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<p><i>List of Contributors ix</i></p> <p><i>Preface xix</i></p> <p><i>About the Companion Website xx</i></p> <p><b>Part 1 Electrophysiology</b></p> <p>1 Measurement of calcium transient ex vivo 3<br /> Kenneth R. Laurita and Bradley N. Plummer</p> <p>2 Confocal imaging of intracellular calcium cycling in isolated cardiac myocytes 12<br /> Søren Grubb, J. Andrew Wasserstrom, and Gary L. Aistrup</p> <p>3 Generating a large animal model of persistent atrial fi brillation 20<br /> Raphaël P. Martins and José Jalife</p> <p>4 Confocal imaging of intracellular calcium cycling in the intact heart 32<br /> Neha Singh, Manvinder Kumar, James E. Kelly, Gary L. Aistrup, and J. Andrew Wasserstrom</p> <p>5 Recording and measurement of action potentials 41<br /> Kenneth S. Ginsburg and Donald M. Bers</p> <p>6 Patch-clamp recordings from isolated cardiac myocytes 50<br /> Kai-Chien Yang, Wei Wang, and Jeanne M. Nerbonne</p> <p>7 Optical mapping of the heart 60<br /> Di Lang, Sarah Gutbrod, Jacob Laughner, and Igor R. Efi mov</p> <p><b>Part 2 Isolation and Maintenance of Primary Stem Cells</b></p> <p>8 Isolation of colony-forming endothelial progenitor cells 71<br /> Haruchika Masuda and Takayuki Asahara</p> <p>9 Cardiac resident stem cells 87<br /> João Ferreira-Martins, Fumihiro Sanada, and Marcello Rota</p> <p>10 Cardiospheres 95<br /> Rachel Ruckdeschel Smith</p> <p>11 Mesenchymal stem cells 104<br /> Jose S. Da Silva and Joshua M. Hare</p> <p>12 Generation and differentiation of human iPS cells 110<br /> Sebastian Diecke, Lei Ye, Sophia Zhang, and Jianyi Zhang</p> <p>13 Isolation of neonatal and adult rat cardiomyocytes 117<br /> Md. Abdur Razzaque and Jeffrey Robbins</p> <p>14 Isolation and culture of vascular smooth muscle cells 125<br /> Milton Hamblin, Lin Chang, and Y. Eugene Chen</p> <p>15 Isolation and culture of cardiac endothelial cells 131<br /> Asish K. Ghosh, Joseph W. Covington, and Douglas E. Vaughan</p> <p>16 Isolation and culture of cardiac fi broblasts 140<br /> Asish K. Ghosh, Joseph W. Covington and Douglas E. Vaughan</p> <p>17 Murine bone marrow transplantation model 146<br /> Prasanna Krishnamurthy, Suresh Kumar Verma, and Raj Kishore</p> <p>18 In vitro differentiation and expansion of vascular endothelial cells derived from mouse embryonic stem cells 149<br /> Anees Fatima, Carey Nassano-Miller, and Tsutomu Kume</p> <p><b>Part 3 Manipulation of the Heart and Vessels in Vivo and ex Vivo</b></p> <p>19 Coronary ligation 157<br /> Alexander R. Mackie and Hossein Ardehali</p> <p>20 Transverse aortic constriction: a model to study heart failure in small animals 164<br /> Suresh Kumar Verma, Prasanna Krishnamurthy, and Raj Kishore</p> <p>21 Pharmacological models of hypertrophy and failure 170<br /> Angela C. deAlmeida, Tariq Hamid, and Sumanth D. Prabhu</p> <p>22 Hindlimb ischemia 177<br /> Jerry C. Lee, Ngan F. Huang, and John P. Cooke</p> <p>23 The Langendorff preparation 187<br /> Hugh Clements-Jewery and Michael J. Curtis</p> <p>24 Myocarditis and other immunological models of cardiac disease 197<br /> Daniela C¡ iháková and Noel R. Rose</p> <p>25 Models of pacing-induced heart failure 203<br /> James A. Shuman, Rupak Mukherjee, and Francis G. Spinale</p> <p>26 Porcine myocardial ischemia models 215<br /> Xian-Liang Tang and Roberto Bolli</p> <p>27 Angiogenesis assays 225<br /> Susmita Sahoo and Douglas W. Losordo</p> <p>28 Immunohistochemical analysis of cardiac tissue 232<br /> Barbara Ogórek, Donato Cappetta, and Jan Kajstura</p> <p>29 A murine model of cardiac arrest by exsanguination 237<br /> Guangming Cheng, Yiru Guo, Harold K. Elias, Carrie M. Quinn, Arash Davani, Yanjuan Yang, Magdy Girgis, Roberto Bolli, and Buddhadeb Dawn</p> <p><b>Part 4 Small Animal Imaging</b></p> <p>30 Blood pressure, telemetry, and vascular measurements in the rodent model 245<br /> Robert S. Danziger</p> <p>31 The setting: imaging conscious, sedated, or anesthetized rodents 250<br /> Gene H. Kim and Roberto M. Lang</p> <p>32 Echocardiography: standard techniques (M-mode, two-dimensional imaging, and Doppler) 254<br /> Gene H. Kim, Lauren Beussink-Nelson, Sanjiv J. Shah, and Roberto M. Lang</p> <p>33 Echocardiography: advanced techniques (tissue Doppler, speckle tracking, and three-dimensional imaging) 275<br /> Lauren Beussink-Nelson, Gene H. Kim, Roberto M. Lang, and Sanjiv J. Shah</p> <p>34 In vivo tomographic cardiac imaging: positron emission tomography and magnetic resonance imaging 287<br /> Bruno C. Huber, Patricia K. Nguyen, and Joseph C. Wu</p> <p>35 In vivo hemodynamics 295<br /> Alexander R. Mackie, Kyle K. Henderson, Sol Misener, and Hossein Ardehali</p> <p><b>Part 5 Metabolism, Mitochondria, and Cell Death</b></p> <p>36 Fractionation of cardiomyocytes and isolation of mitochondria 307<br /> Christopher P. Baines</p> <p>37 Assessment of glucose and fatty acid metabolism ex vivo 313<br /> Darrell D. Belke and E. Dale Abel</p> <p>38 Quantification and characterization of atherosclerotic lesions in mice 320<br /> Abhinav Agarwal, Millicent G. Winner, Srinivas D. Sithu, and Sanjay Srivastava</p> <p>39 Assessment of cell death in the heart 332<br /> Russell S. Whelan, Klitos Konstantinidis, and Richard N. Kitsis</p> <p>40 Assessment of mitochondrial function in isolated cells 343<br /> Amy K. Rines and Hossein Ardehali</p> <p>41 Multinuclear NMR spectroscopy of myocardial energetics and substrate utilization in isolated perfused mouse hearts 351<br /> Stephen C. Kolwicz, Jr. and Rong Tian</p> <p>42 Measurement of reactive oxygen species in cardiovascular disease 359<br /> Mahmood Khan, Fatemat Hassan, Sashwati Roy, and Chandan K. Sen</p> <p>43 Assessing autophagy 371<br /> Roberta A. Gottlieb</p> <p>44 Assessment of cardiomyocyte size 378<br /> A. Martin Gerdes and Alessandro Pingitore</p> <p><b>Part 6 Manipulation of Gene Expression in Vitro and in Vivo</b></p> <p>45 Generation of Cre-loxP mouse models for conditional knockout and overexpression of genes in various heart cells 389<br /> Marisa Z. Jackson and Warren G. Tourtellotte</p> <p>46 Modulation of myocardial genes via use of adenoviral vectors and RNA interference approaches 397<br /> Qianhong Li</p> <p>47 Overexpression and downregulation of proteins in vitro 407<br /> Marina Bayeva and Hossein Ardehali</p> <p>48 In vivo microRNA studies 416<br /> Eva van Rooij</p> <p><b>Part 7 Model Systems</b></p> <p>49 Vascular and cardiac studies in zebrafi sh 425<br /> Hans-Georg Simon, Molly Ahrens, and Brandon Holtrup</p> <p>50 Vascular and cardiac studies in Drosophila 432<br /> Lin Yu, Joseph P. Daniels, and Matthew J. Wolf</p> <p><i>Index 441</i></p>

<p>Edited by<br /> <br /> <b>Hossein Ardehali MD, PhD, FAHA</b>, Associate Professor of Medicine, Molecular Pharmacology and Biological<br /> Chemistry<br /> Feinberg Cardiovascular Research Institute<br /> Northwestern University<br /> Feinberg School of<br /> Medicine, Chicago, IL USA</p> <p><b>Roberto Bolli MD</b><br /> Professor of Medicine Chief, Division of Cardiovascular Medicine<br /> Director, Institute of Molecular Cardiology<br /> University of Louisville<br /> Louisville, KY USA</p> <p><b>Douglas W. Losordo MD</b><br /> Vice President New Therapeutic Development<br /> Baxter Healthcare Corporation<br /> Deerfield, IL USA</p> <p> </p>

<p>While some aspects of clinical and scientific research are common across medicine, research in cardiovascular diseases requires knowledge of an increasing array of procedures, techniques, models and measurements that are highly specialized and unique to this area of investigation. Edited and written by senior investigators, many of whom have been involved in pioneering studies in cardiovascular disease, this new book provides today’s cardiology research professionals with practical how-to guidance on designing, conducting, measuring and reporting their research.</p> <p><i>Manual of Research Techniques in Cardiovascular Medicine</i>:</p> <p>• Reviews the must-know techniques, methods, analyses, models and measurements of cardiovascular research in a practical, “how-to-do-it” approach, resulting in clear-headed guidance to even the most complicated techniques and methods.</p> <p>• Covers each technique or method beginning with an explanation of when and why it should be selected<br /> followed by step-by-step instructions that include why each step is important; what can go wrong; how to trouble shoot and correct errors, and how to analyze and report results.</p> <p>• Helps ensure research success, but also, critically, enables readers to understand why and how some experiments fail.</p> <p>• Contains illustrative figures to reinforce the key components/steps of each method covered.</p> <p>• Includes a companion website with video clips to support the discussion of particularly challenging techniques, for example animal surgical procedures.<br /> <br /> <i>Manual of Research Techniques in Cardiovascular Medicine</i> is an invaluable resource for investigators at all levels, from technicians, students, fellows and other healthcare professionals who are approaching research for the first time, to seasoned investigators in cardiovascular disease, pharmacology, and related areas who need expert guidance on emerging techniques or unfamiliar models or methods.</p>

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