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A Study of the Renal Tubular Absorption and Glycine and Histidine in Dogs Following the Intravenous Infusion of Concetrated Solutions of These Amino Acids Daniel M

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A Study of the Renal Tubular Absorption and Glycine and Histidine in Dogs Following the Intravenous Infusion of Concetrated Solutions of These Amino Acids Daniel M Yale University EliScholar – A Digital Platform for Scholarly Publishing at Yale Yale Medicine Thesis Digital Library School of Medicine 1960 A study of the renal tubular absorption and glycine and histidine in dogs following the intravenous infusion of concetrated solutions of these amino acids Daniel M. Jones Yale University Follow this and additional works at: http://elischolar.library.yale.edu/ymtdl Recommended Citation Jones, Daniel M., "A study of the renal tubular absorption and glycine and histidine in dogs following the intravenous infusion of concetrated solutions of these amino acids" (1960). Yale Medicine Thesis Digital Library. 2755. http://elischolar.library.yale.edu/ymtdl/2755 This Open Access Thesis is brought to you for free and open access by the School of Medicine at EliScholar – A Digital Platform for Scholarly Publishing at Yale. It has been accepted for inclusion in Yale Medicine Thesis Digital Library by an authorized administrator of EliScholar – A Digital Platform for Scholarly Publishing at Yale. For more information, please contact [email protected]. YALE MEDICAL LIBRARY Manuscript Theses Unpublished theses submitted for the Master's and Doctor's degrees and deposited in the Yale Medical Library are to be used only with due regard to the rights of the authors. Bibliographical references may be noted, but passages must not be copied without permission of the authors, and without proper credit being given in subsequent written or published work. This thesis by has been used by the following persons, whose signatures attest their acceptance of the above restrictions. NAME AND ADDRESS DATE Digitized by the Internet Archive in 2017 with funding from The National Endowment for the Humanities and the Arcadia Fund https://archive.org/details/studyofrenaltubuOOjone A STUDY OF THE RENAL TUBULAR ABSORPTION AND EXCRETION OF GLYCINE AND HISTIDINE IN DOGS FOLLOWING THE INTRAVENOUS INFUSION OF CONCENTRATED SOLUTIONS OF THESE AMINO ACIDS DANIEL M. JONES i> * A thesis submitted to the faculty of the Yale University School of Medicine in partial fulfillment of the requirements for the degree Doctor of Medicine DEPARTMENT OF PHYSIOLOGY I960 ( JUN 1960 ) x^libr^ y y i (3 r/A ACKNOWLEDGMENT Dr. Edward L. Alpen, U.S. Naval Radiological Defense Labora¬ tory, San Francisco, California, for his invaluable assis¬ tance and encouragement during the laboratory phase of this study. Dr. Daniel L. Kline, Department of Physiology, Yale Univer¬ sity School of Medicine, for his constructive criticism and guidance in the preparation of this thesis. ! . -• TABLE OF CONTENTS INTRODUCTION 1 STATEMENT OF THE PROBLEM 9 METHODS 11 Animals 11 Experimental Procedure 11 Analytical Methods 1$ Creatinine Analysis 15 Microbiological Amino Acid Assay 16 RESULTS 17 General Response to Glycine Perfusion 17 The Effect of Glycine Plasma Levels on Serine Plasma 21 Levels Factors Involved in Glycine Reabsorption 23 Histidine Infusion 2li DISCUSSION 26 Effect of Glycine and Histidine Tubular Loading on 27 Reabsorption Concurrent Infusion of Amino Acid Mixture and Glycine 28 SUMMARY 30 BIBLIOGRAPHY 31 FIGURES AND TABLES FIGURE 1 The relationship between glycine plasma level, 20 the plasma clearance of glycine by the kidney, and the filtered tubular load of glycine. FIGURE 2 Proportion of glycine reabsorbed at different 21 filtered load levels when infused alone and with a casein hydrolysate solution. FIGURE 3 The influence of glycine plasma level on the 22 plasma level of serine. FIGURE 1| Proportion of histidine reabsorbed at different 23 filtered load levels. TABLE I Glycine kidney clearance studies. 18 TABLE II Histidine kidney clearance studies. 19 '' c . A STUDY OF THE RENAL TUBULAR ABSORPTION AND EXCRETION OF GLYCINE AND HISTIDINE IN DOGS FOLLOWING THE INTRAVENOUS INFUSION OF CONCENTRATED SOLUTIONS OF THESE AMINO ACIDS Introduction and Historical Background The remarkable advances in renal physiology during the last 107 years since the publication of Bowman's monograph (9) on the structure of the glomeruli have been made possible by the formu¬ lation of a satisfactory theory of urine formation. Throughout the period of the first 80 years two principal schools of thought contended for supremacy. One represented the views of Bowman and Iieidenhain that tubular secretory activity accounted for the addition of the urinary solutes to water separated from blood by glomerular filtration, the only function of the water being to carry the solutes through the collecting tubules. The other theory was held by Carl Ludwig and his followers who maintained that an ultra-filtrate of plasma was formed at the glomeruli, and subsequent concentration of the urinary solutes was by water reabsorption in the tubules. In 1917 Cushny in a monograph en¬ titled, "The Secretion of the Urine", endeavored to unite the two conflicting views into a "modern theory" (8). He envisioned the formation of a plasma ultra-filtrate in the glomerulus, but rejected the idea of tubular secretion of solutes or reabsorption of water, and instead postulated the reabsorption by the tubules . -2- of an ideal solution approximating Locke’s Solution in compo¬ sition. It was not until 1923 when A.N. Richards and associates? applying micro-puncture techniques which permitted the sampl¬ ing and analysis of capsular and tubular fluids , that unequi¬ vocal evidence was obtained for glomerular ultra-filtration and tubular reabsorption and excretion. In a definitive paper published in 192H (29) they described their micro-puncture experiments. Since their pioneer work advanced the knowledge of renal physiology so markedly, it would be fitting to describe their ingenious methods in some detail. Essentially, the in situ kidney of a frog, Rana pipiens, was exposed so that trans¬ mitted light could be directed to the dorso-lateral surface, thus making the vascular structures clearly visible under the binocular microscope. A sharp-pointed quartz capillary pipette held in a micrometric apparatus was then manipulated under micro¬ scopic vision in order to pierce Bowman’s capsule and collect a sample of glomerular filtrate. After collection, the fluid was transferred to a long glass capillary tube which was then broken into sections and chemical determinations carried out on each section by various microtechniques. Walker and Oliver (28) performed a similar type of experi¬ ment in rats and guinea pigs in 19Ul. They collected fluid from the various parts of the tubule as well as the glomerulus. Be¬ cause of the fact that fluid was obtained from different parts of the nephron, it was necessary for them to identify the exact location from which fluid was withdrawn. This was accomplished by injecting the point from which fluid was collected with India * -3- ink at the close of the experiment, fixing the kidney in appro¬ priate solutions and then teasing out the entire nephron under a microscope so as to identify the exact site from which fluid had been collected. From the results of these and many other experiments it is now realized that the remarkedly constant composition of the plasma in all higher animals is the product of the kidney's regu¬ latory function of excreting a urine the composition of which is adjusted so as to keep the plasma composition constant within a narrow range. With the realization of function and the recognition that renal disease and nephrotoxins may differentially alter specific functions of the kidney have come the development of many tests to measure the degree and specific site of malfunction. The conception of comparing simultaneous determinations of a common substance in blood and urine as an index of general re¬ nal functional ability is attributed to a French clinician, N. Grehant. In 190U Grehant introduced the use of the urea concen¬ tration ratio, urine/blood, as an index of function (15>). Al¬ though this ratio gave an estimate when the urine flow rate was high enough and constant enough, generally these conditions did not prevail so this index did not find wide acceptance. Ambard and Weill (1) in 1912 first included both urine vol¬ ume and urea output in attempting quantitatively to relate urea excretion to blood concentration. Although the influence of urine volume was not directly investigated by the French physi¬ cians a correction for urine volume appeared in the formula they derived relating urea excretion to blood and urea concentration. , . ■ ■ . -u- Subsequent investigators, however, found a wide range of vari¬ ations in results calculated on normal subjects using the Ambard-Weill formula. D.D. Van Slyke and co-workers in 1921 showed that urinary urea excretion is independent of urine volume only when the urine flow rate is greater than two milliliters per minute (2). They called this minimal urine flow rate the augmentation limit. Their formulas, modifications of the Ambard-Weill formula, took urine flow rate into consideration and have found wide use in clearance determinations. The term "clearance” as used today was first introduced in a paper by Holler, McIntosh, and Van Slyke in 1928 (21) and was defined as, "the volume of blood which one minutes excretion suffices to clear of an excretory substance when the urine volume is large enough to permit maximum output." The formula developed was Cx - UV/B, where Cx = clearance of x in ml. per minute, U = concentration of x in the urine, V = urine output in ml. per min¬ ute, and B = concentration of x in the blood. This formula has served as the basis of all subsequent clearance studies. These clearance techniques, developed mainly by Homer Smith and associates, permitted the precise measurement of the renal functions of glomerular filtration rate, renal blood flow, and tubular absorptive and secretory capacity, thus making it feasible to define and assess quantitatively the various tubular functions. A detailed review of the historical development of modern clear¬ ance techniques can be found in Homer Smith's monograph on the kidney (25)» Briefly, these methods, based ultimately upon the Fick principle3 rely on the following theses.
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