Chapter 4

UCLA (1961-1996, 2001-2006), Animal Behavior, Biological Membranes, Water & Hydrophobic Bonding

Overview
My paper of 1962 did not conclude my publication of material in the Journal Ecology. There followed a detailed paper in 1963, including construction and operation of all devices employed, enclosure environment, descriptions of early behavior in the enclosure, and specific activities and adapted behavior.

Wheel-running studies were reported in Behaviour in 1963, the first of four papers published in this journal in the same series, itself unprecedented. The studies illustrated the potential usefulness of wheel-running for behavior studies, and cast a new light on this activity and on certain other aspects of the behavior of confined animals, topics already mentioned briefly in Chap. I.

It was emphasized that, taken alone, the nature of a specific stimulus or activity is an unreliable guide for interpreting the behavior of small mammals upon whom it is forced or presented unexpectedly. The same holds for behavior of confined animals given control over the initiation or termination of the stimulus or activity, except when great stress is involved.. Stimuli rewarding (or punishing) in certain circumstances, can have the opposite effect in others.

In his tentative acceptance of the paper for Behaviourin 1963, Editor W. H. Thorpe, a world renowned leading behaviorist at Cambridge U., wrote,

I have now looked through the manuscript "Compulsory Regime and Control of Environment in Animal Behavior" which you submitted for publication in Behaviour. I found it both original and stimulating and it seems to me that you have opened up a valuable new technique for the study of volitional behaviour in small mammals.....

A serendipitous event in 1961, led to studies occupying much of my attention for the next 5 or 6 years, and led to my writing 3 books. In a 1962 report in Science, "Countercurrent Streaming in Liquid Surfaces and Its Relevance to Protoplasmic Streaming," I remarked that: movements in surface films counter to the interior stream are brought about by film pressure generated by surface tension gradients. In a similar process, the formation of new interfaces during protoplasmic syntheses could sustain gradients of interfacial tension with resulting protoplasmic movements.

In 1962 and 1963, I followed up with papers on similar topics in other journals but, most notably, by a paper in Nature. Unfortunately, my proposed transition between membrane states was not the one Mother Nature had adopted. More than a few papers I have written generated numbers of reprint requests over 100, several over 200, but for the Nature paper there were a record 1,799 requests, 899 from the U.S. and 900 from abroad. The numbers provide unique data for assessing comparative research interest in biological membranes in 1963.

These studies led to my writing a book in 1965, "Structure and Function in Biological Membranes." This became unwieldy for a single volume, so it was divided into two. The chapter on the physical chemistry of water, however, was of sufficient interest to chemists, physicists, and other scientists to warrant publishing it in 1964 as a separate volume, "Water and Solute-Water Interactions." This was the first book in the English Language on the physical chemistry of water, and became my most cited work. Much very favorable correspondence concerning the books followed. Journal reviews also were very favorable, although there were justified caveats concerning my proposed "disc and pillar" membrane transformation.

In the next few years, I was invited to chair several conferences, be featured speaker, or deliver keynote papers on topics closely related to water, under the mistaken impression that I was an expert, rather than merely a scientific reviewer. Of necessity, I declined. I did, however, feel sufficiently informed and obligated to review two papers for the journal that had published Weiss' and my growth control paper.

Another avenue of interest and investigation opened for me in 1962, following studies of captive white-footed mice. In addition to being able to monitor many activities of small animals in captivity, I planned to use my past experiences in physical programming and transducing techniques to simulate natural conditions for animals in captivity. The obvious first candidate was ambient light, for which I built devices to simulate twilights. 

Emerging from such studies was an unexpected finding. In all previous rhythmicity studies with mammals, investigators simply turned lights on and off to simulate day and night (still being done today - 2007). In my preliminary studies, I had found that when white-footed mice began running in constant light conditions, they customarily 'warmed up' gradually to top speed, just as an athlete does. Not so, however, during twilights!

Using artificial dusk, the animals started at their highest speed. This gradually decreased as dusk progressed. By dusk's end it had reached roughly the value maintained the rest of the 'moonlit' night. If running thereafter was sustained throughout the night, the mice usually ran in a single direction. When dawn began, the animals showed the opposite behavior, with their speed gradually increasing to a peak value, whereupon they stopped abruptly and retired shortly thereafter. In addition to showing these remarkable, light-level-specific changes in running speed  during twilights, the studies gave evidence of compelling influences of twilights on activity phasing.

Investigators had previously concluded that mice could not be synchronized to periods that differed by more than 3-hr from 24-hr. These restrictions were shown to be a limitation of artificial on-off light cycles. With the new techniques, mice were readily synchronized to clear cut 16-hr cycles for long periods. Much later (unpublished) studies were to show that birds could be synchronized to periods as short as 3 hours, with activity literally being turned on and off by light changes.

But were the mice, adapted to 16-hr cycles of wheel-running, also spending the rest of their lives on 16-hr cycles, that is, their eating, drinking, excreting, and sleeping? It emerged unequivocally in the Experientia study that all other activities of the animals also followed the 16-hr period. These other activities, even on 24-hr cycles, typically occurred during daytime excursions from the nest. A popular article summarizing the above results appeared in the Los Angeles Times in Dec., 1964, titled, "Of Mice and Men: All Miss the 'Rat' Race."

At about this time I was studying interfacial phenomena in connection with my membrane studies. My attention was drawn to an article in Scientific American, in which it was stated that certain amino acid side chains "repel water as wax does." Similar statements were being made regularly by biochemists and molecular biologists. Physical chemists know that this statement is incorrect. Nothing repels a water molecule.At any rate, I felt it was time to get something into print in a prominent place to put a stop to such misinformation before it also found its way into elementary texts. Accordingly, I wrote a 'Letter to the Editor' of Scientific American.

Although I received a prompt reply to the effect that they would "very likely publish" my comments, they never did so. Although, I continued to try to get this misinformation corrected in the press, little did I suspect that my numerous efforts would not bear fruit for 30 years. My letter on the topic finally appeared in American Scientist in 1994. Although I never wrote a technical report on hydrophobic bonding, this was done by Prof. David Chandler, for Nature in 2002. 
End Overview

Continuous automatic monitoring of the activities of small captive animals

The paper, "Automatic Multi-Channel Sensing and Recording of Animal Behavior," did not conclude my publication of material in Ecology at this time. There followed my detailed paper of the above title (1963;44:95-110) the following year. This paper includes all details on the construction of the wheel-running enclosure and all its programming and monitoring devices (Fig. 4-1, programming and data reducing panels in 1963), nests, storage chambers, enclosure environment, etc.

In the areas of recording the paper includes event display and readout, digital printout, and analogue recording. There follow descriptions of early behavior in the enclosure, exploration, eating, drinking, and eliminating, wheel-running, nesting, general activity and behavior (Figs. 4-2a & 4-2b, hourly profiles of activity), auto-and-cross correlations between eating, drinking, and eliminating, etc., etc. The following referee comments to the first draft of this paper are of interest.

Referee comments. This full presentation of exciting technical improvements in monitoring of animal activities raises the question of why the author bothered to write and submit the short paper already published in the Winter 1962 issue of Ecology when already this one (based in part on more of the same material) was probably being prepared and, once finished, was submitted to the same journal! It is regrettable that the author did not himself withdraw the earlier item once he had this ready for submittal. In the opinion of this referee he should have done this.

One point should be made clear before further critical comment on this MS. The techniques described and analyzed have such a fundamental bearing on experimental work of comparative psychologists, behaviorists, and ecologists that, in the opinion of this reviewer, this paper should be published in Ecology notwithstanding the earlier appearance of a quite superfluous "announcement" article on the same subject.....

The MS is well written, but leaves an impression left by most MSs - that it can stand cutting. The author is urged to do some pruning. The general interest and impact of his material will be enhanced thereby. Note also, that the summary is to be shortened by exclusion of explanatory material inappropriate to a summary.

In partial explanation to questions raised by the referee, the earlier paper was my first article in this genre, it was intended as an introductory article, it was not intended originally for Ecology but for Science, I did not and do not have 'executive' control over where or when my submitted manuscripts appear and, significantly, editorial changes occurred at Ecology between 1962 and 1963; the Editor who appeared to somewhat deprecate my "gadget for studying animal behavior," though also favorable on the whole, was no longer at the helm. A paper noted earlier, on canyon mice, also was to appear soon in Ecology (1965;46:452-461).

Further studies of wheel-running

Detailed studies were reported in the journal, Behaviour in 1963, Compulsory Regime and Control of Environment In Animal Behavior. I. Wheel-running (20:252-281). This is the first of four papers published in this journal in the same series, which was a precedent. The studies I reported in this paper illustrated the potential breadth of usefulness of wheel-running for behavior studies, and cast a new light on it and on certain other aspects of the behavior of confined animals.

Thus, once deer mice, Peromyscus crinitus, have learned to turn on a motor which drives the wheel (for a set time), they repeatedly do so, and run the motor-driven wheel. The mice were found to have characteristic patterns of spontaneous running activity, from the points of view of distribution in time, and sustained speed and direction. The patterns tend to be maintained on both entirely volitional schedules (Fig. 4-3a), and schedules on which the wheel must be unlocked periodically for free running (Fig. 4-3b; unlocked by mouse every 5 min for 5 min) requiring prior and repeated instrumental acts (pressing levers). The animals sometimes go through a "warm-up" phase before attaining and sustaining their customary running speed (Figs. 4-3a & 4-3b).

Given control over both the starting and stopping of rotation, the mice turn the motor both on and off, themselves, running the wheel for varying periods which they, themselves, determine. But they will only run the motor-driven wheel at times they choose. As often as the motor that drives the wheel is turned on by the experimenter, they doggedly press a lever within seconds, that turns it off, even though the result is that they are not be able to run the wheel at all. The opposite is also true. The animals promptly restart the rotation of wheel they are running of their own choice, after it is stopped by the experimenter. These topics, already mentioned in Chap. 2, under "Behavior of Small Captive Mammals," were treated in detail in this Behaviour paper. Some illustrations of equipment employed have already been seen (Figs. 2-10 through  2-12).

It was emphasized that, taken alone, the nature of a specific stimulus or activity is an unreliable guide for interpreting the behavior of small mammals upon whom it is forced, or to whom it is presented unexpectedly. The same holds for interpreting the behavior of confined animals given control over its initiation or termination, except when great stress is involved.. Stimuli which are rewarding (or punishing) in certain circumstances, can be punishing (or rewarding) in other circumstances.

In his tentative acceptance of the paper for Behaviour, Editor W. H. Thorpe, a world renowned leading behaviorist at Cambridge Univ., wrote:

I have now looked through the manuscript "Compulsory Regime and Control of Environment in Animal Behavior" which you submitted for publication in Behaviour. I found it both original and stimulating and it seems to me that you have opened up a valuable new technique for the study of volitional behaviour in small mammals. I have not *yet had time to criticize the paper in detail but I don't think there are many points which will need alteration. I shall therefore send it off to one of my editorial colleagues and you will hear from either him or me in due course about the question of its acceptance.

S. A. Barnett, a well known British behaviorist wrote as follows on Aug. 14, 1963 regarding this paper:

I am most interested by the work you report recently in Behaviour. Your interpretation of what you have found, in terms of control of the environment by the animal, seems to me most original and has stimulated me to think. One question that has occurred to me concerns the tendency of small mammals to behave in such a way as to increase the amount of change or variation in their environment - up to a point. Some of the most striking behaviour you describe clearly produces such a change - as if the mouse were inducing change for its own sake. This is of course, in one aspect, control of the environment; but I am not certain that this is the sort of thing you had in mind. Perhaps you will be discussing this point in later publications....

Incidentally, I most strongly agree with your emphasis on the importance of studying genetically heterogeneous wild type animals.

Structure and Function In Biological Membranes (1961-1968)

A serendipitous event occurred in 1961 that led to studies that occupied much of my attention for the next 5 or 6 years and led to my writing 3 books. It came about as follows. I was in the habit of emptying the graphite in the wood debris from sharpening pencils in a pencil sharpener into the toilet, instead of into a waste paper basket or the kitchen garbage, which can be messy. After flushing, on one occasion, I witnessed a most fascinating phenomenon. When water began to rise after the flushing, numerous streaming circlets of graphite particles were observed around the confines of the bowl near the water surface. Immediately impressed with the possibility that the phenomenon had implications for protoplasmic streaming. I determined to duplicate the phenomenon in the laboratory.

The next day, I allowed a stream of water to flow down an inclined plane into a pool in a large, shallow container. To my amazement, a countercurrent surface flow pattern, visualized with particles of graphite, developed within the stream and the pool. Surface back flow of water from the pool occurred upward along the periphery of the down flowing stream, even joining the down flowing stream at the nozzle (Fig. 4-4). The phenomenon does not occur in chemically pure water, and could be shown to be dependent on the presence of ubiquitous surfactant particles. On the other hand, some studies have shown that a relatively high dynamic surface tension of the stream is primarily a consequence of its motion [see Science report, below].

In a report in Science, "Countercurrent Streaming in Liquid Surfaces and its Relevance to Protoplasmic Streaming" (1962;136:652-653), I remarked that: movements in surface films counter to the interior stream are brought about by film pressure generated by surface tension gradients. In a similar process, the formation of new interfaces during protoplasmic syntheses could sustain gradients of interfacial tension with resulting protoplasmic movements. Forces at non-equilibrium interfaces also can give rise to vigorous, long lasting interior circulation in fixed fluid bodies. Some papers describing the possible significance of these processes for protoplasmic contractility and transport of cytoplasmic matrix are in preparation. [This report had to be greatly shortened on Editor Graham DuShane's insistence, as "we cannot continually make exceptions for every contribution of an author." I guess I had exceeded the 1,200 word limit in two of my three previous reports, in 1961.]

The above described report appeared in the May 18 issue of Science. No sooner had the issue been received by two subscribers than they both promptly wrote letters dated May 22, 1962, excerpted below.

With excitement I have just read your report in the latest issue of Science, for I have been seeking information about the relevance of interfacial tension driven flows to biological systems for several years. I am anxious to learn more about your research, and I shall be most grateful to have copies of your forthcoming papers as they become available. L. E. Scriven, U. of Minnesota, Chemical Engineering [Scriven subsequently was invited to spend some time in Weiss' lab at Rockefeller (see Chap. 2) - JLK]

Your paper....in Science has shed light on a longstanding puzzle at our leisurely evening coffee hour. I had even contemplated writing a note to Science on it, but fortunately procrastination and my very limited knowledge of surface chemistry prevented me from doing so....

While my observations in the coffee cup will contribute nothing to physiology, they were fun. I certainly was delighted to find them discussed in your paper. From now on my wife and I can enjoy our coffee relieved of the worry about surface chemistry. Werner E. Lipton, Fresno, California [Mr. Lipton is active in Fresno County transportation and land use affairs - JLK]

In the same year, I followed up with papers in Experimental Cell Research (1962;27:595-598), "Cytoplasmic Streaming and Non-equilibrium Interfaces," and Life Sciences (5:177-183), "On the Genesis of Cytoplasmic Streaming," passages from which I quote in the following.

The existence of interfacial stresses in dynamic fluids suggests a role of protoplasmic streaming in addition to those of transport and locomotion. Thus, these stresses in streams of matrix could influence the degree of interaction of matrix with adjacent matter....

Experiments on the effects of local interfacial disturbances in fluid systems, coupled with the known properties of biological membranes, have led to a molecular theory of membrane functions....One consequence of the genesis of movements of cytoplasmic matrix by intramembranous transitions would be the channeling of streams of matrix, for the endoplasmic reticulum commonly consists of membranes organized in parallel or organized into interconnecting networks of tubules and vesicles. Since leaf valves are common in circulatory systems, the membranes also may provide the valves (and pores) for control of sites and directions of incoming and outgoing streams.

These papers were followed in 1963 by papers on the same subjects in Developmental Biology (7:22-37) and Theoretical Biology (4:124-141) but, most notably, by a paper in Nature titled, "Structure and Functions of Biological Membranes," almost the same title as for two of my following books (vols. 1 and 2). Unfortunately, the main thesis of the paper, on membrane transitions, was not the one Nature had adopted. I proposed transitions between cylindrical and disc micellar subunits, whereas Nature had found a better solution, employing individually tailored, polymeric protein pores, specific for different ions and other particles.

More than a few papers I have written generated numbers of reprint requests over 100, several over 200, but for the Nature paper there were a record 1,799 requests, 899 from the U.S. and 900 from abroad. I received more requests from some countries than most researchers receive for a paper in entirety, and more requests from some Univ. Departments or Institutions than from some countries and states. The actual numbers provide a unique opportunity to assess comparative research interest in biological membranes at that time. They are tallied below.

Individual tallies

Africa	6	Australia	37	Austria	1
Belgium	22	Bulgaria	3	Canada	92
Czechoslovakia	96	Denmark	13	England	112
Finland	3	France	41	Germany	119
Holland	45	Hong Kong	1	Hungary	34
India	7	Ireland	8	Israel	33
Italy	22	Japan	15	Lebanon	1
Malaysia	3	Mexico	3	New Zealand	6
Norway	8	Poland	16	Portugal	1
Rumania	7	Russia	15	Scotland	12
South America	34	Spain	5	Sweden	32
Switzerland	22	Wales	3	Yugoslavia	4

 

Tallies by American States

Alabama	1	Arizona	12	Arkansas	3
California	108	Colorado	7	Connecticut	17
Delaware	1	Florida	27	Georgia	8
Hawaii	2	Illinois	41	Indiana	10
Iowa	9	Kansas	12	Kentucky	3
Louisiana	9	Maine	3	Maryland	8
Massachusetts	72	Michigan	38	Minnesota	20
Missouri	11	Montana	1	Nebraska	1
New Hampshire	4	New Jersey	15	New York	121
North Carolina	16	North Dakota	1	Ohio	41
Oklahoma	3	Oregon	16	Pennsylvania	49
Rhode Island	1	South Carolina	1	South Dakota	1
Tennessee	6	Texas	33	Utah	1
Vermont	2	Virginia	5	Washington	24
Wisconsin	28	Puerto Rico	1	Washington, DC	16

 

Departments or Institutes with 5 or more requests

Australian Commonwealth Industrial and Research Organization   5

Boston U. and Massachusetts Memorial Hospitals   9

Charles U. (Czech.), Pediatrics   5

Columbia U. College of Physicians & Surgeons, Neurology   5

Czechoslovakia Academy of Sciences (currently 53 institutes)

      Inst. of Microbiology 5  

      Lab of Physics and Pathology of the Eye   5

      Inst. of Pharmacology   9

      Inst. of Physiology   11

      Inst. of Experimental Biology and Genetics   5

Inst. of Human Nutrition   6

Inst. of Hematology and Blood Transfusion   6

Duke U. Med. Center, Physiol. & Pharmacol.   6

Florida State U., Biological Science   9

                              Genetics Lab.   5

German Academy of Science in Berlin, Biochemistry   5

Hebrew U., Hadassah Medical  School   9

Johns Hopkins Univ.

      School of Medicine

                  Microbiology   5

                  Physiological Chemistry   6

Massachusetts Inst. of Technology

      Biology   5

      Nutrition and Food Service   14

Michigan State Univ., Botany & Plant Physiology   5

New England Medical Center Hospitals   5

Karl Marx Univ.   5

Komensky Univ. (Czech.),   5

Rice Univ., Biology   5

Rockefeller Univ.   9

Southwest Foundation for Res. & Education   5

Tel-Aviv Univ. Medical School, Pathology   6

UNIVERSITY of, etc.:

Alberta, Pharmacology   8

California, Berkeley, Physiology   5

California, Riverside, Life Sciences   9

Cambridge, Pharmacology   5

Frankfort am Main, Plant Physiology   6

Illinois, Burnsides Research Lab.   7

Montreal, Biology   6

Rochester, Radiation Biology   5

Sheffield, Physiology   9

Sloan-Kettering Inst. for Cancer Research   7

Strasbourg, Biochemistry   7

SUNY, Buffalo, Pharmacy   5

Utah College of Med., Pharmacol.   9

Utrecht, Organic Chemistry Lab.   8

Washington, Physiol. & Biophysics   5

Wisconsin

      College of Agriculture, Bacteriology   7

      Med. School, Bardeen Med. Labs., Anatomy   5

U.S. Dept. of Agriculture Plant Industry Station   5

U.S. Public Health Service

      National Institutes of Health   15

      National Heart Inst.   6

      National Cancer Inst.   6

Wenner-Grens Institute   5

The reader may well puzzle over the reasons for such an unprecedented response to this paper in Nature. They apparently were as follows. Firstly, I was the essentially the only researcher proposing a readily understandable membrane structural transformations to account for membrane functions (see my proposed disk pillar transformation in Fig. 4-5a). Secondly, membrane researchers R. R. Dourmashkin and coworkers had just published an electron micrograph of a negatively stained plasma membrane from chicken liver (Fig. 4-5b) in Nature (1962;194:116). This appeared to show the ends of what could have been the pillars in the structural pillar state I had proposed. Additionally, their electron micrograph had received further notice in the widely-read New Scientist (1962;15:158). Also, others had obtained very similar structures from a saponin-treated mixture of lecithin and cholesterol after negative staining.

Although the finding of Dourmashkin et al. turned out to be an artifact, and the particular membrane structure and configurations that I proposed eventually were found to be invalid, they had the beneficial side effect of stimulating me to make detailed studies of all aspects of membrane physics and chemistry. As a result I wrote a book on these topics, "Structure and Function in Biological Membranes." This became unwieldy for a single volume, so it was divided into volumes I and II.

The chapter on the physical chemistry of water, however, was deemed to be of sufficient interest to chemists, physicists, and other scientists to warrant additional publication as a separate volume. "Water and Solute-Water Interactions." This was the first book in the English Language on the physical chemistry of water, and became my most cited work. The water book was published in 1964, and the membrane volumes in 1965, both by Holden Day, Inc. of San Francisco. In subsequent years, these three books, vol. 1, vol. 2, and W&S WI, were cited 204, 180, and 406 times, respectively, all three being among the most cited of my sole author works.

In the preparation of the book I corresponded with many recognized experts in the field, including Pauling, Scheraga. Forslind, Magnusson, Kauzmann, and Kamb. Concerning the reception and utility of these books, I first quote personal communications, followed by excerpts from reviews.

Correspondence concerning the books and chapter on water

May I finally congratulate you on a very fine piece of work. I believe that your treatment of the various aspects of the water structure problem represents a very valuable contribution, indeed, to the general knowledge and that it will provide a great stimulus for future research. E. Forslind, Royal Inst. of Technology, Stockholm

I found this a very stimulating chapter and congratulate you on your marvelous summary of the present state of knowledge....should stimulate your biological readers to do some serious thinking about the role of water in their many phenomena. G. Eisenman, U. of Utah, Medicine

I am enclosing your manuscript [water chapter], with my comments in the margin. I enjoyed reading it....I should very much appreciate it if you would please return the enclosed manuscript to me, after you have noted my comments. It is a nice summary - and I'd like to have it. H. Scheraga, Weizman Inst. of Science

Your chapter on water is an excellent comprehensive review. L.B. Magnusson, Argonne National Lab., Chemistry Division

I have found time to read through, with some care, the page proofs which you sent on "Water and Solute-water Interactions," [W&S-WI] and I must congratulate you; I found the presentation excellent. The material has already been very useful to me in my own work, not only in calling attention to relevant work with which I was unfamiliar, but in clarifying points which previously had been very fuzzy in my mind. Your presentation of conflicting theories and your reluctance to arbitrate among them will, I feel, in the long run make your work more, rather than less, valuable....the excellence of your presentation and the wide audience which I am sure it will enjoy makes me all the more regretful that my own work was published too late to be included. R.A. Horne, Arthur D. Little, Inc.

I am writing, first of all, to tell you how much I enjoyed reading your recent book [W&S-WI]. It is a great convenience to me to have so much of the pertinent information on water and aqueous solutions collected in a single source. J.F. Brandts, U. of Mass., Chemistry

As a worker in the field of non aqueous electrolyte solutions since 1951 may I congratulate you on your recent book [W&S-WI], which fills a very real need in this field. D.N. Glew, Dow Chemical Co. of Canada, Ltd.

I wish to congratulate you on your fine book [W&S-WI]. The lucid and enlightening presentation greatly stimulates the interest of experimental workers who are studying problems relating to the structure of water and of aqueous solution. T. Katsurai, Tokyo, Japan

Your small monograph on [W&S-WI] is indeed most delightful and I am looking forward to seeing the copies we hjave ordered of your two volume study of membranes. W. Drost Hansen, U. of Miami, Inst. of Marine Science

I haven't had a chance to go through your small book, [W&S-WI], in detail, but I have turned the pages fairly slowly. There is no question but that the book is a fine contribution and will be useful to those who want to get some feel for the current series [status?] of water structure. W. Kauzmann, Princeton U., Chemistry

Your small book on the structure and properties of water has proved to be very useful and represents the best compilation of information on the subject I know. R. Lumry, U. of Minnesota, Physical Chemistry

....from a more superficial perusal I got the impression that it is a very useful book. It is quite an achievement to have condensed in such a relatively small volume so much of recent consideration of water structure and the book will obviously be of great value to all interested and perhaps most of all to molecular biologists....J.Th.G. Overbeek Rijksuniversiteit Utrecht, van't Hoff Lab.

I read with great interest the lucid presentation of ice and water structure in your book [W&S-WI], G.J. Malecki, Malecki Labs., Chicago

I have been pleased to read your paper [chapter] on water, which I return herewith. I have found it interesting and I am sure it will be a useful part of your book {on biological membranes]. I am sorry to say, however, that my circumstances with respect to available time such that I am not able to think about your presentation sufficiently carefully to make any suggestions. I have got back [9/5/63] from a tour of Latin America, and have a great pile of work to be done without delay....With best regards, I am, Sincerely yours, Linus Pauling

Reviews of water book [W&S-WI] in Journals in English

In his Introduction the author states that he has presented all major viewpoints, with comprehensive literature citations. This is most certainly true. The book is lucidly written and gives an astounding amount of detail in theory and measurement in a very short space. J. Steigman, J. Polymer Sci, 1965

This approach is to be commended since the use of impressive mathematical formulations usually obscures the physical picture and gives the impression of having given a complete solution to the problem....The author has successfully condensed much in this slim volume. The book will be appreciated by research workers in physics and chemistry for whose benefit it is published and especially by electrochemists whose appreciation of the role of water is not second to that of biologists. M.A.V. Devanathan, J Sci Industrial Res, 1965

In this connexion, the path is not rendered smooth for the reader, but in view of our knowledge of the subject, it is an honest approach. The book is recommended to all those interested in the theories of ionic solutions. The wide coverage of the literature should also prove to be very useful to the reader. A. Maccoll, Chem. Industry, 1965

Because of the fundamental significance of it subject, this little book should arose great general interest....The author's attempt to make his publication as up to date as possible is illustrated by the fact that he reviews even the latest papers, including some dated 1964. G. Schwarz, Angewandte Chemie, 1966

That this was a heroic undertaking is attested to by the more than 370 items in its list of references, and by the acknowledgement in the preface to 14 workers in the field with whom the author corresponded for the purpose of receiving suggestions and comments and/or to make sure that he gave a fair presentation of their points of view. The job was not simplified by the fact that it necessitated excursions into a variety of highly specialized fields, including proton resonance chemical shifts, spin lattice relaxation times, and cold neutron scattering, in many of which the interpretation of experimental data is still often controversial, not to mention the fact that new data are still coming in which sometimes call older inferences in question....The author meets this problem head on and has wisely "not sought to smooth over the complexities of the problems nor the tangle of the many existing discrepancies." All things considered, therefore, it is the opinion of this reviewer that the author has done a good job....even a person who has been interested in water for a long time is likely to find items in its bibliography which he has missed and/or discussion which remind him of unfinished business....This is a good book to have, and, at the comparatively modest price of $5.50, it is a bargain, of which many workers in many branches of science will want to take advantage. H.S. Frank, J Amer Chem Soc, 1965

The book will surely be welcomed by chemists who deal with theories on the structure of water and interactions in water solutions....his clear exposition of the various proposed theories is thoroughly documented with a total of nearly 400 references to journals and other sources....A rich mine of information. Anonymous, Choice, 1965

Prof. Kavanau had all these considerations in mind when he recently completed a monograph on the structure and functions of biological membranes, for he included a substantial discussion of water structure and of molecular and ionic segregation in aqueous solution. His discussion....on the whole has withstood extraction from its original context very well. A certain amount of biological connotation survives, but this is neither intrusive nor irrelevant, and the text can be read with profit by chemists and physicists. The reader....need not be dispirited by Prof. Kavanau's concluding passages, where there is a glimpse ahead to formidable problems concerning the role of water in biochemical processes. E.C. Potter, Nature, 1965

The monograph thus assumes the form of a very complete presentation of facts and theories primarily pointing out the wide latitude of disagreement which exists over the interpretation of the data. This approach is excellent from the point of view of the serious researcher who has a basic familiarity with but need a thorough review of the latest work in this area. P.J. Niebergall, Amer J Pharm Education, 1965

This book contains a very full and valuable collection of references to the measurement of the physical properties of water. Of particular interest are recent x-ray diffraction determinations of its structure. R.J. Watts Tobin, Phil Mag, 1965

There are very good general accounts of nuclear magnetic resonance and its applications to the determination of the structure of liquids and solutions (p. 37 ff.) and of the abnormal mobility of the proton in aqueous solutions (p. 47 ff.)....After a consideration of the electrostatic ion-water interaction, the author sums up its limitations very neatly: "The wide acceptance of this model is a reasonable approximation...is because of the success with which even the simplest formulations of the theory yield order of magnitude agreement with experiment." To sum up, this is a useful compilation, in which most of the current theories of water and electrolyte solutions structure are discussed non-mathematically....But the reader is warned--it is heavy going! W.H. Lee, Tech Book Rev, 1965

As above, for the book on water, I first quote here from correspondence on the volumes on biological membranes, followed by reviews published in journals in English.

Correspondence

It [Vol. II] is indeed a monumental work and full of such detailed discussions of complex literature, and one can only be admiring of your perspicacity and energy....you have collected some excellent and stimulating examples of the electron microscopist 's art. R.G.E. Murray, Univ. Western Ontario, Medicine

In these two volumes you have assembled a wealth of information of an exceedingly important cytological structure and shown how many of the new findings can fit into a meaningful interpretation, and you have done it in a most scholarly way. A.C. Taylor, U. Texas, Medical Center

Please accept my congratulations for a heroic effort. Something like this has been badly needed for a long time. D.P. Agin, U. Chicago, Physiology

I have spent some time going over and enjoying it [Vol. I], and am sending an order for a personal copy. This is the best evidence that you have done a splendid job in thus pulling together so many disparate items and approaches.... You would have been interested in seeing how frequently the two professors and our students referred to it in our Graduate Course on Lipids. H.L. Davis, U. Nebraska, Medicine

It [Vol. I] has already proved very useful to me....Your review of NMR measurements in the chapter on water was just what I needed. I hope your book enjoys the wide circulation it deserves. R.H. Aranow, RIAS

I did also have a very extensive discussion of your views on membrane structure with Torsten Teorell last week, and both he and I are excited by the important implications of this for our views of ion transport phenomena. G. Eisenman, U. Utah, Medicine, Physiology

I wish to congratulate you on this work [Vol. I]. It represents a valuable and unique contribution to the field....Your appraisal of micelle formation and transformations is very thorough and exhaustive. I have picked up a lot of factual information while reading this chapter [Chap. 4]....it provides a most detailed analysis of the monolayers. J. Leja, U. Alberta, Metallurgy

I am impressed and excited by your recent theoretical work on membrane structure and function....and by the prospect of extending this theme to other phenomena such as cyclosis and ameboid movement, as your theory allows. B.S. Guttman, Calif. Inst. Tech.

I am writing this brief note to say that it was a pleasure to see your two volume work arrive on the market. I have had to change many of my ideas about membranes as a consequence of reading your excellent books. R.P. Zimmered, Juniata Coll., Biology

I had the opportunity to read your book [Vol. II]. I was impressed with the tremendous amount of physico chemical data which are compiled therein and related to problems of the membrane structure and function. This book is an invaluable help as a reference in studying the basic factors involved in membrane structure and function. E.J. Stadelmann, U. Minnesota, Plant Path. & Physiol.

Unfortunately, I have by no means read it all but have looked through it carefully and am much impressed. It is really quite a remarkable job, and I don't really see how you did it in such a short time. R. Lumry, U. Minnesota, Physical Chem.

Although time has obviously been too short for a full appreciation, I have spent some pleasant and interesting hours with [Vol. I]. I find the ideas it contains, your ideas on membranes, quite stimulating, the wealth of literature cited very useful for anyone working in the field....J.Th.G. Overbeek, Rijksuniversiteit Utrecht, van't Hoff Laboratorium.

This is to congratulate you on the excellent monograph by you on the "Structure and Function of Biological Membranes" which we had been expecting ever since the announcement about it in Salem's article on Intermolecular Forces in Biological Systems.

I was wondering if you will be attending the Tokyo Physiology Conference. If so, we would appreciate having you visit our Department on your way. We are interested in....the study of Vandenheuvel on the Van der Walls interaction in the Finean model. It will be most stimulating for the Department to have the opportunity of discussing these with you and to listen to you concerning your recent results. R.K. Mishna, Head of the Department of Biophysics, All India Institute of Medical Sciences, Aug. 23, 1965.

A number of us at the U. of North Carolina have been keenly interested in your books on membrane structure and function and, indeed, are using a number of the concepts you present as a guide in selecting agents to facilitate transport in various biological systems. Thus, we are quite interested in meeting you, having you discuss some of the concepts in greater detail, and even giving your opinion on our utilization of the basic concepts....If you could be with us for a portion of two days and present a seminar on each of the days....Albert H. Mattocks, Prof. of Pharmacy, Feb. 25, 1968.

In the next few years, I was invited more than once to chair conferences, be featured speaker, or deliver keynote papers on topics closely related to water under the mistaken impression that I was an expert on the physical chemistry of water, rather than merely a reviewer of the current status of its many aspects. Of necessity I was obliged to respectfully decline.

These invitations included:

To organize and moderate a day's program on the topics of metal catalyzed systems in transport, generally, and the problems of the transport of metal, specifically, at the Gordon Research Conference on Metals and Metal Binding, July 20-24 at the New Hampton School, New Hampton, New Hampshire, by Maynard B. Chenoweth, Conference Chairman, Biochemical Research Laboratory, Dow Chemical Co., Midland, Mich., Oct. 14, 1963.

To be one of the principal speakers at the 1964 Gordon Research Conference on "Ionic Movements and Interactions in Biological, Chemical, and Physical Phenomena," Aug. 31-Sept. 4, at the Tilton School in Tilton, New Hampshire, from Prof. George Eisenman, Chair of The Conference, U. of Utah, Salt Lake City, Nov. 1, 1963.

From J. Kenneth Craver, Chairman of the first Gordon Research Conference on the Physics and Chemistry of Pulp and Paper, to introduce the Conference with a review of the subject of cellulose water interactions during the week of Aug. 15-19, 1966 in the Pacific Northwest, near Seattle.

From Prof. C. Lafargue, Chair of Physics at U. of Bordeaux on Dec. 20, 1967 to present a communication and preside over a workshop at a Colloquium on Properties of Metastable States in March, 1968.

To submit a report on the subject of Water Structure to the XVth International Solvay Conference on Chemistry held in Brussels on June 25-July 1, on the topic "Electrostatic Interactions and the Structure of Water," from Prof. I Prigogine, Director of the National Institutes of Physics and Chemistry, Sept. 22, 1971.

To participate in a 3-day workshop on salt effects on plant structures and processes, to be held in 1976 at The United States Salinity Laboratory under the auspices of the United States Australia Agreement for Scientific and Technical Cooperation, from Dr. Richard H. Nieman of the United States Dept. of Agriculture, on Aug. 29, 1975.

I did, however, feel sufficiently informed and obligated to accept Editor B. C. Abbot's invitation to review two papers for the Journal of General Physiology, which had published Weiss' and my Growth and Growth Control paper, as follows.

Authors: K.M. L., R. D., and R.M. H. (July 13, 1965)

Title: The surface charge of isolated toad bladder epithelial cells. Mobility, effect of pH and divalent ions

 

Comments:

Some corrections of careless errors in this MS. are in order:

1. The authors' dimensional units for mobility are incorrect throughout the text, Tables, and graphs. The careless usage employed by the authors is widespread but is not acceptable in a paper dealing specifically with mobilities. The dimensions of mobility are cm/sec, not cm sec^-1volt^-1cm^-1. Even if the authors' usage were correct, it should be cm²sec^-1volt^-1, not the latter. Mobility is the limiting speed attained in a unit force field. In the case of electrical mobility of charged particle, one takes as the unit of force a unit potential gradient (1 volt/cm) acting upon the particles. Thus, unit potential gradient is implicit in the definition and is not part of the dimensions of the mobility itself.

2. Time should not be designated by T, as on page 5, but by t, particularly when T is used to designate temperature on page 7.

3. The term "radius" is used on page 5 when "hydrated radius" is meant. More importantly one should not use an ancient value for the hydrated radius of K⁺ dating back to 1939 when so much work on hydrated radii has been done since, and when such wide disagreement over the results prevails. A recent value of Padova (J. Chem. Physics 1963;39:1552) is 2.17Å, while values as great as 3.31Å have been proposed (see Nightingale, J Chem Physics 1959;63:181). Since the results depend on the hydrated radius of K⁺, it might be desirable to express them in terms of their dependence, or as a range, depending upon a corresponding range for the value of the hydrated radius employed.

4. The heading on Table I should be (electronic charge/u²) x 10^-4, not x 10⁴. The actual values have been multiplied by 10-4 to obtain the values in the Table, not by 104 [one of the commonest mistakes made in labeling the axes of graphs]. In any case the heading is ambiguous and could mean either (electronic charge)/(u² x 10⁴) or (electronic charge/u²) x 10⁴. Similarly, the ordinate in Fig. 3 is completely ambiguous. The axes in all the graphs should be relabeled, "Mobility (cm/sec) x 10⁵".

5. On page 5, the factor 10⁵ has been omitted from the mobility values.

Author: E. M. (August 30, 1965)

Title: Cellular electrophoretic mobility and the mitotic cycle.

Comments:

The subject of this MS. Is of considerable interest but parts of the Discussion section are vague and weak and would benefit considerably from being thought out more carefully and rewritten.

1. "Shear boundary" or "shear layer" is preferable to "shear plane," "plane of shear," or "slip plane."

2. On page 10, the first sentence is incomplete. In any case, an "increase" is not "formed from" something.

3. Concerning the second sentence on page 10, "preformed surface" cannot be "located in the hydrodynamic slip plane of the parent cell." The shear boundary is the region of contact between the solvated membrane and the "free" solvent. Thus, it is completely inconsistent to refer to preformed membrane or surface being present in this region. If additional membrane is present it must be evaginated, invaginated, folded, etc., or the cell membrane must be compressed or contracted.

4. In the same sentence one cannot assume that a "closer packing of the charged groups" will result in a "higher electrophoretic mobility." The author is implying that AS CHARGED GROUPS are packed together more closely the electric field increased proportionately. While this might be true for a cell surface because the charge density is very low to begin with, it is not necessarily true in general. Dielectric saturation of the solvent can lead to greatly reduced ionization of ionizable groups as the density of these groups is increased. The author should not lead the reader astray but should qualify his assumption and recognize the complexity of the situation. Nor is a greater density of charged groups necessary, but merely a greater number. The number of such groups could, for example, be increased as a result of the formation of blebs, which may be growing regions of the membrane.

5. A repulsion between cells would involve double layer interactions and would not be a simple Coulomb effect as implied on page 11.

Journal reviews in English of Vols. I & II

Whether or not one is in agreement with his hypothesis of "disc and pillar" dynamic lipid micellar structure for natural membranes is relatively unimportant is assessing the value of the book (Vol. I), since less than one fifth of the text is influenced by the hypothesis, and even within these sections (about 45 pages) much other useful information is included....The nearly 80 pages devoted to "water" represent such a good record and review of present knowledge of the influence of the lowly water molecule that it has been reprinted intact as a separate volume...very valuable contribution to the increasingly important field of investigation concerning the natural membrane. A.I. McMullen, J. Pharmaceutical Science, 1965

It (Vol. I) is a detailed, often encyclopedic, non-mathematical review of the basic physical chemistry and chemical physics of the components and conditions included in membrane structure and function....To whom is this book addressed? Primarily it is for those involved in graduate study and research on biological membranes who will read and study Professor Kavanau's second volume. However, this book will also be of interest to advanced students and research workers in the biophysical aspects of the life sciences and in colloid and surface chemistry. E.B. McQuarrie, The Vortex, 1965

An excellent and original contribution to the study of membranes, and essential reading for all in the field. J.F. Danielli, SUNY at Buffalo [Danielli was a leader in the field. JLK]

An extremely interesting study of biological membranes, this book begins with micelle formation and properties, and passes through topics of increasing complexity to a consideration of the structure of membranes. The approach is rigorous and is concerned entirely with the physical and chemical aspects of membrane structure, dealing especially effectively with the areas of protein and mixed monolayers and with the structure of liquid water in the region of other molecules. This book will be of value to students and workers at an advanced level in biochemistry and biophysics. Anonymous, Choice, 1965

Dr. Kavanau has clearly combed the literature very thoroughly....and this is evident in the multiplicity of references cited in the body of the text (Vol. I) and the many excellent illustrations--especially those of molecular models....There is no doubt that practicing colloid and surface chemists will find much here to suggest important topics still waiting to be investigated, and the author deserves our thanks for bringing together subject matter which is widely scattered in the literature. The production is good, the format attractive, and attention to proofreading has obviously been careful. An extensive bibliography and good indexes round off a good production. E. Hutchinson,, J Chem Education, 1965

The book [Vol. I] builds its subject systematically and on a solid basis....The author is a noted biologist, but he has a serious background in mathematics and physics. The latter [physics] is felt all over the book and, in fact, it forms the very basis for it. The former [mathematics], unfortunately did not break through. One can regret that none of the subjects are mathematically treated. It would have enhanced the value of the book. J. De Ley, Hydrobiologia, 1966

Until we know more about the rationale of the living process within cells, books like this [Vol. 2] will help us to assimilate the findings of experimental research and to make mental notes here and there about how the data might fit whatever general theory one imagines....Many and varied functions are described and documented as completely as possible in a subject whose literature is growing rapidly. The electron microscope photographs, elaborate line drawings and careful descriptions give the reader a clear image of the truly astounding devices life has contrived at the submicroscopic level....highly recommended to physicists who want a clear and thoroughly documented summary of current work about the aqueous solid state of living membranes. The format is excellent. J.G. Hoffman, Physics Today, 1966

The author is to be warmly complimented for having performed a rather neat feat of assembling a formidable mass of data and concepts and theories on biomembranes (Vol. II) from the standpoint of structure function relationships. For this he deserves the gratitude of workers whose interests are currently converging towards the discovery of a molecular basis for membrane functions. C.R. Krishna Murti, J Scientific Industrial Research, 1965

While the ideas offered are ingenious [ Vol. I1], there seems very little experimental or theoretical evidence which supports them rather than any of the other current theories. Nevertheless, for the initiated this is a highly imaginative, original, and provocative book. L. Wolpert, Endeavour, 1966

....Chap. 13 [Vol. II], which is entitled Electron Optical Evidence for Membrane Substructure, contains much material of professional interest to colloid and surface chemists. Indeed one might go further and say that this chapter should be of quite general interest to chemists, for it demonstrates how substantial is the progress that has been made in both chemical and structural understanding of natural membranes during the past decade....The extraordinary elegance and detail with which electron microscopy has been exploited is well represented by the carefully chosen and beautiful micrographs with which this chapter is illustrated--some 40 in all, in addition to many in the earlier chapters....The production of Vol. 2 is again first rate. E. Hutchinson, J. Chemical Education, 1965

The second volume fulfills the expectations of the first one. It treats mainly on dynamic aspects. There is a very readable section on protoplasmic streaming in which current views are ably exposed. Fragmentation, coalescence, contraction, expansion, growth and 'degrowth' are extensively reviewed. Very useful also is the discussion on amoeboid movement, useful not only to biochemists, but also to microbiologists and protozoologists, who will find here one of the rare summaries on this subject. Among the other major aspects is the review on permeability phenomena, in particular diffusion, active transport and impulse conduction. The last section on electron optical evidence for membrane substructure is an excellent one from the micro-morphological and anatomical point of view.

Undoubtedly Dr. Kavanau's book is the only one available on the structure and function in membranes. It is therefore very useful for all those interested in the knowledge of the micro-structures of the cell. The extensive reference list is helpful in locating further work....the book is an important review, an approach to the research literature and a 'must' for all those interested in or working on the fascinating boundary of protoplasm. J. De Ley, Hydrobiologia, 1966

....much of the discussion of molecular events associated with the proposed transformation between discs and pillars [Vol. II] is useful and stimulating. The Bibliography provides an important guide to recent research in many fields relevant to the problem of membrane structure and function. Anonymous, Quarterly J. Experimental Physiology, 1966

The author has done an admirable job in assembling recent research information and in discussing current theories on the functioning of cell membranes....The volume [Vol. II] contains many excellent electron micrographs of cell membranes and cell structures....The book should be of value to advanced students and researchers in the area of cell membranes. J.H. Pazur, Records Chemical Progress, 1965

In the two volumes under review the author provides us with a brilliant exposition of the known facts about micelle formation and transformations, the properties of pure and mixed monolayers and bilayers, as well as the structure of biological membranes. Equally the account of biological aspects of such as protoplasmic streaming and amoeboid locomotion is high powered. Anonymous, Food & Cosmetics Toxicology, 1965

Professor Kavanau's work is the first monographic discussion devoted entirely to the structure and function of biological membranes, and at the same time containing a detailed analysis of his own theories concerning certain aspects of the function of membranes....The chapters discussing the physico chemical basis of the biological membrane phenomena, such as micelle formation, mono and bilayers, can be considered most useful. To read these chapters is very profitable for biologists even if a physical chemist may regret the absence of a mathematical analysis. Z. Boszorményi, Acta Chimica Academy Science Hungaricae, 1966

The review of the literature on lipids and water is comprehensive, though rather uncritical. In this it has been one of the author's aims to show the wide latitude of disagreement that exists over the interpretation of even the most elementary phenomena. Those who, like Kavanau, dare venture further into complex biological systems require considerable courage!...While the treatment avoids mathematical notation, a knowledge of molecular forces and principles of surface a nd colloid chemistry is assumed....The books are well produced and in view of current trends will undoubtedly find a place on many library shelves. B.R. Malcolm, Academic Press Tech Book Review, 1965

The complete work is particularly appropriate for advanced courses in the biological topics related to membrane function and cell ultrastructure, as well as possessing broad general interest for advanced students and research workers in biology and medicine. The book is illustrated by a large number of photographs, diagrams, and graphs. Anonymous, The Book Exchange, 1965

If I were to quote the entire reviews rather than just excerpts, they might tell almost as much about the authors as the books. Also, many reviews have been omitted that were in foreign tongues. Two reviewers called me to task for dividing the membrane work into two volumes. One felt that it was justified. Two or three criticized the lengthy Reference list, and even the Subject Indices. One of my pet peeves (others will follow), however, is the failure of many authors to provide adequate indices; many feel that once the text is complete, the essential job is done. For all my books, I lean over backwards to provide thorough coverage in the indices.

I was surprised that several membrane book reviewers bemoaned the absence of mathematical analyses, whereas Devanathan, reviewing the water book (chapter) commended me, "since the use of impressive mathematical formulations usually obscures the physical picture and gives the impression of having given a complete solution to the problem." Mathematical expressions usually merely formalize already well visualized relationships, and provide a means for verifying and predicting. I discerned few topics dealt with in the membrane books that would have benefited from efforts to express them as equations.

Years later, as mentioned earlier, in November, 2003, I attended the weekly Wednesday evening dinner and seminar of Bill Schopf's Center for the Study of Evolution and the Origin of Life. There, I met the speaker, Dave Deamer, of UC Santa Cruz. In a subsequent e-mail he informed me that "[I]t was very nice to finally meet you....your two volume book on membranes inspired my early interest in biophysics. I can still visualize water of hydration surrounding ions...." As the reader will recall from coverage of my papers on "the origin of life," Dave and I also corresponded on that topic.

 

 

Other related journal and book publications

1962. Cytoplasmic streaming and non-equilibrium interfaces. Experimental Cell Research (27:595-598)
1966. Protein conformation in biological membranes. Science (153:213.
1966. Remarks on water structure. Federation Proceedings 25:977-978.
1966. Membrane structure and function. Federation Proceedings 25:1096-1107.
1966. Intracellular transport. Symposia of the International Society for Cell Biology, vol. V, Edited by K.B. Warren, Academic Press, New York.
1968. Membrane structure and function. Papers on Biological Membrane Structure, Edited by D. Branton and R.B. Park, Little Brown, New York.
1968. Anisotropic surfaces and protoplasmic movements. Cellular Dynamics I & II, Edited by L. Peachey, New York Academy of Sciences pp.84-96.
1968. Intracellular transport. Quarterly Review of Biology 43:194.

Highly instrumented studies in animal behavior

Another avenue of interest and investigation opened for me in 1962, following upon my studies of the behavior of captive white-footed mice. In addition to being able to monitor many activities of small animals in captivity, I planned to put my past experiences in physical programming and transducing techniques to work in simulating natural conditions for captive animals. The obvious first candidate for this goal was the ambient light.

To this end I had measured local twilight changes with transducing and recording equipment, and built a simple motor-driven rheostat (coiled resistor) to help simulate them. When inserted in a circuit providing voltage to a lamp, this variable resistor could be programmed to dim or brighten the lamp as desired, giving the effects of dawn and dusk transitions. During these simulations, light intensity roughly halved (or doubled) every 3 minutes. I also simulated moonlight for nighttime conditions with a suitable dim light.

As mentioned earlier, I was monitoring time, direction, and speed of running of deer mice in artificial enclosures during those times. Enclosures were situated in sound-proof, light-proof cabinets kept at constant temperature. Emerging from such studies, when supplemented by artificial twilights, was an unexpected finding. In all previous rhythmicity studies with mammals, investigators simply turned lights on and off to simulate day and night. In my preliminary studies, I had found that when white-footed mice began running in constant light conditions, they customarily 'warmed up' gradually to top speed, just as an athlete does. Not so, however, during twilights!

Using artificial twilights, during dusk, the animals did not 'warm-up' when they began running. Instead they started at their highest speed. This gradually decreased as a dusk progressed. By the end of dusk it had reached roughly the value kept up for the rest of the 'moonlit' night. If running thereafter was sustained throughout the night, the mice usually ran in a single direction. When possible, they faced the 'moon.' When dawn began, 5 hours later, the animals showed the opposite behavior, with their speed gradually increasing to a peak value, whereupon they stopped running abruptly and retired shortly thereafter (Fig. 4-6). In addition to showing these remarkable changes in running speed with changes in light level during twilights, the studies gave evidence of compelling influences of twilights on the animals' activity phasing (Fig. 4-7).

Mice were known to be particularly resistant to being forced into unnatural periodicities. Investigators had previously concluded that they could not be synchronized to periods that differed by more than 3 hours from the natural 24-hour period, that is, not more than 27 hours or less than 21 hours. These restrictions were shown by our artificial twilight studies to be simply a limitation of the on-off light cycles used. With the new techniques, mice were readily synchronized to clear cut 16-hour cycles for long periods, 14 days in one example (Fig. 4-7).

More than that, the dawn periods were much more compelling to the mice than the dusk periods. For example, on the 16-hour hour cycles, a mouse sometimes was still asleep in the nest and did not even start running during dusk, even though outside light conditions might be perceived in the nest. But for any mouse running during dawn, the light changes compelled it to cease activity, but only after running at top speed. These initial results were published in Nature in 1962 (194:1293-1295), under the title, "Twilight Transitions and Biological Rhythmicity." Additional studies appeared in Experientia (1962;18:382-388) titled, "Activity Patterns of Regimes Employing Artificial Twilight Transitions." My later studies, with birds (unpublished), were to show that they could be synchronized to periods as short as 3 hours, perhaps even less, with activity literally being turned on and off by light changes.

But were the mice, adapted to 16-hour cycles of wheel-running activity, spending the rest of their lives on 16-hour cycles, that is, their eating, drinking, excreting, and sleeping? This was investigated in the following Experientia study. Both time of drinking and time of obtaining food pellets were monitored automatically. Presence in the nest or at the bottom of the wheel was determined with infrared electric eyes. It emerged unequivocally that all the other activities of the animals also followed the 16-hr period (Fig. 4-8). These other activities, as is typical of wild mice kept in artificial enclosures, even on 24-hr cycles, typically occurred during 'daytime' excursions from the nest.

It may be of interest that initially, as was my custom, I sought to publish my findings on the influences of artificial twilights in Science. I was, however, unable to surmount the hurdles placed in my path by one reviewer. He or she was more concerned that no, even slightly, inaccurate historical impression be given than to facilitate the dissemination of novel new findings. In consequence, I divided the paper into two parts and published them separately in two journals, as described above. A revealing paragraph from the Science reviewer follows.

The author has to be sure removed some of the statements, indeed all the statements, which seemed to me objectionable in their reference to, or implications in connection with, the existing literature. I do want to emphasize again, as I did in my original review, that it is clear to me that the author has carried out some extremely useful work and that its merit as to publication is beyond question - at least in more extensive, explicit, and revised form. I am emphasizing this again because again I am unable to recommend that the present manuscript is suitable for publication in Science....The paper is surely too long for Science... [Here he assumes the editor's role rather than the reviewer's. Even 17 pages was not too long in my invited Review in Science 5 years later.- JLK]

A popular article summarizing the above results obtained with wild mice appeared in the Los Angeles times on Dec. 27, 1964, under the title, "Of Mice and Men: All Miss the 'Rat' Race."

Nothing repels water molecules! 1964-1994, 2003, 2005-2007, 2010

At about the time I was studying interfacial phenomena in connection with my membrane studies, my attention was drawn to an article by Max Perutz "Scientific American," in which it was stated that the side chain of leucine and the chromophore of phenylalanine "repel water as wax does." Similar statements were being made regularly by biochemists and molecular biologists. Physical chemists know that this statement is incorrect. I am unaware even of the existence of any molecule or intermolecular force, that could repel water. Only if a water droplet were electrostatically charged would it be repelled in the neighborhood of a similarly charged substance.

At any rate, I felt it was time to get something into print in a prominent place to try to put a stop to such statements before they also found their way into elementary texts (I had not covered the topic in my 1964 text on the physical chemistry of water). Accordingly, I wrote a Letter to the Editor of Scientific American to the above effect and received a prompt reply:

Thank you for your kind letter of November 9 [1964]. Your comments on the article by M. F. Perutz are most interesting, and we will very likely publish them."

Unfortunately, they did not publish the letter. I know not why but it can be noted that Perutz had recently (1962) received the Nobel Prize in Chemistry. A good guess is that the editors did not wish to embarrass a recent Nobelist and possible future contributor.

Although, I continued to make efforts (see below) to get this misinformation corrected in the semi popular and popular literature and press, little did I suspect that I would not succeed until 30 years later, and then only partially. In the meantime the error was propagated probably hundreds, perhaps thousands, of times. My efforts and their largely unsuccessful results are of sufficient interest and puzzlement to warrant recording them below.

First, however, it may help clarify matters if I give the successful letter as it appeared in American Scientist (1994;82:405-406), compliments of Managing Editor, David Schoonmaker. Although I never wrote a technical report on hydrophobic bonding [I did submit an unpublished Technical Comment to Science], this finally was done by Prof. David Chandler for Nature (2002;417:491), titled, "Two Faces of Water" (see, also, below). Coincidentally, Prof. Chandler is in the Chemistry Dept. at UC Berkeley, my alma mater, with which I was affiliated early in my career by virtue of my employment at the Lawrence Radiation Laboratory.

Hydrophobic Effect (All Wet?)

To the Editors:

Biochemists, molecular biologists, and science writers commonly misinform readers that saturated hydrocarbon chains "hate," repel or avoid water molecules-most recently in the informative article "Protein Folding" (May June) by David Holtzmann. Therein one finds the statement, "[t]hen there are the water-hating side chains that also attract one another and repel water. These are referred to by scientists as "hydrophobic."

In point of fact, water molecules and hydrocarbon chains attract one another. To a first approximation, the interaction can be elucidated in terms of the work of adhesion, the work required to separate one square centimeter of interface between two liquids into two liquid-gas surfaces. In competition between two moieties of substances interacting physically with water, those with greater work of adhesion with water will be favored in associating with water molecules, whereas those of the other substance will tend to be excluded or expelled (as if by default), not repelled. When moieties of one substance are strongly hydrophilic and of the other only weakly so (as with saturated hydrocarbon chains and 'hydrophobic amino acids), moieties of the latter substance are almost completely excluded. Since water's work of adhesion with saturated hydrocarbon chains (36-48 ergs) is much lower than that with other substances in protoplasm, the chains tend to be excluded from aqueous interfaces.

The hydrophobic effect has its basis in an intrinsically less stable hydrogen bonding of water molecules to one another near a nonpolar solute as compared to the ordered arrangement of the water molecules in pure water (The Hydrophobic Effect, C. Tanford, Wiley, 1973). Specifically, the hydrophobic effect is characterized by the appearance of dangling hydroxyl bonds; about 25 percent of the water molecules at interfaces with several representative hydrophobic substances possess such bonds (Science 1994;264:826-828).

Misinformation concerning the hydrocarbon water interaction derives from the widespread use of the misnomer "hydrophobic." Because the terms "hydrophilic" and "hydrophobic" are already firmly entrenched in the literature, it might be desirable, for the benefit of the general reader, to append the following elucidating parenthetical expressions to them: with hydrophilic, attracts water strongly; with hydrophobic, "attracts water weakly."

J. Lee Kavanau
UCLA (Emeritus)
Los Angeles, California

[The latest words on this subject come from Rezus and Bakker in Phys. Rev. Lett. 2007;95:148301 [and David Chandler (Nature 2007;445:831, titled, "Oil on Troubled. Waters), in which he reviews data on the oil water boundary]. Rezus and Bakker used femtosecond time resolution of a laser probe. The population of more slowly rotating water molecules steadily increased as the number of hydrophobic methyl groups on nitrogen based solutes rose. A correlation of four immobilized OH groups with each methyl group suggests a strong disruption of the solvent dynamics in the hydrophobic vicinity. But the authors emphasize that the structures in which the water molecules are briefly locked remains disordered rather than ice-like.]

I was impractical to record every time the same misinformation was propagated in Scientific American, but I did record the following.

In Aug., 1983, in "The Purification and Manufacture of Human Interferons," it is stated that "....is coated with an organic material that is hydrophobic, or water repellent...."

In Jan., 1992, in "How Cells Absorb Glucose," there occurs "...a double sheet of lipid molecules that repels water...."

On the following Feb. 4, I wrote the editor-in-chief about the continued propagation of misinformation in their pages. I received a postcard of acknowledgement implying that he took my letter of submission as a personal communication, not as a 'Letter to the Editor,' whereupon I wrote again on March 27, clarifying the matter. Receiving no reply, I wrote again on Sept. 10, to the Managing Editor, again without reply.

In Oct., 1992, in Histones as Regulators of Genes, we find: "....twists into tight hydrophobic (water-repelling) helices."

I followed up the Sept. 10 letter on October 7, noting that, "On reading your Oct. 2, 1992 issue, I learned that "because of the volume of mail, letters cannot be acknowledged individually." Since it appears that you are not planning to publish my letter of 10 Sept., I beg your continued indulgence in my effort to persuade you that this matter merits further consideration. " Thereupon I recalled the history of my correspondence with Scientific American and elaborated on the significance of the matter in question, and how useful it would be for Scientific American to publish a correction.

Resistance to publishing a simple Letter to the Editor remained as strong as ever, however, with the following reply elicited from the Managing Editor,

Dear Professor Kavanau,

I have circulated you letter to the editors and writers on the staff who might have occasion to deal with the hydrocarbon water interaction, and we will, of course, keep an eye out for space on the "Letters" page. But as you gathered from the editorial note that appears on that page, the volume of mail is considerable, and the space is extremely limited. Than you for writing to us again.

                                                      Sincerely, Managing Editor

      In other words, correcting this particular egregious misinformation, though it was repeatedly propagated had very low priority with its editors. After that I gave up on Scientific American and continued my efforts elsewhere, as described below. I did take note of an additional instance, where this matter was touched upon. In some sense, this can be regarded as a slight improvement over the above examples. It went as follows:

In Jan., 1995, in Elastic Biomolecular Machines, it is stated inScientific American that:

"Parts of the polymer chain are hydrophobic ("water-fearing" or apolar), and others are hydrophilic ("water-loving" or polar);...."

Misinformation similarly was appearing with regularity in Science. On 11 Sept., 1992 in Meeting Briefs: "User friendly chemistry takes center stage at the ACS meeting," we find:

                                                 "....and plastic is hydrophobic (it repels water)."

On Sept. 25, I wrote to the Editor, "I should like to submit the enclosed Letter [similar to the letter to American Scientist], which might be titled, "Hydrocarbon Water Interactions," for consideration for publication in Science."

On Nov. 4, I received a reply from the Letters Editor as follows:

Thank you for your letter of 25 September pointing out erroneous material in a recent issue of Science. Although we are not planning to publish it, we have forwarded a copy of it to the appropriate writers and editors for their information.

We appreciate your taking the time to write us. Sincerely, Letters Editor.

On April 16, 1993 in Meeting Briefs: "Chemists gather in Denver to get the big picture," it is stated that:

the insides of rings are hydrophobic (water-avoiding).

I had pointed out in my proposed Letter to the Editor of Sept. 25 that "....hydrocarbons and all other substances attract water." Accordingly, they could not be said to be "water-avoiding." I also pointed out that the water molecules were being excluded in competition with more strongly attracted substances.

I wrote again on April 26, 1993, to the same Letters Editor, explaining that the error continued to be propagated and submitted a more comprehensive letter with more extensive documentation, suggesting that it be titled, "Hydrocarbon Water Repulsion Myth." I also suggested that, "Publication of a Letter treating the hydrocarbon water interaction in Science would go very far toward dispelling the widely held misconception referred to above."

The same Letters Editor replied on May 27th, 1993 as follows.

Thank you for your letter of 26 April about the use of the term "hydrophobic" in a recent News article in Science. I regret to say that we are not planning to publish it. We are aware of your earlier letter on this topic; we have forwarded copies of your most recent letter to Mr. A. and his editors for their information. We appreciate your persistence and your taking the time to write to us again. Sincerely, Letters Editor.

Nevertheless on March 18, 1994 under Research News: Molecule makers learn the rules of a crooked game, another reporter states:

"Sets of hydrophilic (water-attracting) and hydrophobic (water-repelling) amino acids alternate...."

I wrote yet again on April 6, 1994 to the Letters Editor recounting my previous attempts to have an explanatory Letter published in Science, and urging the Editor again to publish such a Letter. I received no immediate reply to this letter.

On June 17th 1994, there appeared an informed technical article, higher-order self-assembly of vesicles by site specific binding, by physical chemists and engineers, in which there appeared the following technically correct statement.

The self-assembly of molecules and small aggregates in solution into larger structures is brought about by a number of attractive forces: van der Waals, ion binding, hydrophobic, polymer bridging, and depletion forces, as well as lock and key type interaction [emphasis added].

Despite this clear, correct exposition, one month later, on July 15, in Research News: "self-assembly comes together," we find the repetition of misleading sentences by a senior reporter:

These membranes are largely made up of phospholipid molecules which have balloon like head groups that are attracted to water, and tails made up of long hydrocarbon chains that flee from water's presence. This "split personality" gives these molecules the ability to self assemble.

With valid information published one month in Science, contradicted by misinformation a month later, instead of writing yet again to the Letters Editor, I resolved to submit a Technical Comment, on July 28, to the Deputy Editor for Engineering and Applied Sciences, Philip Abelson, with whom I had had several favorable previous contacts, covering the matter again, but more thoroughly.

I received a reply to both letters, but again from the Letters Editor, as follows.

I had not answered your letters of 6 April and 28 July, believing that there was no more I could say about our News Department's use of the terms "hydrophilic" and "hydrophobic." I am happy to report, however, that Friday the news editors asked for copies of all your letters so that they could be sure they were not erring in an article now being prepared.

I hope that your letters have the desired effect and the article uses the terms correctly.

We appreciate your interest in Science. Yours sincerely, Letters Editor.

With that communication, I also gave up temporarily on Science. There was no let up, however, in Science's dissemination of misinformation on the topic of hydrophobic bonding. For example, there appeared in following months, and this has continued to this day, the following.

On Jan. 20, 1995, in Research News, "Small spheres lead to big ideas," by the same senior reporter mentioned above, we find that:

Hemoglobin molecules are drawn to the surface of these bubbles because the molecules contain a hydrophobic component that is repelled by water.

Only a week later, suggesting that the pace of misinformation flow in Science was actually increasing, in Perspectives: "An Intelligent Channel," it is stated that:

The notion that the general porins may generate a local electric field near the channel constriction, sufficient to orient small hydrophilic molecules and repel hydrophobic ones...."

Again on Nov. 8, 1996, by still a 4th reporter in Research News: "Filling in the blanks in the p53 protein structure," we find:

....a structure called an amphipathic helix, which has the water-loving amino acids lined up on one side and the hydrophobic (fat like or water-hating) amino acids on the other.

I ceased keeping any record of these happenings in Science, but, even as I write, the dissemination of misinformation continues. Thus, in January 17, 2003 issue in the News Section, under "Chemists Concoct Quick Change Surface," we find:

exposing their water-repelling hydrocarbon chains to the surface.

Evidently the senior reporter who wrote this took little or no note of communications from either the News or Letters Editor.

The ink was hardly dry when, in the February 28 issue, a report appeared from researchers in Istanbul, Turkey, titled, "Transformation of a Simple Plastic into a Superhydrophobic Surface." In it we learn that:

Water repellency is important in many industrial and biological processes.

This paper was highlighted with a blurb on an editorial page under the title,

I Really Hate to Get Wet.

In much the same vein, a statement appeared in News of the Week in the May 23 issue (2003;300:1219), under "Tricks for Beating the Heat" "Help Panels See the Light." Under the pen of the same News Writer referred to several times above, we learn that:

They did it by outfitting dye-sensitized solar cells with water-repelling, light-absorbing dyes....."

While on the subject, as I write, in the Editors' Choice section (2005;309:357), we learn from a Senior Editor, reporting on Materials Science, that:

The foam volume depends on how hydrophobic (water-repelling) the nanoparticles are.

In the Dec. 2006 issue of Scientific American, in a News Scan, "Call It Beetle Guard," by Mark Fischetti we learn that:

.....seeded a polymer thin film with rows of silica nanoparticles coated the surface with a fluorinated chemical that aggressively repels water....could neutralize chemical and biological agents as they were repelled by military clothing....Repelled vapor molecules roll across the peaks.

Thus, 42 years after I first wrote an unpublished 'Letter to the Editor' to Scientific American about their publishing misinformation about substances that repel water, they continue to do so. The article was based on one in Nano Letters (2006;6:1213-1217) titled, "Patterned Superhydrophobic Surfaces: toward a Synthetic Mimic of the Namib Desert Beetle," by 7 MIT researchers. While nowhere in the article does the word "repel" or "repulsion" occur, use of the term "superhydrophobicity" in its title (and commonly by materials researchers) not only perpetuates a misnomer, it magnifies it. In correspondence with one of the authors, I learned that:

Water repellant in the context of our work simply means that water droplets placed on the surface will roll off freely at a small angle. This is how we and many others define the superhydrophobic state (contact angle greater than 150° coupled with a small rolling angle).

Thus, the new term "superhydrophobic" which most researchers will interpret to mean "super repulsion of water" is defined in terms of effects of cohesion (surface tension), adhesion, and gravity, all of which are attractive forces. In other words, combined effects of the structure of the surface, the surface tension of the water, the weakness of attraction of the water to the surface (adhesion), and the weakness of the gravitational attraction (in tending to flatten a droplet), allow the water to achieve a sufficiently large contact angle (form an almost spherical shape) that a slight tilt of the surface will allow a droplet to roll off, under the attraction of gravity.

The term superhydrophobic even has entered common usage, the "Lotus effect" being the way in which water drops roll off the superhydrophobic leaves of lotus plants, taking dirt particles away with them.

The significance of surface structure and contact angle of liquids leading to incorrect designations of "water repellency" are further confused in several recent articles. Among these is one in the J. Colloid Interface Science, titled, "Super Water-and Oil-Repellent Surfaces Resulting from Fractal Structure," by Shibuichi et al. (1998;208:287-294). In it we learn that:

Super water-and oil-repellent surfaces have been made utilizing the fractal structure of the solid surfaces. Super water-repellent surfaces showing a contact angle of 160° for water droplets have been made of anodically oxidized aluminum surface treated with....

It seems that the publication of this misinformation about substances repelling water in Science is destined never to end. Just today, reading in the 7th Dec., 2007 issue in "This Week in Science," Stella Hurtley and Phil Szuromi misinform us that:

Several approaches have been used to make superhydrophobic materials that excel at repelling water.

This statement appears in their summation of the article; "Designing Superoleophobic Surfaces," by Anish Tuteja et al. (318:1618-1622).

If workers and writers in this field would simply replace the misnomer "water repellent" with "very weakly water adherent" it would go a long way toward ending the confusion. Likewise, confusion could be ended if workers would take the time to consider, "whence does this so called repulsive force arise?" The only common, effective repulsive forces arise between similarly charged electrostatic or similarly oriented magnetic fields.

Concluding this topic of the properties of hydrophobic molecules, I remain somewhat puzzled. What has been the basis for the repeated, dogged reluctance of editors of leading publications to accept reasonable letters of correction? This is particularly unfortunate when the letters concern published information on topics of great importance, that continue to be dealt with erroneously in article after article on molecular biology and related topics.

I can suggest a remote possibility. Some editors may have checked the matter with physical chemists and found the subject to be of such great complexity that claiming "repulsion by, or of, water," as opposed to giving correct technical explanations, was the lesser of the two evils. Thus, Chandler's technical explication of the topic in , Nature, 2002, includes:

Water and oil famously don't mix: the term hydrophobic (water-fearing) is commonly used to describe substances that, like oil, do not mix with water. Although it may look as if water repels oil, in reality the separation of oil and water in ambient conditions is not due to repulsion between oil and water molecules, but to particularly favorable hydrogen bonding between water molecules.....Hydrogen bonding simply goes around hydrophobic species. In the close vicinity of the oily molecules, the possible configurations of hydrogen bonding may be restricted but the overall amount of hydrogen bonding remains relatively unchanged. Thus, the cost of hydrating a small, hydrophobic solute has more to do with the number of ways in which hydrogen bonds can form than with their strength. That is, the solvation free energy of the system is largely entropic and not enthalpic. However, the geometric picture breaks down for an extended oily region because not all hydrogen bonds can persist.

It seems that the publication of misinformation about substances repelling water is also destined never to end in Scientific American Thus, in the August, 2008, issue there appears an article, "Self-Cleaning Materials," by the science writer Peter Forbes. He also has been misled by the abundant misinformation about the existence of water repellent substances, and the Editors along with him. In a sidebar in the article, the Editors inform us that, "transform its waxy surface into an extremely water repellent, or superhydrophobic material. Although the article is extremely interesting for its factual expositions, it is way off the mark for comments like, "hydrophobic, or water-hating," "the nature of a hydrophobic material is to repel," "examining water repellency," "the lotus leaf's repulsion of water," and "high contact angle needed for strong repellency."

The conclusion of my attempts to dispel the notion that hydrophobic bonding involves repulsion of or by water molecules was swift and unexpected. It began with the appearance in the Nov. 20 , 2009 issue of Science in the This Week section in which the usual misconception was stated, followed by my writing to the Letters Editor, Jennifer Sills, my usual Letter of correction. To my surprise the letter was accepted and included in the 19 Feb. issue (p. 958) together with an illustration of an oily substance dripping into water. Although the text was accurate, its message was greatly obscured by the new title. Instead of that submitted, which was, "Water Molecules Not Repelled," it read, "Oil and Water Do Mix." I communicated promptly with Editor Sills, noting that the new title was erroneous and asking for a correction back to my submitted title. She replied that they were planning on using their own new title, "Opposites Attract." It was clear to me that the Letters Editor was opposed to making the asked for correction to the original title, which apparently would be admitting to too blatant an editorial change, so I decided to accept their change, which was at least not totally false. But I did explain, as follows.

OK, that would be better. I should point out, though, that oil and water are only "opposites" in the sense of having been given inappropriate "opposite-sounding" names --- "hydrophobic" and "hydrophilic". By way of explanation, they are both hydrophilic. But they still do not mix, for reasons touched on by Chandler in his paper. That is, water is not REPELLED by oils, but tends to be EXCLUDED from their neighborhood because of strong hydrogen bonding by the water. In other words, oil molecules can't get close to water molecules because that would entail breaking hydrogen bonds. The attraction between water and oil molecules is not strong enough to do that

The correction finally appeared in the 7 May issue, as follows. ÒOil and Water Do MixÓ by J. L. Kavanau (19 Feb., P. 958). Due to an editorial error, the title was incorrect. It should have been "Opposites Attract."

Meanwhile, a similar misconception had been included in the March-April, 2010 issue of American Scientist. Since I was unable get a Letter of Correction printed with the correct title, "Water Molecules Not Repelled," in Science, I decided to try sending the same Letter, except for the particulars, to American Scientist. To my surprise, the Letter with its correct title was accepted and appeared in the May-June issue. This ends the story begun in 1964 with my very first Letter to Scientific American.