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Flow-Cytometric Analysis of Platelet-Membrane Glycoprotein Expression and Platelet Activation

Flow-cytometric measurement of platelet-surface glycoproteins in unfixed whole blood is a sensitive and quantitative approach that offers many advantages over other methods of platelet analysis for the following reasons:

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Flow-Cytometric Analysis of Mouse Platelet Function

Characterization of platelet-membrane glycoproteins and their intracellular signaling pathways has become essential to understanding platelet function not only in the context of thrombosis and hemostasis, but also other processes like inflammation, tumor metastasis, a ...

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Effects of Anticoagulants Used During Blood Collection on Human Platelet Function

Nearly all the chapters in this book involve some sort of study of platelets (and megakaryocytes) in vitro with a view to gaining insight into the true behaviour of these cells in the circulation. But a word of warning is required. Studies performed in vitro can only approximate the true situation in vivo. ...

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Platelet Microparticles

One of the responses of activated platelets to certain stimuli is the shedding of microparticles. Microparticles released from platelets (PMPs) may play a role in the normal hemostatic response to vascular injury, as these particles exhibit prothrombinase activity (1). It is also possi ...

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Preparation of Washed Platelet Suspensions From Human and Rodent Blood

Citrate is the preferred anticoagulant for blood collection, as EDTA damages platelets and heparin modifies their function (1). Citrate allows the rapid generation of platelet-rich plasma (PRP), with a high yield of platelets; however, this method has certain disadvantages. In partic ...

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Platelet Counting

The assessment of the platelet count is essential within both routine hematology and research laboratories. Accurate and precise enumeration of platelets is not only critical to assist in diagnosis and treatment of various clinical disorders, but it is also a vital research tool particu ...

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Electron Microscopy Methods for Studying Platelet Structure and Function

Platelets are very small—the smallest of cellular elements in circulating blood. They lack a nucleus and their cytoplasm is relatively clear. As a result it was impossible to visualize platelets in the crude instruments used by early microscopists. In fact, platelets were not identified un ...

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Analysis of the Releasable Nucleotides of Platelets

In platelets, two pools of nucleotides have been demonstrated: one is utilized for the metabolic needs of the platelets (1) and the second stores nucleotides in a metabolically inert form in dense granules. Upon activation of platelets, nucleotides are released from the dense granules into t ...

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Isolation of Primary Megakaryocytes and Studies of Proplatelet Formation

Bone marrow megakaryocytes were first described well over one hundred years ago (1,2) and it is almost one hundred years since their role in platelet formation was first appreciated (3,4). Prior to the development of megakaryocyte isolation techniques, a great deal had been learned about mega ...

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Isolation and Culture of Megakaryocyte Precursors

Megakaryocyte (MK) precursor cells correspond to a spectrum of cells extending from an early progenitor to the promegakaryoblast, a 2N cell that switches from a mitotic to an endomitotic process (1). The MK progenitors express the CD34+ antigen and are either CD38– or CD38+, depending upon their m ...

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Thrombopoietin Bioassay

The production of megakaryocytes (MKs) and platelets is a finely controlled process that maintains circulating platelet numbers within a narrow range in normal individuals while providing sufficient reserve capacity for rapid production in emergencies such as bleeding. Forty y ...

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Central Nervous System Inflammation and Prion Disease Pathogenesis

The study of inflammation in the prion diseases is relatively new. Indeed, for a number of years the accepted dogma was that the prion diseases lacked an inflammatory response in the brain (1–3). This persists in spite of a number of studies showing that the pathological hallmarks of the prion diseases ...

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The Electroneuropathology of Prion Disease

Neuropathological studies can reveal a great deal about the appearance of cells and structures within the nervous system during the course of a disease, but they cannot determine which neurons were working properly when the samples were taken or in the period before death. Neurons may look norm ...

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HER-2/neu Oncogene Amplification Determined by Fluorescence In Situ Hybridization

The proto-oncogene HER-2/neu (C-erbB-2) has been localized to chromosome 17q and encodes a transmembrane tyrosine kinase growth factor receptor. The name for the HER-2 protein is derived from human epidermal growth factor receptor (EGFR) because it features substantial homology with ...

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A Nested Reverse Transcription-Polymerase Chain Reaction Assay to Detect BCR/abl

Chronic myelogenous leukemia (CML), a clonal myeloproliferative disorder in adults, and some pediatric and adult acute lymphoblastic leukemias (ALLs) are characterized by the presence of a Philadelphia chromosome, t(9;22)(q34;q11) (1). In this chromosomal translocation, exons ...

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Methods for Studying Voltage-Gated Sodium Channels in Heterologous Expression Systems

The discovery that oocytes of the frog Xenopus laevis can be induced to express working membrane ion channels by introducing channel mRNA into their cytoplasm (heterologous expression) has greatly impacted the field of ion channel physiology. With the addition of site-directed mutag ...

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Methods for Isolation of Endothelial and Smooth Muscle Cells and In Vitro Proliferation Assays

Angiogenesis, the formation of new blood vessel from pre-existing blood vessel, occurs in a variety of normal and pathological conditions. It is complex morphogenetic process involving the coordinate migration, invasion, and reorganization of several cell types including endot ...

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Applications of Adenoviral Vector-Mediated Gene Transfer in Cardiovascular Research

Cardiovascular disease is a leading cause of morbidity and mortality worldwide. New studies are needed to explore novel therapeutic options for patients that are refractory to existing therapies. Gene transfer using adenoviral vectors has shown promising results in animal studie ...

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Efficient Transfection of Primary Cells Relevant for Cardiovascular Research by nucleofection

Cell types that are important for cardiovascular research, e.g., cardiomyocytes, endothelial cells, or adult stem cells, are often hard to isolate, culture, and transfect. Low-transfection efficiencies are a major limitation because, in many cases, results achieved with surrogate m ...

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Murine and HIV-Based Retroviral Vectors for In Vitro and In Vivo Gene Transfer

The success of experimental gene therapy is dependent on the ability to safely and efficiently introduce transgenes into the target cell or tissue. Retroviral-based vectors, notably those derived from Moloney murine leukemia virus (MLV) and lentiviral vectors derived from HIV, have p ...

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