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Figure 3: Long-Term, High-Dose Treatment of 24-Week-Old

Helena M. Viola, Victoria P.A. Johnstone, Abbie M. Adams, Susan Fletcher & Livia C. Hool
(A) Representative ratiometric JC-1 fluorescence recorded in myocytes from a PMO-treated mouse and an untreated mdx mouse before and after exposure to 10 μM BayK(–) ± 15 μM nisoldipine (Nisol) under calcium-free conditions (0 mM Ca2+). Arrow indicates addition of drugs. (B) Mean ± SEM of JC-1 fluorescence for all myocytes exposed to drugs as indicated. (C) Representative traces of flavoprotein fluorescence recorded in myocytes from a PMO-treated mouse and an untreated mdx mouse before...

Figure 4: HFpEF Is Aggravated in Cardiac Myocyte-Specific Fstl1 KO Mice

, María Valero-Muñoz, Richard M. Wilson, Eric E. Essick, Conor T. Fowler, Kazuto Nakamura, Maurice Van Den Hoff, Noriyuki Ouchi & Flora Sam
(A) Tail cuff systolic blood pressure (BP) recorded weekly for 4 weeks in wild-type (WT) and cFslt1-KO mice infused with aldosterone (HFpEF) or saline (Sham) (n = 10-12 mice per group). (B) Heart weight/body weight ratio and (C) wet lung/dry lung ratio in WT and cardiac myocyte-specific Fstl1 knockout (cFslt1-KO) mice (n = 5 to 10 mice/group). Diastolic function assessed 4 weeks after aldosterone (HFpEF) or saline (Sham) infusion in WT and cFslt1-KO mice (n...

Figure 6: In NRVMs, Dabuzalgron Regulates Activators of Apoptosis and Protects Mitochondrial Membrane Potential After Treatment With DOX

Ju Youn Beak, Wei Huang, Joel S. Parker, Sean T. Hicks, Cam Patterson, Paul C. Simpson, , Jian Jin & Brian C. Jensen
NRVMs were treated for 4 h with doxorubicin 2 μmol/l in the presence and absence of dabuzalgron 10 μmol/l. (A and B) Mitochondrial membrane potential was assessed using JC-1, and fluorescent intensity was quantified using a plate reader. Red indicates intact mitochondrial membrane potential; green indicates compromised mitochondrial membrane potential. Representative images (A) and summary findings (B) are presented. (C) NRVM lysates were blotted for selected regulators of apoptosis and mitochondrial cell death effectors. Representative...

Visual Abstract - Intracoronary Poloxamer 188 Prevents Reperfusion Injury in a Porcine Model of ST-Segment Elevation Myocardial Infarction: 10.1016/j.jacbts.2016.04.001

Jason A. Bartos, Timothy R. Matsuura, Adamantios Tsangaris, Matthew Olson, Scott H. McKnite, Jennifer N. Rees, Karen Haman, , Matthias L. Riess, Frank S. Bates, Joseph M. Metzger &
• STEMI remains a significant cause of in-hospital mortality, and up to 20% of people go on to develop heart failure.
• P188 is a nonionic triblock copolymer believed to prevent cellular injury after ischemia and reperfusion. The CORE trial examined P188 for STEMI patients showing no benefit when it was infused through a peripheral IV catheter approximately 30 min after revascularization with thrombolytic therapy.
• STEMI was induced in pigs using endovascular coronary artery occlusion to...

Visual Abstract -Adenosine Receptor Activation in the “Trigger” Limb of Remote Pre-Conditioning Mediates Human Endothelial Conditioning and Release of Circulating Cardioprotective Factor(s): 10.1016/j.jacbts.2016.06.002

Hussain Contractor, Rasmus Haarup Lie, Colin Cunnington, Jing Li, Nicolaj B. Støttrup, Cedric Manlhiot, Hans Erik Bøtker, Michael R. Schmidt, , , Andrew Redington & Rajesh K. Kharbanda
• Pre-conditioning has emerged as a potentially powerful means of reducing ischemia-reperfusion injury.
• Several animal models have implicated adenosine in pre-conditioning pathways, but its role in human physiology is unknown.
• In human volunteers, the authors demonstrate that adenosine receptor activation in “trigger” tissue is an important step in initiating a pre-conditioning signal, but adenosine receptor blockade in “target” tissue does not block the protection afforded by pre-conditioning.
• The authors also demonstrate that pre-conditioning elaborates a...

Visual Abstract - Serial Coronary Imaging of Early Atherosclerosis Development in Fast-Food-Fed Diabetic and Nondiabetic Swine: 10.1016/j.jacbts.2016.08.006

Nienke S. Van Ditzhuijzen, Mieke Van Den Heuvel, Oana Sorop, Alexia Rossi, Timothy Veldhof, Nico Bruining, Stefan Roest, Jurgen M.R. Ligthart, Karen Th. Witberg, Marcel L. Dijkshoorn, Koen Nieman, Monique T. Mulder, Felix Zijlstra, Dirk J. Duncker, Heleen M.M. Van Beusekom & Evelyn Regar
In swine with and without diabetes mellitus fed a fast-food diet:
• OCT, NIRS, CCTA, vascular function testing, and histology can be consecutively and longitudinally performed to assess gradual coronary atherosclerosis development
• OCT and NIRS enabled detailed assessment of early coronary atherosclerosis development, whereas CCTA was not able to detect discrete early atherosclerotic changes.
• OCT, NIRS, vascular function testing, and histology demonstrated no differences in early atherosclerosis development.

Visual Abstract - A Myocardial Slice Culture Model Reveals Alpha-1A-Adrenergic Receptor Signaling in the Human Heart: 10.1016/j.jacbts.2016.03.005

, , Patrick M. Cowley, , Megan D. Montgomery, Philip M. Swigart, Teresa De Marco, Anthony J. Baker & Paul C. Simpson
• Model for translational studies
• Human LV slices
• Simple, high throughput, viable
• Assays signaling and contraction
• Supports viral transduction
• Useful for proof-of-concept in man
• Shows the α1A-AR functions in human heart

Visual Abstract - Microtubule-Mediated Misregulation of Junctophilin-2 Underlies T-Tubule Disruptions and Calcium Mishandling in mdx Mice: 10.1016/j.jacbts.2016.02.002

Kurt W. Prins, Michelle L. Asp, , Wang Wang & Joseph M. Metzger
• Decreased junctophilin-2 levels are associated with cardiac t-tubule derangements in mdx mice, the mouse model of Duchenne muscular dystrophy (DMD).
• Reduced junctophilin-2 protein levels correlate with increases in total microtubule content in mdx hearts.
• Colchicine-mediated microtubule depolymerization increases junctophilin-2 protein levels and improves localization patterns which, in turn, are associated with t-tubule reorganization and reduced calcium sparks.
• This study identifies microtubule-mediated misregulation of junctophilin-2 as a novel molecular mechanism in Duchenne cardiomyopathy.

Visual Abstract - Chemotherapeutic Efficacy of Phosphodiesterase Inhibitors in Chagasic Cardiomyopathy: 10.1016/j.jacbts.2016.04.005

Jian-Jun Wen, , John Thacker & Nisha Jain Garg
• Mice infected with T. cruzi control acute parasitemia but develop chronic chagasic cardiomyopathy.
• Treatment with SIL (a phosphodiesterase inhibitor) during a therapeutic window of indeterminate phase provided powerful cardioprotective effects against chronic development of cardiomyopathy and LV dysfunction.
• SIL normalized the cGMP-dependent protein kinase activity and mitochondrial oxidative metabolism, and established the oxidant/antioxidant balance in chagasic myocardium.
• SIL prevented the oxidative/inflammatory adducts that precipitate cardiomyocytes death and cardiac remodeling in CCM.

Visual Abstract - Real-Time Magnetic Resonance Imaging Guidance Improves the Diagnostic Yield of Endomyocardial Biopsy: 10.1016/j.jacbts.2016.05.007

Toby Rogers, , , Adrienne E. Campbell-Washburn, William H. Schenke, Jonathan R. Mazal, , Anthony Z. Faranesh & Robert J. Lederman
• The diagnostic yield of endomyocardial biopsy is low, particularly in diseases that affect the myocardium in a nonuniform distribution.
• It is feasible to perform real-time MRI-guided endomyocardial biopsy using a dedicated active-visualization bioptome.
• In an animal model of focal myocardial disease, real-time MRI guidance improved the diagnostic yield compared with x-ray fluoroscopy.

Visual Abstract - Pre-Operative Left Ventricular Torsion, QRS Width/CRT, and Post-Mitral Surgery Outcomes in Patients With Nonischemic, Chronic, Severe Secondary Mitral Regurgitation: 10.1016/j.jacbts.2016.04.006

Yuichi Notomi, Tadashi Isomura, Shunichi Kanai, Masami Maeda, Joji Hoshino, Taichi Kondo, Yasuhisa Fukada & Koji Furukawa
• Determining which patients with NICSMR will benefit from MS is a clinical dilemma.
• LV torsion (which is a shear strain, not volume strain such as ejection fraction and originates in LV myocardial architectures) may reveal the myopathic conditions and reflect intra-LV electrical conduction.
• The LV torsional profile predicted post-MS outcomes in NICSMR patients with a narrow QRS but not in those with a wide QRS.
• The findings may help to resolve the clinical dilemma...

Visual Abstract - A Transcriptomic and Epigenomic Comparison of Fetal and Adult Human Cardiac Fibroblasts Reveals Novel Key Transcription Factors in Adult Cardiac Fibroblasts: 10.1016/j.jacbts.2016.07.007

Malin K.B. Jonsson, Robin J.G. Hartman, Matthew Ackers-Johnson, Wilson L.W. Tan, Bing Lim, Toon A.B. Van Veen & Roger S. Foo
• The interplay between cardiomyocytes and cardiac fibroblasts is increasingly being recognized as important in cardiac disease.
• Fetal and adult cardiac fibroblasts influence their neighboring cardiomyocytes in different ways. A genome-wide comparison of the 2 reveals that they share >80% of gene transcripts.
• Motif analysis of empirical regulatory elements located next to differentially expressed genes led to identification of key differential regulators of fibroblast identity.
• STAT1 and PLAGL1 were identified and validated as key transcription...

Visual Abstract - Serial Coronary Imaging of Early Atherosclerosis Development in Fast-Food-Fed Diabetic and Nondiabetic Swine: 10.1016/j.jacbts.2016.08.006

Nienke S. Van Ditzhuijzen, Mieke Van Den Heuvel, Oana Sorop, Alexia Rossi, Timothy Veldhof, Nico Bruining, Stefan Roest, Jurgen M.R. Ligthart, Karen Th. Witberg, Marcel L. Dijkshoorn, Koen Nieman, Monique T. Mulder, Felix Zijlstra, Dirk J. Duncker, Heleen M.M. Van Beusekom & Evelyn Regar
In swine with and without diabetes mellitus fed a fast-food diet:
• OCT, NIRS, CCTA, vascular function testing, and histology can be consecutively and longitudinally performed to assess gradual coronary atherosclerosis development
• OCT and NIRS enabled detailed assessment of early coronary atherosclerosis development, whereas CCTA was not able to detect discrete early atherosclerotic changes.
• OCT, NIRS, vascular function testing, and histology demonstrated no differences in early atherosclerosis development.

Figure 5: CTGF mAb Activates JNK and Reduces Collagen Production in Cultured Human Cardiac Fibroblasts

Laura E. Vainio, Zoltán Szabó, , Johanna Ulvila, Raisa Yrjölä, Tarja Alakoski, Jarkko Piuhola, Walter J. Koch, Heikki Ruskoaho, Shaun D. Fouse, Todd W. Seeley, , Pierre Signore, Kenneth E. Lipson, Johanna Magga & Risto Kerkelä
Cultured human fibroblasts were treated with 10 μg/ml CTGF mAb or control IgG, and co-treated with TGF-β1 (1 ng/ml) or inhibitor of JNK inhibitor (JNKi) [(L)-Form, 2 μM)], where indicated. (A) Western blot analysis for phosphorylated JNK2, p-ERK, phosphorylated SMAD2, phosphorylated focal adhesion kinase (p-FAK), smooth muscle alpha actin (α-SMA), and collagen 1 (Col1) is shown. Vinculin was used as loading control. (B) Quantitative analysis for collagen production and fibroblast proliferation. Data are presented as...

Figure 3: CTGF mAb Protects Against Post-MI LV Hypertrophy and Fibrosis (Study II)

Laura E. Vainio, Zoltán Szabó, , Johanna Ulvila, Raisa Yrjölä, Tarja Alakoski, Jarkko Piuhola, Walter J. Koch, Heikki Ruskoaho, Shaun D. Fouse, Todd W. Seeley, , Pierre Signore, Kenneth E. Lipson, Johanna Magga & Risto Kerkelä
Mice were subjected to MI and 1 week after surgery randomly divided to receive IgG vehicle or CTGF mAb for 6 weeks. (A) Analysis for heart weight to body weight (HW/BW) ratio, LV mass, left atrial end-diastolic area (LAA;d), and EF. (B) Analysis of cardiomyocyte cross-sectional area from Masson trichrome−stained LV sections. (C) Analysis of mean capillary cross-sectional size and the number of capillaries per cardiomyocyte in the nonischemic myocardium from CD31 staining, (D) Analysis...

Figure 4: Acute Overexpression of S16D-Hsp20 in Cardiomyocytes Promotes Upregulation of the

George T. Gardner, Joshua G. Travers, Jiang Qian, , Kobra Haghighi, Nathan Robbins, Min Jiang, , Guo-Chang Fan, Jack Rubinstein, & Evangelia G. Kranias
(A) Assessment of the following cytokine gene expression levels in Ad.GFP and Ad.S16D cardiomyocytes Ccl2, Ccl3, TNF-α, and IL-6. TGFβ1 and IL-1β were not detectable in either group. Values were normalized to those of the internal control, 18 S, and expressed as fold-changes relative to those of Ad.GFP. (B) Quantification of IL-6 secretion into the conditioned medium from Ad.GFP and Ad.S16D cardiomyocytes was determined by using an ELISA assay. The number of samples (n) per...

Figure 4: Acute Overexpression of S16D-Hsp20 in Cardiomyocytes Promotes Upregulation of the

George T. Gardner, Joshua G. Travers, Jiang Qian, , Kobra Haghighi, Nathan Robbins, Min Jiang, , Guo-Chang Fan, Jack Rubinstein, & Evangelia G. Kranias
(A) Assessment of the following cytokine gene expression levels in Ad.GFP and Ad.S16D cardiomyocytes Ccl2, Ccl3, TNF-α, and IL-6. TGFβ1 and IL-1β were not detectable in either group. Values were normalized to those of the internal control, 18 S, and expressed as fold-changes relative to those of Ad.GFP. (B) Quantification of IL-6 secretion into the conditioned medium from Ad.GFP and Ad.S16D cardiomyocytes was determined by using an ELISA assay. The number of samples (n) per...

Figure 1: Renal Denervation Procedure and Histological Assessment

Song-Yan Liao, Zhe Zhen, Yuan Liu, , , Anita Tsang &
(A) Renal angiogram of left renal artery in anteroposterior view before renal denervation (RD). (B) A 5-F quadripolar multiple electrode array ablation catheter (EnligHTNTM, St. Jude Medical, St. Paul, Minnesota) that consists of an expandable basket with 4 electrodes was used for RD. (C) An example of acute gross histology after RD to show the placement of lesions inside the renal artery. (D) Percentage area of left ventricular infarct as measured by histological examination in...

Figure 3: Implantation of Bioactive IL-4 Surfaces in Subcutaneous Mouse Model

Richard P. Tan, Alex H.P. Chan, Simon Wei, Miguel Santos, Bob S.L. Lee, , Behnam Akhavan, Marcela M. Bilek, Martin K.C. Ng, Yin Xiao & Steven G. Wise
(A) Experimental design schematic. (B) Immunohistological quantification of macrophages at the implant surface: total CD68+ macrophages (top), CD206+ M2 polarization (middle), and M2/M1 (CD206+/ major histocompatibility complex (MHC) Class II+) ratio (bottom). (C) Representative images of M2 (top row) and M1 (bottom row) macrophages. CD68 stained in green, CD206 stained in orange, and MHC Class II stained in purple. Dotted lines represent the interface between tissue (above) and implant (below). Scale bar represents 60μm; n...

Figure 2: Blockade of the CD47-SIRP-1α Pathway Specifically Enhances CM Phagocytosis by Mϕs in Culture

Shuang Zhang, Xin-Yi Yeap, Matthew DeBerge, , Kevin Wang, , Jane E. Wilcox, Steven M. White, John P. Morrow, Paul W. Burridge, Daniel Procissi, Evan A. Scott, William Frazier & Edward B. Thorp
(A) Cocultivations of macrophages (Mϕs) and dying adult mouse ventricular CMs with or without anti-CD47 (a47) versus isotype (C). The CMs were labeled with red R18 dye. The Mϕs were labelled with calcein-AM green. (Left) Scale bar, 300 μm. (Right) Scale bar is 20 μm. (B) Quantification of percent phagocytosis in CMs versus cardiofibroblasts. Zoom is 600%. *p = 0.01 relative to CM “c”. aCD47, blocking antibody; c, isotype control. (C) Percent phagocytosis of Cd47+/+...

Figure 2: Blockade of the CD47-SIRP-1α Pathway Specifically Enhances CM Phagocytosis by Mϕs in Culture

Shuang Zhang, Xin-Yi Yeap, Matthew DeBerge, , Kevin Wang, , Jane E. Wilcox, Steven M. White, John P. Morrow, Paul W. Burridge, Daniel Procissi, Evan A. Scott, William Frazier & Edward B. Thorp
(A) Cocultivations of macrophages (Mϕs) and dying adult mouse ventricular CMs with or without anti-CD47 (a47) versus isotype (C). The CMs were labeled with red R18 dye. The Mϕs were labelled with calcein-AM green. (Left) Scale bar, 300 μm. (Right) Scale bar is 20 μm. (B) Quantification of percent phagocytosis in CMs versus cardiofibroblasts. Zoom is 600%. *p = 0.01 relative to CM “c”. aCD47, blocking antibody; c, isotype control. (C) Percent phagocytosis of Cd47+/+...

Figure 2: Retrospective Cohort Study Design

Meghan A. Bowler, Michael A. Raddatz, Camryn L. Johnson, Brian R. Lindman &
The retrospective clinical analysis described here was designed based on the approval and clinical trial history of celecoxib. Celecoxib was approved on December 31, 1998, and clinical trial results revealed a potential cardiovascular risk in early 2005, defining our drug surveillance period. Body mass index (BMI) data were collected within 1 year of January 1, 2005. The aortic stenosis surveillance period extended from January 1, 2005, to January 27, 2018. APC = Adenoma Prevention With...

Figure 3: In Vivo CD47 Blockade Versus CD47 Induction Regulates CM Phagocytosis Efficiency and Cardiac Inflammation Resolution

Shuang Zhang, Xin-Yi Yeap, Matthew DeBerge, , Kevin Wang, , Jane E. Wilcox, Steven M. White, John P. Morrow, Paul W. Burridge, Daniel Procissi, Evan A. Scott, William Frazier & Edward B. Thorp
(A) Flow cytometric measure of Mϕ engulfment of transgenic mCherry CMs. Gating strategy is shown gating CD11b myeloid cells that are mCherry+ and Ly6g low (non-neutrophil phagocytes or Mϕs). Mice were subjected to 45 minutes of ischemia followed by injection of isotype IgG control (ctrl) versus CD47 monoclonal antibodies in phosphate-buffered saline, just before unoccluding the left anterior descending coronary artery. Right image depicts sorted Mϕs with internalized CM mCherry. (B) Dot plots, histogram, and...

Figure 2: Retrospective Cohort Study Design

Meghan A. Bowler, Michael A. Raddatz, Camryn L. Johnson, Brian R. Lindman &
The retrospective clinical analysis described here was designed based on the approval and clinical trial history of celecoxib. Celecoxib was approved on December 31, 1998, and clinical trial results revealed a potential cardiovascular risk in early 2005, defining our drug surveillance period. Body mass index (BMI) data were collected within 1 year of January 1, 2005. The aortic stenosis surveillance period extended from January 1, 2005, to January 27, 2018. APC = Adenoma Prevention With...

Figure 3: In Vivo CD47 Blockade Versus CD47 Induction Regulates CM Phagocytosis Efficiency and Cardiac Inflammation Resolution

Shuang Zhang, Xin-Yi Yeap, Matthew DeBerge, , Kevin Wang, , Jane E. Wilcox, Steven M. White, John P. Morrow, Paul W. Burridge, Daniel Procissi, Evan A. Scott, William Frazier & Edward B. Thorp
(A) Flow cytometric measure of Mϕ engulfment of transgenic mCherry CMs. Gating strategy is shown gating CD11b myeloid cells that are mCherry+ and Ly6g low (non-neutrophil phagocytes or Mϕs). Mice were subjected to 45 minutes of ischemia followed by injection of isotype IgG control (ctrl) versus CD47 monoclonal antibodies in phosphate-buffered saline, just before unoccluding the left anterior descending coronary artery. Right image depicts sorted Mϕs with internalized CM mCherry. (B) Dot plots, histogram, and...

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