Whole Organ Approaches to Cellular Metabolism

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Authors and discussants in the planning conference, Whole Organ Approaches to Cellular Metabolism, held at the Montreal General Hospital, July 14-16, 1995. Left to right, using informal names: Back row, standing: Nicole Siauve, Hans van Beek, Sasha Popel, Andi Deussen, Moise Bendayan, Jan Schnitzer, Eugenio Rasio, Tom Harris, Mel Silverman, Rick Haselton, Said Audi, Chris Dawson, Colin Rose, and Dick Effros. Front row, sitting: Fernando Vargas, Sandy Pang, Jim Bassingthwaighte, Francis Chinard, Carl Goresky, Jack Linehan, Andreas Schwab, Dick Weisiger, Harry Goldsmith. (Absent: Keith Kroll.)

James B. Bassingthwaighte Carl A. Goresky John H. Linehan Editors

Whole Organ Approaches to Cellular Metabolism Permeation, Cellular Uptake, and Product Formation

With 190 Illustrations

, Springer

James B. Bassingthwaighte Department of Bioengineering University of Washington Seattle, WA 98195, USA

John H. Linehan Biomedical Engineering Department Marquette University Milwaukee, WI 53233-1881,USA

Carl A. Goresky (deceased) formerly, Division of Gastroenterology Department of Medicine McGill University School of Medicine Montreal, Quebec H3G Canada

Library of Congress Cataloging-in-Publication Data Bassingthwaighte, James.

Whole organ approaches to cellular metabolism : permeation, cellular uptake, and product formation / James B. Bassingthwaighte, Carl A. Goresky, John H. Linehan.

p. cm. Includes bibliographical references and index.

I. Metabolism. 2. Cell metabolism. 3. Endothelium. 4. Capillaries. 1. Goresky, Carl A., 1932-1996. II. Linehan, John H. III. Title. QPI7l.B37 1998 572',4-dc21 97-19015

Printed on acid-free paper.

© 1998 Springer-Verlag New York Inc. Softcover reprint of the hardcover 1st edition 1998 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Springer-Verlag New York, Inc., 175 Fifth Avenue, New York. NY 1001 0, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation. computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use of general descriptive names, trade names, trademarks, etc., in this publication, even if the former are not especially identified, is not to be taken as a sign that such names, as understood by the Trade Marks and Merchandise Marks Act, may accordingly be used freely by anyone.

Production managed by Terry Kornak; manufacturing supervised by Joe Quatela. Typeset by Princeton Editorial Associates, Scottsdale, AZ. and Roosevelt, NJ.

9 8 7 6 5 432 I ISBN-13: 978-1-4612-7449-0 e-ISBN-13: 978-1-4612-2184-5 DOl: 10.1007/ 978-1-4612-2184-5

Carl Arthur Goresky August 25th, 1932 to March 21st, 1996

Carl Goresky was the epitome of the physician-scientist, and even more. Two dozen scientists gathered at the Montreal General Hospital in July 1995 to give tribute to Carl's scientific contributions; they met in admiration, respect, and love for the man, rather than the symbol of science. They met to plan this book on the methods and approaches to making discoveries about cellular metabolism in the intact organ. This is part of the issue of carrying forward the information from genomics, proteomics, and molecular and cellular biology into physiological phenotyping and an understanding of the behavior of an intact organ and organ­ism. Such research can be undertaken only by studying intact systems, an ap­proach Carl pioneered and promoted.

Carl grew up in CastIegar, in the mountains of British Columbia, where his father was the town physician. Carl played the piano so well that he could have made a career of it; he climbed mountains, hunted, collected minerals, and worked as a stevedore on the Columbia River barges. At 16 he went to McGill, and by 22 had completed a B.Sc. and his M.D. As a part of a medical residency at Johns Hopkins Medical School he spent 2 years with Dr. Francis Chinard. Francis had pioneered the multiple indicator dilution technique for estimating solute transport and volumes of distribution (Chinard et aI., 1955). Carl brought the technology,

v

VI Carl Arthur Goresky

including a sample collecting system and many ideas, back to McGill, where he completed a Ph.D. His advisor was an encouraging, brilliant man, Arnold Burgen, whose policy was to give free reign to such a "student," which was just as well because Dr. Burgen left for Oxford before the thesis was complete.

The first part of the thesis was the hallmark 1963 paper (Goresky. 1963). It demonstrated that a set of solutes passing through the liver following simultane­ous bolus injection into the portal vein emerged into the hepatic vein in a charac­teristic way. The shapes of their outflow dilution curves were identical, relative to their mean transit time, and could be superimposed upon each other by scaling the time axis by their individual mean transit times. The observation that the curves superimposed defined all the solutes to be flow-limited in their exchange between blood and tissue: RBC, plasma protein, sucrose, sodium, and water. This concep­tual step was based on the deeper idea that the capillary-tissue exchange unit was axially distributed, not a lumped compartment or mixing chamber. These two ideas, coupled with Christian Crone's demonstration that the bolus injection tech­nique could be used to measure capillary permeability (Crone, 1963), set the stage for the use of the multiple indicator dilution technique to elucidate substrate transmembrane transport and intracellular metabolism. Carl's paper on sul­fobromophthalein published in 1964, the remainder of the thesis, did exactly that. A refinement of the analysis to correct for catheter delay was published the same year with Carl's first student Mel Silverman, who worked later with Francis Chinard.

Kenneth Zierler. Chinard's compatriot as an undergraduate and colleague as a faculty member at Hopkins, had watched Carl's development in Francis' labora­tory in 1958-59, and his excellent performance as chief medical resident the next year. As a reviewer of the 1964 papers for Circulation Research he saw the brilliance of these: "There was so much meat in it, so creative." Of the 1963 work he said, "Carl made at least three very important points in this paper, which was obviously technically meticulous." The first point concerned the axially dis­tributed geometry of the capillary, which Carl called a "linear two-compartment system," but which Ken preferred to call a linear two-component system to dis­tinguish it from the mixing chamber idea associated with the word compartment. His second point was Carl's simple diagram of the system of partial. rather than ordinary, differential equations. The third was the flow-limited behavior de­scribed above.

By "technically meticulous" I think Ken was referring not only to the experi­mental methods but also the methods of analysis. From his first paper onward, Carl used mathematical phrasing, and characterized the biology in terms of the parameters of a precisely hypothesized physiological system. The wealth of pa­pers that followed over 34 years had his mathematical mark upon them. Each advanced the field another step. The flow-limited transport idea applied to gasses carried by erythrocytes, the "red cell carriage effect" (Gore sky et al., 1975). The use of Michaelis-Menten expressions for saturable transformation appeared in the 1964 papers. Crone demonstrated this for transport across the brain capillary membrane barrier for glucose a year later (Crone, 1965).

Carl Arthur Goresky vii

The general, model-free mass balance expressions were laid out by Zierler (Meier and Zierler, 1954; Zierler, 1962a, 1962b), but Carl had developed the next stages through model-dependent analyses of the observations: (1) passive barrier limitation (Goresky et aI., 1970); (2) concentrative transport (Goresky et aI., 1973); (3) carrier-mediated transport (Silverman and Goresky, 1965); (4) intra­tissue diffusion (Goresky and Goldsmith, 1973); (5) intraorgan flow heterogeneity (Rose and Goresky, 1976); (6) transport limitations by two barriers in series (Rose et aI., 1977; Rose and Goresky, 1977); (7) reaction via intracellular enzymes (Gore sky et aI., 1983); (8) receptor binding (Cousineau et aI., 1986); and (9) oxygen transport (Rose and Goresky, 1985).

As Carl unraveled the mysteries of increasingly complex systems, he main­tained the purity, even if not the simplicity, of the mathematics he used. He believed in finding the analytical solutions to the partial differential equations, and while getting advice from Glen Bach of the Department of Mechanical Engineering, fought his way through each new method of solution. He didn't really trust the accuracy of numerical methods, I suspect, or didn't feel that they offered so much benefit that mathematical elegance could be sacrificed. I like numerical methods for the freedom of concept that they offer, and for speed of solution, but these were secondary issues for him. Carl was strongly principled.

Carl maintained close relationships with many colleagues inside and outside of McGill over his career. Foremost among these were Francis Chinard, his early mentor, and Ken Zierler, Mel Silverman, Arnold Burgen, and others. My relation­ship with Carl began in 1960 when Carl came to the Mayo Clinic to see his classmate Andy Engel; Carl and I were both beginning our independent studies using indicator dilution methods. Thereafter we met regularly not only at scien­tific meetings but also at each other's homes and institutions, sharing our efforts to sort out what we didn't understand. Carl made everyone feel a partner in these explorations; while the average guru tells one how it is, Carl helped everyone to reason their way toward an answer.

Carl's qualities as a teacher were seldom equalled. He was patient, careful, and kind, and led the residents and fellows through a topic. The GI residents loved him; when he died in the Montreal General, they all came as a group to his bedside to pay their respects. But when presenting a new topic at a scientific meeting he didn't always think of himself as a teacher but as the presenter of the information, in all its glory. Some presentations were difficult for the general aUdience, though great for the cogniscenti; Carl was modest to a fault, in the sense that he seemed to think that everyone was as smart and quick as he was. At McGill and on many occasions elsewhere he was a magnificent teacher. One of the best lectures I have ever heard, Carl gave out of the blue; he was asked to explain indicator dilution methods to an evening meeting of the National Academy of Engineering in Washington, D.C. Knowing that the biology was unknown to his audience, but that quantitative approaches were known, he gave a most erudite comprehensive review of the concepts and applications in a half hour, with just chalk and blackboard.

Carl provided leadership in the medical sciences. He edited the journal Clinical

viii Carl Arthur Goresky

and Investigative Medicine throughout his last 12 years. He headed the Division of Gastroenterology at the two McGill hospitals, the Royal Victoria and the Montreal General, having brought their two gastroenterology divisions into the first merger between the two hospitals. His efforts in science and medicine were recognized for the impact he had on both. He received the Landis Award of the Microcirculatory Society, the Gold Medal of the Canadian Liver Foundation, the Distinguished Achievement Award of the American Association for the Study of Liver Diseases, and many others. In 1995 he was named officer of the Order of Canada, equivalent to a knighthood in the United Kingdom.

Behind him he leaves many colleagues who will carry on his efforts. Harry Goldsmith and Andreas Schwab, his close friends and colleagues in the research unit, Colin Rose in Cardiology, Phil Gold and Doug Kinnear in Medicine, all at the Montreal General, Eugenio Rasio and Moise Bendayan at the University of Montreal, Jocelyn Dupuis at the Montreal Heart Institute, Mel Silverman and Sandy Pang at the University of Toronto, and others scattered around the globe, continue, like myself, to learn from him and to build upon his ideas. Gone he may be, but never to be forgotten.

James B. Bassingthwaighte

References

Chinard, F. P., G. J. Vosburgh, and T. Enns. Transcapillary exchange of water and of other substances in certain organs of the dog. Am. J. Physiol. 183:221-234, 1955.

Crone, C. The permeability of capillaries in various organs as determined by the use of the "indicator diffusion" method. Acta Physiol. Scand. 58:292-305, 1963.

Crone, C. Facilitated transfer of glucose from blood into brain tissue. J. Physiol. 181: 103-113,1965.

Meier, P., and K. L. Zierler. On the theory of the indicator-dilution method for measurement of blood flow and volume. J. Appl. Physiol. 6:731-744, 1954.

Zierler, K. L. Circulation times and the theory of indicator-dilution methods for determin­ing blood flow and volume. In: Handbook of Physiology, Sect. 2: Circulation, Wash­ington, D.C.: American Physiological Society, 1962, pp. 585-615.

Zierler, K. L. Theoretical basis of indicator-dilution methods for measuring flow and volume. Circ. Res. 10:393-407, 1962.

Preface

The field of capillary-tissue exchange physiology has been galvanized twice in the past 25 years. A 1969 conference at the National Academy of Sciences in Copenhagen resulted in the book Capillary Permeability: The Transfer of Mole­cules and Ions Between the Capillary Blood and the Tissue (Crone and Lassen, 1970). It focused on the physiochemical aspects of transcapillary water and solute transport. The field has matured considerably since. This volume was designed as the successor to the 1970 book, and was created at a gathering of the authors at McGill University. It too captures the breadth of a field that has been dramatically enriched by numerous technical and conceptual advances. In 1970 it was already known that the capillary wall was not merely a "cellophane bag" exerting steric hindrances on solute particles. Instead, the endothelial surface was recognized as the site of binding reactions and permeation by passive or carrier-mediated trans­port. Furthermore, the cells of the blood could traverse evanescent wide openings in the "zippered" clefts. Today, research priorities have turned more to cell-cell interactions, toward understanding the utility of the gap junctional connections between endothelial cells and neighboring smooth muscle cells, neuronal twigs, and the parenchymal cells of organs. New discoveries in the past few years have revealed the critical importance of the close relationships between the endothelial cells and the parenchymal cells. Endothelial cell transporters, enzymes, and recep­tors play critical roles in substrate transport to the parenchymal cells of the organ, and in receptor-mediated responses related both to vasoregulation and to the functions of the parenchymal cells of the organ. Thus the focus has shifted away from permeation mechanisms and toward cellular metabolism.

This book brings together contributions from prominent researchers in the kinetics of blood-tissue exchange processes, in endothelial biochemistry and metabolism, and in cellular to whole body imaging, around the central theme of endothelial and parenchymal cellular function. The planning meeting "Whole Organ Approaches to Cellular Metabolism" was sponsored by the Commission on Bioengineering in Physiology of the International Union of Physiological Sci­ences, and supported generously by the Whitaker Foundation. Harry Goldsmith organized a setting conducive to group discussion at the Montreal General Hospi­tal. There was a focus on the interpretation of high-resolution data which provide

IX

x Preface

insight into cellular function using simulation analysis applied to physiological systems. This is the only workable approach for whole animal and human studies using nuclear magnetic resonance, positron emission tomography, and X-ray computed tomography-imaging modes that are well suited for acquisition of data in situations where modeling is essential to understanding of cellular func­tion. Examples are studies of cancerous growth processes, myocardial and cerebral ischemia, and the stages of recovery from injury. Positron emission tomography is particularly useful for examining the distribution of receptors or the dynamics of changing states of flow and metabolism. Noninvasive imaging methods are the key to the identification of the local densities of receptors and the assessment of their normal functions. The whole organ analytical approach pro­vides the mechanism for integrating knowledge from all of these areas and relat­ing them to a common set of underlying processes.

As this book was being brought together Carl Goresky died of renal adeno­carcinoma. He worked strenuously to the end, and on his last day worked on Chapter 1, the principles. The book is dedicated to his memory, to the many ideas he pioneered, and to the leadership he provided in science and medicine.

Another colleague has been lost just as his career was blossoming. Keith Kroll, who was born on December 9, 1948 and died on July 15, 1997, had the same spirit of perseverance and dedication as did Carl as he struggled with a devastatingly rapid progression of gastric adenocarcinoma. His last two years saw him emerge as a leader in the understanding of cellular energy balance in the heart.

Carl Goresky and Keith Kroll were determined, brilliant scholars, kindly teachers, and wonderful colleagues. While we try to follow in their footsteps, we cannot do what they would have done.

James B. Bassingthwaighte John H. Linehan

Contents

Preface IX

Contributors xv Introduction xix

1. Introduction 1. Modeling in the Analysis of the Processes of Uptake and

Metabolism in the Whole Organ 3 James B. Bassingthwaighte, Carl A. Goresky, and John H. Linehan

2. Mechanisms of Endothelial Transport, Exchange, and Regulation

2. Transport Functions of the Glycocalyx, Specific Proteins, and Caveolae in Endothelium 31 Jan E. Schnitzer

3. Study of Blood Capillary Permeability with the Rete Mirabile 71 Eugenio A. Rasio, Moise Bendayan, and Carl A. Goresky

4. Interactions Between Bovine Adrenal Medulla Endothelial and Chromaffin Cells 91 Fernando F. Vargas, Soledad Calvo, Raul Vinet, and Eduardo Rojas

5. Studies of the Glomerular Filtration Barrier: Integration of Physiologic and Cell Biologic Experimental Approaches 109 Melvin Silverman

6. Endothelial Barrier Dynamics: Studies in a Cell-Column Model of the Microvasculature 135 Frederick R. Haselton

3. Metabolism in the Heart and Skeletal Muscle 7. Strategies for Uncovering the Kinetics of Nucleoside Transport and

Metabolism in Capillary Endothelial Cells 163 James B. Bassingthwaighte, Keith Kroll, Lisa M. Schwartz, Gary M. Raymond, and Richard B. King

Xl

xii Contents

8. Norepinephrine Kinetics in Normal and Failing Myocardium: The Importance of Distributed Modeling 189 Colin P. Rose

9. Metabolic Response Times: A Generalization of Indicator Dilution Theory Applied to Cardiac O2 Consumption Transients 205 Johannes HG.M. van Beek

10. Quantitative Assessment of Sites of Adenosine Production in the Heart 235 Andreas Deussen

11. Role of Capillary Endothelial Cells in Transport and Metabolism of Adenosine in the Heart: An Example of the Impact of Endothelial Cells on Measures of Metabolism 261 Keith Kroll and James B. Bassingthwaighte

12. Distribution of Intravascular and Extravascular Resistances to Oxygen Transport 277 Aleksander S. Popel, Tuhin K. Roy, and Abhijit Dutta

4. Metabolism in the Liver

13. Liver Cell Entry In Vivo and Enzymic Conversion 297 Carl A. Goresky, Glen G. Bach, Andreas J. Schwab, and K. Sandy Pang

14. Probing the Structure and Function of the Liver with the Multiple-Indicator Dilution Technique 325 K. Sandy Pang, Carl A. Goresky, Andreas J. Schwab, and Wanping Geng

15. A Generalized Mathematical Theory of the Multiple-Indicator Dilution Method 369 Andreas J. Schwab

16. Impact of Extracellular and Intracellular Diffusion on Hepatic Uptake Kinetics 389 Richard A. Weisiger

5. ~etabolism in the Lung 17. The Uptake and Metabolism of Substrates by Endothelium in the

Lung 427 John H Linehan, Said H Audi, and Christopher A. Dawson

18. Pulmonary Endothelial Surface Reductase Kinetics 439 Christopher A. Dawson, Robert D. Bongard, David L. Roerig, Marilyn P. Merker, Yoshiyuki Okamoto, Said H Audi, Lars E. Olson, Gary S. Krenz, and John H Linehan

19. Water and Small Solute Exchanges in the Lungs 455 Francis P. Chinard

Contents XIll

20. Pulmonary Perfusion and the Exchange of Water and Acid in the Lungs 469 Richard M. Ejfros, Julie Biller, Elizabeth Jacobs, and Gary S. Krenz

21. The Transport of Small Molecules Across the Microvascular Barrier as a Measure of Permeability and Functioning Exchange Area in the Normal and Acutely Injured Lung 495 Thomas R. Harris

22. Lipophilic Amines as Probes for Measurement of Lung Capillary Transport Function and Tissue Composition Using the Multiple-Indicator Dilution Method 517 Said H. Audi, John H. Linehan, Gary S. Krenz, David L. Roerig, Susan B. Ahlf, and Christopher A. Dawson

Publications of Carl A. Goresky 545 Index 557

Contributors

Susan B. Ahlf, Department of Veteran Affairs Medical Center, Milwaukee, WI 53295, USA

Said H. Audi, Research Service, Physiology, Veterans Administration Medical Center, Milwaukee, WI 53295, USA

Glen G. Bach, Department of Mechanical Engineering, McGill University, Montreal, Quebec, Canada

James B. Bassingthwaighte, Center for Bioengineering, University of Wash­ington, Seattle, WA 98195, USA

Moise Bendayan, Department of Anatomy, University of Montreal, Montreal, Quebec, Canada

Julie Biller, Department of Pulmonary and Critical Care, Medical College of Wis­consin, Milwaukee, WI 53226, USA

Robert D. Bongard, Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA

Soledad Calvo, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892

Francis P. Chinard, Department of Medicine, New Jersey Medical School, New­ark, NJ 07103-2714, USA

Christopher A. Dawson, Department of Physiology, Medical College of Wis­consin, and Veterans Administration Medical Center, Milwaukee, WI 53295-1000, USA

xv

xvi Contributors

Andreas Deussen, Institute fur Physiologie, Medizinische FakuItat Karl Gustav Karus, Technische Universitat Dresden, Dresden, D-01307, Germany

Abhijit Dutta, Fluent, Inc., Centerra Resource Park, Lebanon, NH 03766-1442, USA

Richard M. Effros, Department of Pulmonary and Critical Care Medicine, MCW Clinic at Froedtert, Milwaukee, WI 53226, USA

Wanping Geng, Bioavail Corporation International, Toronto, Ontario MIL 4S4, Canada

Carl A. Goresky, formerly Division of Gastroenterology, Department of Medi­cine, McGill University School of Medicine, Montreal, Quebec H3G, Canada

Thomas R. Harris, Biomedical Engineering Department, Vanderbilt University, Nashville, TN 37203, USA

Frederick R. Haselton, Department of Biomedical Engineering, Vanderbilt Uni­versity, Nashville, TN 37235, USA

Elizabeth Jacobs, Pulmonary Division, Medical College of Wisconsin, Mil­waukee, WI 53226, USA

Richard B. King, Department of Bioengineering, University of Washington, Seattle, WA 98195-7962, USA

Gary S. Krenz, Department of Mathematics, Statistics, and Computer Science, Marquette University, Milwaukee, WI 53201-1881, USA

Keith Kroll, formerly Center for Bioengineering, University of Washington, Seattle, WA 98195, USA

John H. Linehan, Biomedical Engineering Department, Marquette University, Milwaukee, WI 53233-1881, USA

Marilyn P. Merker, Departments of Anesthesiology and Pharmacology, Medical College of Wisconsin, Milwaukee, WI 53226, USA

Yoshiyuki Okamoto, Department of Chemistry, Polytechnic University, Brooklyn, NY 11201, USA

Lars E. Olson, Department of Biomedical Engineering, Marquette University, Milwaukee, WI 53201-1881, USA

K. Sandy Pang, Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 2S2, Canada

Contributors XVll

Aleksander S. Popel, Department of Biomedical Engineering, School of Medi­cine, Johns Hopkins University, Baltimore, MD 21205, USA

Eugenio A. Rasio, Department of Nutrition, Notre Dame Hospital, Montreal, Quebec H2L, Canada

Gary M. Raymond, Department of Bioengineering, University of Washington, Seattle, WA 98195-7962, USA

David L. Roerig, Departments of Anesthesiology and Pharmacology, Medical College of Wisconsin, Milwaukee, WI 53226, USA

Eduardo Rojas, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA

Colin P. Rose, University Medical Clinic, Montreal General Hospital, Montreal, Quebec H3G 1 A4, Canada

Tukin K. Roy, Departments of Anesthesiology and Critical Care Medicine, Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA

Jan E. Schnitzer, Department of Pathology, Harvard University Medical School, and Beth Israel Hospital, Boston, MA 02215, USA

Andreas J. Schwab, Department of Medicine, McGill University School of Medi­cine, and Montreal General Hospital, Montreal. Quebec H3G 1A4, Canada

Lisa M. Schwartz. Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA

Melvin Silverman, Department of Medicine, University of Toronto School of Medicine, Toronto, Ontario M5S lA8, Canada

Johannes H.G.M. van Beek, Laboratorium voor Fysiologie, Vrije Universiteit, 1081 BT Amsterdam, The Netherlands

Fernando F. Vargas, Department of Human Physiology, University of California Davis School of Medicine, Davis, CA 95616-8644, USA

Raul Vinet, Department of Pharmacology, Faculty of Medicine, University of Chile, Santiago, Chile

Richard A. Weisiger, Department of Medicine, University of California, San Fran­cisco, San Francisco, CA 94143-0538, USA

Introduction

Whole Organ Approaches to Cellular Metabolism are based on making obser­vations via a variety of techniques at varied resolution. Whole organ data are interpreted in terms of the structures and behavior of tissues: cells of different types, subcellular structures and processes, and the physical chemistry of molecu­lar motions, reactions, and surface phenomena. Often one obtains some data at the suborgan level to aid in the process. This book is structured so as to give some insight first into the general theory of mass balance and conservation principles, then into the more biophysical and molecular aspects of the field, and finally into a succession of applications to various organs. There is no attempt to provide complete coverage of the organs of the body or of full ranges of solutes, sub­strates, hormones, or pharmaceuticals. The principles developed and illustrated should be adaptable to the study of any organ.

The book is divided into five sections, the first two of which cover the basic fundamentals and the general mechanisms involved in transport. The last three sections are focused on particular organ systems.

Section I provides a general background of the principles and practice of indicator dilution methods in the study of cellular metabolism. They are mainly based on mass balance: the expectation that what goes in is retained or comes out.

Section 2 concerns the physical chemistry of transport mechanisms: the interac­tion between convection, diffusion, permeation, and reaction. These five chapters provide background for the physiological behavior of endothelial cells in their interactions with cells in the blood, with smooth muscle cells, and with the organ's parenchymal cells. Research initiated in the 1940s and still vital includes influ­ences of the glycocalyx, of pH, surface charges, and of zeta potentials on the interactions of solutes and ions to surfaces, on the apparent affinity of receptors, and on the asymmetries of transport rates. Electrophysiology of membrane chan­nels and cell-to-cell conductance of small solutes are topics related to broader phenomena such as calcium cycling. Shear-dependent channel activation of NO release illustrates how endothelial cells can communicate with others.

Section 3 focuses in the first five chapters on the role of endothelial cellular biochemistry in cardiac metabolism. Intraendothelial reactions can have re-

xix

xx Introduction

markable influences on solute fluxes. A classic example is the finding that after an isolated heart is perfused with solution containing tracer adenosine for 30 min­utes, more than 90% is to be found in endothelial cells, not in the myocytes (Sparks et aI., 1984). Why is the endothelial capacity for purine so high, when presumably it is the myocytes that have the high ATP turnover? Although this section uses purine handling as an example of the kind of interactions that will be found for other solutes and other signalling pathways, the relationship between purine and energy production from oxygen utilization is of prime importance in cardiac research. The final chapter of this section (Chapter 12) details the theory and experimental results on oxygen transport and metabolism, and although it emphasizes events in skeletal muscle, elucidates the processes occurring in all organs.

Section 4, on the liver, exemplifies how one may examine cellular metabolism in vivo. The absence of a hindering endothelial barrier in the sinusoid facilitates the interpretation of metabolic transformations inside hepatocytes. These chapters range over normal metabolic and pharmacokinetic processes, and into the intra­cellular diffusional processes that must playa role in the liver's excretory func­tions. The multiple indicator dilution technique has been the key technology leading to enhanced understanding of hepatic function at the whole organ level. Weisiger's studies (Chapter 16), using optical methods to examine solute con­centrations at the cellular level, illustrate that the techniques of cell biology are essential in interpreting whole organ data to a more refined level.

Section 5, on the metabolic functions of the lung, illustrates the power of the multiple tracer indicator dilution approach to dissect events occurring along the pulmonary capillary endothelium, regions a fraction of a micrometer thick. New insight is provided into the complex processes of water transport, which underlie all those processes concerning solutes. Molecular interactions at the surfaces and composite processes occurring within the blood-to-air barrier are all explored to create new insight into barrier function.

It is not fortuitous that the applications are mainly in the heart, lung, and liver, for these are the organs studied most extensively. However, Chinard's pioneering contributions (e.g., Chinard et al., 1955, 1997) on the kidney, Crone's in the brain (Crone, 1963, 1965), Renkin's (Renkin, 1959a, 1959b) and Zierler's (Andres et aI., 1954; Meier and Zierler, 1954) in skeletal muscle, and Yudilevich's (Yudilevich and Martin de Julian, 1965; Yudilevich et aI., 1979) in the salivary gland demonstrated that the techniques of experimentation and analysis are gen­eral. Although the multiple indicator dilution techniques are most easily applied to organs with a single inflow and single outflow, they can be used in more complex organs with multiple inflows and outflows, as suggested by theory (Perl et aI., 1969), and applied to the interpretation of brain image sequences (Raichle et aI., 1978). For those who wish to determine the metabolic status of intact tissues and organs, this book provides a take-off point for the future.

James B. Bassingthwaighte

Introduction xxi

References

I. Andres, R., K. L. Zierler, H. M. Anderson, W. N. Stainsby, G. Cader, A. S. Ghrayyib, and 1. L. Lilienthal, Jr. Measurement of blood flow and volume in the forearm of man; with notes on the theory of indicator-dilution and on production of turbulence, hemo­lysis, and vasodilation by intra-vascular injection. 1. Clin. Invest. 33:482-504, 1954.

2. Chinard, F. P., G. 1. Vosburgh, and T. Enns. Transcapillary exchange of water and of other substances in certain organs of the dog. Am. 1. Physiol. 183:221-234, 1955.

3. Chinard, F. P., G. Basset, W. O. Cua, G. Saumon, F. Bouchonnet, R. A. Garrick, and V. Bower. Pulmonary distribution of iodoantipyrine: Temperature and lipid solubility effects. Am. J. Physiol. 272 (Heart Circ. Physio. 41):H2250-H2263, 1997.

4. Crone, C. The permeability of capillaries in various organs as determined by the use of the "indicator diffusion" method. Acta Physiol. Scand. 58:292-305. 1963.

5. Crone, C. Facilitated transfer of glucose from blood into brain tissue. J. Physiol. 181:103-113,1965.

6. Crone, C. and N. A. Lassen. Capillary Permeability: The Transfer of Molecules and Ions Between Capillary Blood and Tissues. Copenhagen: Munksgaard, 1970, 681 pp.

7. Meier, P., and K. L. Zierler. On the theory of the indicator-dilution method for measure­ment of blood flow and volume. 1. Appl. Physiol. 6:731-744, 1954.

8. Perl, w., R. M. Effros, and F. P. Chinard. Indicator equivalence theorem for input rates and regional masses in multi-inlet steady-state systems with partially labeled input. J. Theor. BioI. 25:297-316,1969.

9. Raichle, M. E., M. 1. WeIch, R. L. Grubb, C. S., Higgins, M. M. Ter-Pogossian, and K. B. Larson. Measurement of regional substrate utilization rates by emission tomogra­phy. Science 199:986-987, 1978.

10. Renkin, E. M. Transport of potassium-42 from blood to tissue in isolated mammalian skeletal muscles. Am. 1. Physiol. 197: 1205-1210, 1959a.

II. Renkin, E. M. Exchangeability of tissue potassium in skeletal muscle. Am. J. Physiol. 197:1211-I2I5,1959b.

12. Sparks, H. V. Jr., M. W. Gorman, F. L. Belloni, and B. Fuchs. Endothelial uptake of adenosine: implications for vascular control. The Peripheral Circulation. edited by S. Hunyor, 1. Ludbrook, 1. Shaw, and M. McGrath. Amsterdam: North Holland, 1984, pp.23-32.

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