Life at the Cell and Below-Cell Level. The Hidden History of a Fundamental Revolution in Biology
Gilbert N. Ling, Ph.D.
Pacific Press
ISBN 0-9707322-0-1

"Dr. Ling is one of the most inventive biochemist I have ever met."
Prof. Albert Szent-Györgyi, Nobel Laureate

Chapter 9.

Troshin's Sorption Theory for Solute Distribution
(p. 43-46)

In 1956, A. S. Troshin published the Russian version of his monograph,90 which was to be translated into three other languages (German,91 Chinese, and English). The English version bears the title: "Problems of Cell Permeability."92 Interestingly enough, the contents of these monographs do not focus on what the title of the book tells us, permeability, which is a rate process. Instead, they deal primarily with solute distribution, which in the view of Troshin and others including myself, represents an equilibrium phenomenon. A possible interpretation of Troshin's choice of the title may be as follows: Troshin knew how the proponents of the membrane (pump) theorywho are by far the majority in this fieldmisinterpretated these equilibrium distribution problems as problems of membrane permeability, but he chose to refer to these underlying phenomenon as "permeability" problems to be in touch with a larger audience.

In the Preface to the original Russian edition and reprinted in the English edition of Troshin's book, Nasonov wrote: "We have come to the conclusion that this theory (the membrane theory) gives a completely false idea of the structure of the cell and the nature of the substance contained in the protoplasm. Furthermore, as a result of the apparent simplicity of the scheme it offers in explanation of the many problematic phenomena, the membrane theory has acquired great popularity among physiologists and has, in our opinion, induced them to follow a false line in their theoretical researches."92 p xiii

And this is how Troshin himself introduced his own book: "According to the theory developed by Lepeschkin, Nasonov and Fischer, and also certain other workers, the greater or lesser permeability of cell for any substance is to be explained not by the greater or lesser penetration of the substance through the cell membrane, but by the difference in the solubility of the substance in the protoplasm and the surrounding aqueous medium and by the adsorption or chemical binding by the cell colloids of the matter penetrating the wall."92 p 3

This self-effacing declaration notwithstanding, I believe that Troshin deserves the lion's share of credit for the "Sorption Theory" for solute distribution in living cells. It is true that Moore, Roaf, Fischer, Lepeschkin, Nasonov and others had pronounced these basic ideas earlierwhich is certainly of prime importance and each of these pioneers should be acknowledged for their prioritybut it was Troshin who transformed these ideas summarily expressed into quantitative data expressed in a rigorous equation form. To introduce Troshin's work, I shall begin with the earlier work of I. Ye. Kamnev.

In 1938 Kamnev published in the Russian-language journal, Archives of Anatomy, Histology and Embryology, an article entitled, "The permeability for sugars of striated frog muscle."34 This is a simple but defining paper. When frog muscles were immersed in a Ringer's solution containing sucrose or galactose, both sugars readily entered the muscle cells and eventually reached constant levels, which were below their respective levels in the bathing solutions (Figure 2). In dead cells, the level of sucrose and galactose rose to levels similar to those in the bathing medium.

Kamnev's conclusion that sucrose and galactose enter the muscle cells rests squarely upon the accuracy of his estimate that frog sartorius muscle has an extracellular space equal to 9% of its total muscle weight86 p 114 (and that sucrose or galactose found beyond what is contained in this extracellular space has entered the muscle cells). Since extracellular space as high as 35% has been reported in the literature,see 336 p 677 confirmation of the 9% figure is vital to establishing the validity of Kamnev's conclusion.

Work from my own laboratory supports Kamnev. Thus between 1967 and 1975, my associates and I obtained from five independent methods, four of which were entirely new (low concentration inulin probe method, 10.3%; poly-glutamate probe method, 8.9%; single fiber sucrose space method, 9%; Br86 efflux-analysis method, 8.2%; the centrifugation method, 9.4%)49 p 136 an average value of 9.2% 0.69% (mean S.D.) for the extracellular space of frog sartorius muscle, which lies close to Kamnev's 9% figure. My evidence that sucrose enters frog muscle cells as shown in Table 1 above support Kamnev's conclusion from a different angle.

Kamnev concluded that the steady level of sugar reached in the muscle cells does not depend on a membrane mechanism but on the solubility of these sugars in the muscle sarcoplasm, extending the idea Martin Fischer first suggested [7]. Kamnev expressed the belief that the sarcoplasm behaves as a phase with different solvent properties from water in the surrounding medium.

Ten years after Kamnev's publication, Troshin resumed this line of study, indicating that various other nonelectrolytes behaved just like galactose and sucrose in reaching steady levels in cell water lower than in the surrounding medium.93 It is the rapid and seemingly equal rates of entry of these solutes into frog muscle and other living cells {see [16.6(3.2)]}, which apparently led Troshin's mentor, Nasonov to the conclusion that there is no cell membrane.86 p 164 (As pointed out earlier, I do not agree with Nasonov on this specific point. I also believe that to no small measure is this disagreement a consequence of my access to the powerful radioactive-tracer technology, apparently unavailable at their times to Nasonov and Troshin.)

Following Bungenberg de Jong,321 Lepeschkin,324 Duclaux, Guilliermond, Oparin and others,824; 92 p 58 Troshin suggested that living cells are complex coacervates. Quoting work from Bungenberg de Jong's laboratory and his own, Troshin showed that, like water in living cells, the water in a simple gelatin coacervate accommodates various solutes at levels lower than that in the surrounding medium. Neither Kamnev nor Troshin offered a molecular explanation how water in living cells differs from normal liquid water, nor a molecular mechanism why sucrose, galactose have low solvency in cell water. Nor did they offer an explanation why urea and ethylene glycol distribute equally across the muscle cell surface [5.1].

On the role of metabolism on solute distribution, Troshin wrote: "Thanks to metabolism, the sorptional activity of the protoplasm is maintained in a definite level.....Upon breakdown of metabolism, this level changes: the solubility of substances in the protoplasm increases and the binding by cell colloids of some substances is depressed...."92 pp 373-374 No mechanism was offered why breakdown of metabolism leads to increased solubility and decreased binding. Four years earlier, Nasonov concluded his section on "phase theory of bioelectric potentials and cellular metabolism" in these words: "energy is evidently necessary for the maintenance of certain labile chemical compounds (including the structure of proteins)."86 p 202 But he too offered no explanation of how energy maintains the structure of proteins and other labile compounds.

 In harmony with the ideas first introduced by Martin Fischer (and partially also by Moore and Roaf), Troshin believed that solutes inside the cells in general fall into two categories: adsorbed (or otherwise bound) and dissolved in cell water. He then introduced a two-term equation (Equation Al in Appendix 1), containing a linear term representing solute dissolved in cell water following Henry's Law (or more correctly, in my opinion, the Berthelot-Nernst Distribution Law of which Henry's Law is a special case for solubility of gases in solvents13 pp 696-697), and a "hyperbolic adsorbed" fraction following Langmuir's adsorption isotherm.117 Troshin also showed how the equilibrium distribution of a variety of solutes in living cells as well as coacervate models can be described by the two-term equation. I have repeatedly suggested that this two-term equation be named "Troshin equation" in honor of the author, who died of cancer in 1982.173 p 20; 174 p 285

 After Troshin's death, the spirit and philosophy of protoplasm-oriented cell physiology at the Leningrad Institute of Cytology began to wane, flicker. That is, until Professor Vladimir Matveev, formerly at the Institute of Marine Biology, Academy of Science USSR at Vladivostok, arrived. Ever since he has been valiantly doing everything possible to bring that spirit and philosophy back again.554 However, to resume our story, we must turn back the clock to the year 1951 when young Troshin published a series of five short papers on solute distribution in coacervate models and in living cells.93 In the same year, I published a short article describing the essence of what was to be known as Ling's Fixed Charge Hypothesis.94

"Life at the Cell and Below-Cell Level.
The Hidden History of a Fundamental Revolution in Biology":

Contents (PDF 218 Kb)
Preface (
PDF 155 Kb)
Answers to Reader's Queries (Read First!) (
PDF 120 Kb)

1. How It Began on the Wrong Foot---Perhaps Inescapably
2. The Same Mistake Repeated in Cell Physiology
3. How the Membrane Theory Began
4. Evidence for a Cell Membrane Covering All Living Cells
5. Evidence for the Cell Content as a Dilute Solution
6. Colloid, the Brain Child of a Chemist
7. Legacy of the Nearly Forgotten Pioneers
8. Aftermath of the Rout
9. Troshin's Sorption Theory for Solute Distribution
10. Ling's Fixed Charge Hypothesis (LFCH)
11. The Polarized Multilayer Theory of Cell Water
12. The Membrane-Pump Theory and Grave Contradictions
13. The Physico-chemical Makeup of the Cell Membrane
14. The Living State: Electronic Mechanisms for its Maintenance and Control
15. Physiological Activities: Electronic Mechanisms and Their Control by ATP, Drugs, Hormones and Other Cardinal Adsorbents
16. Summary Plus
17. Epilogue 

A Super-Glossary
List of Abbreviations
List of Figures, Tables and Equations
References (
PDF 193 Kb)
Subject Index
About the Author

"Life at the Cell and Below-Cell Level..."
"Gilbert Ling"