Pilot Proteomic Profile of Differentially Regulated Proteins in Right Atrial Appendage Before and After Cardiac Surgery Using Cardioplegia and Cardiopulmonary Bypass

Abstract

Background-Although highly protective, cardiac surgery using cardioplegia and cardiopulmonary bypass (CP/CPB) subjects myocardium to hypothermic reversible ischemic injury that can impair cardiac function which results in a greatly enhanced risk of mortality. Acute changes in myocardial contractile activity are likely regulated via protein modifications. We performed the following study to determine changes in the protein profile of human myocardium following CP/CPB. Methods and Results-Right atrial appendage was collected from 8 male patients pre and post-CP/CPB. Atrial tissue lysates were subjected to 2-dimensional electrophoresis, total protein staining, gel averaging, and quantitative densitometry. Ten prominent spots regulated in response to CP/CPB were identified using mass spectrometry. Two hundred twenty-five and 256 protein spots were reliably detected in 2D-gels from pre-and post-CP/CPB patients, respectively. Five unique (ie, not detected post-CP/CPB) and 17 significantly increased spots were detected pre-CP/CPB. Thirty-four unique and 25 significantly increased spots were detected in the post-CP/CPB group. Identified proteins that changed after CP/CPB included: MLC-2a, ATP-synthase delta chain and Enoyl-CoenzymeA hydratase, glutathione-s-transferase omega, ␣-1-acid-glycoprotein, and phosphatidylethanolamine-binding protein. Conclusions-Cardiac

Microarray and proteomic analysis of the cardioprotective effects of cold blood cardioplegia in the mature and aged male and female

“…The importance of the mitochondrion and the mitochondrial proteome have been previously demonstrated; our data agree with the proteomic data of others who have shown changes in energy metabolism pathways following ischemia and reperfusion (6). In particular our data would agree with that of White et al (44) who have shown that ischemia-reperfusion in the New Zealand White rabbit results in changes in the proteome and includes changes in contractile fiber and energy production, similar to our findings.…”

p38 MAPK-dependent small HSP27 and αB-crystallin phosphorylation in regulation of myocardial function following cardioplegic arrest

“…SDS-PAGE and immunoblotting were performed using standard methodology as previously described (6). Antibodies for immunoblotting and confocal microscopy were as follows: phospho-specific and total HSP27, total HSP25, and cryAB antibodies were obtained from Stressgen (Vancouver, BC); phosphoand total p38 MAPK were obtained from Cell Signaling (Danvers, MA), and anti-MLC-2v was obtained from Synaptic Systems (Göt-tingen, Germany).…”

Isolation of Mitochondrial DNA from Single, Short Hairs without Roots Using Pressure Cycling Technology

“…6,7 Several groups have shown that PCT increased both the amount and number of extracted proteins from cells. [8][9][10][11] The utility of PCT for extracting DNA from lowtemplate forensic samples, 12,13 including hair, 14,15 bone fragments, 16 skin and blood stains, 15 sexual assault samples, 17 and fecal matter, 18 has also received some attention, due to the foreseeable advantages of an automated approach for reducing casework backlogs and potentially improving DNA recovery. In addition to reducing backlogs, using PCT may decrease the risk of cross-contamination and sample mix-ups, especially in cases where hairs submitted for DNA analysis arrive at the laboratory on an item of clothing.…”