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== Affiliations and support ==
:::: Garcia-Souza LF(1,2), Cizmarova B(2,3), Sumbalova Z(2,4), Menz V(1), Burtscher M(1), Gnaiger E(2,5)
::::#Inst Sport Science, Univ Innsbruck, Austria
::::#Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck, Austria
::::#Dept Medical Clinical Biochem, Fac Medicine, Pavol Jozef Šafárik Univ Košice, Slovakia
::::#Pharmacobiochemical Lab, 3rd Dept Internal Medicine, Fac Medicine, Comenius Univ, Bratislava, Slovakia
::::#Oroboros Instruments, Innsbruck, Austria. - firstname.lastname@example.org
::::::Supported by [[K-Regio MitoFit]]. Contribution to European Union Framework Programme Horizon 2020 COST Action CA15203 [[MitoEAGLE]].
== Figure 1 ==
[[File:Protocol Intact PLT Luiz.png|left|650px]]
'''Figure 1. Representative experiment of extended substrate-uncoupler-inhibitor titration (SUIT) protocol.''' Blue line corresponds to oxygen concentration (µM) in the chamber and red line corresponds to oxygen flow of intact platelets (pmol •s<sup>-1</sup>•10<sup>-8</sup> cells). The titration events and marks are represented as follows: ROUTINE respiration (R) is initially measured in intact cells (1ce) in mitochondrial respiration medium (MiR05-Kit); single titration of pyruvate (2P, 10 mM) to supplement MiR05 medium with oxidative substrate; uncoupler (3U*, titration of CCCP) is used for determination of ET capacity; glucose (4Glc, 11 mM) to compare Crabtree effect with culture media; malate (5M, 2 mM) stimulation is observed only in permeabilized cells; rotenone (6Rot, 0.5 mM) inhibiting Complex I (CI) in intact cells; succinate (7S, 10 mM) stimulation of mt-respiration is observed in non-viable cells only; digitonin (8Dig) is used for permeabilization of the eintire cell population, providing a reference state of succinate-stimulated ET capacity; cytochrome ''c'' (8c) titration test provides information about outer mt-membrane integrity; antimycin A (Ama, 2.5 mM) inhibiting Complex III; ascorbate 2 mM & TMPD 0.5 mM (10AsTm) stimulation of Complex IV plus chemical background; azide (11Azd, 200 mM) as an inhibitor of cytochrome c oxidase provides information about the oxygen consumption chemical background subtracted from flux in 10AsTm.
== References ==
::::#Straub A, Breuer M, Wendel HP, Peter K, Dietz K, Ziemer G (2007) Critical temperature ranges of hypothermia-induced platelet activation: possible implications for cooling patients in cardiac surgery. Thromb Haemost 97:608-16.
::::#Karabatsiakis A, Böck C, Salinas-Manrique J, Kolassa S, Calzia E, Dietrich DE, Kolassa IT (2014) Mitochondrial respiration in peripheral blood mononuclear cells correlates with depressive subsymptoms and severity of major depression. Transl Psychiatry 4:e397.
::::#Johnson L, Tan S, Wood B, Davis A, Marks DC (2016) Refrigeration and cryopreservation of platelets differentially affect platelet metabolism and function: a comparison with conventional platelet storage conditions. Transfusion 56:1807-18.
::::#Johnson L, Coorey CP, Marks DC (2014) The hemostatic activity of cryopreserved platelets is mediated by phosphatidylserine‐expressing platelets and platelet microparticles. Transfusion 54:1917-26.
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