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Secondary oxidants in human serum exposed to singlet oxygen: the influence of hemolysis

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Abstract

Singlet oxygen (1O2) is produced by leucocytes during inflammatory reactions, various biochemical reactions and during photoreactions. It deactivates by reacting with a number of targets to produce reactive oxygen species (ROS) and peroxides (that in turn produce ROS). To verify whether serum had the same capability to deactivate secondary oxidants after exposure to 1O2, we provoked a photoreaction using rose bengal added to sera of 53 healthy donors and, after light delivery, reduced 2’,7’-dichlorofluorescein (DCFH) was added at the end of irradiation and fluorescence of the oxidized derivative (DCF) was recorded. To avoid optical artifacts, we analyzed the influence of hemolysis. Deactivation capability of secondary oxidants after exposure to 1O2 was stable over a long period of time, slightly different between men and women, but standard biochemistry parameters had little influence. Hemolysis, age and platelet number reduced deactivation of 1O2-induced secondary oxidants. Addition of lysed cancer cells had no influence. Blood sampling in clot act tubes gave a better signal than in heparinized tubes. Red blood cells (RBCs) loaded with antioxidants strongly decreased deactivation of secondary oxidants. Assays are in progress to evaluate the clinical implications of these findings.

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Abbreviations

DCF:

Dichlorofluorescein (oxidized, fluorescent)

DCFH:

Dichlorofluorescein (reduced)

DCF-DA:

Dichlorofluorescein diacetate

FCS:

Fetal calf serum

HS:

Human serum

OD:

Optical density

PDT:

Photodynamic therapy

RB:

Rose bengal

ROS:

Reactive oxygen species

RPMI:

Roswell Park Memorial Institute

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Authors and Affiliations

  1. Laboratoire de Photobiologie des Cancers, Département Laser, 44093, Nantes, France

    David Olivier, Samuel Douillard, Isabelle Lhommeau & Thierry Patrice

  2. Biochemistry (E.B), Hôpital Laënnec, CHU de Nantes, 44093, Nantes, France

    Edith Bigot

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  1. David Olivier
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Correspondence to Thierry Patrice.

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Olivier, D., Douillard, S., Lhommeau, I. et al. Secondary oxidants in human serum exposed to singlet oxygen: the influence of hemolysis. Photochem Photobiol Sci 8, 1476–1486 (2009). https://doi.org/10.1039/b9pp00032a

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