5, Page(s) 579–587

    Abstract: Background: Trauma patients with abnormal fibrinolysis have increased morbidity and mortality. Knowledge of mechanisms differentiating fibrinolytic phenotypes is important to optimize treatment. We hypothesized that subjects with abnormal fibrinolysis ... ...

    Abstract Background: Trauma patients with abnormal fibrinolysis have increased morbidity and mortality. Knowledge of mechanisms differentiating fibrinolytic phenotypes is important to optimize treatment. We hypothesized that subjects with abnormal fibrinolysis identified by whole blood viscoelastometry can also be distinguished by plasma thrombin generation, clot structure, fibrin formation, and plasmin generation measurements.
    Methods: Platelet-poor plasma (PPP) from an observational cross-sectional trauma cohort with fibrinolysis shutdown (% lysis at 30 minutes [LY30] < 0.9, n = 11) or hyperfibrinolysis (LY30 > 3%, n = 9) defined by whole blood thromboelastography were studied. Noninjured control subjects provided comparative samples. Thrombin generation, fibrin structure and formation, and plasmin generation were measured by fluorescence, confocal microscopy, turbidity, and a fluorescence-calibrated plasmin assay, respectively, in the absence/presence of tissue factor or tissue plasminogen activator (tPA).
    Results: Whereas spontaneous thrombin generation was not detected in PPP from control subjects, PPP from hyperfibrinolysis or shutdown patients demonstrated spontaneous thrombin generation, and the lag time was shorter in hyperfibrinolysis versus shutdown. Addition of tissue factor masked this difference but revealed increased thrombin generation in hyperfibrinolysis samples. Compared with shutdown, hyperfibrinolysis PPP formed denser fibrin networks. In the absence of tPA, the fibrin formation rate was faster in shutdown than hyperfibrinolysis, but hyperfibrinolysis clots lysed spontaneously; these differences were masked by addition of tPA. Tissue plasminogen activator-stimulated plasmin generation was similar in hyperfibrinolysis and shutdown samples. Differences in LY30, fibrin structure, and lysis correlated with pH.
    Conclusion: This exploratory study using PPP-based assays identified differences in thrombin generation, fibrin formation and structure, and lysis in hyperfibrinolysis and shutdown subgroups. These groups did not differ in their ability to promote tPA-triggered plasmin generation. The ability to characterize these activities in PPP facilitates studies to identify mechanisms that promote adverse outcomes in trauma.
    Level of evidence: Prognostic/Epidemiological; Level III.
    MeSH term(s) Humans ; Tissue Plasminogen Activator ; Fibrinolysis ; Fibrinolysin ; Thrombin ; Thromboplastin ; Cross-Sectional Studies ; Blood Coagulation Disorders/diagnosis ; Blood Coagulation Disorders/etiology ; Fibrin
    Chemical Substances Tissue Plasminogen Activator (EC 3.4.21.68) ; Fibrinolysin (EC 3.4.21.7) ; Thrombin (EC 3.4.21.5) ; Thromboplastin (9035-58-9) ; Fibrin (9001-31-4)
    Language English
    Publishing date 2022-06-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2651070-4
    ISSN 2163-0763 ; 2163-0755
    ISSN (online) 2163-0763
    ISSN 2163-0755
    DOI 10.1097/TA.0000000000003723
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  8. Article ; Online: Guidelines on the use of sex and gender in cardiovascular research.

    Usselman, Charlotte W / Lindsey, Merry L / Robinson, Austin T / Habecker, Beth A / Taylor, Chloe E / Merryman, W David / Kimmerly, Derek / Bender, Jeffrey R / Regensteiner, Judith G / Moreau, Kerrie L / Pilote, Louise / Wenner, Megan M / O'Brien, Myles / Yarovinsky, Timur O / Stachenfeld, Nina S / Charkoudian, Nisha / Denfeld, Quin E / Moreira-Bouchard, Jesse D / Pyle, W Glen /
    DeLeon-Pennell, Kristine Y

    American journal of physiology. Heart and circulatory physiology

    2023  Volume 326, Issue 1, Page(s) H238–H255

    Abstract: In cardiovascular research, sex and gender have not typically been considered in research design and reporting until recently. This has resulted in clinical research findings from which not only all women, but also gender-diverse individuals have been ... ...

    Abstract In cardiovascular research, sex and gender have not typically been considered in research design and reporting until recently. This has resulted in clinical research findings from which not only all women, but also gender-diverse individuals have been excluded. The resulting dearth of data has led to a lack of sex- and gender-specific clinical guidelines and raises serious questions about evidence-based care. Basic research has also excluded considerations of sex. Including sex and/or gender as research variables not only has the potential to improve the health of society overall now, but it also provides a foundation of knowledge on which to build future advances. The goal of this guidelines article is to provide advice on best practices to include sex and gender considerations in study design, as well as data collection, analysis, and interpretation to optimally establish rigor and reproducibility needed to inform clinical decision-making and improve outcomes. In cardiovascular physiology, incorporating sex and gender is a necessary component when optimally designing and executing research plans. The guidelines serve as the first guidance on how to include sex and gender in cardiovascular research. We provide here a beginning path toward achieving this goal and improve the ability of the research community to interpret results through a sex and gender lens to enable comparison across studies and laboratories, resulting in better health for all.
    MeSH term(s) Female ; Humans ; Male ; Sex Characteristics ; Cardiovascular System ; Cardiology ; Biomedical Research
    Language English
    Publishing date 2023-11-24
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 603838-4
    ISSN 1522-1539 ; 0363-6135
    ISSN (online) 1522-1539
    ISSN 0363-6135
    DOI 10.1152/ajpheart.00535.2023
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  9. Article ; Online: Increased histone-DNA complexes and endothelial-dependent thrombin generation in severe COVID-19.

    Bouchard, Beth A / Colovos, Christos / Lawson, Michael A / Osborn, Zachary T / Sackheim, Adrian M / Mould, Kara J / Janssen, William J / Cohen, Mitchell J / Majumdar, Devdoot / Freeman, Kalev

    Vascular pharmacology

    2021  Volume 142, Page(s) 106950

    Abstract: Coagulopathy in severe COVID-19 is common but poorly understood. The purpose of this study was to determine how SARS-CoV-2 infection impacts histone levels, fibrin structure, and endogenous thrombin potential in the presence and absence of endothelial ... ...

    Abstract Coagulopathy in severe COVID-19 is common but poorly understood. The purpose of this study was to determine how SARS-CoV-2 infection impacts histone levels, fibrin structure, and endogenous thrombin potential in the presence and absence of endothelial cells. We studied individuals with SARS-CoV-2 infection and acute respiratory distress syndrome at the time of initiation of mechanical ventilation compared to healthy controls. Circulating histone-DNA complexes were elevated in the plasma of COVID-19 patients relative to healthy controls (n=6, each group). Using calibrated automated thrombography, thrombin generation was altered in COVID-19 patient plasma samples. Despite having increased endogenous thrombin potential, patient plasma samples exhibited prolonged lag times and times to peak thrombin in the presence of added tissue factor and PCPS. Strikingly different results were observed when endothelial cells were used in place of tissue factor and PCPS. While healthy control plasma samples did not generate measurable thrombin after 60 min, plasma samples from COVID-19+ patients formed thrombin (mean lag time ~20 min). Consistent with the observed alterations in thrombin generation, clots from COVID-19 subjects exhibited a denser fibrin network, thinner fibers and lower fibrin resolvability. Elevated histones, aberrant fibrin formation, and increased endothelial-dependent thrombin generation may contribute to coagulopathy in COVID-19.
    MeSH term(s) COVID-19 ; DNA ; Endothelial Cells ; Histones ; Humans ; SARS-CoV-2 ; Thrombin
    Chemical Substances Histones ; DNA (9007-49-2) ; Thrombin (EC 3.4.21.5)
    Language English
    Publishing date 2021-12-03
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2082846-9
    ISSN 1879-3649 ; 1537-1891 ; 1879-3649
    ISSN (online) 1879-3649 ; 1537-1891
    ISSN 1879-3649
    DOI 10.1016/j.vph.2021.106950
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  10. Article ; Online: Creating an EBP framework on a journey to becoming an EBP agency: pioneers in the field of children's mental health.

    Archer-Kuhn, Beth / Bouchard, Terrance Thomas / Greco, Adelle

    Journal of evidence-based social work

    2014  Volume 11, Issue 1-2, Page(s) 2–17

    Abstract: Agencies servicing children, youth, and families have been particularly pressured to demonstrate service effectiveness and accountability by government funders. The human service fields have not fully embraced research evidence into the organizational ... ...

    Abstract Agencies servicing children, youth, and families have been particularly pressured to demonstrate service effectiveness and accountability by government funders. The human service fields have not fully embraced research evidence into the organizational culture creating a challenge of introducing research evidence into agencies. Gaps in knowledge have been identified when agencies attempt to travel down the path of introducing evidence-based practice into organizational culture. The paradigm shift of introducing research into practice was the journey taken by one mid-sized agency in southwestern Ontario, Canada. A framework for assessing evidence-based practice programs in services was created as part of their journey.
    MeSH term(s) Adolescent ; Canada ; Child ; Evidence-Based Practice/organization & administration ; Evidence-Based Practice/standards ; Humans ; Mental Health Services/organization & administration ; Mental Health Services/standards ; Needs Assessment ; Organizational Objectives ; Program Development ; Program Evaluation ; Social Work/organization & administration ; Social Work/standards
    Language English
    Publishing date 2014
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2219118-5
    ISSN 1543-3722 ; 1543-3714
    ISSN (online) 1543-3722
    ISSN 1543-3714
    DOI 10.1080/15433714.2013.837338
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