The emphasis of this book is upon protocols, and if no other protocol books for molecular biology or gene therapy had been published, it would be an excellent value; however, many such books already exist. Even the idea of having a book that describes molecular techniques unique to the cardiovascular system is overplayed, because many of the protocols described here (and already published elsewhere) have been used for other systems and require little adaptation for the study of vascular disease.
Nevertheless, the authors of the individual chapters have made every effort to relate their particular techniques to the study of vascular tissues. To this end, the editor has assembled a broad base of well established authors to describe their protocols. Inevitably, there is much repetition of some procedures (descriptions of RT-PCR methodology, for example).
This book is presumably directed towards physician-scientists working in the vascular field and certainly will provide an excellent “recipe book” for laboratory technical staff—especially those without a lot of background knowledge—to use in carrying out a vast array of experiments. Each chapter begins with an introduction that explains the usefulness of the technique(s) to be described. These introductions vary in their degree of explicitness, but this is in keeping with the function of this book, which is simply to provide protocols that, if followed rigorously, will yield answers. To that extent, this volume comprises an excellent package of information for a clinical laboratory, where very often the principal investigator lacks the time (or detailed knowledge) to provide information for the technical staff.
The book is divided into 7 sections. The 1st of these deals with methods of studying the genetics of vascular disease (detection of mutations and polymorphisms, heteroduplex analysis of genetic variations, and mapping of smooth-muscle-restricted genes). The introductions to the 3 chapters in this section are succinct, but the protocols are very detailed. The 2nd section (7 chapters) covers mRNA isolation methodology and gene cloning, as well as variations of the techniques used for these purposes. This section concludes with an example of how these techniques enabled identification of specific protein kinases for vascular cells. For many clinical laboratories working on vascular disease processes, this may be the most useful section of the book.
Section 3 deals with transcriptional control of vascular cells and promoter activity, and section 4 covers mRNA analysis by such methods as Northern blotting, RT-PCR, in situ hybridization, and RNase protection assays. Including these techniques makes the book a complete package, but these methods (and several others) have been presented in excruciating detail in laboratory manuals for more than a decade and so could have been truncated here.
Section 5 contains methods related to apoptosis and phenotypic modulation of vascular cells. It is good that these topics are more than adequately covered because they have received much attention recently. Chapter 21 in this section provides detailed information on the use of two-dimensional electrophoresis to identify phenotypic specific proteins. The information provided on the use of protein (rather than nucleic acid) databases should be of value to someone venturing for the 1st time into the relatively new area called proteomics. It is a pity that more information is not presented here because this is going to be an important field of research for the physician-scientist in coming years. It is all very well to identify a piece of DNA that codes for something, and there are many examples of these finds; however, the function (or impaired function) of the discovered gene product (protein) is now becoming the key issue in our understanding of disease in general and vascular pathogenesis in particular. Admittedly, the field of proteomics is relatively new, but the stature of this book could have been considerably enhanced by providing more information regarding this emerging subject. Perhaps section 4 could have been reduced in length to make room for protocols involving work with proteins.
Sections 6 and 7 can be called the gene therapy sections, since they cover in vitro and in vivo gene transfer. This is the part of the book that focuses most specifically on vascular tissue because of the unique extracellular matrix that is associated with vascular cells and that influences their susceptibility to transfection. These sections emphasize viral mediated gene transfer, and information for these methods is provided in detail, together with some good examples of successful applications. Although a number of techniques for the introduction of foreign genes into tissues or cells are covered, at least 3 much-used methods for in vitro transfection are not mentioned: the use of starburst polydendrimer molecules (Super-Fect,TM for example), electroporation, and, most importantly, the Helios geneg-un. The latter has seen some application in vitro. In fact, there are claims suggesting that in animal models the use of the geneg-un yields transfection efficiencies comparable to those obtained with adenovirus gene transfer.
In this gene therapy section, the editor probably should have thought of including some information on the use of genetically modified cells as linings for cardiovascular prostheses. This is a rapidly expanding area of research, which started slowly about a decade ago. Perhaps this omission can be excused, since this research crosses disciplines to include work by bioengineers as well as clinical and basic scientists. The editor clearly could not cover every conceivable use of gene therapy. Even so, this is an area that is truly applicable to cardiovascular disease and that uses molecular biology and gene therapy protocols, the subject matter of this book.
In summary, it should be stressed that this is a good recipe book. As in many recipe books, there are sections that deal with relatively simple stuff (cooking potatoes) and sections that cover more sophisticated cuisine (Peking duck stuffed with pâté de foie gras?) such as the identification of novel protein kinases in vascular cells, as described in chapter 10. For the price, this book may prove worthwhile in helping to ease the load on the typical physician-scientist trying to care for patients and at the same time direct his or her laboratory research program.