Peripheral detection of S100β during cardiothoracic surgery: what are we really measuring? Vincent Fazio, MS, Sunil K Bhudia, MD, Nicola Marchi, PhD,

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Presentation transcript:

Peripheral detection of S100β during cardiothoracic surgery: what are we really measuring? Vincent Fazio, MS, Sunil K Bhudia, MD, Nicola Marchi, PhD, Barbara Aumayr, BS, Damir Janigro, PhD The Annals of Thoracic Surgery Volume 78, Issue 1, Pages 46-52 (July 2004) DOI: 10.1016/j.athoracsur.2003.11.042

Fig 1 (A and B) The significant difference (p < 0.01) between the S100β measured in fractionated samples (MW <50 kDa and MW >50 kDa) from both venous (A) and cavity (B) samples obtained during CPB surgery. The S100β values are expressed in mass units (ng · mL−1) in order to emphasize the relative quantity of protein measured by ELISA. (C and D) The relative difference of S100β measured in fractionated samples during 3 time points during CPB surgery (prepump, pump, and postpump) for both venous (C) and cavity (D) samples. Note that a significant difference in S100β was only measured in serum (venous) samples at pump and postpump time points (p < 0.05). (Note different axes ranges in A and B). White bar = <50 kDa; hatched bar = >50 kDa. (Ave = average; CPB = cardiopulmonary bypass; ELISA = enzyme linked immunosorbent assay; kDa = kiloDaltons; min = minutes; MW = molecular weight; SE = standard error; vs = versus.) The Annals of Thoracic Surgery 2004 78, 46-52DOI: (10.1016/j.athoracsur.2003.11.042)

Fig 2 (Top) Western blot analysis (nondenatured preparation) of cavity and peripheral samples during CPB surgery. Values at the bottom indicate the relative concentrations of S100β as measured by ELISA. (Bottom) A second Western blot analysis of high MW (MW >50 kDa) venous fraction (peripheral) conducted under denaturing conditions utilizing the same S100β primary antibody utilized to quantitate the protein with the commercially available ELISA kit. (C = cavity sample; CPB = cardiopulmonary bypass; ELISA = enzyme linked immunosorbent assay; kDa = kiloDaltons; MW = molecular weight; P = peripheral sample.) The Annals of Thoracic Surgery 2004 78, 46-52DOI: (10.1016/j.athoracsur.2003.11.042)

Fig 3 Left panels represent comparative 2D-PAGE scans of samples from cavity (top) and venous (bottom) collection during CPB surgery using Coomassie blue, total protein stain. The right panels display the same samples analyzed by Western blot technique for S100β protein (using Sangtec 100 kit antibodies). The circled and arrow markings indicate corresponding areas of varied protein detection. (2D-PAGE = two-dimensional polyacrylamide gel electrophoresis; CPB = cardiopulmonary bypass.) The Annals of Thoracic Surgery 2004 78, 46-52DOI: (10.1016/j.athoracsur.2003.11.042)