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Althaus K, Hoepner G, Zieger B, Prüller F, Pavlova A, Boeckelmann D, Birschmann I, Müller J, Rühl H, Sachs U, Kehrel B, Streif W, Bugert P, Zaninetti C, Cooper N, Schulze H, Knöfler R, Bakchoul T, Jurk K. The Diagnostic Assessment of Platelet Function Defects - Part 2: Update on Platelet Disorders. Hamostaseologie 2025. [PMID: 39870108 DOI: 10.1055/a-2404-0216] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2025] Open
Abstract
Congenital platelet disorders are rare and targeted treatment is usually not possible. Inherited platelet function disorders (iPFDs) can affect surface receptors and multiple platelet responses such as defects of platelet granules, signal transduction, and procoagulant activity. If iPFDs are also associated with a reduced platelet count (thrombocytopenia), it is not uncommon to be misdiagnosed as immune thrombocytopenia. Because the bleeding tendency of the different platelet disorders is variable, a correct diagnosis of the platelet defect based on phenotyping, function analysis, and genotyping is essential, especially in the perioperative setting. In the case of a platelet receptor deficiency, such as Bernard-Soulier syndrome or Glanzmann thrombasthenia, not only the bleeding tendency but also the risk of isoimmunization after platelet transfusions or pregnancy has to be considered. Platelet granule disorders are commonly associated with either intrinsically quantitative or qualitative granule defects due to impaired granulopoiesis, or granule release defects, which can also affect additional signaling pathways. Functional platelet defects require expertise in the clinical bleeding tendency in terms of the disorder when using antiplatelet agents or other medications that affect platelet function. Platelet defects associated with hematological-oncological diseases require comprehensive information about the patient including the clinical implication of the genetic testing. This review focuses on genetics, clinical presentation, and laboratory platelet function analysis of iPFDs with or without reduced platelet number. As platelet defects affecting the cytoskeleton usually show thrombocytopenia, but less impaired or normal platelet functional responses, they are not specifically addressed.
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Affiliation(s)
- Karina Althaus
- Medical Faculty of Tübingen, Institute for Clinical and Experimental Transfusion Medicine, Tübingen, Germany
- Center for Clinical Transfusion Medicine, University Hospital of Tübingen, Tübingen, Germany
| | - Gero Hoepner
- Center for Clinical Transfusion Medicine, University Hospital of Tübingen, Tübingen, Germany
- Department of Anaesthesiology and Intensive Care, University Hospital Tübingen, Tübingen, Germany
| | - Barbara Zieger
- Department of Pediatrics and Adolescent Medicine, University Medical Center Freiburg, Freiburg, Germany
| | - Florian Prüller
- Klinisches Institut für Medizinische und Chemische Labordiagnostik, Medizinische Universität Graz, Graz, Austria
| | - Anna Pavlova
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Doris Boeckelmann
- Department of Pediatrics and Adolescent Medicine, University Medical Center Freiburg, Freiburg, Germany
| | - Ingvild Birschmann
- Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Institut für Laboratoriums- und Transfusionsmedizin, Bochum, Germany
| | - Jens Müller
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Heiko Rühl
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Ulrich Sachs
- Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany
| | - Beate Kehrel
- Department of Anaesthesiology and Intensive Care, Experimental and Clinical Haemostasis, University-Hospital Munster, Münster, Germany
| | - Werner Streif
- Kinder- und Jugendheilkunde, Innsbruck Medical University, Innsbruck, Austria
| | - Peter Bugert
- Medical Faculty Mannheim, Institute of Transfusion Medicine and Immunology, Heidelberg University, Mannheim, Germany
| | - Carlo Zaninetti
- Institute of Immunology and Transfusion Medicine, Universitätsmedizin Greifswald, Greifswald, Germany
| | - Nina Cooper
- Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany
| | - Harald Schulze
- Institute of Experimental Biomedicine, Universitätsklinikum Würzburg, Würzburg, Germany
| | - Ralf Knöfler
- Department of Paediatric Haemostaseology, Dresden University Hospital, Dresden, Germany
| | - Tamam Bakchoul
- Medical Faculty of Tübingen, Institute for Clinical and Experimental Transfusion Medicine, Tübingen, Germany
- Center for Clinical Transfusion Medicine, University Hospital of Tübingen, Tübingen, Germany
| | - Kerstin Jurk
- Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
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Hoepner G, Althaus K, Müller J, Zieger B, Pavlova A, Boeckelmann D, Knöfler R, Bugert P, Kehrel B, Streif W, Birschmann I, Rühl H, Sachs U, Prüller F, Zaninetti C, Schulze H, Cooper N, Jurk K, Bakchoul T. The Diagnostic Assessment of Inherited Platelet Function Defects - Part 1: An Overview of the Diagnostic Approach and Laboratory Methods. Hamostaseologie 2025. [PMID: 39870109 DOI: 10.1055/a-2436-5318] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2025] Open
Abstract
In this article, our goal is to offer an introduction and overview of the diagnostic approach to inherited platelet function defects (iPFDs) for clinicians and laboratory personnel who are beginning to engage in the field. We describe the most commonly used laboratory methods and propose a diagnostic four-step approach, wherein each stage requires a higher level of expertise and more specialized methods. It should be noted that our proposed approach differs from the ISTH Guidance on this topic in some points. The first step in the diagnostic approach of iPFD should be a thorough medical history and clinical examination. We strongly advocate for the use of a validated bleeding score like the ISTH-BAT (International Society on Thrombosis and Haemostasis Bleeding Assessment Tool). External factors like diet and medication have to be considered. The second step should rule out plasmatic bleeding disorders and von Willebrand disease. Once this has been accomplished, the third step consists of a thorough platelet investigation of platelet phenotype and function. Established methods consist of blood smear analysis by light microscopy, light transmission aggregometry, and flow cytometry. Additional techniques such as lumiaggregometry, immune fluorescence microscopy, and platelet-dependent thrombin generation help confirm and specify the diagnosis of iPFD. In the fourth and last step, genetic testing can confirm a diagnosis, reveal novel mutations, and allow to compare unclear genetics with lab results. If diagnosis cannot be established through this process, experimental methods such as electron microscopy can give insight into the underlying disease.
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Affiliation(s)
- Gero Hoepner
- Center for Clinical Transfusion Medicine Tuebingen, Tuebingen, Germany
- Department of Anaesthesiology and Intensive Care, University Hospital Tuebingen, Tuebingen, Germany
| | - Karina Althaus
- Center for Clinical Transfusion Medicine Tuebingen, Tuebingen, Germany
- Medical Faculty of Tuebingen, Institute for Clinical and Experimental Transfusion Medicine, Tuebingen, Germany
| | - Jens Müller
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Barbara Zieger
- Department of Pediatrics and Adolescent Medicine, University Medical Center Freiburg, Freiburg, Germany
| | - Anna Pavlova
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Doris Boeckelmann
- Department of Pediatrics and Adolescent Medicine, University Medical Center Freiburg, Freiburg, Germany
| | - Ralf Knöfler
- Department of Paediatric Haemostaseology, Dresden University Hospital, Dresden, Germany
| | - Peter Bugert
- Medical Faculty Mannheim, Institute of Transfusion Medicine and Immunology, Heidelberg University, Mannheim, Germany
| | - Beate Kehrel
- Department of Anaesthesiology and Intensive Care, Experimental and Clinical Haemostasis, University-Hospital Munster, Münster, Germany
| | - Werner Streif
- Kinder- und Jugendheilkunde, Innsbruck Medical University, Innsbruck, Austria
| | - Ingvild Birschmann
- Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Institut für Laboratoriums- und Transfusionsmedizin, Bochum, Germany
| | - Heiko Rühl
- Institute of Experimental Haematology and Transfusion Medicine (IHT), University Hospital Bonn, Bonn, Germany
| | - Ulrich Sachs
- Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany
- Center for Transfusion Medicine and Haemotherapy, Department of Thrombosis and Haemostasis, European Comprehensive Care Center for Haemophilia at Giessen and Marburg University Hospital, Giessen, Germany
| | | | - Carlo Zaninetti
- Institute of Immunology and Transfusion Medicine, Universitätsmedizin Greifswald, Greifswald, Germany
| | - Harald Schulze
- Institute of Experimental Biomedicine, Universitätsklinikum Würzburg, Würzburg, Germany
| | - Nina Cooper
- Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany
- Center for Transfusion Medicine and Haemotherapy, Department of Thrombosis and Haemostasis, European Comprehensive Care Center for Haemophilia at Giessen and Marburg University Hospital, Giessen, Germany
| | - Kerstin Jurk
- Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
| | - Tamam Bakchoul
- Center for Clinical Transfusion Medicine Tuebingen, Tuebingen, Germany
- Medical Faculty of Tuebingen, Institute for Clinical and Experimental Transfusion Medicine, Tuebingen, Germany
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Stépanian A, Fischer F, Flaujac C, Eschwège V, Delassasseigne C, Leflem L, Loridon F, Voisin S, Lasne D. Light transmission aggregometry for platelet function testing: position paper on current recommendations and French proposals for accreditation. Platelets 2024; 35:2427745. [PMID: 39555668 DOI: 10.1080/09537104.2024.2427745] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2024] [Revised: 10/17/2024] [Accepted: 11/05/2024] [Indexed: 11/19/2024]
Abstract
Light transmission aggregometry (LTA) is a method used to investigate platelet functions in platelet-rich plasma (PRP), notably when screening for platelet disorders. Various national guidelines and recommendations help in setting up the LTA test in specialized laboratories. However, due to the nature of the sample matrix and its subsequent specificities, more accurate positions are needed to achieve LTA accreditation according to the standard NF EN ISO 15 189. We reviewed guidelines and recommendations as they can be useful in the accreditation process, and we conducted a survey on LTA practice among members of the Société Française de Thrombose et d'Hémostase (SFTH) in 2021. We formulated 28 proposals, which have been approved by vote within the SFTH. All aspects to take into consideration for the proper conduct of LTA assays and their accreditation have been covered. Notably, preanalytical, analytical and postanalytical aspects are depicted, including blood sampling, PRP preparation, instruments, agonists, performance assessment, personnel training and data interpretation. This document, essentially representing a French position paper on the current recommendations and subsequent proposals for LTA accreditation, might prove useful also outside France for relevant laboratories and auditors involved in LTA accreditation.
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Affiliation(s)
- Alain Stépanian
- Service d'Hématologie Biologique, PhyMedExp UMR UM - CNRS 9214 - Inserm U1046, CHU de Montpellier, Université de Montpellier, Montpellier, France
| | | | - Claire Flaujac
- Laboratoire de biologie médicale, secteur hémostase, centre hospitalier de Versailles (André Mignot), Le Chesnay-Rocquencourt, France
| | | | | | - Léna Leflem
- Laboratoire Eurofins, Ivry-sur-Seine, France
| | | | - Sophie Voisin
- Laboratoire d'Hématologie, CHU de Toulouse, Toulouse, France
| | - Dominique Lasne
- Laboratoire d'Hématologie Générale, Hôpital Universitaire Necker-Enfants Malades, AP-HP, Paris, France
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Khourssaji M, Bareille M, Alberio L, Borgel D, Fouassier M, Béné MC, Lecompte T, Mullier F. Mepacrine Flow Cytometry Assay for the Diagnosis of Platelet δ-granule Defects: Literature Review on Methods-Towards a Shared Detailed Protocol. Thromb Haemost 2024. [PMID: 39260401 DOI: 10.1055/a-2413-2870] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 09/13/2024]
Abstract
Accurate assessment of platelet secretion is essential for the diagnosis of inherited or acquired platelet function disorders and more specifically in identifying δ-storage pool disease. Mepacrine, a fluorescent dye, specifically accumulates in platelet δ-granules. The mepacrine flow cytometry (mepacrine FCM) assay has been used for more than half a century in the clinical laboratory as a diagnostic tool for platelet δ-granule disorders. The assay requires a small volume of blood, can be performed in thrombocytopenic patients, provides rapid assessment of δ-granule content and secretion, and, thus, enables differentiation between storage and release defects. There is however a broad heterogeneity in methods, reagents, and equipment used. Lack of standardization and limited data on analytical and clinical performances have led the 2022 ISTH SSC (International Society on Thrombosis and Haemostasis Scientific and Standardization Committee) Subcommittee on Platelet Physiology expert consensus to rate this assay as simple but of uncertain value. Yet, the data used by experts to formulate the recommendations were not discussed and even not mentioned. Guidance for laboratory studies of platelet secretion assay would be very helpful for clinical laboratories and health authorities especially considering the implications of the new In Vitro Diagnostic Regulation in Europe. The purpose of the present work was to review the reported methodologies for the mepacrine FCM assay and to offer an example of detailed protocol. This would help standardization and pave the way for more rigorous comparative studies.
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Affiliation(s)
- Mehdi Khourssaji
- Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium
- Institut de Recherche Expérimentale et Clinique (IREC) - Pôle Mont, Université catholique de Louvain, Yvoir, Belgium
| | - Marion Bareille
- Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium
- Institut de Recherche Expérimentale et Clinique (IREC) - Pôle Mont, Université catholique de Louvain, Yvoir, Belgium
| | - Lorenzo Alberio
- Division of Haematology and Central Haematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland
| | - Delphine Borgel
- Service d'Hématologie Biologique, Hôpital Necker AP-HP, Paris, France
| | - Marc Fouassier
- Centre de Ressources et de Compétences - Maladies Hémorragiques Constitutionnelles, CHU de Nantes, Nantes, France
| | | | - Thomas Lecompte
- Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium
- Hematology Department and Grand East Competence Center on Inherited Platelet Disorders, CHU Nancy, Nancy, France
| | - François Mullier
- Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium
- Institut de Recherche Expérimentale et Clinique (IREC) - Pôle Mont, Université catholique de Louvain, Yvoir, Belgium
- Université de Namur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), Namur, Belgium
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Mecca R, Senthil N, Lakshmanan S, Anna Anil A. Clinical Management of a Rare Hereditary Bleeding Disorder in an Adult: Glanzmann Thrombasthenia. Cureus 2024; 16:e72391. [PMID: 39588442 PMCID: PMC11586241 DOI: 10.7759/cureus.72391] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 10/24/2024] [Indexed: 11/27/2024] Open
Abstract
Glanzmann thrombasthenia (GT) is a rare autosomal recessive bleeding disorder affecting the megakaryocyte lineage and is associated mainly with mucocutaneous bleeding. We herein report a woman in her early 40s, with no known comorbidities, who presented with severe gingival bleeding and severe fatigue. Past history revealed recurrent gingival bleeding, bruising, and heavy menstrual bleeding. Platelet aggregation studies revealed normal aggregation. Finally, a diagnosis of GT with microcytic hypochromic anemia was established after ruling out all the other differential diagnoses. The patient was transfused with multiple units of packed cells (PCs), random donor platelets (RDPs), and single donor platelets (SDPs) and treated with intravenous (IV) and topical tranexamic acid. Our patient presented with a rare bleeding disorder very late in life. GT can be potentially life-threatening and is difficult to diagnose in the early stages. Therefore, it is imperative to highlight the importance of early diagnosis of this rare condition to provide supportive care for a good prognosis.
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Affiliation(s)
- Reya Mecca
- Internal Medicine, Sri Ramachandra Institute of Higher Education and Research, Chennai, IND
| | - N Senthil
- Internal Medicine, Sri Ramachandra Institute of Higher Education and Research, Chennai, IND
| | - Suja Lakshmanan
- Internal Medicine, Sri Ramachandra Institute of Higher Education and Research, Chennai, IND
| | - Archa Anna Anil
- Internal Medicine, Sri Ramachandra Institute of Higher Education and Research, Chennai, IND
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Kashyap AK, Saikrishna B, Duggal B. Optimized Methodology to Produce Platelet-Rich Plasma and Perform Platelet Aggregation in Patients With Coronary Artery Disease. Cureus 2024; 16:e69032. [PMID: 39391413 PMCID: PMC11464479 DOI: 10.7759/cureus.69032] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 09/09/2024] [Indexed: 10/12/2024] Open
Abstract
BACKGROUND Inter-individual differences in clopidogrel metabolism and platelet counts within platelet-rich plasma (PRP) intrigued us to optimize light transmittance aggregometry (LTA) assay in coronary artery disease (CAD) patients administered with clopidogrel and ticagrelor. The objective of the study was to optimize PRP preparation using separating gel PRP tubes to perform LTA among CAD patients on clopidogrel and ticagrelor. METHODOLOGY Initially, we optimized PRP preparation and platelet aggregation (PA) on healthy controls. To validate the protocol, we recruited 10 healthy controls and 28 CAD patients, comprising 16 on clopidogrel and 12 on ticagrelor regimen. Bio-X, India, supplied PRP tubes (9 mL) with 3.2% sodium citrate. PRP and autologous platelet-poor plasma (PPP) were prepared by centrifugation at 151 g for seven minutes and 3,780 g for 10 minutes, respectively. LTA was performed using platelet aggregometer TA-4V (Stago, Asnières-sur-Seine, France). Adenosine diphosphate (ADP) (10 µM) was used as an agonist. RESULTS The mean maximum platelet aggregation (MPA) among controls, clopidogrel patients, and ticagrelor patients were 55.161±13.69%, 53.17±22%, and 35.84±20.79% (p=0.042), respectively. The mean platelet volumes among groups were 9.34±2.51 fL, 7.60±0.85 fL, and 10.99±1.62 fL (p≤0.01), respectively. CONCLUSION We successfully optimized PRP preparation for the LTA assay using a PRP tube. Its application to measure PA routinely in a cardiology clinic seems propitious.
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Affiliation(s)
- Atul K Kashyap
- Cardiology, All India Institute of Medical Sciences, Rishikesh, Rishikesh, IND
| | - Bonu Saikrishna
- Cardiology, All India Institute of Medical Sciences, Rishikesh, Rishikesh, IND
| | - Bhanu Duggal
- Cardiology, All India Institute of Medical Sciences, Rishikesh, Rishikesh, IND
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Flaujac C, Delassasseigne C, Hurtaud-Roux MF, Delahousse B, Boissier E, Desconclois C. Stability of Hemostasis Parameters in Whole Blood, Plasma, and Frozen Plasma: Literature Review and Recommendations of the SFTH (French Society of Thrombosis and Haemostasis). Semin Thromb Hemost 2024. [PMID: 39214147 DOI: 10.1055/s-0044-1788901] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 09/04/2024]
Abstract
Preanalytical sample management is critical for a proper assessment of hemostasis parameters, and may differ depending on prescribed tests or additional tests considered to be necessary after initial results. Although there is quite vast literature on this issue, the Working Group of the French Society of Thrombosis and Haemostasis (SFTH) deemed it necessary to make an in-depth literature review and propose recommendations for the proper handling of samples prior to hemostasis assays. This extensive assessment is accessible on-line in French at the SFTH website. Here, a more synthetic view of these recommendations is proposed, supported by easy-to-use tables. The latter respectively deal with the stability of whole blood or fresh plasma, frozen samples, and proper handling of samples forwarded on dry ice. Procedures are classified as recommended, acceptable, not conformed and lacking data. This work involved the retrieval of 125 references, first screened by a working group of 6 experts, then reviewed by 20 other experts in the field. The highly detailed conditions summarized in these tables will hopefully help hemostasis laboratories to secure the conditions recommended for sample collection and transportation. Moreover, as some conditions clearly lacked recommendations, this review can open new fields of investigation for hemostasis preanalytics.
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Affiliation(s)
- Claire Flaujac
- Service de Biologie (secteur Hémostase), Centre Hospitalier de Versailles André Mignot, Le Chesnay, France
| | - Céline Delassasseigne
- Service d'Hématologie, Centre Hospitalier Universitaire de Bordeaux, Bordeaux, France
| | | | - Benedicte Delahousse
- Service d'Hématologie - Hémostase, Centre Hospitalier Universitaire Hôpital Trousseau, Tours, France
| | - Elodie Boissier
- Service d'Hématologie Biologique, Centre Hospitalier Universitaire de Nantes, Nantes, France
| | - Céline Desconclois
- Service d'Hématologie Biologique, Centre Hospitalier Universitaire Antoine Béclère, Université Paris-Saclay, Clamart, France
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Gebetsberger J, Prüller F. Classic Light Transmission Platelet Aggregometry: Do We Still Need it? Hamostaseologie 2024; 44:304-315. [PMID: 38065556 DOI: 10.1055/a-2117-4614] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 08/18/2024] Open
Abstract
For more than 50 years, light transmission aggregometry has been accepted as the gold standard test for diagnosing inherited platelet disorders in platelet-rich plasma, although there are other functional approaches performed in whole blood. In this article, several advantages and disadvantages of this technique over other laboratory approaches are discussed in the view of recent guidelines, and the necessity of functional assays, such as light transmission aggregometry in the era of molecular genetic testing, is highlighted.
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Affiliation(s)
| | - Florian Prüller
- Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
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Villar P, Carreño S, Moro S, Díez Galindo I, Bernardo Á, Gutiérrez L. Platelet function testing: Update on determinant variables and permissive windows using a platelet-count-based device. Transfus Apher Sci 2024; 63:103930. [PMID: 38644062 DOI: 10.1016/j.transci.2024.103930] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/23/2024]
Abstract
While there are various aspects of platelet biology that can be studied in the lab (i.e. adhesion, degranulation, integrin activation), the master test for platelet function is that which gives a measure of the platelet aggregation capacity upon stimulation with an agonist. Platelet function testing is necessary for the diagnosis of platelet disorders and the monitoring of patients receiving anti-platelet treatments. Furthermore, it becomes relevant in the quality control of platelet concentrates for transfusion purposes, especially considering the global concern about long term storage, other forms of storage (i.e. cryopreservation, lyophilization), and the impact of Pathogen Reduction Treatments (PRTs) on platelet performance upon transfusion. However, it has been acknowledged as technically difficult and demanding, since a fine platelet function test must be carried out under specific conditions. Still, there might be occasions that preclude the platelet function testing abiding to the gold standard requirements, thus, leaving us with the necessity to redefine which variables may condition or limit the analysis of platelet function testing. In the present manuscript, we test different variables (such as the anticoagulant used or the time elapsed since extraction) and the possibility to reconstitute blood prior to platelet function analysis. This study aims to provide windows of action at the diagnostics lab, especially when not all of the recommended procedures and conditions can be followed: for example, when a sample is sent from a long distance, when there is a limitation on blood extraction volume or when certain parameters (platelet count) preclude reliable test results.
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Affiliation(s)
- Patricia Villar
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain
| | - Sofía Carreño
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain
| | - Sara Moro
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain
| | - Inés Díez Galindo
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain
| | - Ángel Bernardo
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain; Hematology Department and AGC Laboratory of Medicine, Hospital Universitario Central de Asturias (HUCA), Oviedo, Spain
| | - Laura Gutiérrez
- Platelet Research Lab, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain; Department of Medicine, University of Oviedo, Oviedo, Spain.
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10
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El Assil A, Benkirane S, El Kettani Y, Cherif Chefchaouni A, Mamad H, Rahali Y, Masrar A. Turnaround Time of the Hematology Results of Cancer Patients During the COVID-19 Pandemic: An Opportunity to Initiate a Quality Improvement Process. Cureus 2024; 16:e61149. [PMID: 38933641 PMCID: PMC11200148 DOI: 10.7759/cureus.61149] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/26/2024] [Indexed: 06/28/2024] Open
Abstract
INTRODUCTION Turnaround time (TAT) is a crucial clinical parameter that reflects the performance of a laboratory especially in the context of oncology and the COVID-19 pandemic. Based on the Lean Six Sigma methodology, we performed a retrospective analysis of the TAT of the complete blood count (CBC) of cancer patients with the aim of reducing this delay in the future. MATERIALS AND METHODS Over one month of the COVID-19 pandemic, a retrospective evaluative audit was carried out on the TAT of the CBC in an oncology department. The root causes of failures of the overall analysis process were detected. The initiation of an improvement approach was implemented through the creation of an improvement flowchart and a new request form. The hospital information system (HIS) data were exported to Microsoft Excel® (Microsoft Corporation, Redmond, Washington, United States). Using the collected data, the mean, standard deviation, median, and interquartile range were calculated using IBM SPSS Statistics for Windows, Version 23, (Released 2015; IBM Corp., Armonk, New York, United States). All time intervals were expressed in minutes. RESULTS Among 263 intra-laboratory TATs analyzed, the median intra-lab TAT was 56 minutes (interquartile range (IQR): 36-80 minutes). A total of 82% of the analyses were performed in less than 90 minutes with a predominance of the interval 30-59 at 42.9%. The main causes of failures were essentially the lack of time stamping of the samples as well as the lack of real-time communication between the biologists and the clinicians. The proposed improvement model is currently being approved by all practitioners whose main items are as follows: At the clinical department level, distinguish the request forms but also the labels of the samples of the oncology hospital by a particular color, indication of clinical signs and sampling time on the request forms and on the HIS. At the laboratory level, create a specific chain for oncology department samples, alarm notification on the HIS, and rapid telecommunication of results for vital situations. CONCLUSION The intra-lab TAT of our study is biologically acceptable. Because our work is limited by the phases outside the control of the laboratory, it should lead to a continuous improvement project.
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Affiliation(s)
- Asmae El Assil
- Central Laboratory Hematology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Souad Benkirane
- Central Laboratory Hematology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Yasmine El Kettani
- National Institute of Oncology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Ali Cherif Chefchaouni
- National Institute of Oncology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Hassane Mamad
- Central Laboratory Hematology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Younes Rahali
- National Institute of Oncology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
| | - Azlarab Masrar
- Central Laboratory Hematology, Ibn Sina University Hospital Center, Rabat, MAR
- Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, MAR
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11
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Sokou R, Parastatidou S, Konstantinidi A, Tsantes AG, Iacovidou N, Piovani D, Bonovas S, Tsantes AE. Contemporary tools for evaluation of hemostasis in neonates. Where are we and where are we headed? Blood Rev 2024; 64:101157. [PMID: 38016836 DOI: 10.1016/j.blre.2023.101157] [Citation(s) in RCA: 1] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2023] [Revised: 11/20/2023] [Accepted: 11/21/2023] [Indexed: 11/30/2023]
Abstract
The assessment of hemostatic disorders in neonates is crucial, but remains challenging for clinicians. Although the concept of developmental hemostasis is widely accepted among hemostasis specialists globally, it is probably under-recognized by clinicians and laboratory practitioners. In parallel with age-dependent hemostatic status maturation, comprehension of the differences between normal values is crucial for the accurate diagnosis of potential hemorrhagic and thrombotic disorders of the vulnerable neonatal population. This review outlines the basics of developmental hemostasis and the features of the available coagulation testing methods, with a focus on novel tools for evaluating the neonatal hemostatic profile. Common errors, issues, and pitfalls during the assessment of neonatal hemostasis are discussed, along with their impact on patient management. Current knowledge gaps and research areas are addressed. Further studying to improve our understanding of developmental hemostasis and its reflection on everyday clinical practice is warranted.
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Affiliation(s)
- Rozeta Sokou
- Neonatal Intensive Care Unit, "Agios Panteleimon" General Hospital of Nikea, Piraeus, Greece.
| | | | | | - Andreas G Tsantes
- Laboratory of Haematology and Blood Bank Unit, "Attiko" Hospital, School of Medicine, National and Kapodistrian University of Athens, Greece
| | - Nicoletta Iacovidou
- Neonatal Department, National and Kapodistrian University of Athens, Aretaieio Hospital, Athens, Greece
| | - Daniele Piovani
- Department of Biomedical Sciences, Humanitas University, Milan, Italy; IRCCS Humanitas Research Hospital, Milan, Italy
| | - Stefanos Bonovas
- Department of Biomedical Sciences, Humanitas University, Milan, Italy; IRCCS Humanitas Research Hospital, Milan, Italy
| | - Argirios E Tsantes
- Laboratory of Haematology and Blood Bank Unit, "Attiko" Hospital, School of Medicine, National and Kapodistrian University of Athens, Greece
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12
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Chan MV, Chen MH, Thibord F, Nkambule BB, Lachapelle AR, Grech J, Schneider ZE, Wallace de Melendez C, Huffman JE, Hayman MA, Allan HE, Armstrong PC, Warner TD, Johnson AD. Factors that modulate platelet reactivity as measured by 5 assay platforms in 3429 individuals. Res Pract Thromb Haemost 2024; 8:102406. [PMID: 38813256 PMCID: PMC11135030 DOI: 10.1016/j.rpth.2024.102406] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2024] [Accepted: 04/05/2024] [Indexed: 05/31/2024] Open
Abstract
Background Assessment of platelet function is key in diagnosing bleeding disorders and evaluating antiplatelet drug efficacy. However, there is a prevailing "one-size-fits-all" approach in the interpretation of measures of platelet reactivity, with arbitrary cutoffs often derived from healthy volunteer responses. Objectives Our aim was to compare well-used platelet reactivity assays. Methods Blood and platelet-rich plasma obtained from the Framingham Heart Study (N = 3429) were assayed using a range of agonists in 5 platelet assays: light transmission aggregometry, Optimul aggregometry, Multiplate impedance aggregometry (Roche Diagnostics), Total Thrombus-Formation Analysis System, and flow cytometry. Using linear mixed-effect models, we determined the contribution of preanalytical and technical factors that modulated platelet reactivity traits. Results A strong intra-assay correlation of platelet traits was seen in all assays, particularly Multiplate velocity (r = 0.740; ristocetin vs arachidonic acid). In contrast, only moderate interassay correlations were observed (r = 0.375; adenosine diphosphate Optimul Emax vs light transmission aggregometry large area under the curve). As expected, antiplatelet drugs strongly reduced platelet responses, with aspirin use primarily targeting arachidonic acid-induced aggregation, and explained substantial variance (β = -1.735; P = 4.59 × 10-780; variance proportion = 46.2%) and P2Y12 antagonists blocking adenosine diphosphate responses (β = -1.612; P = 6.75 × 10-27; variance proportion = 2.1%). Notably, female sex and older age were associated with enhanced platelet reactivity. Fasting status and deviations from standard venipuncture practices did not alter platelet reactivity significantly. Finally, the agonist batch, phlebotomist, and assay technician (more so for assays that require additional sample manipulation) had a moderate to large effect on measured platelet reactivity. Conclusion Caution must be exercised when extrapolating findings between assays, and the use of standard ranges must be medication-specific and sex-specific at a minimum. Researchers should also consider preanalytical and technical variables when designing experiments and interpreting platelet reactivity measures.
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Affiliation(s)
- Melissa V. Chan
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Ming-Huei Chen
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Florian Thibord
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Bongani B. Nkambule
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Amber R. Lachapelle
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Joseph Grech
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Zoe E. Schneider
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | | | - Jennifer E. Huffman
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
| | - Melissa A. Hayman
- Centre for Immunobiology, the Blizard Institute, Faculty of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom
| | - Harriet E. Allan
- Centre for Immunobiology, the Blizard Institute, Faculty of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom
| | - Paul C. Armstrong
- Centre for Immunobiology, the Blizard Institute, Faculty of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom
| | - Timothy D. Warner
- Centre for Immunobiology, the Blizard Institute, Faculty of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom
| | - Andrew D. Johnson
- Population Sciences Branch, National Heart, Lung, and Blood Institute, Framingham, Massachusetts, USA
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13
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Nissen PH, Mikkelsen TS, Højskov CS, Højbjerg JA. A case of platelet δ-granule defect identified by decreased CD63 expression and decreased serotonin release measured by flow cytometry and liquid chromatography tandem mass spectrometry. Scand J Clin Lab Invest 2024; 84:68-70. [PMID: 38300114 DOI: 10.1080/00365513.2024.2309613] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2023] [Revised: 12/21/2023] [Accepted: 01/21/2024] [Indexed: 02/02/2024]
Affiliation(s)
- Peter H Nissen
- Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus Denmark
- Department of Clinical Medicine, Aarhus University, Aarhus, Denmark
| | | | | | - Johanne Andersen Højbjerg
- Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus Denmark
- Department of Clinical Medicine, Aarhus University, Aarhus, Denmark
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14
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Regan IE, Cox D, Kelleher ST, Nolan B, Shaw K, Smith OP, McMahon CJ. Towards a greater understanding of reduced response to aspirin in children with congenital heart disease post-cardiac surgery using immature platelet fraction. Thromb Res 2024; 233:101-108. [PMID: 38039722 DOI: 10.1016/j.thromres.2023.11.014] [Citation(s) in RCA: 4] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/04/2023] [Revised: 10/10/2023] [Accepted: 11/13/2023] [Indexed: 12/03/2023]
Abstract
OBJECTIVE A high platelet turnover rate may produce a population of platelets that confers an inadequate response to aspirin. We aimed to investigate the relationship between residual platelet aggregation and platelet turnover in paediatric cardiology patients on aspirin monotherapy by evaluating the fraction of immature platelets as a marker for turnover and secondly to test the predictive value of the immature platelet fraction (IPF) to classify patients as responsive or non-responsive to aspirin. METHODS Sixty patients divided into two age categories (≤90 days, >90 days of age) were included in this prospective observational study. Patients were then stratified into tertiles using their IPF level. Platelet studies included thromboelastography with platelet mapping (TEGPM). RESULTS The overall incidence of 'inadequate response to aspirin' was 38 % in our patient cohort recently post-cardiac surgery a consequence that warrants further study. The frequency of inadequate response to aspirin was higher in the upper tertile of IPF when compared to the lower tertile, (88 %) versus (4 %) respectively (p < 0.05). The 'cut off' for IPF was determined to be 3.9 % with a sensitivity of 95.7 %, and a specificity of 92.9 % (area under the curve of 0.955 [CI 0.896-1.014, p < 0.05]). CONCLUSION This study demonstrates that inadequate response to aspirin occurs in approximately 38 % of patients undergoing specific high-risk congenital cardiac procedures using the dosing practice of a national centre. This study supports the hypothesis that an elevated platelet turnover may result in aspirin being less effective in patients who are recently post cardiac surgery. These data are of direct translational relevance.
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Affiliation(s)
- Irene E Regan
- Department of Coagulation/Haematology, Children's Health Ireland at Crumlin, Dublin, Ireland; School of Medicine, University College Dublin, Belfield, Dublin 4, Ireland; National Children's Research Centre, Children's Health Ireland, Dublin, Ireland
| | - Dermot Cox
- School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons Ireland, Dublin, Ireland
| | - Sean T Kelleher
- Department of Paediatric Cardiology, Children's Health Ireland at Crumlin, Dublin, Ireland
| | - Beatrice Nolan
- Department of Coagulation/Haematology, Children's Health Ireland at Crumlin, Dublin, Ireland
| | - Kathryn Shaw
- Department of Paediatric Pharmacy, Children's Health Ireland at Crumlin, Dublin, Ireland
| | - Owen P Smith
- Department of Coagulation/Haematology, Children's Health Ireland at Crumlin, Dublin, Ireland
| | - Colin J McMahon
- Department of Paediatric Cardiology, Children's Health Ireland at Crumlin, Dublin, Ireland; School of Medicine, University College Dublin, Belfield, Dublin 4, Ireland; School of Health Professions Education (SHE), Maastricht University, Maastricht, Netherlands; National Children's Research Centre, Children's Health Ireland, Dublin, Ireland.
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15
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Kornya MR, Abrams-Ogg ACG, Blois SL, Wood RD. Validation of storage and shipping of feline blood samples for analysis on the Platelet Function Analyzer-200 for determining the effect of clopidogrel. Vet Clin Pathol 2023; 52:588-595. [PMID: 37488077 DOI: 10.1111/vcp.13265] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2022] [Revised: 04/02/2023] [Accepted: 04/24/2023] [Indexed: 07/26/2023]
Abstract
BACKGROUND The Platelet function analyzer-200 (PFA-200) can determine the effect of clopidogrel in cats, but analysis traditionally must be performed at point-of-care (POC). The ability to ship samples of blood to a laboratory would allow widespread access. OBJECTIVES We aimed to validate the shipping of blood samples for PFA-200 analysis in cats to determine the effect of clopidogrel. METHODS Twenty healthy cats and 10 cats receiving clopidogrel were recruited. Blood was collected from cats and aliquoted into two samples, one was analyzed at POC within 2 hours using the PFA-200, and the other was packaged and transported to a location 4 km away, stored, and transported back to the lab for analysis the following day. RESULTS Median closure times (CTs) with the collagen/adenosine diphosphate (COL/ADP) cartridge in healthy cats were 51.5 seconds (POC) and 78.8 seconds (shipped), which were significantly different (P < 0.001), and for cats on clopidogrel, median CTs were 147.5 seconds (POC) and 190 seconds (shipped), which were not significantly different (P = 0.131). Median CTs with the P2Y cartridge in healthy cats were 50.5 seconds (POC) and 64.9 seconds (shipped), which were significantly different (P = 0.03), and in cats receiving clopidogrel, median CTs were 300 seconds (POC) and 300 seconds (shipped) which were not significantly different (P = 1.000). Reference intervals for CTs differed for COL/ADP at POC (19.8-89.7 seconds) and shipped (50.9-161.6 seconds) and for P2Y at POC (35.5-118.8 seconds) and shipped (35.1-108.9 seconds). Receiver operating characteristics showed similar areas under the curve (AUCROCs) regarding the effect of clopidogrel for COL/ADP at POC (0.994 seconds) and shipped (0.932) and for P2Y at POC (0.904 seconds) and shipped (0.975 seconds). When classifying for the presence of clopidogrel effects, Cohen's Kappa was 0.62 for COL/ADP and 1.00 for P2Y. CONCLUSIONS Shipping blood samples for PFA analysis are feasible with similar performance to POC analyses for determining the effect of clopidogrel in cats.
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Affiliation(s)
- Matthew R Kornya
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada
| | - Anthony C G Abrams-Ogg
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada
| | - Shauna L Blois
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada
| | - R Darren Wood
- Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada
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16
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Muraoka WT, Nair PM, Darlington DN, Wu X, Bynum JA, Cap AP. A novel, quantitative clot retraction assay to evaluate platelet function. Platelets 2023; 34:2254403. [PMID: 37700390 DOI: 10.1080/09537104.2023.2254403] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/06/2023] [Revised: 08/24/2023] [Accepted: 08/25/2023] [Indexed: 09/14/2023]
Abstract
Blood platelets are crucial to prevent excessive bleeding following injury to blood vessels. Platelets are crucial for the formation of clots and for clot strength. Platelet activation involves aggregation, attachment to fibrin and clot retraction. Most assays that address platelet function measure platelet aggregation, not clot retraction. Here, we describe a 96-well-based clot retraction assay that requires a relatively short runtime and small sample volume. The assay involves continuous optical density monitoring of platelet-rich plasma that is activated with thrombin. The data can be analyzed using time-series analytical tools to generate quantitative information about different phases of clot formation and clot retraction. The assay demonstrated good repeatability and reproducibility and was robust to different calcium concentrations. Impairment of platelet bioenergetics, actin polymerization, fibrin interaction, and signaling significantly affected clot retraction and was detected and showed good agreement with light transmission aggregometry, suggesting that clot retraction is predictive of platelet function. Using this microplate clot retraction assay, we showed a significant difference in platelets stored in autologous plasma compared with platelet additive solution after 7 days of room temperature storage.
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Affiliation(s)
- Wayne T Muraoka
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
| | - Prajeeda M Nair
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
| | - Daniel N Darlington
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
- The Department of Surgery, University of Texas Health, San Antonio, TX, USA
| | - Xiaowu Wu
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
- The Department of Surgery, University of Texas Health, San Antonio, TX, USA
| | - James A Bynum
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
- The Department of Surgery, University of Texas Health, San Antonio, TX, USA
| | - Andrew P Cap
- U.S. Army Institute of Surgical Research, Fort Sam Houston, TX, USA
- The Department of Surgery, University of Texas Health, San Antonio, TX, USA
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17
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Babuty A, Debord C, Drillaud N, Eveillard M, Trossaert M, Ternisien C, Sigaud M, Cador E, Béné MC, Fouassier M. Prothrombin consumption as an indicator of hemorrhagic phenotype in mild platelet function disorders. Eur J Haematol 2023; 111:787-795. [PMID: 37553915 DOI: 10.1111/ejh.14079] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/17/2023] [Revised: 07/28/2023] [Accepted: 07/31/2023] [Indexed: 08/10/2023]
Abstract
BACKGROUND The bleeding risk of patients with mild platelet function disorders is difficult to assess and their phenotype remains ill-explored. AIM This study was designed to establish a comprehensive biological phenotype of patients with mild platelet function disorders. METHODS Twenty patients were included with persistent abnormal light transmission aggregometry (LTA). The ISTH bleeding assessment tool (ISTH-BAT) was assessed to identify laboratory analyses associated with an abnormal hemorrhagic score. RESULTS The majority of patients had defects that might affect Gαi protein signaling pathways or minor abnormalities. No LTA nor flow cytometry parameters were associated with an above-normal hemorrhagic score. However, prothrombin consumption, which corresponds to the ratio of serum residual factor II to plasma residual factor II, was significantly higher (p = .006) in the abnormal ISTH-BAT group (mean = 14%, SD = 6) compared with the normal ISTH-BAT group (mean = 8%, SD 4). Prothrombin consumption was significantly associated with ISTH-BAT score (r = .5287, IC 95% 0.0986-0.7924, p = .0165). CONCLUSION In this group of patients, there was an association between a pathological bleeding score and increased prothrombin consumption. This test could be used as an additional indicator of platelet function abnormality liable to be related to bleeding risk.
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Affiliation(s)
- Antoine Babuty
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
| | - Camille Debord
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
| | - Nicolas Drillaud
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
| | - Marion Eveillard
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
| | - Marc Trossaert
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
| | - Catherine Ternisien
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
| | - Marianne Sigaud
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
| | - Emmanuelle Cador
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
| | - Marie C Béné
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
| | - Marc Fouassier
- Nantes Université, CHU Nantes, Service d'Hématologie Biologique, Nantes, France
- Nantes Université, CHU Nantes, Centre de Ressource et de Compétence-Maladies Hémorragiques Constitutionnelles, Nantes, France
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18
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Jourdi G, Ramström S, Sharma R, Bakchoul T, Lordkipanidzé M. Consensus report on flow cytometry for platelet function testing in thrombocytopenic patients: communication from the SSC of the ISTH. J Thromb Haemost 2023; 21:2941-2952. [PMID: 37481072 DOI: 10.1016/j.jtha.2023.07.006] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2023] [Revised: 07/04/2023] [Accepted: 07/05/2023] [Indexed: 07/24/2023]
Abstract
BACKGROUND Platelet count alone does not reliably predict bleeding risk, suggesting platelet function is important to monitor in patients with thrombocytopenia. There is still an unmet need for improved platelet function diagnostics in patients with low platelet count in many clinical situations. Flow cytometry is a promising tool allowing reliable platelet function study in this setting. OBJECTIVES The goal of this joint project between the International Society on Thrombosis and Haemostasis (ISTH) Scientific Standardization Committee (SSC) Subcommittees on Platelet Physiology and Platelet Immunology is to provide expert consensus guidance on the use of flow cytometry for the evaluation of platelet function, particularly activation, in patients with low platelet counts. METHODS A literature review was performed to identify relevant questions and areas of interest. An electronic expression of interest form was thereafter announced on the ISTH webpage, followed by a survey encompassing 37 issues regarding preanalytical, analytical, postanalytical, and performance aspects. Areas of disagreement or uncertainty were identified and formed the basis for 2 focus group discussions. RESULTS Consensus recommendations relative to patient sample collection, preanalytical variables, sample type, platelet-count cutoff, any potential specific modification of the standard flow cytometry protocol, and results expression and reporting are proposed based on the current practices of experts in the field as well as on literature review. CONCLUSION The proposed consensus recommendations would allow standardization of protocols in upcoming clinical studies. The clinical utility of platelet function testing using flow cytometry to predict bleeding risk still needs rigorous multicenter outcome studies in patients with thrombocytopenia.
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Affiliation(s)
- Georges Jourdi
- Research Center, Montreal Heart Institute, Montreal, Quebec, Canada; Faculty of Pharmacy, Université de Montréal, Montreal, Quebec, Canada; Université Paris Cité, INSERM, Innovative Therapies in Haemostasis, Paris, France; Service d'Hématologie Biologique, AP-HP, Hôpital Lariboisière, Paris, France
| | - Sofia Ramström
- Cardiovascular Research Centre, School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden.
| | - Ruchika Sharma
- Versiti Blood Center of Wisconsin Pediatric Hematology/Oncology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA; Division of Hematology/Oncology/BMT, UT Southwestern Medical Center, Dallas, Texas, USA
| | - Tamam Bakchoul
- Institute for Clinical and Experimental Transfusion Medicine, Medical Faculty of Tuebingen, University Hospital of Tuebingen, Tuebingen, Germany
| | - Marie Lordkipanidzé
- Research Center, Montreal Heart Institute, Montreal, Quebec, Canada; Faculty of Pharmacy, Université de Montréal, Montreal, Quebec, Canada
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19
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Hindle MS, Cheah LT, Yates DM, Naseem KM. Preanalytical conditions for multiparameter platelet flow cytometry. Res Pract Thromb Haemost 2023; 7:102205. [PMID: 37854456 PMCID: PMC10579537 DOI: 10.1016/j.rpth.2023.102205] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2023] [Revised: 08/02/2023] [Accepted: 08/30/2023] [Indexed: 10/20/2023] Open
Abstract
Background Flow cytometry is an important technique for understanding multiple aspects of blood platelet biology. Despite the widespread use of the platform for assessing platelet function, the optimization and careful consideration of preanalytical conditions, sample processing techniques, and data analysis strategies should be regularly assessed. When set up and designed with optimal conditions, it can ensure the acquisition of robust and reproducible flow cytometry data. However, these parameters are rarely described despite their importance. Objectives We aimed to characterize the effects of several preanalytical variables on the analysis of blood platelets by multiparameter fluorescent flow cytometry. Methods We assessed anticoagulant choice, sample material, sample processing, and storage times on 4 distinct and commonly used markers of platelet activation, including fibrinogen binding, expression of CD62P and CD42b, and phosphatidylserine exposure. Results The use of suboptimal conditions led to increases in basal platelet activity and reduced sensitivities to stimulation; however, the use of optimal conditions protected the platelets from artifactual stimulation and preserved basal activity and sensitivity to activation. Conclusion The optimal preanalytical conditions identified here for the measurement of platelet phenotype by flow cytometry suggest a framework for future development of multiparameter platelet assays for high-quality data sets and advanced analysis.
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Affiliation(s)
- Matthew S. Hindle
- Discovery and Translational Science Department, Leeds Institute of Cardiovascular & Metabolic Medicine, University of Leeds, UK
- Centre for Biomedical Science Research, School of Health, Leeds Beckett University, UK
| | - Lih T. Cheah
- Discovery and Translational Science Department, Leeds Institute of Cardiovascular & Metabolic Medicine, University of Leeds, UK
| | - Daisie M. Yates
- Discovery and Translational Science Department, Leeds Institute of Cardiovascular & Metabolic Medicine, University of Leeds, UK
| | - Khalid M. Naseem
- Discovery and Translational Science Department, Leeds Institute of Cardiovascular & Metabolic Medicine, University of Leeds, UK
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20
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Larsen JB, Hvas AM, Hojbjerg JA. Platelet Function Testing: Update and Future Directions. Semin Thromb Hemost 2023; 49:600-608. [PMID: 36384230 DOI: 10.1055/s-0042-1757898] [Citation(s) in RCA: 16] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022]
Abstract
Platelets play a key role in maintaining normal hemostasis and are also recognized as partners in the development of arterial thrombosis. Today, platelet function testing is used for very different clinical purposes; first, for investigation of platelet dysfunction in acute bleeding and diagnosis of platelet disorders in patients with long-lasting bleeding tendency, and second, for testing the efficacy of antiplatelet therapy in patients with increased thromboembolic risk. Moreover, it has been discussed whether platelet function testing can be used for prediction of bleeding risk (e.g., prior to major surgery). Ever since light transmission aggregometry was introduced, laboratories around the world have worked on testing platelet function, and during the last decades a wide range of new methods has emerged. Besides the clinical utility of platelet function testing, the present review summarizes the test principles and advantages and disadvantages of the different methods, depending on the purpose for which it is to be used. A critical step in investigation of platelet function is the preanalytical factors that can substantially affect test results. Therefore, this review also provides an overview of preanalytical variables that range from patient-related factors such as smoking, coffee, and exercise prior to blood sampling to selection of anticoagulant, needle gauge, and time from blood sampling to analyses. Finally, this review outlines further perspectives on platelet function testing for clinical practice and for research purposes.
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Affiliation(s)
- Julie Brogaard Larsen
- Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark
- Department of Clinical Medicine, Aarhus University, Aarhus, Denmark
| | | | - Johanne Andersen Hojbjerg
- Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark
- Department of Clinical Medicine, Aarhus University, Aarhus, Denmark
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21
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Wilson S, Joseph J, Danta M, Rabbolini DJ. Viscoelastometry to Manage Bleeding in Liver Disease. Cureus 2023; 15:e41401. [PMID: 37546051 PMCID: PMC10402654 DOI: 10.7759/cureus.41401] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 07/05/2023] [Indexed: 08/08/2023] Open
Abstract
A state of "re-balanced haemostasis" describes complex coagulation changes that arise in patients with liver disease. Changes include alterations in procoagulant and anticoagulant proteins, platelets and von Willebrand factor, as well as the fibrinolytic system. Various circumstances including infection, trauma, or surgery may disrupt this balance and predispose an individual to bleeding or thrombosis. The prothrombin time, international normalised ratio, and activated partial thromboplastin time are conventional coagulation screening tests that are routinely employed by clinicians to investigate unexplained bleeding, monitor anticoagulation, and inform preoperative assessments of bleeding risk. These standard coagulation tests assess quantitative defects in procoagulant clotting factors and are insensitive to levels of natural anticoagulants, which together with procoagulant factors, are often perturbed in liver disease. Therefore, the prolongation of clotting times measured by these tests often does not reflect the multifaceted alterations of haemostasis in these patients. Viscoelastic testing (VET) provides a more encompassing assessment of clotting function by recording real-time viscoelastic changes in whole blood and includes parameters that provide information on coagulation factor function, platelet contribution to clot formation, as well as fibrinolysis. To date, VET has been employed to predict and inform transfusion support in obstetric, trauma, and cardiac surgical fields, and its use in patients undergoing liver transplantation is well established. The ability of VET to accurately predict bleeding risk and precisely guide transfusion algorithms for patients with liver disease undergoing other invasive procedures or experiencing bleeding complications has been the topic of research over the last decade. This review is a critical summary of this data and provides a detailed snapshot of the position of VET as a clinical tool in patients with liver disease.
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Affiliation(s)
- Samantha Wilson
- Gastroenterology and Hepatology, School of Clinical Medicine, St. Vincent's Healthcare Campus, Faculty of Medicine, University of New South Wales, Sydney, AUS
| | - Joanne Joseph
- Hematology, School of Clinical Medicine, St. Vincent's Healthcare Campus, Faculty of Medicine, University of New South Wales, Sydney, AUS
- Hematology, St. Vincent's Centre for Applied Medical Research, St Vincent's Hospital, Sydney, AUS
| | - Mark Danta
- Gastroenterology and Hepatology, School of Clinical Medicine, St. Vincent's Healthcare Campus, Faculty of Medicine, University of New South Wales, Sydney, AUS
- Gastroenterology and Hepatology, St. Vincent's Hospital, Sydney, AUS
| | - David J Rabbolini
- Faculty of Medicine and Health, University of Sydney, Sydney, AUS
- Haematology, Oxford University Hospitals NHS Foundation Trust, Oxford, GBR
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22
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Aranda E, Iha S, Solari S, Rodríguez D, Romero V, Villarroel L, Pereira J, Panes O, Mezzano D. Serotonin secretion by blood platelets: accuracy of high-performance liquid chromatography-electrochemical technique compared with the isotopic test and use in a clinical laboratory. Res Pract Thromb Haemost 2023; 7:102156. [PMID: 37601022 PMCID: PMC10439442 DOI: 10.1016/j.rpth.2023.102156] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/14/2023] [Revised: 06/07/2023] [Accepted: 07/06/2023] [Indexed: 08/22/2023] Open
Abstract
Background Mild secretion defects are the most frequent and challenging blood platelet disorders to diagnose. Most δ-granule secretion tests lack validation, are not quantitative, or have unreliable response to weak platelet agonists. Objectives To compare platelet serotonin secretion by HPLC-electrochemical detection technique (HPLC-ECD) with the reference isotopic test (3H-5-HT), evaluating its performance in clinical laboratories. Methods The assay validation followed STARD-2015 recommendations. HPLC-ECD measured the nonsecreted serotonin remaining in platelet pellets after aggregation, comparing it with the reference 3H-5-HT assay. We studied subjects with inherited and aspirin-induced blood platelet disorders and assessed the HPLC-ECD operation for routine clinical diagnosis. Results Calibration curves were linear (R2 = 0.997), with SD for residuals of 3.91% and analytical sensitivity of 5ng/mL. Intra- and interassay imprecision bias ranged between -8.5% and 2.1% and -9% and 3.1%, respectively. Serotonin recovery and stability were >95%, and the variability range of measurements was -5.5% to 4.6%. Statistical differences detected between tests were biologically irrelevant, with bias of 1.48% (SD, 8.43) and CI agreement of -18% to 15%. Both assays distinctly detected platelet secretion induced by 10 μM epinephrine and 4 μmM adenosine diphosphate. However, HPLC-ECD is quantitative and more sensitive to low serotonin content in blood platelets. Reference cutoffs for each agonist were determined in 87 subjects. Initially, the HPLC-ECD requires relatively expensive equipment and trained operators but has remarkably cheap running costs and a turn-around time of 24-36 hours. We have used this diagnostic tool routinely for >8 years. Conclusion HPLC-ECD assay for platelet serotonin secretion is highly accurate, has advantages over the reference 3H-5-HT test, and is suitable as a clinical laboratory technique.
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Affiliation(s)
- Eduardo Aranda
- Department of Hematology-Oncology, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Seiki Iha
- Department of Clinical Laboratory, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Sandra Solari
- Department of Clinical Laboratory, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - David Rodríguez
- Department of Clinical Laboratory, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Viviana Romero
- Department of Hematology-Oncology, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Luis Villarroel
- Department of Public Health, School of Medicine, Pontificia Universidad Católica de Chile
| | - Jaime Pereira
- Department of Hematology-Oncology, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Olga Panes
- Department of Hematology-Oncology, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
| | - Diego Mezzano
- Department of Hematology-Oncology, School of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile
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23
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Pruthi RK. Testing strategies used in the diagnosis of rare inherited bleeding disorders. Expert Rev Hematol 2023:1-15. [PMID: 37144355 DOI: 10.1080/17474086.2023.2211257] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/06/2023]
Abstract
INTRODUCTION Rare Bleeding Disorders have a low population prevalence and may not be recognized by most clinicians. In addition, knowledge gaps of the indicated laboratory tests and their availability add to the potential for delayed diagnosis or misdiagnosis. The lack of widely available commercial, regulatory body approved esoteric tests limit them to reference laboratories, thus limiting easy access for patients. AREAS COVERED A literature search of Pubmed, Medline, Embase and review of international society guidelines was performed. Additional references from published articles were reviewed. A patient-centered approach to recognition and evaluation of RBD is discussed. EXPERT OPINION Recognition of RBD relies on obtaining a detailed patient personal and family hemostatic history. Inquiry into a history of involvement of other organ systems is important and if present should lead to suspicion of an inherited platelet disorder or a variant of Ehlers Danlos Syndrome. Multiple factors contribute to the complexity of development of efficient algorithms for diagnostic testing. Limitations in diagnostic sensitivity and specificity of screening tests, diagnostic tests, and esoteric tests further compound the complexity of establishing a diagnosis. Educational efforts focusing on clinician awareness of RBDs and available testing options are vital for optimal management of such patients.
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Affiliation(s)
- Rajiv K Pruthi
- Mayo Comprehensive Hemophilia Center, Division of Hematology, Department of Internal Medicine, Mayo Clinic College of Medicine, Rochester, MN, USA
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24
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Sanfilippo S, Buisson L, Rouabehi H, Dujaric ME, Donnet T, de Raucourt E, Dumont B, Peynaud-Debayle E. The qLabs® FIB system, a novel point-of-care technology for a rapid and accurate quantification of functional fibrinogen concentration from a single drop of citrated whole blood. Thromb Res 2023; 226:159-164. [PMID: 37178638 DOI: 10.1016/j.thromres.2023.03.018] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2022] [Revised: 03/13/2023] [Accepted: 03/28/2023] [Indexed: 04/08/2023]
Abstract
Hypofibrinogenemia is often associated with excessive bleeding and requires immediate treatment. The qLabs FIB® is a handheld and easy-to-use point-of-care (POC) device designed for the rapid measurement of functional fibrinogen concentration from a single drop of citrated whole blood. The aim of this study was to evaluate the analytical performances of the qLabs FIB system. Fibrinogen concentrations from 110 citrated whole blood specimen were measured by both the qLabs FIB and the Clauss laboratory reference methods (STA®-Liquid Fib assay on STA-R® Max from Stago). A three-laboratories comparison study was conducted to assess reproducibility and repeatability of the qLabs FIB using plasma quality control material. In addition, single-site assays were conducted to assess the repeatability from citrated whole blood specimen covering the qLabs FIB reportable range. A very strong correlation between the qLabs FIB and the Clauss laboratory reference method was observed (r = 0.95). Using a clinical cut-off value of 2.0 g/L, the area under the receiver operating characteristic curve (ROC) of citrated whole blood was 0.99 and sensibility and specificity were 100 % and 93.5 %, respectively. Percent CVs for reproducibility and repeatability assessed from quality control material, were both <5 %. Repeatability assessed from citrated whole blood specimen showed a CV of 2.6 to 6.5 %. In conclusion, the qLabs FIB system enables a rapid and reliable measurement of functional fibrinogen levels from citrated whole blood and exhibits a strong prediction power at the 2 g/L clinical cut-off when compared to the Clauss laboratory reference. Further clinical studies should demonstrate its ability to quickly confirm the diagnosis of acquired hypofibrinogenemia and help identify patients who may benefit from targeted hemostatic treatment.
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25
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Sidonio RF, Bryant PC, Di Paola J, Hale S, Heiman M, Horowitz GS, Humphrey C, Jaffray J, Joyner LC, Kasthuri R, Konkle BA, Kouides PA, Montgomery R, Neeves K, Randi AM, Scappe N, Tarango C, Tickle K, Trapane P, Wang M, Waters B, Flood VH. Building the foundation for a community-generated national research blueprint for inherited bleeding disorders: research priorities for mucocutaneous bleeding disorders. Expert Rev Hematol 2023; 16:39-54. [PMID: 36920856 DOI: 10.1080/17474086.2023.2171983] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/16/2023]
Abstract
BACKGROUND Excessive or abnormal mucocutaneous bleeding (MCB) may impact all aspects of the physical and psychosocial wellbeing of those who live with it (PWMCB). The evidence base for the optimal diagnosis and management of disorders such as inherited platelet disorders, hereditary hemorrhagic telangiectasia (HHT), hypermobility spectrum disorders (HSD), Ehlers-Danlos syndromes (EDS), and von Willebrand disease (VWD) remains thin with enormous potential for targeted research. RESEARCH DESIGN AND METHODS National Hemophilia Foundation and American Thrombosis and Hemostasis Network initiated the development of a National Research Blueprint for Inherited Bleeding Disorders with extensive all-stakeholder consultations to identify the priorities of people with inherited bleeding disorders and those who care for them. They recruited multidisciplinary expert working groups (WG) to distill community-identified priorities into concrete research questions and score their feasibility, impact, and risk. RESULTS WG2 detailed 38 high priority research questions concerning the biology of MCB, VWD, inherited qualitative platelet function defects, HDS/EDS, HHT, bleeding disorder of unknown cause, novel therapeutics, and aging. CONCLUSIONS Improving our understanding of the basic biology of MCB, large cohort longitudinal natural history studies, collaboration, and creative approaches to novel therapeutics will be important in maximizing the benefit of future research for the entire MCB community.
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Affiliation(s)
- Robert F Sidonio
- Department of Pediatrics, Aflac Cancer and Blood Disorders, Atlanta, Georgia, USA.,Hemophilia of Georgia Center for Bleeding and Clotting Disorders, Atlanta, Georgia, USA
| | - Paulette C Bryant
- Pediatric Hematology Oncology, St. Jude Affiliate Clinic at Novant Health Hemby Children's Hospital, Charlotte, North Carolina, USA.,National Hemophilia Foundation, New York, New York, USA
| | - Jorge Di Paola
- Department of Pediatrics, Washington University in St. Louis, St. Louis, Missouri, USA.,Hematology/Oncology Department, Washington University in St. Louis, St. Louis, Missouri, USA
| | - Sarah Hale
- Takeda Pharmaceuticals U.S.A, Lexington, Massachusetts, USA
| | - Meadow Heiman
- Indiana Hemophilia and Thrombosis Center, Indianapolis, Indiana, USA
| | | | | | - Julie Jaffray
- Department of Pediatrics, Children's Hospital Los Angeles, Los Angeles, California, USA.,Keck School of Medicine, University of Southern California, Los Angeles, California, USA
| | - Lora C Joyner
- East Carolina University Hemophilia Treatment Center, Greenville, North Carolina, USA
| | - Raj Kasthuri
- Division of Hematology, UNC Blood Research Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
| | - Barbara A Konkle
- Washington Center for Bleeding Disorders, Seattle, Washington, USA
| | | | - Robert Montgomery
- Blood Center of Wisconsin, Versiti, Milwaukee, Wisconsin, USA.,Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
| | - Keith Neeves
- Hemophilia and Thrombosis Center, University of Colorado Denver, Denver, Colorado, USA.,Department of Bioengineering, University of Colorado Denver, Denver, Colorado, USA.,Department of Pediatrics, University of Colorado Denver, Denver, Colorado, USA.,Department of pediatrics, Anschutz Medical Campus, Aurora, Colorado, USA
| | - Anna M Randi
- National Heart and Lung Institute, Imperial College, London, UK
| | - Nikole Scappe
- National Hemophilia Foundation, New York, New York, USA
| | - Cristina Tarango
- Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.,Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA
| | - Kelly Tickle
- Department of Pediatrics, Aflac Cancer and Blood Disorders, Atlanta, Georgia, USA.,Hemophilia of Georgia Center for Bleeding and Clotting Disorders, Atlanta, Georgia, USA.,Children's Healthcare of Atlanta, Atlanta, Georgia, USA
| | - Pamela Trapane
- Division of Pediatric Genetics, Department of Pediatrics, University of Florida College of Medicine-Jacksonville, Jacksonville, Florida, USA
| | - Michael Wang
- Department of pediatrics, Anschutz Medical Campus, Aurora, Colorado, USA
| | | | - Veronica H Flood
- Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
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26
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Chan MV, Lordkipanidzé M, Warner TD. Assessment of Platelet Function by High-Throughput Screening Light Transmission Aggregometry: Optimul Assay. Methods Mol Biol 2023; 2663:627-636. [PMID: 37204741 DOI: 10.1007/978-1-0716-3175-1_41] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/20/2023]
Abstract
Platelet function testing is critical in the diagnosis of bleeding disorders and allows monitoring of antiplatelet therapy. The gold standard assay, light transmission aggregometry (LTA), was developed 60 years ago and remains widely used worldwide. It requires, however, access to expensive equipment and is time-consuming, and the interpretation of results requires evaluation by an experienced investigator. It also suffers from a lack of standardization, resulting in widely variable results between laboratories. 96-well plate-based Optimul aggregometry utilizes the same principles of LTA and aims to standardize agonist concentrations with the development of 96-well plates which are precoated with 7 concentrations of each lyophilized agonist (arachidonic acid, adenosine diphosphate, collagen, epinephrine, TRAP-6 amide, and U46619) and stored at ambient room temperature (20-25 °C) for up to 12 weeks. For platelet function testing, 40 μL of platelet-rich plasma is added to each well, and the plate is placed onto a plate shaker, after which platelet aggregation is determined by changes in light absorbance. This method reduces the blood volume required and allows for in-depth platelet function analysis without specialist training, or the need to purchase expensive, dedicated equipment.
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Affiliation(s)
- Melissa V Chan
- National Heart, Lung and Blood Institute's The Framingham Heart Study, Framingham, MA, USA.
- National Heart, Lung and Blood Institute, Population Sciences Branch, Framingham, MA, USA.
| | - Marie Lordkipanidzé
- Research Center, Montreal Heart Institute, Montreal, QC, Canada
- Faculty of Pharmacy, Université de Montréal, Montreal, QC, Canada
| | - Timothy D Warner
- Centre for Immunobiology, The Blizard Institute, Faculty of Medicine & Dentistry, Queen Mary University of London, London, UK
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27
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Gosselin RC. Preparing Surrogate Abnormal Quality Control Samples for Platelet Function Studies and Viscoelastic Testing. Methods Mol Biol 2023; 2663:637-645. [PMID: 37204742 DOI: 10.1007/978-1-0716-3175-1_42] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/20/2023]
Abstract
In the United States, published options for clinical laboratories to perform quality control (QC) procedures less stringent than the regulatory requirements (Clinical and Laboratory Improvement Act, CLIA) based on risk assessment, although the laboratory must perform to manufacturer's minimum requirements. The US requirements for internal quality control requires at least two levels of control material every 24 h of patient testing. For some coagulation testing, the recommended quality control may be a normal sample or commercial controls that do not address all reporting components of the test. Additional circumstances and difficulties in achieving this minimum QC requirement can be due to either (1) nature of the sample type (i.e., whole blood sample requirements), (2) lack of commercial or suitable control material, or (3) unusual or rare samples. The purpose of this chapter is to provide provisional guidance for laboratory sites to prepared samples to verify the performance of reagents and testing performance of platelet function studies and viscoelastic measurements.
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Affiliation(s)
- Robert C Gosselin
- Thrombosis & Hemostasis Center, University of California, Davis Health System, Sacramento, CA, USA
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28
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Gosselin RC. Preanalytical Variables in Hemostasis Testing. Methods Mol Biol 2023; 2663:39-50. [PMID: 37204702 DOI: 10.1007/978-1-0716-3175-1_2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/20/2023]
Abstract
Hemostasis testing performed in clinical laboratories are critical for assessing hemorrhagic and thrombotic disorders. The assays performed can be used to provide the information required for diagnosis, risk assessment, efficacy of therapy, and therapeutic monitoring. As such, hemostasis tests should be performed to the highest level of quality, including the standardization, implementation, and monitoring of all phases of the testing, which include the preanalytical, analytical, and post-analytical phases. It is well established that the preanalytical phase is the most critical component of the testing process, being the hands-on activities, including patient preparation for blood collection, as well as the actual blood collection, including sample identification and the post-collection handling to include sample transportation, processing, and storage of samples when testing is not performed immediately. The purpose of this article is to provide an update to the previous edition of coagulation testing-related preanalytical variables (PAV) and, when properly addressed and performed, can reduce the most common causes of errors in the hemostasis laboratory.
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Affiliation(s)
- Robert C Gosselin
- Hemostasis & Thrombosis Center, University of California, Davis Health System, Sacramento, CA, USA
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29
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Sarkar MK, Hinz C. Assessment of Platelet Function by Automated Light Transmission Aggregometry. Methods Mol Biol 2023; 2663:611-625. [PMID: 37204740 DOI: 10.1007/978-1-0716-3175-1_40] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/20/2023]
Abstract
Light transmission aggregometry (LTA) has long been the historical "gold standard" of platelet function testing and is typically performed in specialized hemostasis laboratories due to its manual and labor intensive process. However, newer automated testing provides a means of standardization and ability to perform the testing in routine laboratories. Here we describe the measurement of platelet aggregation in the CS-Series™ (Sysmex Corporation, Kobe, Japan) and CN-Series™ (Sysmex Corporation, Kobe, Japan) routine blood coagulation analyzers. Differences in the methods for both analyzers are further described. For the CS-5100™ analyzer, the final diluted concentrations of the agonists are prepared by manual pipetting from reconstituted agonist solutions. These prepared dilutions are eight times concentrated with respect to the final working concentration of the agonists and appropriately diluted within the analyzer to achieve the desired concentration of agonists prior to testing. For the CN-6000™ analyzer, the dilutions of agonists and the final working concentrations are automatically prepared by the auto-dilution feature in the analyzer.
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30
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Diagnosing Czech Patients with Inherited Platelet Disorders. Int J Mol Sci 2022; 23:ijms232214386. [PMID: 36430862 PMCID: PMC9695320 DOI: 10.3390/ijms232214386] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2022] [Revised: 11/13/2022] [Accepted: 11/15/2022] [Indexed: 11/22/2022] Open
Abstract
A single-center study was conducted on 120 patients with inherited disorders of primary hemostasis followed at our hematological center. These patients presented a variety of bleeding symptoms; however, they had no definitive diagnosis. Establishing a diagnosis has consequences for the investigation of probands in families and for treatment management; therefore, we aimed to improve the diagnosis rate in these patients by implementing advanced diagnostic methods. According to the accepted international guidelines at the time of study, we investigated platelet morphology, platelet function assay, light-transmission aggregometry, and flow cytometry. Using only these methods, we were unable to make a definitive diagnosis for most of our patients. However, next-generation sequencing (NGS), which was applied in 31 patients, allowed us to establish definitive diagnoses in six cases (variants in ANKRD26, ITGA2B, and F8) and helped us to identify suspected variants (NBEAL2, F2, BLOC1S6, AP3D1, GP1BB, ANO6, CD36, and ITGB3) and new suspected variants (GFI1B, FGA, GP1BA, and ITGA2B) in 11 patients. The role of NGS in patients with suspicious bleeding symptoms is growing and it changes the diagnostic algorithm. The greatest disadvantage of NGS, aside from the cost, is the occurrence of gene variants of uncertain significance.
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31
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Rinaldi I, Muthalib A, Wijayadi T, Sutedja B, Susanto N, Magdalena L, Tandaju JR, Wardhana IL, Winston K. Surgical Complications in Myeloproliferative Neoplasm Patient with Essential Thrombocythemia: A Case Report. Int Med Case Rep J 2022; 15:491-497. [PMID: 36120702 PMCID: PMC9480603 DOI: 10.2147/imcrj.s375777] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/25/2022] [Accepted: 08/10/2022] [Indexed: 11/23/2022] Open
Abstract
Introduction Essential thrombocythemia (ET) is a myeloproliferative neoplasm (MPN) which could complicate surgical procedures due to thrombosis and spontaneous bleeding. However, currently, there is neither concrete guideline nor prerequisite for ET patients who underwent operations. Case Report A 48 year-old-female was admitted to the emergency unit on 21 February 2020 due to vomiting and inability to pass gas/stool. The patient previously had an operation for uterine myoma two weeks before which showed thrombocyte count of 688,000/mm3. The patient was previously diagnosed with essential thrombocythemia with positive JAK2V617 point mutation on 24 June 2019. Laboratory examination showed thrombocyte 1,134,000/mm3 and leukocyte 22,700/mm3 suggestive of neutrophilia. CT scan showed fluid collection with blood density in the abdomen and pelvis. She was then diagnosed with obstructive ileus due to abdominal abscess and intestine adhesion. Adhesiolysis by laparoscopy was performed on 29 February 2020 with thrombocyte count of 727,000/mm3. Patient was able to pass flatus and defecate three days post-surgery. However, a decrease of hemoglobin to 8.2 g/dL on 3 March 2020 with thrombocyte count of 700,000/mm3 was suggestive of internal bleeding. She was discharged three weeks post-surgery after improvement of clinical condition with thrombocyte count of 850,000/mm3. She was given hydroxyurea 1000 mg once every two days, aspirin 80 mg OD, anagrelide 1 mg OD, and amlodipine 10 mg OD. Conclusion Myeloproliferative disease patients with high thrombocyte count are subjected to increased risk of thrombotic complications in perioperative settings, thus perioperative management and risk assessment are important to improve quality of life and prevent complications. Surgery in MPN patients with elevated thrombocytes may be considered if the benefits outweigh the risks. More studies in this field should be conducted in-order to provide more data for a guideline or systematic review/meta-analyses.
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Affiliation(s)
- Ikhwan Rinaldi
- Division of Hematology and Medical Oncology, Department of Internal Medicine, Cipto Mangunkusumo National General Hospital, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
- Department of Internal Medicine, Gading Pluit Hospital, Jakarta, Indonesia
- Correspondence: Ikhwan Rinaldi, Email
| | - Abdul Muthalib
- Division of Hematology and Medical Oncology, Department of Internal Medicine, Cipto Mangunkusumo National General Hospital, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
- Department of Internal Medicine, Gading Pluit Hospital, Jakarta, Indonesia
| | - Teguh Wijayadi
- Department of Internal Medicine, Gading Pluit Hospital, Jakarta, Indonesia
| | - Barlian Sutedja
- Department of Surgery, Gading Pluit Hospital, Jakarta, Indonesia
| | - Nelly Susanto
- Department of Radiology, Gading Pluit Hospital, Jakarta, Indonesia
| | - Lingga Magdalena
- Department of Radiology, Gading Pluit Hospital, Jakarta, Indonesia
| | | | | | - Kevin Winston
- Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
- Hospital Medicine, Bhakti Medicare Hospital, Sukabumi, Indonesia
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32
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Brusilovskaya K, Simbrunner B, Lee S, Eichelberger B, Bauer D, Zinober K, Schwabl P, Mandorfer M, Panzer S, Reiberger T, Gremmel T. Peripheral versus central venous blood sampling does not influence the assessment of platelet activation in cirrhosis. Platelets 2022; 33:879-886. [PMID: 35294323 DOI: 10.1080/09537104.2021.2007868] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Cirrhotic patients have an increased risk of bleeding and thromboembolic events, with platelets being involved as key players in both situations. The impact of peripheral versus central blood sampling on platelet activation remains unclear. In 33 cirrhotic patients, we thus analyzed platelet function in peripheral (P) and central (C) blood samples. Platelet surface expression of P-selectin, activated glycoprotein (GP) IIb/IIIa, and leukocyte-platelet aggregate formation were measured by flow cytometry in response to different agonists: thrombin receptor-activating peptide-6, adenosine diphosphate, collagen-related peptide (CrP), epinephrine, AYPGKF, Pam3CSK4, and lipopolysaccharide. Unstimulated platelet surface expression of P-selectin (p = .850) and activated GPIIb/IIIa (p = .625) were similar in peripheral and central blood samples. Stimulation with various agonists yielded similar results of platelet surface expression of P-selectin and activated GPIIb/IIIa in peripheral and central samples, except for CrP-inducible expression of activated GPIIb/IIIa (median fluorescence intensity, MFI in P: 7.61 [0.00-24.66] vs. C: 4.12 [0.00-19.04], p < .001). The formation of leukocyte-platelet aggregate was similar in central and peripheral blood samples, both unstimulated and after stimulation with all above-mentioned agonists. In conclusion, peripheral vs. central venous blood sampling does not influence the assessment of platelet activation by flow cytometry in cirrhosis.Abbreviations: ACLD: advanced chronic liver disease; ADP: adenosine diphosphate; ALD: alcoholic liver disease; AYPGKF: PAR-4 agonist AYPGKF; CrP: collagen related protein; EPI: epinephrine; FACS: fluorescence-activated cell sorting; GP: glycoprotein; HVPG: hepatic venous pressure gradient; IQR: interquartile range; LPS: lipopolysaccharide; LSM: liver stiffness measurement; MFI: median fluorescence intensity; NAFLD: nonalcoholic fatty liver disease; PAM: lipopeptide Pam3CSK4; PAR: protease-activated receptor; PBS: phosphate-buffered saline; PH: portal hypertension; TIPS: transjugular intrahepatic portosystemic stent shunt; TLR: toll-like receptor; TRAP-6: thrombin receptor-activator peptide-6; vWF: von Willebrand factor.
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Affiliation(s)
- Ksenia Brusilovskaya
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Christian-Doppler Laboratory for Portal Hypertension and Liver Fibrosis, Medical University of Vienna, Vienna, Austria
| | - Benedikt Simbrunner
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Christian-Doppler Laboratory for Portal Hypertension and Liver Fibrosis, Medical University of Vienna, Vienna, Austria
| | - Silvia Lee
- Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria
| | - Beate Eichelberger
- Department of Blood Group Serology and Transfusion Medicine, Medical University of Vienna, Vienna, Austria
| | - David Bauer
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria
| | - Kerstin Zinober
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Christian-Doppler Laboratory for Portal Hypertension and Liver Fibrosis, Medical University of Vienna, Vienna, Austria
| | - Philipp Schwabl
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Christian-Doppler Laboratory for Portal Hypertension and Liver Fibrosis, Medical University of Vienna, Vienna, Austria
| | - Mattias Mandorfer
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria
| | - Simon Panzer
- Department of Blood Group Serology and Transfusion Medicine, Medical University of Vienna, Vienna, Austria
| | - Thomas Reiberger
- Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Vienna Hepatic Hemodynamic Lab (HEPEX), Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria.,Christian-Doppler Laboratory for Portal Hypertension and Liver Fibrosis, Medical University of Vienna, Vienna, Austria
| | - Thomas Gremmel
- Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria.,Department of Internal Medicine I, Cardiology and Intensive Care Medicine, Landesklinikum Mistelbach-Gänserndorf, Mistelbach, Austria.,Institute of Antithrombotic Therapy in Cardiovascular Disease, Karl Landsteiner Society, St. Pölten, Austria
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Hsu H, Chan MV, Armstrong PC, Crescente M, Donikian D, Kondo M, Brighton T, Chen V, Chen Q, Connor D, Joseph J, Morel-Kopp MC, Stevenson WS, Ward C, Warner TD, Rabbolini DJ. A pilot study assessing the implementation of 96-well plate-based aggregometry (Optimul) in Australia. Pathology 2022; 54:746-754. [PMID: 35750510 DOI: 10.1016/j.pathol.2022.03.012] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2021] [Revised: 03/09/2022] [Accepted: 03/21/2022] [Indexed: 11/19/2022]
Abstract
Identification of disordered platelet function is important to guide peri-operative bleeding management as well as long term treatment and prognostic strategies in individuals with platelet bleeding disorders. Light transmission aggregometry (LTA), the current gold standard diagnostic test of platelet function is a time consuming technique almost exclusively performed in specialised laboratories and almost universally unavailable in regional centres in Australia, where there is an unmet need for access to specialised platelet function diagnostic services. 96-well plate-based aggregometry (Optimul, UK), has been utilised in research laboratories as a novel platform to investigate platelet function. We evaluated the Optimul assay at two centres in Australia, one regional and one tertiary metropolitan, to assess its feasibility as a screening test applicable to remote regional centres. Concentration-response curves were established from 45 healthy volunteers at the participating regional hospital and from 31 healthy volunteers at the tertiary institution. Optimul successfully detected anti-platelet effects in individuals taking aspirin (n=4), NSAID (n=2), clopidogrel (n=2) and dual therapy with aspirin and clopidogrel (n=1). When tested in parallel to LTA in individuals referred for the evaluation of abnormal bleeding symptoms there was overall a very good level of agreement between Optimul and LTA [Cohen's kappa (k2)=0.84], supporting its role as a useful screening tool in the assessment of platelet function. Optimul assay performance was quick and the methodology simple, requiring no specialised training or resources to be implemented at either the regional or metropolitan laboratory. Widespread implementation, particularly in regional laboratories within Australia where specialised platelet function testing is unavailable, has the potential to drastically improve the inequity of access to such services.
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Affiliation(s)
- Hannah Hsu
- Prince of Wales Hospital, Sydney, NSW, Australia
| | - Melissa V Chan
- Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK; National Heart, Blood and Lung Institute, Population Sciences Branch, Framingham, MA, USA
| | - Paul C Armstrong
- Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
| | - Marilena Crescente
- Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
| | - Dea Donikian
- Haematology NSW Health Pathology Randwick, Sydney, NSW, Australia
| | - Mayuko Kondo
- Haematology NSW Health Pathology Randwick, Sydney, NSW, Australia
| | - Timothy Brighton
- Haematology NSW Health Pathology Randwick, Sydney, NSW, Australia; Prince of Wales Hospital, Sydney, NSW, Australia
| | - Vivien Chen
- Haematology, Concord Repatriation General Hospital and NSW Health Pathology, Sydney, NSW, Australia; ANZAC Research Institute and University of Sydney, Sydney, NSW, Australia
| | - Qiang Chen
- Northern Blood Research Centre, Kolling Institute, University of Sydney, Sydney, NSW, Australia; Department of Haematology and Transfusion Medicine, Royal North Shore Hospital, Sydney, NSW, Australia
| | - David Connor
- St Vincent's Centre for Applied Medical Research, Sydney, NSW, Australia; St Vincent's Clinical School, University of New South Wales, Sydney, Australia
| | - Joanne Joseph
- St Vincent's Centre for Applied Medical Research, Sydney, NSW, Australia; St Vincent's Clinical School, University of New South Wales, Sydney, Australia; St Vincent's Hospital, Sydney, NSW, Australia
| | - Marie-Christine Morel-Kopp
- Northern Blood Research Centre, Kolling Institute, University of Sydney, Sydney, NSW, Australia; Department of Haematology and Transfusion Medicine, Royal North Shore Hospital, Sydney, NSW, Australia
| | - William S Stevenson
- Northern Blood Research Centre, Kolling Institute, University of Sydney, Sydney, NSW, Australia; Department of Haematology and Transfusion Medicine, Royal North Shore Hospital, Sydney, NSW, Australia
| | - Christopher Ward
- Northern Blood Research Centre, Kolling Institute, University of Sydney, Sydney, NSW, Australia; Department of Haematology and Transfusion Medicine, Royal North Shore Hospital, Sydney, NSW, Australia
| | - Timothy D Warner
- Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
| | - David J Rabbolini
- Northern Blood Research Centre, Kolling Institute, University of Sydney, Sydney, NSW, Australia; Northern Clinical School and the Rural Clinical School (Northern Rivers), Faculty of Medicine and Health, University of Sydney, Sydney, NSW, Australia; Lismore Base Hospital, Lismore, NSW, Australia.
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Aarnink EW, Huijboom MF, Bor WL, Maarse M, Zheng KL, ten Cate H, Ten Berg JM, Boersma LV. Hemostatic biomarkers and antithrombotic strategy in percutaneous left atrial interventions: State-of-the-art review. Thromb Res 2022; 215:41-51. [DOI: 10.1016/j.thromres.2022.05.009] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/21/2022] [Revised: 04/29/2022] [Accepted: 05/16/2022] [Indexed: 11/24/2022]
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Kaiser T, Liebscher K, Scholz U, Pfrepper C, Netto J, Drogies T, Tiebel O, Knöfler R, Krause M. Influencing factors and differences in Born aggregometry in specialized hemostaseological centers – results of a multi-center laboratory comparison. TH OPEN 2022; 6:e213-e220. [PMID: 36046201 PMCID: PMC9395241 DOI: 10.1055/a-1827-7025] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/24/2021] [Accepted: 02/14/2022] [Indexed: 12/02/2022] Open
Abstract
Introduction
Light transmission aggregometry (LTA) is regarded as the gold standard in platelet function diagnostics. However, there is a relevant degree of interlaboratory variability in practical applications.
Objective
The aim of the present study was to develop a practicable laboratory comparison on LTA and to analyze differences and influencing factors in regard to standardization in five specialized hemostaseological centers.
Methods
The study was performed on 30 patients in total. Each center performed LTA on blood samples from six healthy volunteers (three men and three women) using the inductors collagen (Col), adenosine diphosphate (ADP), arachidonic acid (ARA), and ristocetin. The LTA was performed three times using different methods as follows: (1) International Society on Thrombosis and Haemostasis recommendations with identical reagents, (2) in-house protocols and the identical reagents; and (3) in-house protocols and in-house reagents.
Results
A total of 396 measurements of 30 probands were performed. Even after standardization of the protocol and using identical reagents, there were significant differences between the centers regarding the final and maximum aggregation (
p
= 0.002 and <0.001) and further significant differences in the maximum and final aggregation according to the wavelength of the device used to measure the LTA (PAP-8: 430 nm, APACT 4004: 740 nm [
p
< 0.001 each]). Using identical reagents but individual inductor concentrations and laboratory protocols also resulted in different maximum and final aggregation. The largest differences were seen with Col and ristocetin; there were significant influences from the reagents' manufacturers in the results of aggregometry for the inductor Col (
p
< 0.01) but not for ADP, ARA, and ristocetin.
Conclusion
In this study, we proved that there are significant influences from the used aggregometers, inductors concentrations, and manufacturers. These results illustrate the challenges and importance of standardization of LTA.
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Affiliation(s)
- Thorsten Kaiser
- Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University of Leipzig, Leipzig, Germany, Leipzig, Germany
| | - Karin Liebscher
- Institute of Transfusion Medicine and Clinical Hemostaseology, Klinikum St. Georg gGmbH, Leipzig, Germany
| | | | | | - Jeffrey Netto
- Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University of Leipzig, Leipzig, Germany, Leipzig, Germany
| | - Tim Drogies
- Medical Central Laboratory Altenburg, Altenburg, Germany
| | - Oliver Tiebel
- Institute of Clinical Chemistry and Laboratory Medicine, Universitätsklinikum Carl Gustav Carus, Dresden, Germany
| | - Ralf Knöfler
- 10. Department of Pediatric Hemostaseology, Univ. Children Hospital, Dresden, Germany
| | - Michael Krause
- MVZ Laboratory Reising-Ackermann MD and Colleagues, Leipzig, Germany
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Heubner L, Mirus M, Vicent O, Güldner A, Tiebel O, Beyer-Westendorf J, Fries D, Spieth PM. Point of care coagulation management in anesthesiology and critical care. Minerva Anestesiol 2022; 88:615-628. [PMID: 35416466 DOI: 10.23736/s0375-9393.22.16380-7] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Point of care (POC) devices are increasingly used in the ICU and in anesthesia. Besides POC-devices for blood gas analysis, several devices are available for coagulation measurements. Although basic principles for thromboelastographic measurements are not novel, some promising developments were made during the last decade improving both user-friendliness and measurement reliability. For instance, POC measurements of activated clotting time (ACT) for heparin monitoring is still regarded as standard-of-care in cardiac interventions and surgery. In the field of anesthesia and intensive care medicine, POC-devices for thromboelastographic and platelet aggregation measurements are widely used. Their impact in case of bleeding and patient blood management for cardiothoracic and trauma surgery is well known. Moreover, there are promising concepts for anticoagulation monitoring including new oral anticoagulant drugs. Coagulation POC-devices may also identify patients at specific risk for thromboembolic events quickly. On the other hand, benefits of POC-devices need to be balanced against limitations, which include technical restrictions and operator related errors, mainly affecting reproducibility and interpretation of results. Therefore, it is recommendable to consider results of POC-coagulation testing in comparison to standard laboratory tests (SLT). Nevertheless, in urgent or emergency situations POC results enable fast decision making to optimize patient care.
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Affiliation(s)
- Lars Heubner
- Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Dresden, Germany. -
| | - Martin Mirus
- Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
| | - Oliver Vicent
- Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
| | - Andreas Güldner
- Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
| | - Oliver Tiebel
- Institute of Clinical Chemistry, University Hospital Carl Gustav Carus, Dresden, Germany
| | - Jan Beyer-Westendorf
- Thrombosis Research Unit, Division of Hematology and Hemostasis, Department of Medicine I, University Hospital Carl Gustav Carus, Dresden, Germany
| | - Dietmar Fries
- Department for General and Surgical Critical Care Medicine, Innsbruck Medical University, Innsbruck, Austria
| | - Peter M Spieth
- Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
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Reichert L, Wallner S, Burkhardt R, Offner R, Ahrens N, Hähnel V. Triple apheresis platelet concentrate quality after pneumatic tube system, conveyor box, and courier transport: An observational study. Health Sci Rep 2022; 5:e596. [PMID: 35425867 PMCID: PMC8989271 DOI: 10.1002/hsr2.596] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2021] [Revised: 03/22/2022] [Accepted: 03/24/2022] [Indexed: 11/10/2022] Open
Affiliation(s)
- Lena Reichert
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
| | - Stefan Wallner
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
| | - Ralph Burkhardt
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
| | - Robert Offner
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
| | - Norbert Ahrens
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
- MVZ for Laboratory Medicine Raubling, amedes Labor Raubling Germany
| | - Viola Hähnel
- Institute for Clinical Chemistry and Laboratory Medicine University Hospital Regensburg Regensburg Germany
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Bourguignon A, Tasneem S, Hayward CP. Screening and diagnosis of inherited platelet disorders. Crit Rev Clin Lab Sci 2022; 59:405-444. [PMID: 35341454 DOI: 10.1080/10408363.2022.2049199] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
Inherited platelet disorders are important conditions that often manifest with bleeding. These disorders have heterogeneous underlying pathologies. Some are syndromic disorders with non-blood phenotypic features, and others are associated with an increased predisposition to developing myelodysplasia and leukemia. Platelet disorders can present with thrombocytopenia, defects in platelet function, or both. As the underlying pathogenesis of inherited thrombocytopenias and platelet function disorders are quite diverse, their evaluation requires a thorough clinical assessment and specialized diagnostic tests, that often challenge diagnostic laboratories. At present, many of the commonly encountered, non-syndromic platelet disorders do not have a defined molecular cause. Nonetheless, significant progress has been made over the past few decades to improve the diagnostic evaluation of inherited platelet disorders, from the assessment of the bleeding history to improved standardization of light transmission aggregometry, which remains a "gold standard" test of platelet function. Some platelet disorder test findings are highly predictive of a bleeding disorder and some show association to symptoms of prolonged bleeding, surgical bleeding, and wound healing problems. Multiple assays can be required to diagnose common and rare platelet disorders, each requiring control of preanalytical, analytical, and post-analytical variables. The laboratory investigations of platelet disorders include evaluations of platelet counts, size, and morphology by light microscopy; assessments for aggregation defects; tests for dense granule deficiency; analyses of granule constituents and their release; platelet protein analysis by immunofluorescent staining or flow cytometry; tests of platelet procoagulant function; evaluations of platelet ultrastructure; high-throughput sequencing and other molecular diagnostic tests. The focus of this article is to review current methods for the diagnostic assessment of platelet function, with a focus on contemporary, best diagnostic laboratory practices, and relationships between clinical and laboratory findings.
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Affiliation(s)
- Alex Bourguignon
- Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Canada
| | - Subia Tasneem
- Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Canada
| | - Catherine P Hayward
- Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Canada.,Department of Medicine, McMaster University, Hamilton, Canada
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Dave RG, Geevar T, Chellaiya GK, Mammen JJ, Vijayan R, Samuel A, Gowri M, Nair SC. Stability and utility of flow cytometric platelet activation tests: A modality to bridge the gap between diagnostic demand and supply. Platelets 2022; 33:1043-1051. [DOI: 10.1080/09537104.2022.2042232] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Affiliation(s)
- Rutvi Gautam Dave
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
| | - Tulasi Geevar
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
| | | | - Joy John Mammen
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
| | - Ramya Vijayan
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
| | - Ashok Samuel
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
| | - Mahasampath Gowri
- Department of Biostatistics, Christian Medical College Vellore, Vellore, India
| | - Sukesh Chandran Nair
- Department of Transfusion Medicine and Immunohematology, Christian Medical College Vellore, Vellore, India
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40
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Zhang Y, Jiang F, Chen Y, Ju LA. Platelet Mechanobiology Inspired Microdevices: From Hematological Function Tests to Disease and Drug Screening. Front Pharmacol 2022; 12:779753. [PMID: 35126120 PMCID: PMC8811026 DOI: 10.3389/fphar.2021.779753] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2021] [Accepted: 12/28/2021] [Indexed: 12/30/2022] Open
Abstract
Platelet function tests are essential to profile platelet dysfunction and dysregulation in hemostasis and thrombosis. Clinically they provide critical guidance to the patient management and therapeutic evaluation. Recently, the biomechanical effects induced by hemodynamic and contractile forces on platelet functions attracted increasing attention. Unfortunately, the existing platelet function tests on the market do not sufficiently incorporate the topical platelet mechanobiology at play. Besides, they are often expensive and bulky systems that require large sample volumes and long processing time. To this end, numerous novel microfluidic technologies emerge to mimic vascular anatomies, incorporate hemodynamic parameters and recapitulate platelet mechanobiology. These miniaturized and cost-efficient microfluidic devices shed light on high-throughput, rapid and scalable platelet function testing, hematological disorder profiling and antiplatelet drug screening. Moreover, the existing antiplatelet drugs often have suboptimal efficacy while incurring several adverse bleeding side effects on certain individuals. Encouraged by a few microfluidic systems that are successfully commercialized and applied to clinical practices, the microfluidics that incorporate platelet mechanobiology hold great potential as handy, efficient, and inexpensive point-of-care tools for patient monitoring and therapeutic evaluation. Hereby, we first summarize the conventional and commercially available platelet function tests. Then we highlight the recent advances of platelet mechanobiology inspired microfluidic technologies. Last but not least, we discuss their future potential of microfluidics as point-of-care tools for platelet function test and antiplatelet drug screening.
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Affiliation(s)
- Yingqi Zhang
- School of Biomedical Engineering, Faculty of Engineering, The University of Sydney, Sydney, NSW, Australia
- Charles Perkins Centre, The University of Sydney, Camperdown, NSW, Australia
- Heart Research Institute, Newtown, NSW, Australia
| | - Fengtao Jiang
- School of Biomedical Engineering, Faculty of Engineering, The University of Sydney, Sydney, NSW, Australia
| | - Yunfeng Chen
- The Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, TX, United States
- The Department of Pathology, The University of Texas Medical Branch, Galveston, TX, United States
| | - Lining Arnold Ju
- School of Biomedical Engineering, Faculty of Engineering, The University of Sydney, Sydney, NSW, Australia
- Charles Perkins Centre, The University of Sydney, Camperdown, NSW, Australia
- Heart Research Institute, Newtown, NSW, Australia
- *Correspondence: Lining Arnold Ju,
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41
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Stasko J, Holly P, Kubisz P. A new decade awaits sticky platelet syndrome: where are we now, how do we manage and what are the complications? Expert Rev Hematol 2022; 15:53-63. [PMID: 35034520 DOI: 10.1080/17474086.2022.2030217] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
INTRODUCTION Sticky platelet syndrome is a less known platelet function disorder with a familiar occurrence and likely genetic background. Clinically, it is characterized by an increased risk of venous and arterial thromboembolic events and obstetric placenta-mediated complications. The increased aggregation after low-dose ADP and/or epinephrine is its distinctive laboratory feature. Though described for almost 40 years, several issues regarding its etiology, involved pathomechanisms, genetic background, optimal diagnostic and treatment approach remain controversial. AREAS COVERED The work aims to summarize published studies, the actual definition of the syndrome, and point out its drawbacks. A literature search on Medline, Embase, and archives from EHA congresses was performed (terms: 'sticky platelet syndrome' - 'platelet hyperreactivity' - 'platelet hyperaggregability'). The authors added in their unpublished data. The introductory overview of the present understanding is followed by the discussion of the pathophysiologic, diagnostic, and therapeutic problems. EXPERT OPINION Despite the growing evidence provided by case reports and series, the lack of robust studies limits the decision-making on diagnostics and management. The diagnostic issues, particularly the standardization of light transmission aggregometry, represent the crucial problem for the broader acceptance of the syndrome.
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Affiliation(s)
- Jan Stasko
- Department of Hematology and Transfusion Medicine, National Center of Hemostasis and Thrombosis, Jessenius Faculty of Medicine in Martin of the Comenius University in Bratislava, University Hospital in Martin, Martin, Slovakia
| | - Pavol Holly
- Department of Hematology and Transfusion Medicine, National Center of Hemostasis and Thrombosis, University Hospital in Martin, Martin, Slovakia
| | - Peter Kubisz
- Department of Hematology and Transfusion Medicine, National Center of Hemostasis and Thrombosis, Jessenius Faculty of Medicine in Martin of the Comenius University in Bratislava, University Hospital in Martin, Martin, Slovakia
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42
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Xu Y, Yu G, Nie R, Wu Z. Microfluidic systems toward blood hemostasis monitoring and thrombosis diagnosis: From design principles to micro/nano fabrication technologies. VIEW 2022. [DOI: 10.1002/viw.20200183] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022] Open
Affiliation(s)
- Yi Xu
- Soft Intelligence Lab State Key Laboratory of Digital Manufacturing Equipment and Technology School of Mechanical Science and Engineering Huazhong University of Science and Technology Wuhan China
| | - Guang Yu
- Experimental Medicine Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China
| | - Ruqiong Nie
- Department of Cardiology Sun Yat‐Sen Memorial Hospital Sun Yat‐Sen University Guangzhou China
| | - Zhigang Wu
- Soft Intelligence Lab State Key Laboratory of Digital Manufacturing Equipment and Technology School of Mechanical Science and Engineering Huazhong University of Science and Technology Wuhan China
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Naylor-Adamson L, Chacko AR, Booth Z, Caserta S, Jarvis J, Khan S, Hart SP, Rivero F, Allsup DJ, Arman M. Bruton's Tyrosine Kinase Inhibitors Impair FcγRIIA-Driven Platelet Responses to Bacteria in Chronic Lymphocytic Leukemia. Front Immunol 2021; 12:766272. [PMID: 34912339 PMCID: PMC8667317 DOI: 10.3389/fimmu.2021.766272] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/28/2021] [Accepted: 10/25/2021] [Indexed: 01/01/2023] Open
Abstract
Bacterial infections are a major cause of morbidity and mortality in chronic lymphocytic leukemia (CLL), and infection risk increases in patients treated with the Bruton’s tyrosine kinase (Btk) inhibitor, ibrutinib. Btk and related kinases (like Tec) are expressed in non-leukemic hematopoietic cells and can be targeted by ibrutinib. In platelets, ibrutinib therapy is associated with bleeding complications mostly due to off-target effects. But the ability of platelets to respond to bacteria in CLL, and the potential impact of ibrutinib on platelet innate immune functions remain unknown. FcγRIIA is a tyrosine kinase-dependent receptor critical for platelet activation in response to IgG-coated pathogens. Crosslinking of this receptor with monoclonal antibodies causes downstream activation of Btk and Tec in platelets, however, this has not been investigated in response to bacteria. We asked whether ibrutinib impacts on FcγRIIA-mediated activation of platelets derived from CLL patients and healthy donors after exposure to Staphylococcus aureus Newman and Escherichia coli RS218. Platelet aggregation, α-granule secretion and integrin αIIbβ3-dependent scavenging of bacteria were detected in CLL platelets but impaired in platelets from ibrutinib-treated patients and in healthy donor-derived platelets exposed to ibrutinib in vitro. While levels of surface FcγRIIA remained unaffected, CLL platelets had reduced expression of integrin αIIbβ3 and GPVI compared to controls regardless of therapy. In respect of intracellular signaling, bacteria induced Btk and Tec phosphorylation in both CLL and control platelets that was inhibited by ibrutinib. To address if Btk is essential for platelet activation in response to bacteria, platelets derived from X-linked agammaglobulinemia patients (lacking functional Btk) were exposed to S. aureus Newman and E. coli RS218, and FcγRIIA-dependent aggregation was observed. Our data suggest that ibrutinib impairment of FcγRIIA-mediated platelet activation by bacteria results from a combination of Btk and Tec inhibition, although off-target effects on additional kinases cannot be discarded. This is potentially relevant to control infection-risk in CLL patients and, thus, future studies should carefully evaluate the effects of CLL therapies, including Btk inhibitors with higher specificity for Btk, on platelet-mediated immune functions.
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Affiliation(s)
- Leigh Naylor-Adamson
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Anisha R Chacko
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Zoe Booth
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Stefano Caserta
- Department of Biomedical Sciences, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Jenna Jarvis
- Department of Biomedical Sciences, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Sujoy Khan
- Department of Immunology & Allergy, Queens Centre, Castle Hill Hospital, Hull University Teaching Hospitals NHS Trust, Cottingham, United Kingdom
| | - Simon P Hart
- Respiratory Research Group, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - Francisco Rivero
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
| | - David J Allsup
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom.,Department of Haematology, Queens Centre for Oncology and Haematology, Castle Hill Hospital, Hull University Teaching Hospitals NHS Trust, Cottingham, United Kingdom
| | - Mònica Arman
- Centre for Atherothrombosis and Metabolic Disease, Hull York Medical School, Faculty of Health Sciences, University of Hull, Hull, United Kingdom
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Kim CJ, Kim J, Sabaté Del Río J, Ki DY, Kim J, Cho YK. Fully automated light transmission aggregometry on a disc for platelet function tests. LAB ON A CHIP 2021; 21:4707-4715. [PMID: 34752594 DOI: 10.1039/d1lc00708d] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/13/2023]
Abstract
Platelet function tests, a group of assays that measure the ability of platelets to aggregate and promote clotting in a sample of blood, are performed in various medical fields to assess inherited platelet function disorders and monitor antiplatelet therapies. Light transmission aggregometry (LTA) is considered the gold standard for platelet function assessment. However, the lack of a standardized protocol is a major drawback when applied at the point of care. Moreover, it is a time-consuming and labor-intensive assay that requires a large volume of blood. Here, we describe the design, fabrication, and operation of a centrifugal microfluidic disc that can perform a fully automated LTA assay from a small volume of a whole blood sample (<1 mL), achieving highly reproducible results (3.2% coefficient of variation) within a short period (<25 min). The assays performed with this device yield more precise and accurate results than traditional LTA because of the automation of the reaction steps, minimal human operation, robust detection strategy via the distinctive structure of the microfluidic chamber, and quick analysis that minimizes the adverse effects of platelet instability.
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Affiliation(s)
- Chi-Ju Kim
- Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
| | - Jungmin Kim
- Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
| | - Jonathan Sabaté Del Río
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
| | - Dong Yeob Ki
- Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
| | - Junyoung Kim
- Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
| | - Yoon-Kyoung Cho
- Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
- Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea
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Rajsic S, Breitkopf R, Bachler M, Treml B. Diagnostic Modalities in Critical Care: Point-of-Care Approach. Diagnostics (Basel) 2021; 11:diagnostics11122202. [PMID: 34943438 PMCID: PMC8700511 DOI: 10.3390/diagnostics11122202] [Citation(s) in RCA: 26] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2021] [Revised: 11/23/2021] [Accepted: 11/24/2021] [Indexed: 02/07/2023] Open
Abstract
The concept of intensive care units (ICU) has existed for almost 70 years, with outstanding development progress in the last decades. Multidisciplinary care of critically ill patients has become an integral part of every modern health care system, ensuing improved care and reduced mortality. Early recognition of severe medical and surgical illnesses, advanced prehospital care and organized immediate care in trauma centres led to a rise of ICU patients. Due to the underlying disease and its need for complex mechanical support for monitoring and treatment, it is often necessary to facilitate bed-side diagnostics. Immediate diagnostics are essential for a successful treatment of life threatening conditions, early recognition of complications and good quality of care. Management of ICU patients is incomprehensible without continuous and sophisticated monitoring, bedside ultrasonography, diverse radiologic diagnostics, blood gas analysis, coagulation and blood management, laboratory and other point-of-care (POC) diagnostic modalities. Moreover, in the time of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, particular attention is given to the POC diagnostic techniques due to additional concerns related to the risk of infection transmission, patient and healthcare workers safety and potential adverse events due to patient relocation. This review summarizes the most actual information on possible diagnostic modalities in critical care, with a special focus on the importance of point-of-care approach in the laboratory monitoring and imaging procedures.
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Affiliation(s)
- Sasa Rajsic
- General and Surgical Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Medical University Innsbruck, 6020 Innsbruck, Austria; (S.R.); (M.B.)
| | - Robert Breitkopf
- Transplant Surgical Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Medical University Innsbruck, 6020 Innsbruck, Austria;
| | - Mirjam Bachler
- General and Surgical Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Medical University Innsbruck, 6020 Innsbruck, Austria; (S.R.); (M.B.)
| | - Benedikt Treml
- General and Surgical Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Medical University Innsbruck, 6020 Innsbruck, Austria; (S.R.); (M.B.)
- Correspondence:
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Gomez K, Anderson J, Baker P, Biss T, Jennings I, Lowe G, Platton S. Clinical and laboratory diagnosis of heritable platelet disorders in adults and children: a British Society for Haematology Guideline. Br J Haematol 2021; 195:46-72. [PMID: 34435350 DOI: 10.1111/bjh.17690] [Citation(s) in RCA: 20] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Affiliation(s)
- Keith Gomez
- Haemophilia Centre and Thrombosis Unit, Royal Free London NHS Foundation Trust, London
| | - Julia Anderson
- Haemophilia Thrombosis and Immunology Centre, Royal Infirmary, NHS Lothian, Edinburgh
| | - Peter Baker
- Haemophilia and Thrombosis Centre, Oxford University Hospitals NHS Foundation Trust, Oxford
| | - Tina Biss
- Haemophilia Comprehensive Care Centre, Royal Victoria Infirmary, Newcastle-upon-Tyne Hospitals NHS Foundation Trust, Newcastle-upon-Tyne
| | - Ian Jennings
- UK NEQAS for Blood Coagulation, Sheffield Teaching Hospitals NHS Foundation Trust, Sheffield
| | - Gillian Lowe
- Haemophilia Comprehensive Care Centre, Queen Elizabeth Hospital, University Hospitals Birmingham NHS Foundation Trust, Birmingham, UK
| | - Sean Platton
- Haemophilia Centre, The Royal London Hospital, Barts Health NHS Trust, London, UK
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Kitchen S, Adcock DM, Dauer R, Kristoffersen AH, Lippi G, Mackie I, Marlar RA, Nair S. International Council for Standardization in Haematology (ICSH) recommendations for processing of blood samples for coagulation testing. Int J Lab Hematol 2021; 43:1272-1283. [PMID: 34581008 DOI: 10.1111/ijlh.13702] [Citation(s) in RCA: 30] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/21/2021] [Revised: 07/27/2021] [Accepted: 08/21/2021] [Indexed: 11/29/2022]
Abstract
This guidance document has been prepared on behalf of the International Council for Standardization in Haematology (ICSH). The aim of the document is to provide guidance and recommendations for the processing of citrated blood samples for coagulation tests in clinical laboratories in all regions of the world. The following areas are included in this document: Sample transport including use of pneumatic tubes systems; clots in citrated samples; centrifugation; primary tube storage and stability; interfering substances including haemolysis, icterus and lipaemia; secondary aliquots-transport, storage and processing; preanalytical variables for platelet function testing. The following areas are excluded from this document, but are included in an associated ICSH document addressing collection of samples for coagulation tests in clinical laboratories; ordering tests; sample collection tube and anticoagulant; preparation of the patient; sample collection device; venous stasis before sample collection; order of draw when different sample types are collected; sample labelling; blood-to-anticoagulant ratio (tube filling); influence of haematocrit. The recommendations are based on published data in peer-reviewed literature and expert opinion.
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Affiliation(s)
- Steve Kitchen
- Sheffield Haemophilia and Thrombosis Centre, Sheffield, UK
| | - Dorothy M Adcock
- Laboratory Corporation of America Holdings, Burlington, North Carolina, USA
| | - Ray Dauer
- Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia
| | - Ann-Helen Kristoffersen
- Department of Medical Biochemistry and Pharmacology, Haukeland University Hospital, Bergen, Norway.,Norwegian Organization for Quality Improvement of Laboratory Examinations (Noklus), Haraldsplass Deaconess Hospital, Bergen, Norway
| | - Giuseppe Lippi
- Section of Clinical Biochemistry, University of Verona, Verona, Italy
| | - Ian Mackie
- Research Department of Haematology, University College London, London, UK
| | - Richard A Marlar
- Department of Pathology, University of New Mexico, Albuquerque, New Mexico, USA
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Abstract
Upon activation, platelets release a plethora of factors which help to mediate their dynamic functions in hemostasis, inflammation, wound healing, tumor metastasis and angiogenesis. The majority of these bioactive molecules are released from α-granules, which are unique to platelets, and contain an incredibly diverse repertoire of cargo including; integral membrane proteins, pro-coagulant molecules, chemokines, mitogenic, growth and angiogenic factors, adhesion proteins, and microbicidal proteins. Clinically, activation of circulating platelets has increasingly been associated with various disease states. Biomarkers indicating the level of platelet activation in patients can therefore be useful tools to evaluate risk factors to predict future complications and determine treatment strategies or evaluate antiplatelet therapy. The irreversible nature of α-granule secretion makes it ideally suited as a marker of platelet activation. This review outlines the release and contents of platelet α-granules, as well as the membrane bound, and soluble α-granule cargo proteins that can be used as biomarkers of platelet activation.
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Affiliation(s)
- Christopher W Smith
- Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, B15 2TT, Birmingham, UK
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Racine-Brzostek SE, Asmis LM. Assessment of platelet function utilizing viscoelastic testing. Transfusion 2021; 60 Suppl 6:S10-S20. [PMID: 33089932 DOI: 10.1111/trf.16081] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2020] [Revised: 08/29/2020] [Accepted: 08/29/2020] [Indexed: 12/19/2022]
Affiliation(s)
- Sabrina E Racine-Brzostek
- Department of Pathology and Laboratory Medicine, New York-Presbyterian Hospital, Weill Cornell Medicine, New York, New York, USA
| | - Lars M Asmis
- Centre for Perioperative Thrombosis and Haemostasis, Zurich, Switzerland
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Abstract
Snake venoms have evolved primarily to immobilize and kill prey, and consequently, they contain some of the most potent natural toxins. Part of that armory is a range of hemotoxic components that affect every area of hemostasis, which we have harnessed to great effect in the study and diagnosis of hemostatic disorders. The most widely used are those that affect coagulation, such as thrombin-like enzymes unaffected by heparin and direct thrombin inhibitors, which can help confirm or dispute their presence in plasma. The liquid gold of coagulation activators is Russell's viper venom, since it contains activators of factor X and factor V. It is used in a range of clotting-based assays, such as assessment of factor X and factor V deficiencies, protein C and protein S deficiencies, activated protein C resistance, and probably the most important test for lupus anticoagulants, the dilute Russell's viper venom time. Activators of prothrombin, such as oscutarin C from Coastal Taipan venom and ecarin from saw-scaled viper venom, are employed in prothrombin activity assays and lupus anticoagulant detection, and ecarin has a valuable role in quantitative assays of direct thrombin inhibitors. Snake venoms affecting primary hemostasis include botrocetin from the jararaca, which can be used to assay von Willebrand factor activity, and convulxin from the cascavel, which can be used to detect deficiency of the platelet collagen receptor, glycoprotein VI. This article takes the reader to every area of the diagnostic hemostasis laboratory to appreciate the myriad applications of snake venoms available in diagnostic practice.
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Affiliation(s)
- Gary William Moore
- Department of Haematology, Specialist Haemostasis Unit, Cambridge University Hospitals NHS Foundation Trust, Cambridge, United Kingdom.,Faculty of Science and Technology, Middlesex University London, London, United Kingdom
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