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Preparation of Washed Human Platelets for Quantitative Metabolic Flux Studies.

J Vis Exp

January 2025

Depeartment of Chemical and Biological Engineering, Colorado School of Mines; Quantitative Biosciences and Engineering, Colorado School of Mines;

Platelets are blood cells that play an integral role in hemostasis and the innate immune response. Platelet hyper- and hypoactivity have been implicated in metabolic disorders, increasing risk for both thrombosis and bleeding. Platelet activation and metabolism are tightly linked, with the numerous methods to measure the former but relatively few for the latter.

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Background: Asthma is a prevalent respiratory disease, and its management remains largely unsatisfactory. Mesenchymal stem cells (MSCs) have been demonstrated to be efficacious in reducing airway inflammation in experimental allergic diseases, representing a potential alternative treatment for asthma. Migrasomes are recently identified extracellular vesicles (EVs) generated in migrating cells and facilitate intercellular communication.

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In recent decades, electrokinetic handling of microparticles and biological cells found many applications ranging from biomedical diagnostics to microscale assembly. The integration of electrokinetic handling such as dielectrophoresis (DEP) greatly benefits microfluidic point-of-care systems as many modern assays require cell handling. Compared to traditional pump-driven microfluidics, typically used for DEP applications, centrifugal CD microfluidics provides the ability to consolidate various liquid handling tasks in self-contained discs under the control of a single motor.

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Proteases are overexpressed at various stages of conditions such as cancers and thus can serve as biomarkers for disease diagnosis. Electrochemical techniques to detect the activity of extracellular proteases have gained attraction due to their multiplexing capability. Here we employ an electrochemical approach based on a 3 × 3 gold (Au) microelectrode array (MEA) functionalized with (2-aminoethyl)ferrocene (AEF) tagged specific peptide substrates to monitor cathepsin B (CB) protease activity.

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Challenges for complement functional assays in the clinical laboratory: From test validation to clinical interpretation.

J Immunol Methods

January 2025

Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States of America. Electronic address:

Complement functional assays are essential first-tier tests for a gamut of disorders spanning from inborn errors of the immune system which lead to recurrent severe infections, to angioedema attacks, presentation of autoimmune disease, thrombotic microangiopathies and rare kidney disorders. These assays evaluate the activity of the three complement pathways and specific complement components, which helps in differential diagnosis and monitoring disease progression. The rising use of complement inhibitors for treating complement-mediated thrombotic microangiopathies has heightened the demand for personalized treatment plans and laboratory assessment of complement blockage.

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