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Apoptosis pp 131–139Cite as

Application of In Vivo EPR for Tissue pO2 and Redox Measurements

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 559))

Summary

The technique of electron paramagnetic resonance (EPR) spectroscopy is more than 50 years old, but only recently it has been used for in vivo studies. Its limited application in the past was due to the problem of high nonresonant dielectric loss of the exciting frequency because of high water content in biological samples. However, with the development of spectrometers working at lower frequencies (1,200 MHz and below) during the last 15 years, it is now possible to conduct in vivo measurements on a variety of animals and isolated organs. This is further facilitated by the development of new resonators with high sensitivity and appropriate stability for in vivo applications. It now has become feasible to obtain new insights into the complex aspects of physiology and pathophysiology using in vivo EPR. Among several important applications of this technique, the in vivo tissue pO2 (partial pressure of oxygen) and redox measurements seem to be the most extensive use of this technique. In this chapter, we describe the procedure for in vivo pO2 and redox measurements in animal models.

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References

  1. Gutteridge JM. (1993). Free radicals in disease processes: a compilation of cause and consequence. Free Radical Res Commun 19, 141–158.

    Article  CAS  Google Scholar 

  2. Ceconi C, Boraso A, Cargnoni A and Ferrari R. (2003). Oxidative stress in cardiovascular disease: myth or fact? Arch Biochem Biophys 420, 217–221.

    Article  PubMed  CAS  Google Scholar 

  3. Warnholtz A, Wendt M, August M and Munzel T. (2004). Clinical aspects of reactive oxygen and nitrogen species. Biochem Soc Symp 71, 121–133.

    PubMed  CAS  Google Scholar 

  4. Nozue M, Lee I, Yuan F, Teicher BA, Brizel DM, Dewhirst MW, Milross CG, Milas L, Song CW, Thomas CD, Guichard M, Evans SM, Koch CJ, Lord EM, Jain RK and Suit HD. (1997). Interlaboratory variation in oxygen tension measurement by Eppendorf “Histograph” and comparison with hypoxic marker. J Surg Oncol 66, 30–38.

    Article  PubMed  CAS  Google Scholar 

  5. Hunjan S, Zhao D, Constantinescu A, Hahn EW, Antich PP and Mason RP. (2001). Tumor oximetry: demonstration of an enhanced dynamic mapping procedure using fluorine-19 echo planar magnetic resonance imaging in the Dunning prostate R3327-AT1 rat tumor. Int J Radiat Oncol Biol Phys 49, 1097–1108.

    Article  PubMed  CAS  Google Scholar 

  6. Young WK, Vojnovic B and Wardman P. (1996). Measurement of oxygen tension in tumors by time-resolved fluorescence. Br J Cancer Suppl 27, S256–S259.

    PubMed  CAS  Google Scholar 

  7. Swartz HM, Dunn JF (eds). (2002). Measurements of Oxygen in Tissues: Overview and Perspectives on Methods to Make the Measurements. Oxygen Transport to Tissue XXII. Pabst Science, Lengerich.

    Google Scholar 

  8. Dunn JF and Swartz HM. (2003). In vivo electron paramagnetic resonance oximetry with particulate materials. Methods 30, 159–166.

    Article  PubMed  CAS  Google Scholar 

  9. Swartz HM. (2003). The measurement of oxygen in vivo using EPR techniques. In Vivo EPR (ESR): Theory and Applications, LJ Berliner (Ed.). Plenum, New York, NY.

    Google Scholar 

  10. Liu KJ, Gast P, Moussavi M, Norby SW, Vahidi N, Walczak T, Wu M and Swartz HM. (1993). Lithium phthalocyanine: a probe for electron paramagnetic resonance oximetry in viable biological systems. Proc Natl Acad Sci USA 90, 5438–5442.

    Article  PubMed  CAS  Google Scholar 

  11. Afeworki M, Miller NR, Devasahayam N, Cook J, Mitchell JB, Subramanian S and Krishna MC. (1998). Preparation and EPR studies of lithium phthalocyanine radical as an oxymetric probe. Free Radical Biol Med 25, 72–78.

    Article  CAS  Google Scholar 

  12. Khan N, Williams BB and Swartz HM. (2006). Clinical applications of in vivo EPR: rationale and initial results. Appl Magn Reson 30, 185–199.

    Article  Google Scholar 

  13. Swartz HM, Khan N, Buckey J, Comi R, Gould L, Grinberg O, Hartford A, Hopf H, Hou H, Hug E, Iwasaki A, Lesniewski P, Salikhov I and Walczak T. (2004). Clinical applications of EPR: overview and perspectives. NMR Biomed 17, 335–351.

    Article  PubMed  CAS  Google Scholar 

  14. Swartz HM. (2002). The measurement of oxygen in vivo using EPR techniques. Biological Magnetic Resonance – Volume 20: In vivo EPR (ESR): Theory and Applications, LJ Berliner (Ed.). Plenum, New York, NY.

    Google Scholar 

  15. Swartz HM and Halpern H. (1998). EPR studies of living animals and related model systems (in vivo EPR). Biol Magn Reson 14, 367–404.

    Article  CAS  Google Scholar 

  16. Grinberg OY, Smirnov AI and Swartz HM. (2001). High spatial resolution multi-site EPR oximetry. The use of convolution-based fitting method. J Magn Reson 152, 247–258.

    PubMed  CAS  Google Scholar 

  17. Smirnov AI, Norby SW, Clarkson RB, Walczak T and Swartz HM. (1993). Simultaneous multi-site EPR spectroscopy in vivo. Magn Reson Med 30, 213–220.

    Article  PubMed  CAS  Google Scholar 

  18. Williams BB, Hou H, Grinberg O, Demidenko E and Swartz H. (2007). High spatial resolution multi-site EPR oximetry of transient focal cerebral ischemia in the rat. Antioxid Redox Signal 9, 1691–1698.

    Article  PubMed  CAS  Google Scholar 

  19. Grinberg OY, Friedman BJ and Swartz HM. (1997). Intramyocardial pO2 measured by EPR. Adv Exp Med Biol 428, 261–268.

    Article  PubMed  CAS  Google Scholar 

  20. Grinberg OY, Grinberg SA, Friedman BJ and Swartz HM. (1997). Myocardial oxygen tension and capillary density in the isolated perfused rat heart during pharmacological intervention. Adv Exp Med Biol 411, 171–181.

    Article  PubMed  CAS  Google Scholar 

  21. Helisch A, Wagner S, Khan N, Drinane M, Wolfram S, Heil M, Ziegelhoeffer T, Brandt U, Pearlman JD, Swartz HM and Schaper W. (2006). Impact of mouse strain differences in innate hind limb collateral vasculature. Arterioscler Thromb Vasc Biol 26, 520–526.

    Article  PubMed  CAS  Google Scholar 

  22. Hou H, Grinberg O, Grinberg S and Swartz H. (2005). Cerebral tissue oxygenation in reversible focal ischemia in rats: multi-site EPR oximetry measurements. Physiol Meas 26, 131–141.

    Article  PubMed  Google Scholar 

  23. Swartz HM, Taie S, Miyake M, Grinberg OY, Hou H, el-Kadi H and Dunn JF. (2003). The effects of anesthesia on cerebral tissue oxygen tension: use of EPR oximetry to make repeated measurements. Adv Exp Med Biol 530, 569–575.

    Article  PubMed  Google Scholar 

  24. James PE, Bacic G, Grinberg OY, Goda F, Dunn JF, Jackson SK and Swartz HM. (1996). Endotoxin-induced changes in intrarenal pO2, measured by in vivo electron paramagnetic resonance oximetry and magnetic resonance imaging. Free Radical Biol Med 21, 25–34.

    Article  CAS  Google Scholar 

  25. James PE, Goda F, Grinberg OY, Szybinski KG and Swartz HM. (1997). Intrarenal pO2 measured by EPR oximetry and the effects of bacterial endotoxin. Adv Exp Med Biol 411, 557–568.

    Article  PubMed  CAS  Google Scholar 

  26. Towner RA, Sturgeon SA, Khan N, Hou H and Swartz HM. (2002). In vivo assessment of nodularin-induced hepatotoxicity in the rat using magnetic resonance techniques (MRI, MRS and EPR oximetry). Chem-Biol Interact 139, 231–250.

    Article  PubMed  CAS  Google Scholar 

  27. Abramovic Z, Sentjurc M, Kristl J, Khan N, Hou H and Swartz HM. (2006). Influence of different anesthetics on skin oxygenation studied by electron paramagnetic resonance in vivo. Skin Pharmacol Physiol 20, 77–84.

    Article  PubMed  Google Scholar 

  28. Hou H, Khan N, O’Hara JA, Grinberg OY, Dunn JF, Abajian MA, Wilmot CM, Demidenko E, Lu S, Steffen RP and Swartz HM. (2005). Increased oxygenation of intracranial tumors by efaproxyn (efaproxiral), an allosteric hemoglobin modifier: In vivo EPR oximetry study. Int J Radiat Oncol Biol Phys 61, 1503–1509.

    Article  PubMed  CAS  Google Scholar 

  29. O’Hara JA, Khan N, Hou H, Wilmo CM, Demidenko E, Dunn JF and Swartz HM. (2004). Comparison of EPR oximetry and Eppendorf polarographic electrode assessments of rat brain PtO2. Physiol Meas 25, 1413–1423.

    Article  PubMed  Google Scholar 

  30. Chen K, Glockner JF, Morse PD 2nd and Swartz HM. (1989). Effects of oxygen on the metabolism of nitroxide spin labels in cells. Biochemistry 28, 2496–2501.

    Article  PubMed  CAS  Google Scholar 

  31. Kocherginsky N and Swartz HM. (1995). Nitroxide Spin Labels, Reactions in Biology and Chemistry. CRC, Boca Raton, FL.

    Google Scholar 

  32. Khan M, Kutala VK, Vikram DS, Wisel S, Chacko SM, Kuppusamy ML, Mohan IK, Zweier JL, Kwiatkowski P and Kuppusamy P. (2007). Skeletal myoblasts transplanted in the ischemic myocardium enhance in situ oxygenation and recovery of contractile function. Am J Physiol Heart Circ Physiol 293, H2129–H2139.

    Article  PubMed  CAS  Google Scholar 

  33. Das S, Khan N, Mukherjee S, Bagchi D, Gurusamy N, Swartz H, Das DK. (2008). Redox regulation of resveratrol-mediated switching of death signal into survival signal. Free Radical Biol Med 44, 82–90.

    Article  CAS  Google Scholar 

  34. Swartz HM, Chen K, Pals M, Sentjurc M and Morse PD II. (1986). Hypoxia-sensitive NMR contrast agents. Mag Reson Med 3, 169–174.

    Article  CAS  Google Scholar 

  35. Miura Y, Utsumi H and Hamada A. (1992). Effects of inspired oxygen concentration on in vivo redox reaction of nitroxide radicals in whole mice. Biochem Biophys Res Commun 182, 1108–1114.

    Article  PubMed  CAS  Google Scholar 

  36. Gomi F, Utsumi H, Hamada A and Matsuo M. (1993). Aging retards spin clearance from mouse brain and food restriction prevents its age-dependent retardation. Life Sci 52, 2027–2033.

    Article  PubMed  CAS  Google Scholar 

  37. Utsumi H, Takeshita K, Miura Y, Masuda S and Hamada A. (1993). In vivo EPR measurement of radical reaction in whole mice: Influence of inspired oxygen and ischemia-reperfusion injury on nitroxide reduction. Free Radical Res Commun 19, S219–S225.

    Article  CAS  Google Scholar 

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Acknowledgements

NIH grants CA118069 and CA120919.

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Authors and Affiliations

  1. EPR Center for Viable Systems, Dartmouth Medical School, Hanover, NH, USA

    Nadeem Khan

  2. Cardiovascular Research Center, University of Connecticut, School of Medicine, Farmington, CT, USA

    Dipak K. Das

Authors
  1. Nadeem Khan
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  2. Dipak K. Das
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Editors and Affiliations

  1. Boston Biomedical Research Institute, Grove Street 64, Watertown, 02472, U.S.A.

    Peter Erhardt

  2. Boston Biomedical Research Institute, Grove Street 64, Watertown, 02472, U.S.A.

    Ambrus Toth

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Khan, N., Das, D.K. (2009). Application of In Vivo EPR for Tissue pO2 and Redox Measurements. In: Erhardt, P., Toth, A. (eds) Apoptosis. Methods in Molecular Biology, vol 559. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-017-5_10

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  • DOI: https://doi.org/10.1007/978-1-60327-017-5_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-016-8

  • Online ISBN: 978-1-60327-017-5

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