1. Oxidative Stress and Nerve Function After Cardiopulmonary Bypass in Patients With Diabetes 
Robina Matyal, MD, Sruthi Sakamuri, BS, Thomas Huang, BA, Khurram Owais, MD, Samir Parikh, MD, Kamal Khabbaz, MD, Angela Wang, BA, Frank Sellke, MD, Feroze Mahmood, MD 
The Annals of Thoracic Surgery 
Volume 98, Issue 5, Pages (November 2014)
DOI: /j.athoracsur
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

2. Fig 1
Representative immunoblots of human atrial tissue for oxidative stress factors. (A) Immunoblotting quantification showed significant enhanced expression of CPT1-M in patients without diabetes compared with patients with controlled diabetes (CD) and patients with uncontrolled diabetes (UD) before cardioplegic cardiopulmonary bypass (CP/CPB). (B) ADIPOR1 was expressed significantly more in patients without diabetes compared with patients with UD in the atrial tissue before CP/CPB (Pre-CP/CPB). (C) Levels of MnSOD were significantly decreased in patients with UD after CP/CPB (Post-CP/CPB). (D) Nuclear factor kappa beta (NF-κβ) was insignificantly increased in patients with UD compared with patients without diabetes and patients with CD. (E) Total protein carbonyl content was significantly increased in patients with UD compared with patients with CD. (F) Representative Western blot bands of corresponding proteins. (G) Protein carbonyl content immunoblotting. Data are mean ± standard error of the mean (*p < 0.05 versus before CP/CPB; †p < 0.05 versus patients without diabetes before CP/CPB; ¶p < 0.05 versus patients with CD after CP/CPB). (CDPR = controlled diabetics pre-CP/CPB; CDPO = controlled diabetics post-CP/CPB; ND = no diabetes; NDPR = non-diabetics pre-CP/CPB; NDPO = non-diabetics post-CP/CPB.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

3. Fig 2
Representative immunoblots of human atrial tissue for nerve growth markers. (A) The levels of neurotrophin-3 (NT-3) were significantly lower in patients with uncontrolled diabetes (UD) compared with patients without diabetes (no diabetes [ND]) before atrial tissue samples obtained before cardioplegic cardiopulmonary bypass (Pre-CP/CPB). (B) The levels of nerve growth factor (NGF) were significantly decreased in patients with UD compared with patients with controlled diabetes (CD) and patients without diabetes after CP/CPB (Post-CP/CPB). (C) Levels of endothelin-1 (ET-1) before CP/CPB were significantly decreased in patients with CD and those with UD compared with patients without diabetes. In patients with CD, ET-1 levels after bypass were significantly higher than before bypass. (D) Representative immunoblotting bands for corresponding proteins are shown with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the loading control. (E) Representative immunohistochemistry slides costaining for neuron-specific protein gene product 9.5 (PGP9.5) (green) and nuclei with (4′6′-diamidino-2-phenylindole [DAPI]; blue) in atrial tissue (original magnification ×20). There was a marked increase in expression of PGP9.5 in patients without diabetes compared with patients with UD. Data are mean ± standard error of the mean (*p < 0.05 versus Pre-CP/CPB, †p < 0.05 versus Pre-CP/CPB ND; Øp < 0.05 versus Post-CP/CPB ND; ¶p < 0.05 versus Post-CP/CPB CD).
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

4. Fig 3
Representative immunoblots of human atrial tissue for angiogenic proteins. (A) Platelet-derived growth factor (PDGF)-β was significantly increased in patients without diabetes (no diabetes [ND]) compared with patients with undiagnosed diabetes (UD) in atrial tissue after cardioplegic cardiopulmonary bypass (Post-CP/CPB) and reperfusion. (B) Thrombospondin-2, an antiangiogenic protein, was not significantly decreased in patients with UD, but (C) angiostatin was significantly elevated in patients with UD in atrial tissue after CP/CPB and reperfusion. (D) Immunoblots of corresponding proteins are displayed. Data are mean ± standard error of the mean (*p < 0.05 versus Pre-CP/CPB; †p < 0.05 versus Pre-CP/CPB ND; ‡p < 0.05 versus Post-CP/CPB ND; Øp < 0.05 versus post-CP/CPB ND).
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

5. Fig 4
Representative immunohistochemistry slides for collagen staining. (A) Sirius red staining of myocardium for collagen levels from patients without diabetes (no diabetes [ND]), from patients with controlled diabetes (CD), and from patients with uncontrolled diabetes (UD) after CPB (original magnification ×5). (B) There is significantly increased fibrosis in the control group compared with the patients without diabetes as well as significantly increased fibrosis in atrial tissue from patients with UD compared with patients with CD. Data are mean ± standard error of the mean (Øp < 0.05 versus in patients without diabetes after cardioplegic cardiopulmonary bypass [Post-CP/CPB]).
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

6. Fig 5
Summary of the proposed mechanism for observed altered mitochondrial function, nerve expression, fibrosis, and angiogenesis in right atrial tissue from patients with uncontrolled chronic diabetes. Increased advanced glycosylation end products and hexosamine and polyol pathways cause impaired fatty acid metabolism. The use of glucose for sorbitol formation exhausts nicotinamide adenine dinucleotide phosphate (NADPH), a substrate for glutathione reductase, an enzyme that prevents oxidative stress. Similarly, there are decreased levels of antioxidant manganese superoxide dismutase (MnSOD). The increased reactive oxygen species lead to upregulation of inflammation and increased fibrosis in the cardiomyocytes. Importantly, there are decreased levels of nerve growth factor (NGF) and neurotrophin (NT)-3 (neurotrophins) from cardiomyocytes effecting nerve growth and differentiation angiogenesis. The inflammatory markers, decreased sympathetic nerves, and decreased levels of neurotrophins cause increased antiangiogenic markers (angiostatin) and decreases angiogenic protein platelet-derived growth factor (PDGF).
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions