Main description:

Experimental techniques are the life blood of science. The better the methodology is, the more reliable and accurate the results will be. Ultimately, this will lead to a clearer interpretation of those results and firmer conclusions from any set of experiments. Experimental methodology in the area of cardiovascular biochemistry and molecular biology has advanced considerably in the last decade. Because of these factors, it was thought that a focused issue of Molecular and Cellular Biochemistry dedicated to the novel, latest technological advances in the field was warranted. We must thank Dr Naranjan S. Dhalla, Editor-in-Chief of Molecular and Cellular Biochemistry, for his willingness to publish an issue with such a focus. We have attracted some of the leaders in the field of cardiovascular biology to submit articles describing some of the most novel, significant techniques currently in use in their laboratories. The purpose of the manuscripts was not to describe the recent experimental findings from each laboratory as is done in most conventional manuscripts. Instead, the purpose of the articles found within this focused volume of Molecular and Cellular Biochemistry was to describe how the technique is performed on the laboratory bench so that others less familiar with the technique may be able to use it in their own labs. The subjects described in this volume can be generally subdivided into three categories: molecular biology, cell biology and basic biochemistry. The methods cover wide areas including various DNA and RNA expression technologies, transfection techniques, quantification of ion flux movement, measurements of lipid metabolism, advances in the culture of specific cardiovascular cell populations, and the use of confocal microscopy to examine cell structure and function. We thank all of the authors who have contributed so much of their time and efforts and, most importantly, shared the `secrets' of these valuable techniques with the rest of the cardiovascular research community.

Contents:

Preface. 1. DNA Transfer Into Vascular Smooth Muscle Using Fusigenic Sendai Virus (HVJ)-Liposomes; M.J. Mann, et al. 2. Adenovirus Mediated Gene Transfer Into Cardiomyocytes; L.A. Kirshenbaum, M.D. Schneider. 3. Retroviral Techniques for Studying Organogenesis, with a Focus on Heart Development; J. Hyer, T. Mikawa. 4. Novel Methods for Adenovirus-Mediated Gene Transfer to Blood Vessels in Vivo; H. Ooboshi, et al. 5. Use of a Hammerhead Ribozyme with Captionic Liposomes to Reduce Leukocyte type 12-lipoxygenase Expression in Vascular Smooth Muscle; Jia-Li, et al. 6. Immunodetection of Activated Mitogen-Activated Protein Kinase in Vascular Tissues; L. Yau, P. Zahradka. 7. Identification of Proteins that Interact with a Protein of Interest: Applications of the Yeast Two-Hybrid System; R.D. Gietz, et al. 8. Construction of a Human Heart CDNA Library and Identification of Cardiovascular Based Genes (CVBest); C.C. Liew, et al. 9. Competitive RT-PCR for Studying Gene Expression in Micro Biopsies; R. Hullin, et al. 10. The Nuclear Membrane Integrity Assay; M.P. Czubryt, et al. 11. Transfer of Macromolecules Into Living Adult Cardiomyocytes by Microinjection; M. Bartoli, W.C. Claycomb. 12. Differential Display: Identifying Genes Involved in Cardiomyocyte Proliferation; S.C. Regard, et al. 13. Analysis of Inositol Phosphates in Heart Tissue Using Anion-Exchange High-Performance Liquid Chromatography; E.A. Woodcock. 14. Analysis of Phospholipid Molecular Species; A. Vecchini, et al. 15. Measurements of Fatty Acid and Carbohydrate Metabolism inthe Isolated Working Rat Heart; G.D. Lopaschuk, R.L. Barr. 16. Fura-2 Fluorescent Technique for the Assessment of Ca2+ Homeostasis in Cardiomyocytes; Yan-Jun Xu, et al. 17. Spectroscopic Determination of Sarcoplasmic Reticulum Ca2+ Uptake and Ca2+ Release; J.S.C. Gilchrist, et al. 18. The Use of Confocal Microscopy in the Investigation of Cell Structure and Function in Heart, Vascular Endothelium and Smooth Muscle Cells; G. Bkaily, et al. 19. A Simple Method for Preparation of Cultured Cardiac Fibroblasts From Adult Human Ventricular Tissue; A.E. Agocha, M. Eghbali-Webb. 20. DNA Antisense Strategies in the Study of Receptors for Vasoactive Peptides, and of Growth and Wound-Healing Factors; P. D'Orleans-Juste. 21. Potent and Selective Gene Inhibition Using Antisense Oligodeoxynucleotides; W.M. Flanagan, R.W. Wagner. 22. Immunohistochemical Analysis of the Adaptation of Adult Guinea-Pig Cardiomyocytes in Long-Term Cultures and in Cocultures with Cardiac Neurons: A Novel Model for Studies of Myocardial Function; M. Horackova, et al.