Jacques Loeb and Hardolf Wasteneys. Is narcosis due to asphyxiation? 
Reprinted from J. Biol. Chem, vol. xiv (1913): 517-523https://digitalcommons.rockefeller.edu/collection-of-reprints-loeb/1019/thumbnail.jp

Loeb, Jacques

Wasteneys, Hardolph

English

The Rockefeller University

Rockefeller University Digital Commons @ RU Collection of Reprints by Jacques Loeb Special Collections 6-1913 Is Narcosis Due to Asphyxiation? Jacques Loeb Hardolph Wasteneys Follow this and additional works at: https://digitalcommons.rockefeller.edu/collection-of-reprints-loeb Recommended Citation Loeb, Jacques and Wasteneys, Hardolph, "Is Narcosis Due to Asphyxiation?" (1913). Collection of Reprints by Jacques Loeb. 17. https://digitalcommons.rockefeller.edu/collection-of-reprints-loeb/17 This Book is brought to you for free and open access by the Special Collections at Digital Commons @ RU. It has been accepted for inclusion in Collection of Reprints by Jacques Loeb by an authorized administrator of Digital Commons @ RU. For more information, please contact nilovao@rockefeller.edu. f IS-NARCOSIS DUE TO ASPHYXIATION? BY JACQUES LO:Jj;B AND HARDOLPH WASTENEYS-(FROM THE ROCKEFELLER INSTITUTE FOR MEDICAL RESEARCH, NEW YORK) FROM . JOURNAL OF BIOLOGICAL CHEMISTRY VoL. XIV, No. 5, JuNE, 1913 Reprinted from THE JOURNAL OF BIOLOGICAL CHEMISTRY, VoL. XIV, No. 5, 1913. IS NARCOSIS DUE TO ASPHYXIATION? BY JACQUES LOEB AND HARDOLPH WASTENEYS .. (From the Rockefeller Institute for Medical Research, New York.) (Receiv�d for publication, April 30, 1913.)1. The idea. that the phenomenon of narcosis might be causedby an interference with oxidation is an old one. Recently it has been advocated by Verworn who tried to support it by indirect . evidence.1 Yet it is obvious that if we wish to ascertain whether or not narcosis is due to a diminution in the rate of oxidations (or, as Verworn expresses it, to an asphyxiation) there is only one way to decide the question, namely by comparing the rate of oxi­dations in narcotized organisms with that in non-narcotized. If we wish to .C,?a�ry out such experiments we must keep in mind that organisms which are capable of muscular action cannot well be used for this purpose since we know that muscular activity can easily raise the rate of oxidation in the animal body one hundred per cent. Since the narcotics cause muscular action to cease, a narcotized animal will consume less oxygen than a free­moving animal; but it would be-a mistake to state that this diminu­tion in the consumption of oxygen was the cause of narcosis. It is the effect and not the cause of narcosis. If we wish to test the asphyxiation hypothesis of narcosis we must use organisms which possess no muscles and in which therefore this source of error does not interfere with the result. The best material for this purpose is offered in the fertilized eggs. Loeb had shown in 1895 that the eggs of the sea urchin and fish cannot develop if the oxygen is with­drawn2 and this result has since been extended to a number of eggs of different forms. The same author showed that the addition of a small amount of KCN also inhibits segmentation. We shall see later that KCN only has this effect if it depresses the rate of oxidations in the egg below a certain level. The suppression of 1 Verworn: The Harvey Lectures, 1911-12, p. 52. 2 Loeb: Pfiuger's Archiv, !xii, p. 249, 1895. ' If J;,1, 5r7 518 Is Narcosis Due to Asphyxiation? segmentation by lack of oxygen as well as by KCN is a reversible process. It is also well known that anaesthetics inhibit 'the segmenta­tion of the fertilized sea urchin egg reversibly. It was, therefore, an easy task to compare the rate of oxidations in equal quantities of fertilized eggs with and without the presence of narcotics. War­burg had already observed that phenyl urethane in the concen­tration in which it suppressed nuclear and cell division did not diminish the rate of oxidations. Only if an excessive amount of phenyl urethane was added, was a diminution in the rate of oxida­tions noticeable; but, as he correctly adds, the question is not whether it is at all possible to iower the rate of oxidations by add­ing narcotics but whether that concentration which suffices for narcosis lowers the rate of oxidations.3 We undertook a series of experiments on the effect of various narcotics upon the rate of oxidations in the newly fertilized eggs of Strongylocentrotus purpuratus. Since the method of procedure and the degree of reliability of our method h�ve been described in previous papers no recapitulation of these points is needed. 2.- Experiments with KCN. We determined the minimum amount of KCN necessary to suppress cell division permanently in the newly fertilized eggs of S. purpuratus and found that the addition of 0.7 cc. of 0.01 per cent KCN to 50 cc. of sea water was the minimum amount required. We then determined the influ­ence of various amounts of KCN .. upon the rate of oxidations. Newly fertilized eggs were divided into equal lots and the amount of oxygen consumed in one hour was determined. Temperature 14°c. TABLE I. 1. Normal sea water .......................... . 2. 50 cc. sea water + 0.7 cc. 0.01 per cent KCN3. 50- cc. sea water + 0.9 cc. 0.01 per cent KCNOXYGEN CONSUMED mgm. 0.51 0.17 0.15 RATE OF OXIDATIONS 1.00 0.33 0.29 In solution 1, the eggs segmented; in solutions2 and 3 no segmen­tation occurred. The eggs were transferred from solutions 2 and 3 0. Warburg: Zeitschr. f. physiol. Chem., lxvi, p. 305, 1910.• J. Loeb and H. Wasteneys 519 3 to normal sea water and all developed into normal larvae. The amount of ·KCN necessary to suppress cell division lowers the rate of oxidations to on�-third of the normal amount. A repeti­tion of the experiment confirmed the result. TABLE II. 1. Normal sea water. . . ................. . 2. 50 cc. sea water + 0.4 cc. 0.01 per cent KCN3. 50 cc. sea water + 0.7 cc. 0.01 per cent KCN4. 50 cc. sea water + 1.0 cc. 0.01 per cent KCNOXYGEN RATE OF CONSUMED OXIDATIONS mgm. 0.65 0._34 0.21 0.18 1.00 0.52 0.32 0.28 The addition of 0.4 cc. of KCN only retarded the development but did not suppress it. The addition of 0.7 cc. of KCN sup­_pressed cell division, and the rate of oxidations was again exactly one-third of the normal. Similar results had previously been obtained by us for the eggs of Arbacia.4 We can, therefore, state that 1f the rate of oxidations of the egg 'is lowered to one-third of the normal amount found at 14°C., the eggs cease to segment. • If the prevention of segmentation by. nar-" cotics were due to the same influence we should have to expect also a lowering of the rate of oxidations to one-third of the normaL rate found at 14 °C. 3. We tried a number of narcotics, various alcohols, chloral.hydrate, chloroform and ethyl urethane. The minimum amount required 'to suppress cell division permanently was ascertained for each of these narcotics. The minimal dose varies slightly for the eggs of various individuals. a. Chloral hydrate. The minimum amount required to suppresscell division is 4.2 cc. of 0.5 per cent solution (made up in -�- NaCl + KCl + CaCl2) to 45.8 cc. of sea water. Temperature 15°C.• Loeb and Wasteneys: Biochem. Zeitschr., xxxvi, p. 355, 1911..,. ' 520 Is Narcosis Due to Asphyxiation? TABLE III. Normal sea water .............................. . 2.4 cc. 0.5 per cent chloral hydrate in 50 cc. sea water ...................................... . _4.2 cc. 0.5 per cent chloral hydrate in 50 cc. sea water ....... ............................... . 6.0 cc. 0.5 per cent chloral hydrate in 50 cc. sea water ........... ........................... . OXYGEN CONSUMED mgm. 0.65 0.60 0.57 0.57 RATE OF OXIDATIONS 1.00 0.92 0.88 0.88 In the solution with 4.2 cc. and 6.0 cc. of chloral hydrate the eggs were no longer able to segment; but they segmented and devel-" oped promptly when transferred to normal sea water. In the solution with 2.4 cc. of chloral hydrate the eggs segmented. The effect of the chloral hydrate upon oxidations was practically nil. The experiment was repeated . TABLE IV. 11Normal sea water...... . ................. . 4.2 cc·. 0.5 per cent chloral hydrate in 50 cc. sea water ...................................... . OXYGEN CONSUMED mum.-0.51 0.42 RATE OF OXIDATIONS 1.00 0.89 Although the segmentations were completely suppressed in this solution the influence of the chloral hydrate was practically negli­gible. We can say with certainty: The narcotic effect of chloral hydrate upon the' egg is not due to asphyxiation or a ·di�inution in the rate of oxidations. b. Ethyl urethane. Three cubic centimeters of a 10 per centsolution (in � NaCl + KCl + CaCb) in 50 cc. of sea water are suffi­cient to suppress cell division. TABLE V. Normal sea water .............................. . 3 cc. 10 per cent ethyl urethane in 50 cc. sea water OXYGEN CONSUMED mgm. 0.51 0.46 RATE OF OXIDATIONS 1.00 0.98 , J. Loe? and H. Wasteneys 521 With a few exceptions no eggs segmented in ethyl urethane, but all segm!=mted when put back into normal sea water. c. Chlorof arm. Seven cubic centimeters of 0.5 per cent CHCl3 in + NaCl + KCl + CaCl2 were required to suppress cell division. TABLE VI. Normal sea water. . . . . ........... . 43 cc. sea water + 7 cc. 0.5 per cent chloroform. OXYGEN CONSUMED mgm,. 0.47 0.41 R,\.TE OF OXIDATIONS 1.00 0.87 No segmentation occurred in the eggs in chloroform, but they segmented and developed normally when transferred to normal sea water. The narcotic effect of chloroform is produced without any considerable lowering of the rate of oxidations. The slight lowering observed is not quite but almost within the limits of error and cannot be considered. d. Propyl alcohol. Minimum amount necessary to suppresscell division is 3.5 cc. of 2 M propyl alcohol in 50 cc. of sea water. In order to avoid lowering of concentration of salts the 2 M i;;olu­tion of alcohol was made up in -r NaCl + KCl + CaCb. TABLE VIL In normal sea water ........................... . 2. 0 cc. 2 M propyl alcohol in 50 cc. sea water ... .3. 5 cc. 2 M propyl alcohol in 50 cc. sea water ... .5.0 cc. 2 M propyl alcohol in 50 cc. sea water ... .'OXYGEN CONSUMED mgm. 1.00 0.90 0.92 0.93 RATE OF OXIDATIONS 1.00 0 90 0.92 0.93 In 2 cc. of propyl alcohol a few of the eggs (less than I per cent) segmented. In 3.5 cc. and 5 cc. of propyl alcohol no egg segmented. A few underwent cytolysis, but as Warburg has shown, and as we were able to confirm, this does not alter the rate of oxidations in the fertilizeq egg. We therefore see that the narcotic effect of propyl alcohol is not accompanied by any lowering of the rate of oxidations in the egg. e. Various other alcohols. The narcotic effect of various alcoholsfollows the rule that each successive alcohol of a series is two or 522 Is Narcosis Due to AsP,hyxiation? three times as efficient for narcosis as the previous one. The amount necessary for the suppression of segmentation in t .he egg of S. purpuratus found for the various alcohols was as follows:RATE OF EFFICIENCY COMPARED WITH METHYL ALCOHOL. Methyl alcohol, 4.0 cc. 10 M in 50 cc. sea water..... . . . . . . . . . . . 1 Ethyl alcohol, 5.0 cc. 4 Min 50 cc. sea water.................. 2 Propyl alcohol, 3.5 cc. 2 Min 50 cc. sea water................. 6 Butyl alcohol, 3.5 cc. Min 50 cc. sea water .................... 12 TABLE VIII. Normal sea water .............................. . 4 cc: 10 M methyl alcohol in 50 cc. sea water .... . 5 cc. 4 Methyl alcohol in 50 cc. sea water ....... . 7 cc. -'i-- butyl alcohol in 50. cc. sea water ....... . OXYGEN CONSUMED mgm. 0.87 0.83 0.73 0.66 RA.TE OF OXIDATIONS 1.00 0.95 0.84 0.76 No egg_s segmented in the solutions containing alcohol except in the methyl alcohol in which 25 per cent of the eggs segmented. When transferred to normal sea water all the eggs segmented. The rate of oxidations in these experime�ts is too high to account for the narcotic effect on the basis of asphyxiation. The experi­ment with butyl alcohol was repeated. TABLE IX. N orznal sea water., ............................ . 6.5 cc. -�- butyl alcohol in 50 cc. sea water .. .... . OXYGEN CONSUMED mgm. 0.84 0.66 RATE OF OXJDATIONS 1.00 0.79 4. Theoretical remarks. The maximal lowering of the rate ofoxidations found under the influence of narcotics was 20 per cent; in the majority of cases it was less than this. In the case of propyl alcohol it was less than 10 per cent; in the case of chioral hydrate it was about 10 per cent. Since the temperature coefficient· for the rate of oxidations in the eggs is about 2 for 10°0.,5 we can pro-6 Loeb and Wasteneys: Biochem. Zeitschr., xxxvi, p. 345, 191 . ,/ , . . J. Loeb and FI. Wasteneysduce a lowering of the rate of oxidations of 20 per cent by lowering the temperature two or three degrees, e.g., putting the eggs into sea water of 12° instead of into 15° as in these experiments. The previous experiments of Loeb have shown that the eggs of S'. pur­puratus segment not only at _12° but even at 3°C., when the rate of oxidations is less than one-half of that observed at 15°C. In addition, the experiments with KCN also show that this substance does not suppress cell division until the rate of oxidations is reduced to one-third of the normal value. From these facts we can state with certainty that the effect of narcotics upon the eggs of the sea urchin' is not due t'o asphyxiation; a conclusion which Warburg reached also in his experiments with phenyl urethane. Kisch6 has recently published experiments which show that the photodynamic effect due to oxidations is raised instead of being diminished if narcotics are added to the medium. SUMMARY. It is shown that chloral hydrate, ethyl urethane, chloroform and various alcohols produce complete narcosis in the fertilized eggs of the 11ea urchin without practically lowering the rate of oxidations in tYie egg. 6 Kisch: Zeitschr. f. Bio!., ix, p. 399, 1913. •

Is  Narcosis  Due  to  Asphyxiation?

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Is Narcosis Due to Asphyxiation?

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