Display options
Cite
Kumae T. Development of a simultaneous multiple measurement method for superoxide generation from phagocytes using the cytochrome c reduction method. Environ Health Prev Med. 1996;1(2):80-6doi: 10.1007/BF02931195.
Kumae, T. (1996). Development of a simultaneous multiple measurement method for superoxide generation from phagocytes using the cytochrome c reduction method. Environmental health and preventive medicine, 1(2), 80-6. https://doi.org/10.1007/BF02931195
Kumae, T. "Development of a simultaneous multiple measurement method for superoxide generation from phagocytes using the cytochrome c reduction method." Environmental health and preventive medicine vol. 1,2 (1996): 80-6. doi: https://doi.org/10.1007/BF02931195
Kumae T. Development of a simultaneous multiple measurement method for superoxide generation from phagocytes using the cytochrome c reduction method. Environ Health Prev Med. 1996 Jul;1(2):80-6. doi: 10.1007/BF02931195. PMID: 21432427.
Copy Download .nbib Format: NLM
Share it on

Environ Health Prev Med. 1996 Jul;1(2):80-6. doi: 10.1007/BF02931195.

Development of a simultaneous multiple measurement method for superoxide generation from phagocytes using the cytochrome c reduction method.

Environmental health and preventive medicine

T Kumae

Affiliations

  1. The Section of Occupational Disease, Department of Industrial Health, The Institute of Public Health, Shirokanedai 4-6-1, 108, Minato-Ku, Tokyo, Japan.

PMID: 21432427 DOI: 10.1007/BF02931195

Abstract

It is important to measure superoxide generation in biological samples in research concerning reactive oxygen, because most reactive oxygen species biologically generated are derived from superoxides.This paper reports the establishment of a simultaneous multiple measurement method for detection of superoxides generated from phagocytes, human peripheral neutrophils and rat alveolar macrophages, using cvtochrome c reduction with stirring at 37°C. Using this method, the following results were obtained.1. For more objective presentation of obtained results, velocities of cytochrome c reduction were calculated by the linear regression method at each measurement point and new indicators of cytochrome c reduction, peak value (maximum reduction velocity) and peak time (time when peak value was observed), were introduced.2. Cytochrome c reduction was measured by dual wavelengths, 550nm and 492nm, under the conditions mentioned below.Final sample volume was 300 μl and phagocytes, 1.5 x 10(5) human neutrophils or 3.0 x 10(5) rat macrophages, were mixed with 2.0mg/ml of opsonized zymosan in the presence of 150 μM cytochrome c.3. In the case of human neutrophils, a small amount of cytochrome c reduction was induced by reactions with the microplate surface, whereas in the case of rat macrophages, which can phagocytize materials without opsonization, it was expected that substantial cytochrome c reduction would be observed. However, in comparison with the results of opsonized zymosan stimulation, peak value reached only 40% and peak time was significantly delayed.4. Peak value showed significant linear changes corresponding with changes of the opsonized zymosan concentration and phagocyte numbers when measurements were carried out 37°C.These results suggest that differences of phagocyte activity and serum opsonic activity, which are difficult to measure using previous methods, are clearly detected by the simultaneous measurement method and the indicators introduced in this paper.

References

  1. J Immunol Methods. 1991 Aug 28;142(1):95-104 - PubMed
  2. Fed Proc. 1987 May 15;46(7):2402-6 - PubMed
  3. Am J Pathol. 1992 Sep;141(3):601-10 - PubMed
  4. Acta Physiol Scand. 1991 Jun;142(2):275-81 - PubMed
  5. J Immunol Methods. 1988 Jul 22;111(2):227-32 - PubMed
  6. Mech Ageing Dev. 1987 Nov;41(1-2):125-37 - PubMed
  7. Arch Toxicol. 1987;60(1-3):144-9 - PubMed
  8. Am J Pathol. 1983 Mar;110(3):297-309 - PubMed
  9. J Appl Physiol (1985). 1992 Nov;73(5):1805-9 - PubMed
  10. Free Radic Biol Med. 1987;3(6):405-21 - PubMed
  11. Proc Soc Exp Biol Med. 1979 Oct;162(1):139-45 - PubMed
  12. J Clin Invest. 1977 Nov;60(5):963-9 - PubMed
  13. Immunology. 1978 Oct;35(4):613-8 - PubMed
  14. J Clin Invest. 1989 Apr;83(4):1326-35 - PubMed
  15. Life Sci. 1992;50(5):335-9 - PubMed
  16. Methods Enzymol. 1984;105:370-8 - PubMed
  17. Am J Physiol. 1989 Dec;257(6 Pt 1):L411-20 - PubMed
  18. J Immunol Methods. 1981;46(2):211-26 - PubMed
  19. Am J Respir Cell Mol Biol. 1994 Oct;11(4):426-31 - PubMed
  20. Arch Surg. 1991 Jan;126(1):104-5 - PubMed
  21. J Toxicol Environ Health. 1993 Mar;38(3):273-92 - PubMed
  22. Mol Cell Biochem. 1992 Apr;111(1-2):103-8 - PubMed
  23. J Lab Clin Med. 1983 Nov;102(5):732-42 - PubMed
  24. Methods Enzymol. 1990;186:567-75 - PubMed
  25. Nihon Ketsueki Gakkai Zasshi. 1987 Aug;50(5):949-57 - PubMed
  26. J Appl Physiol (1985). 1986 Feb;60(2):353-69 - PubMed

Publication Types