Differentiating between intra- and extracellular
chemiluminescence in diluted whole-blood samples

J 2012

Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples

RÁJECKÝ, Michal, Antonín LOJEK a Milan ČÍŽ

Základní údaje

Originální název

Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples

Autoři

RÁJECKÝ, Michal, Antonín LOJEK a Milan ČÍŽ

Vydání

International Journal of Laboratory Hematology, MALDEN, WILEY-BLACKWELL, 2012, 1751-5521

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.293

DOI

http://dx.doi.org/10.1111/j.1751-553X.2011.01370.x

UT WoS

000300695600004

Klíčová slova anglicky

Chemiluminescence; isoluminol; luminol; phagocyte; whole blood

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 14. 5. 2014 14:20, Mgr. Michal Rájecký, Ph.D.

Anotace

V originále

Introduction: The differentiation between extra- and intracellular production of reactive oxygen species (ROS) in whole blood was measured by luminol- and isoluminol-enhanced chemiluminescence (CL). Methods: Azide (total CL inhibition), azide + horseradish peroxidase (HRP, restoring extracellular CL), superoxide dismutase + catalase (depleting extracellular ROS) and HRP (enhancing extracellular CL) were used to modulate luminol-and isoluminol-enhanced CL (10(-6)-10(-3) M luminophores) of 125x diluted whole blood which was activated by both calcium ionophore A23187 (Ca-I) and opsonized zymosan particles (OZP) separately. Results: Both activators stimulated intra-and extracellular production of ROS. Luminol-enhanced CL of Ca-I-activated samples detected the intracellular ROS, and with the addition of HRP detected the extracellular CL as well. CL enhanced with isoluminol in concentrations of 10(-4) M or less was mostly extracellular. There was a mixture of intra-and extracellular CL in OZP-activated samples, probably because of the ingestion of luminophore molecules. Conclusion: Measurement of Ca-I-activated CL enhanced with 10(-4) M luminol is recommended for the detection of intracellular ROS. The addition of HRP leads to the detection of overall ROS production while the OZP-activated system with its addition of HRP can only be used to detect overall ROS production. Ca-I-activated CL enhanced with 10(-4) M isoluminol and with addition of HRP is recommended for the detection of extracellular CL.

Citovat

RÁJECKÝ, Michal, Antonín LOJEK a Milan ČÍŽ. Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples. International Journal of Laboratory Hematology. MALDEN: WILEY-BLACKWELL, 2012, roč. 34, č. 2, s. 136-142. ISSN 1751-5521. Dostupné z: https://dx.doi.org/10.1111/j.1751-553X.2011.01370.x.

@article{1080654,
   author = {Rájecký, Michal and Lojek, Antonín and Číž, Milan},
   article_location = {MALDEN},
   article_number = {2},
   doi = {http://dx.doi.org/10.1111/j.1751-553X.2011.01370.x},
   keywords = {Chemiluminescence; isoluminol; luminol; phagocyte; whole blood},
   language = {eng},
   issn = {1751-5521},
   journal = {International Journal of Laboratory Hematology},
   title = {Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples},
   volume = {34},
   year = {2012}
}

TY  - JOUR
ID  - 1080654
AU  - Rájecký, Michal - Lojek, Antonín - Číž, Milan
PY  - 2012
TI  - Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples
JF  - International Journal of Laboratory Hematology
VL  - 34
IS  - 2
SP  - 136-142
EP  - 136-142
PB  - WILEY-BLACKWELL
SN  - 17515521
KW  - Chemiluminescence
KW  - isoluminol
KW  - luminol
KW  - phagocyte
KW  - whole blood
N2  - Introduction: The differentiation between extra- and intracellular production of reactive oxygen species (ROS) in whole blood was measured by luminol- and isoluminol-enhanced chemiluminescence (CL). Methods: Azide (total CL inhibition), azide + horseradish peroxidase (HRP, restoring extracellular CL), superoxide dismutase + catalase (depleting extracellular ROS) and HRP (enhancing extracellular CL) were used to modulate luminol-and isoluminol-enhanced CL (10(-6)-10(-3) M luminophores) of 125x diluted whole blood which was activated by both calcium ionophore A23187 (Ca-I) and opsonized zymosan particles (OZP) separately. Results: Both activators stimulated intra-and extracellular production of ROS. Luminol-enhanced CL of Ca-I-activated samples detected the intracellular ROS, and with the addition of HRP detected the extracellular CL as well. CL enhanced with isoluminol in concentrations of 10(-4) M or less was mostly extracellular. There was a mixture of intra-and extracellular CL in OZP-activated samples, probably because of the ingestion of luminophore molecules. Conclusion: Measurement of Ca-I-activated CL enhanced with 10(-4) M luminol is recommended for the detection of intracellular ROS. The addition of HRP leads to the detection of overall ROS production while the OZP-activated system with its addition of HRP can only be used to detect overall ROS production. Ca-I-activated CL enhanced with 10(-4) M isoluminol and with addition of HRP is recommended for the detection of extracellular CL.
ER  -

RÁJECKÝ, Michal, Antonín LOJEK a Milan ČÍŽ. Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples. \textit{International Journal of Laboratory Hematology}. MALDEN: WILEY-BLACKWELL, 2012, roč.~34, č.~2, s.~136-142. ISSN~1751-5521. Dostupné z: https://dx.doi.org/10.1111/j.1751-553X.2011.01370.x.

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