Histidine-Tryptophan-Ketoglutarate or Celsior: Which Is More Suitable for Cold Preservation for Cardiac Grafts From Older Donors? Sungsoo Lee, MD, Chien-Sheng.

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Histidine-Tryptophan-Ketoglutarate or Celsior: Which Is More Suitable for Cold Preservation for Cardiac Grafts From Older Donors? Sungsoo Lee, MD, Chien-Sheng Huang, MD, Tomohiro Kawamura, MD, Norihisa Shigemura, MD, Timothy R. Billiar, MD, Atsunori Nakao, MD, Yoshiya Toyoda, MD The Annals of Thoracic Surgery Volume 91, Issue 3, Pages 755-763 (March 2011) DOI: 10.1016/j.athoracsur.2010.11.024 Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Myocardial energy levels. Adenosine triphosphate (ATP) levels measured bioluminescently in cardiac grafts after 0, 6, 12, and 18 hours ischemic time from either young or older donors (A) and (B). In histidine-tryptophan-ketoglutarate (HTK)-treated grafts compared with saline and Celsior (CEL), myocardium contained higher ATP levels. Values are means ± SD; n = 3, *p < 0.05 vs CEL. (— = HTK; − − − = CEL; —- = saline.) The Annals of Thoracic Surgery 2011 91, 755-763DOI: (10.1016/j.athoracsur.2010.11.024) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Regulator of myocardial energy metabolism. Immunoblots of myocardial protein extracts for analysis of AMP-activated protein kinase (AMPK) and phosphorylated (p-) AMPK Thr 172 obtained from hearts from older donors and treated with saline, Celsior (CEL), and histidine-tryptophan-ketoglutarate (HTK) and subjected to 6 hours of cold ischemia and 3 hours of reperfusion. Image is representative of three independent experiments. The Annals of Thoracic Surgery 2011 91, 755-763DOI: (10.1016/j.athoracsur.2010.11.024) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Ischemia-reperfusion induced cell infiltration and apoptosis. Detection of naphthol, ED-1, and dUTP nick-end labeling (TUNEL)-positive cells (arrows) in grafts from older donors after 6 hours of cold ischemia and 12 hours of reperfusion (A). Neutrophil infiltration in the cardiac grafts stored in saline, CEL, and HTK solution was assessed by naphthol staining (original magnification 400×). Macrophage infiltration in the cardiac grafts stored in saline, Celsior (CEL), and histidine-tryptophan-ketoglutarate (HTK) solution was assessed by ED-1 staining (original magnification 400×). Apoptosis in the cardiac grafts stored in saline, CEL, and HTK solution was assessed by TUNEL staining (original magnification 400×). Immunoblots for cleaved caspase-3 and β-actin on myocardial protein extracts obtained from hearts treated with saline, CEL, and HTK and subjected to 6 hours of cold ischemia and 12 hours of reperfusion (B). Images are representative of three independent experiments. The Annals of Thoracic Surgery 2011 91, 755-763DOI: (10.1016/j.athoracsur.2010.11.024) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions