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ABSTRACT OF The Goulstonian Lectures ON THE CHEMICAL PHYSIOLOGY OF THE ANIMAL CELL.
MARCH 25, 1893. the same in both. I have tried a good many experiments with salted plasma, and found, like Pekelharing, that if the nucleo-albumen is previously gently warmed with a little calcium chloride solution it acts like fibrin ferment and causes a rapid formation of fibrin ; calcium chloride alone does not ON THE produce this result. But if the treatment with calcium is omitted the nucleo-albumen has little or no activity THE chloride in this direction ; it may even hinder the coagulation in comparison with control specimens of the same salted plasma ANIMAL CELL. which had been diluted with water. I feel quite unable to Delivered at the Royal College of Physicians on, Tuesday, offer an adequate explanation of this difficulty. March 14th, 3. If fibrin ferment behaves as Pekelharing considers, its , action differs from that of all other ferments with which we BY W. D. are familiar. The usual action of a ferment is to act as a F.R.C.P. water-carrier, and, having given water to the substance on which it acts, splitting into new products takes place PROFESSOR OF PHYSIOLOGY, KING’S COLLEGE, LONDON. There is of course no reason why ferments should not act as calcium-carriers, but if we include such LECTURE III. an action we must either extend the meaning of the term THE COAGULATION OF THE BLOOD. fermentation or abandon the use of the word altogether in of blood coagulation. Of all ferment actions that [IN commencing his last lecture Dr. Halliburton expressed speaking of rennet in causing milk to curdle is most nearly akin to Ms indebtedness to his fellow worker Mr. T. G. Brodie, This resemblance has been a matter of bloodclotting. of at and referred Demonstrator Physiology King’s College, remark from the time of Andrew Buchanan onwards. In to the researches of W. L. Dickinson, E. H. Hankin, Brileke, each case we have the conversion of a soluble into an inHammarsten, Green, Ringer and Sainsbury, Arthus and soluble proteid, and in each case the presence of a calcium Pages, and proceeded at greater length to criticise the salt is necessary. Hammarsten showed that the process of milk curdling consists of two distinct stages, and moie results obtained by Pekelharing as follows :] has confirmed this observation. It appears The substances which have at various times received the recently Ringer to me quite probable that in fibrin formation there is fibrin ferment, cellnames of fibrinoplastic substance, double the action of the calcium a similar globulin fJ, serum fibrinogen, fibrinogen. A tissue-fibrinogen coming in at the endprocess, in causing the separation of the clot. and so forth are all varieties of one substance, which is a But it is difficult to prove or disprove this idea. As for mucleo-albumen ; and, further, it is in all cases a nucleo- milk it is very to boil it without coagulating the proteid, possible albumen which in cooperation with calcium compounds brings and one can thus destroy the ferment. With blood plasma about coagulation in the blood. The question then arises, this is impossible, for the blood proteids are coagulated by Does this definitely, once and for all, settle the long vexed and so one would destroy both ferment and fermentable subject of blood coagulation7 I am afraid it does not ; it heat, substance. I shall, however, not pretend to be able to decide helps us a little nearer the truth, but the final solution of the this question ; sufficient data have not yet accumulated for a problem may be a long way off yet. The observations which definite opinion to be pronounced one way or the other, have been made by Mr. Brodie and myself were attempts to and, therefore, I shall for the present retain the use of the answer some of the points left undecided by Pekelharing’s conventional term 11 fibrin ferment." work, and I wish that it had fallen to my lot to announce 4. The last difficulty standing in the way of our full that all the difficulties had disappeared, but as we went on of Pekelharing’s theories is that it takes no acceptance what we found was that new and previously unsuspected of the part that carbonic acid plays in blood cognisance difficulties made their appearance. I will first point out Clinical evidence supports physiological excoagulation. what I conceive to be the weak links in Pekelharing’s chain in that it is the venous blood which is most liable to periments of evidence. coagulated. We can hardly doubt from such evidence that 1. The theory that 6 °peptone"hinders coagulation because be either the quantity or more probably the tension of carbonic of its great affinity for calcium salts is a very tempting acid has an influence in hindering or promoting hypothesis. It is supported by the fact that injection coagulation. important I must again confess myself unable to see the This full 0f a soluble calcium salt counteracts its action. of such facts, and I also think that in the present fact we have confirmed. We used anaesthetised rabbits statebearing of our knowledge it would not be safe to attempt any in our experiments and injected about 10 cc. of 10 per cent. theorising as to the way in which they are to be fitted into "peptone" solution into the external jugular vein ; after Pekelharing’s hypothesis. about a minute a small quantity of blood was withI may now pass to a more systematic account of the exdrawn from the carotid artery ; this took about twenty which Mr. Brodie and I have been making during minutes to coagulate. Then 50 cc. of a 1 per cent. solution of periments the last two years ; they are very numerous, and all I shall calcium chloride was injected, and another sample of blood is a summary of their chief results. They were comwithdrawn ; this clotted in a few seconds. The same result! attempt menced with the object of determining whether the nucleowas obtained when ’’ leech extract " was used instead of albumens obtained by the sodium chloride method had the peptone ; leech extract probably owes its activity to a pro- same physiological action as the nucleo-albumens obtained by teose and is much more effective in hindering coagulation in acetic-acid method and termed by him "tissue the case of the rabbit than commercial peptone. The analogy Wooldridge’s The fact that both produce intra-vasoular of "peptone "and soap, which has already been pointed fibrinogens." formed part of the chain of evidence that the two coagulation out-, also lends support to Pekelharing’s views. But I have materials were practically the same, upon which I dwelt heard the objection raised that other substances whichl in my second lecture. The next point taken up was whether undoubtedly have a great affinity for calcium do not act in the lecithin, which I regard as an impurity, but to which this way, and I have been asked whether potassium oxalate attached considerable importance, was really the acts like peptone. We have put this to the test of experi- Wooldridge active substance, or whether the nucleo-albumen itself proment and have proved that the injection of a few cubic centiduces the result. metres of a weak (0 4 per cent.) solution of this salt Having shown by experiment that that substance produces ’enders the blood when it is subsequently drawn off its effect vzzaus its lecithin, whichever of the two methods is In one experiment a sample very slow in coagulating. .nf blood, which was drawn off before the injection, clotted adopted for its preparation, the greater number of our in less than a minute ; after this injection another sample subsequent experiments were performed with the material the sodium chloride method from the did not clot for seventeen minutes. We thus have an prepared by thymus. We adopted this method because it is more additional confirmation of Pekelharing’s hypothesis. and the results obtained were more certain. The 2. The second difficulty may be put in the form of a rapid material prepared by Wooldridge’s method often produces do not to nucleo-albumens cause question: Why coagulation no intra-vascular clotting at all ; this was recognised by occur in extra-vascular as readily as they do in intrahimself, and he spoke of it as the " negative vascular plasma ?7 The amount of calcium salts is presumably Wooldridge phase " in his experiments, because in the dog, which was the animal he chiefly used, the blood was not only free from 1 The term "peptone" includes the proteoses with which it is clot. but it clotted with tardiness after removal from the generally mixed.
ABSTRACT OF
The Goulstonian Lectures
CHEMICAL PHYSIOLOGY OF
HALLIBURTON, M.D., LOND., F.R.S.,
(hydrolysis).
,
, .
.
’
’
l
y ,
’
-
;,
No. 3630.
_
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-
640
body. I shall later have to return to this question of the "negative phase." Our failures to produce intra-vascular coagulation by the nucleo-albumen prepared by the sodium chloride method have been very few and are in part to be explained by the fact that the solution had been kept some days before it was used. Most solutions which are active This difficulty when fresh lose their activity when kept. we have now to a large extent obviated by rapidly drying the solution in a vacuum over sulphuric acid. The powdered substance, which contains both the proteid and the sodium carbonate used to dissolve it, is readily soluble in water. The solution is viscid, but after filtration it is perfectly In all our expericlear and free from solid particles. ments we have paid attention to this point ; all our solutions have been thoroughly freed from solid particles by filtration. I may perhaps now describe an experiment with its ordinary
during these processes was by no means improbable, especially as Dr. Starling called our attention to the fact that, Wooldridge found that his "tissue fibrinogens" lost their activity after purification. Now, repeated treatment with acetic acid (which is Wooldridge’s method) would perhaps. result in displacing a weak acid like nucleic acid, and there is no a priori reason why repeated treatment with water and sodium chloride should not do the same. Accordingly we put the hypothesis to the test of experiment. We have not yet had time to perform many analyses, but so far The so-called pure the result has been again negative. material contains the same percentage of phosphorus as the
active substance. If, therefore, we wish to avoid the unsatisfactory conclusion that what we have removed is an. intangible and shadowy substance known as a ferment, it is. necessary to fall back on the other hypothesis and consider The animal, and we have used rabbits almost whether in the process of purification we have added someresults. exclusively, is anaesthetised with ether and fixed in a holder thing to the nucleo-albumen. A few days ago at the Physioin the usual way. The external jugular vein is exposed and logical Society Dr. Haycraft suggested that by the repeated! use of salt we have obtained an inactive sodium chloride a cannula inserted ; through this the solution is injected and about 10 cc. of the solution (which would contain approxi- compound of the proteid. This will have to stand or fall by mately 0 05 gramme of proteid) is usually sufficient to the test of analysis, but it appears to me to be a very probable: cause death. The animal dies within a few seconds, explanation. the respiration suddenly stops ; there is no dyspnoea as All observers are agreed, with the exception of Dr. a rule, but there may be a slight stretching movement. Wooldridge, that these corpuscles play an important part ill> Another marked symptom is extreme exophthalmos and dila- the coagulation process, and we can hardly doubt that thetation of the pupils. The post-mortem examination is per- fibrin ferment or nucleo-albumen which causes blood to clot formed as rapidly as possible and the thrombosis found is when it is dried originates from the leucocytes. And here I usually limited to the venous blood, but in many cases there would emphasise the importance of comparisons between veris extensive clotting in the arteries also. We have not often tebrate blood and the blood of the crustacea. Hardyl has found a clot in the portal vein. The heart is still beating, recently shown that here certain corpuscles which he very. the right side full of dark clot, the left usually empty, or it appropriately designates the "explosive" corpuscles yie]
globulicidal,
641 urea, which produce leucolysis. In rabbits, howseldom sees the negative phase, and perhaps our reagents he found which acted in this way were-albumose, results differ from Wright’s because he has chiefly used dogs. peptone, pepsin, nucleic acid, nuclein, leech extract, pyo- We have searched for peptone in blood and urine by Devoto’s sanin, tuberculin, curare, urea, uric acid, and sodium urate. modification of the ammonium sulphate method, and in some Now Lowit is a strong supporter of Schmidt’s views on the cases by the use of the trichloracetic acid method. This reagent preponderating influence of disintegrated leucocytes in precipitates all the proteids except proteoses and peptone. bringing about blood coagulation. At the same time he Our results are as follows : (1) After the injection of nucleostrongly contests Lilienfeld’s view that the nuclei partici- albumen, peptone was never found either in the blood or urine ; pate in the process. He therefore asks why it is that (2) There "is no difficulty by either method of recognising if all these reagents cause leucolysis they do not cause " peptone after intra-vascular injection of small quantities thrombosis, and concludes that what is wanting is the of that substance. (3) After the injection of" non-proteid calcium. He accordingly injected calcium chloride either leucolytic agents there is also no "peptone discoverable before or after the injection of these other substances and by Devoto’s method. There were, however, in some cases states that he never failed to produce intra-vascular clotting traces formed by the trichloracetic acid method after injecin this way. In our repetition of these experiments we have tion of curare and of calcium chloride. I must confess that followed L6wit’s directions very closely ; we have not been these experiments do not throw much light on the subject, through his entire list, but we have used Witte’s but as far as they go they tend to support the hypothesis that peptone, Griibler’s peptone, leech extract, curare, urea, the "peptone" found by other observers originates rather We have confirmed his from the destruction of leucocytes than from the foreign uric acid and sodium urate. results in so far that calcium chloride alone does not material introduced into the circulation. Since writing the above, a paper has been sent to me by produce intra-vascular coagulation; the blood when shed, We found that the Lilienfeld,5 which bears on this same question. He takes however, coagulates very rapidly. same result followed when the calcium chloride was used in much the same view of the matter as Wright. He calls the conjunction with peptone, leech extract and the other sub- substances I have called nucleo-albumens nucleo-histons, on With one exception, in some twenty account of the supposed resemblance between the proteid stances just named. experiments we never got a trace of intra-vascular clotting. moiety of the substance and histon," the peptone-like The one exceptional case was when we used a peptone solu- material obtained from nuclei by Kossel. He lays it down tion that had undergone putrefaction, and the result might as a general rule that substances which hinder intra-vascular therefore be attributed to the nucleo-albumen in the bacteria coagulation do so either in virtue of the ’histon " they contain themselves. We, however, failed to repeat the experiment or the histon they produce by the disintegration of leucocytes; in other rabbits with the same solution. That introduces us andtbat substances which produce intra-vascular coagulation do to another difficultv we have met with-namely, the idiosyn- so in virtue of the nuclein they contain or because they cause crasies of the rabbit tribe. A solution which produces the liberation of nuclein by the disintegration of leucocytes. coagulation in one animal may fail to do so in another, If both substances are liberated together the nuclein is the and it appears that the dark-coloured rabbits are more more powerful as a rule and produces coagulation in histonprone to undergo intra-vascular coagulation than albinos. plasma. From our own experiments I can fully confirm In connexion with these same experiments I may mention Lilienfeld’s views as to the importance of nuclein. I do not that we found that substances like leech extract which pre- regard it as at all necessary, however, to imagine that the vent blood coagulation are directly antagonised by solutions nuclein must originate from the nucleus, as the cell protoof nucleo-albumen ; and after the injection of the first the plasm also contains nuclein in the form of nucleo-albumer.. latter still causes intra-vascular coagulation. The blood-platelets have long been known to have some irnOur work, then, has not confirmed that of Lowit, and portant bearing on blood coagulation. If the clotting of a shows that something else besides leucolysis must occur in microscopic preparation of blood be watched, the fibrin order to produce thrombosis. Wright has more recently strands are seen to start from groups of the platelets. found that extensive destruction of the leucocytes follows What blood platelets are has been the subject of much peptone injection. A possible objection to these observations discussion and innumerable theories. I have no wish is that the leucocytes may not be actually destroyed, but to add another theory, but will be content with they run together and get packed away in some organ like pointing out that the experiments I described on the the spleen. This view is confirmed by the fact that if one influence of nucleo-algumens in destroying white corperforms the experiment of allowing peptone solution to run puscles, and forming f10m them what look like platelet’, into a drop of freshly drawn blood on a microscope slide no favour the theory that these little bodies are the disintegradestruction of leucocytes is observable. But the phenomena tive products of the leucocytes. Lilienfeld takes the same of extra-vascular blood are often so different from what view, and his micro-chemical method, which shows they are occurs in the vessels that it is not safe to draw inferences composed of a phosphorus-rich substance, and his further from one to the other. We must await further experiments work,in which by gastric digestion he has separated them into nuclein and albumen, adds strong confirmation to this before this matter can be considered settled. To return now to a question which I have already men- idea. Thus we are brought back once more to the subject of tioned-namely, the so called "negative phase " which some- the disintegration of leucocytes, and I will conclude what I times follows the injection of nucleo-albumen. It was first have to say on the subject of blood coagulation by a few and more recently confirmed by general remarks on that question. observed by It was Schmidt who originally taught the importance of the is not coagulated by the injection that if the blood Wright, destruction of leucocytes as the main cause of coagulation. it undergoes clotting when shed with great slowness. hypothesis to explain this is that the nucleo-albumen injected These corpuscles are exceedingly prone to undergo changes consists of two parts, one the nuclein, which produces the when exposed to altered conditions ; but a considerable change clotting, and the other is a peptone-like substance which has come over our notions on the subject since that view was hinders clotting. The negative phase is explained by sup- first promulgated. Rauschenbach, one of Schmidt’s pupils, posing that the "peptone" moiety is split off in the circula- recognising that numbers of corpuscles do not disintegrate, tion and produces there the well-known effects of the intra- spoke of a leucocytes which are acted upon and disintegrated " vascular injection of peptone. This is supported by the by the plasma when the blood is shed, and j3 leucocytes which observation that in these animals "peptone" is found remain unaltered. Some very remarkable experiments pubin the urine, and further supported by the experiments of lished within the last few months by Dr. Sherrington,7 who Pekelharing, who demonstrated the existence of peptone in has succeeded in keeping leucocytes alive outside the body the blood under similar circumstances. for several weeks in oxalate plasma, have been very strongly In a previous paper4urged that it was not proven that against the idea that they are specially prone to disintegrate. the"peptone "actually originates from the foreign proteid The word "disintegration" does not, however, mean cominjected, because the nucleo-albumens fail entirely to give plete destruction and disappearance of the corpuscles in any of the peptone or proteose reactions. I think it much question. It signifies merely retrograde changes that end in more probable that the peptone originates from the disintethe death of th% corpuscles. Haycraft regards these changes gration of leucocytes. Peptonuria ordinarily occurs in sup- as being due to mechanical stimulation of living and naked purative diseases where there is a great deal of disintegration protoplasm by foreign solid matter, for if the fluid after being of leucocytes, and I thought one might obtain some light on 5 the subject by the injection of non-proteid materials, such as Verhandlungen der Physiologischen Gesellschaft, Berlin, 1891-92,
primary result leucolysis or destruction of the corpuscles, especially of the multi-nucleated ones. The
an immediate
curare or
white
ever,
one
Wooldridge,
Wright’s
No. 16.
4 Journal of
Physiology, 1892, p. 820.
7
6 Chemisehes Centralblatt, 1892, ii, 82. International Physiological Congress, Liége, 1892.
642 shed is surrounded by a fluid-like oil coagulation is prevented. A CASE OF ACROMEGALY. By watching the corpuscles under the microscope he has shown that they become flattened and irregular ; they then lose their granules, or these retire to one part BY J. MACKIE WHYTE, M.A., M.B. EDIN., M.R.C.S. ENG., PATHOLOGIST TO THE DUNDEE ROYAL INFIRMARY. Such changes are of the cell, leaving the rest clear. better marked in the coarsely granular corpuscles than in the finely granular ones. He regards the shedding out of the THE subject of the following notes first came under the fibrin ferment which accompanies these changes rather as a observation of Dr. Alexander Camphell and myself in process of metabolism than of disintegration. I am inclined to adhere, however, to the old word disintegration, as what September, 1890, on account of ingrowth of nails on both goes on is something different from what occurs in normal her great toes. My description was taken down at that time, llletaboJic processes. As I pointed out some years ago, the but, as it has recently been verified and slightly added to, I disappearance of the granules from the cells inevitably shall speak of her as she is now, merely premising that no suggests a comparison between the process and secretion, in alteration of any importance has occurred during the past which the shedding out of a ferment by secreting cells is two She is a domestic servant, single, aged twentyaccompanied by the disappearance of the granules which seven,years. and good health, being strong and fit for her enjoyed indicate the existence of its zymogen, or mother substance. throws no light on the, This idea has since been verified bv the discoverv of the work till 1886. Her family history The mother is alive and healthy, of slight build, and case. of fibrin the ferment, and so strongly has zymogen medium size. The father died of paralysis five years resemblance to secreting cells been recognised that some below the have gone so far as to speak of the coarsely granular or ago, aged seventy. The maternal grandfather lived to be eighty of age ; the maternal grandmother died of "dropsy," eosinophile cells as unicellular glands having a difEerent years aged fifty. The paternal grandparents died in old age. A origin and function from the neutrophile or finely granular sisterandtwo brothers are alive and healthy ; the youngest cells which appear to be the principal phagocytes. brother died of typhoid fever twelve years ago. From her Hankin has arrived at the conclusion that the alexines or history syphilis and alcoholism can be absolutely protective proteids which confer bacteria-killing power on personal her surroundings have always been favourable blood serum originate from the eosinophile cells, and this excluded, and to health. Fig. 1, engraved from a photograph taken in 1884, bears upon our present question because of the similarity, or it may be identity, between fibrin ferment and the alexines. FIG. 1. There is, however, evidence that cells which are not eosinoin fibrin The cells crusferment. phile may yield explosive tacean blood are hyaline, not eosinophile. (Hardy.) According to Sherrington, too, the eosinophile cells are not fibrino-plastic. I think it would be premature to lay down any definite rule as to which group of corpuscles fibrin ferment comes from ; and further, seeing that substances which produce coagulation can be obtained from cellsas diverse as those in the thymus, brain, liver, testis and kidney, it seems very probable that both kinds of leucocytes in the blood may give rise to fibrin ferment ; they both consist of protoplasm, and the main proteid constituent of all protoplasm is nucleo-albumen. During these lectures I have from time to time stepped aside from my subject proper in order to point out certain practical hearings which I considered to be of importance. I wish that I had had time, and still more the necessary experience, to have done so more fully. If, however, any grains of usefulness are scattered through what I have said, there is no doubt that others will pick them up and work out their practical side. .!!n connexion with blood coagulation and thrombosis I need not stop to point out the immense importance of a correct knowledge of this subject to the pathologist, and I will content myself with two remarks only on its practical aspects. The first of these is to point out the great resemblance between the phenomena observed in the animals in the experiments described and those cases of pulmonary thrombosis which are occasionally seen in the human subject. One can hardly doubt that here we have to deal with poisoning by the same material. Professor Gerald Yeo was witnessing some of our experiments a few days since, and he described to me a Engraved from a photograph taken in 1884 before onset of illness.: It was one of case he had had in King’s College Hospital.9 amputation of the breast for cancer, and the wound was shows her to have been then rather slim in build, with a’ perfectly aseptic ; but ten days after the operation, without bright expression, thin lips and other features, and small, any warning, the patient died suddenly with great dyspnoea. well-shaped hands. About the time of onset of her present Examination of the brain revealed no lesion, but at the illness, namely in 1886, she was subject to certain depressing necropsy a firm fibrinous clot was drawn like a tree with its emotions. In that year she observed that she became more easily branches from the pulmonary artery. The dyspncea in this fatigued, and in the course of a few months great physical and similar cases points to the cause of death in the lungs changes occurred. Menstruation, which had begun at fourteen themselves rather than in the respiratory centre; this and been normal hitherto, became scanty and in six months difference is easily explicable on the ground that the dose of finally ceased. Her body became stouter, so that with her poison enters the circulation more slowly than in our experi- ordinary clothing breathing was greatly impeded. To get up ments where it is injected. The second practical point from the kneeling posture she had sometimes to roll over on which I wish to emphasise is a word of warning. When I her side. Her face changed to such an extent that friends have seen animals die in a few seconds, as though by a who had not seen her for ()JllC time failed to recognise her: lightning flash, after a dose of five cubic centimetres of this The size of her gloves increased aendily from No. 63to No. 8, poison, which could not have contained more than a few and her shoemaker wastroubled with the difficulty of procenti;rammes’of‘ the material, I feel very loth to urge that curing "tops," remarking with somewhat ungallant candour any practical use of such substances should be made in the that he had never known a woman to have so large a foot. human subject, either for the stoppage of hemorrhage or The actual increase in the size is given from No. 4 to No. 7. for the production of a clot in aneurysms. The cure would Profuse perspirations, violent headaches occipital and frontal, in such instances be worse than the disease. and a feeling of stiffness as if she was encased in starch were amongst her troubles. The various changes seem 8 Centralblatt fur Bacteriologie und Parasitenkunde, Bd. xii., Nos. 22 to have been coincident, although the muscular weakness and 23. first attracted her own attention. In spite of her physical dis9 Many similar cases will be found in Sir J. Fayrer’s Clinical and in India. Observations ability she managed to keep her places," being possessed Pathological
.
ABSTRACT OF
The Goulstonian Lectures
CHEMICAL PHYSIOLOGY OF
HALLIBURTON, M.D., LOND., F.R.S.,
(hydrolysis).
,
, .
.
’
’
l
y ,
’
-
;,
No. 3630.
_
_
_
_
-
640
body. I shall later have to return to this question of the "negative phase." Our failures to produce intra-vascular coagulation by the nucleo-albumen prepared by the sodium chloride method have been very few and are in part to be explained by the fact that the solution had been kept some days before it was used. Most solutions which are active This difficulty when fresh lose their activity when kept. we have now to a large extent obviated by rapidly drying the solution in a vacuum over sulphuric acid. The powdered substance, which contains both the proteid and the sodium carbonate used to dissolve it, is readily soluble in water. The solution is viscid, but after filtration it is perfectly In all our expericlear and free from solid particles. ments we have paid attention to this point ; all our solutions have been thoroughly freed from solid particles by filtration. I may perhaps now describe an experiment with its ordinary
during these processes was by no means improbable, especially as Dr. Starling called our attention to the fact that, Wooldridge found that his "tissue fibrinogens" lost their activity after purification. Now, repeated treatment with acetic acid (which is Wooldridge’s method) would perhaps. result in displacing a weak acid like nucleic acid, and there is no a priori reason why repeated treatment with water and sodium chloride should not do the same. Accordingly we put the hypothesis to the test of experiment. We have not yet had time to perform many analyses, but so far The so-called pure the result has been again negative. material contains the same percentage of phosphorus as the
active substance. If, therefore, we wish to avoid the unsatisfactory conclusion that what we have removed is an. intangible and shadowy substance known as a ferment, it is. necessary to fall back on the other hypothesis and consider The animal, and we have used rabbits almost whether in the process of purification we have added someresults. exclusively, is anaesthetised with ether and fixed in a holder thing to the nucleo-albumen. A few days ago at the Physioin the usual way. The external jugular vein is exposed and logical Society Dr. Haycraft suggested that by the repeated! use of salt we have obtained an inactive sodium chloride a cannula inserted ; through this the solution is injected and about 10 cc. of the solution (which would contain approxi- compound of the proteid. This will have to stand or fall by mately 0 05 gramme of proteid) is usually sufficient to the test of analysis, but it appears to me to be a very probable: cause death. The animal dies within a few seconds, explanation. the respiration suddenly stops ; there is no dyspnoea as All observers are agreed, with the exception of Dr. a rule, but there may be a slight stretching movement. Wooldridge, that these corpuscles play an important part ill> Another marked symptom is extreme exophthalmos and dila- the coagulation process, and we can hardly doubt that thetation of the pupils. The post-mortem examination is per- fibrin ferment or nucleo-albumen which causes blood to clot formed as rapidly as possible and the thrombosis found is when it is dried originates from the leucocytes. And here I usually limited to the venous blood, but in many cases there would emphasise the importance of comparisons between veris extensive clotting in the arteries also. We have not often tebrate blood and the blood of the crustacea. Hardyl has found a clot in the portal vein. The heart is still beating, recently shown that here certain corpuscles which he very. the right side full of dark clot, the left usually empty, or it appropriately designates the "explosive" corpuscles yie]
globulicidal,
641 urea, which produce leucolysis. In rabbits, howseldom sees the negative phase, and perhaps our reagents he found which acted in this way were-albumose, results differ from Wright’s because he has chiefly used dogs. peptone, pepsin, nucleic acid, nuclein, leech extract, pyo- We have searched for peptone in blood and urine by Devoto’s sanin, tuberculin, curare, urea, uric acid, and sodium urate. modification of the ammonium sulphate method, and in some Now Lowit is a strong supporter of Schmidt’s views on the cases by the use of the trichloracetic acid method. This reagent preponderating influence of disintegrated leucocytes in precipitates all the proteids except proteoses and peptone. bringing about blood coagulation. At the same time he Our results are as follows : (1) After the injection of nucleostrongly contests Lilienfeld’s view that the nuclei partici- albumen, peptone was never found either in the blood or urine ; pate in the process. He therefore asks why it is that (2) There "is no difficulty by either method of recognising if all these reagents cause leucolysis they do not cause " peptone after intra-vascular injection of small quantities thrombosis, and concludes that what is wanting is the of that substance. (3) After the injection of" non-proteid calcium. He accordingly injected calcium chloride either leucolytic agents there is also no "peptone discoverable before or after the injection of these other substances and by Devoto’s method. There were, however, in some cases states that he never failed to produce intra-vascular clotting traces formed by the trichloracetic acid method after injecin this way. In our repetition of these experiments we have tion of curare and of calcium chloride. I must confess that followed L6wit’s directions very closely ; we have not been these experiments do not throw much light on the subject, through his entire list, but we have used Witte’s but as far as they go they tend to support the hypothesis that peptone, Griibler’s peptone, leech extract, curare, urea, the "peptone" found by other observers originates rather We have confirmed his from the destruction of leucocytes than from the foreign uric acid and sodium urate. results in so far that calcium chloride alone does not material introduced into the circulation. Since writing the above, a paper has been sent to me by produce intra-vascular coagulation; the blood when shed, We found that the Lilienfeld,5 which bears on this same question. He takes however, coagulates very rapidly. same result followed when the calcium chloride was used in much the same view of the matter as Wright. He calls the conjunction with peptone, leech extract and the other sub- substances I have called nucleo-albumens nucleo-histons, on With one exception, in some twenty account of the supposed resemblance between the proteid stances just named. experiments we never got a trace of intra-vascular clotting. moiety of the substance and histon," the peptone-like The one exceptional case was when we used a peptone solu- material obtained from nuclei by Kossel. He lays it down tion that had undergone putrefaction, and the result might as a general rule that substances which hinder intra-vascular therefore be attributed to the nucleo-albumen in the bacteria coagulation do so either in virtue of the ’histon " they contain themselves. We, however, failed to repeat the experiment or the histon they produce by the disintegration of leucocytes; in other rabbits with the same solution. That introduces us andtbat substances which produce intra-vascular coagulation do to another difficultv we have met with-namely, the idiosyn- so in virtue of the nuclein they contain or because they cause crasies of the rabbit tribe. A solution which produces the liberation of nuclein by the disintegration of leucocytes. coagulation in one animal may fail to do so in another, If both substances are liberated together the nuclein is the and it appears that the dark-coloured rabbits are more more powerful as a rule and produces coagulation in histonprone to undergo intra-vascular coagulation than albinos. plasma. From our own experiments I can fully confirm In connexion with these same experiments I may mention Lilienfeld’s views as to the importance of nuclein. I do not that we found that substances like leech extract which pre- regard it as at all necessary, however, to imagine that the vent blood coagulation are directly antagonised by solutions nuclein must originate from the nucleus, as the cell protoof nucleo-albumen ; and after the injection of the first the plasm also contains nuclein in the form of nucleo-albumer.. latter still causes intra-vascular coagulation. The blood-platelets have long been known to have some irnOur work, then, has not confirmed that of Lowit, and portant bearing on blood coagulation. If the clotting of a shows that something else besides leucolysis must occur in microscopic preparation of blood be watched, the fibrin order to produce thrombosis. Wright has more recently strands are seen to start from groups of the platelets. found that extensive destruction of the leucocytes follows What blood platelets are has been the subject of much peptone injection. A possible objection to these observations discussion and innumerable theories. I have no wish is that the leucocytes may not be actually destroyed, but to add another theory, but will be content with they run together and get packed away in some organ like pointing out that the experiments I described on the the spleen. This view is confirmed by the fact that if one influence of nucleo-algumens in destroying white corperforms the experiment of allowing peptone solution to run puscles, and forming f10m them what look like platelet’, into a drop of freshly drawn blood on a microscope slide no favour the theory that these little bodies are the disintegradestruction of leucocytes is observable. But the phenomena tive products of the leucocytes. Lilienfeld takes the same of extra-vascular blood are often so different from what view, and his micro-chemical method, which shows they are occurs in the vessels that it is not safe to draw inferences composed of a phosphorus-rich substance, and his further from one to the other. We must await further experiments work,in which by gastric digestion he has separated them into nuclein and albumen, adds strong confirmation to this before this matter can be considered settled. To return now to a question which I have already men- idea. Thus we are brought back once more to the subject of tioned-namely, the so called "negative phase " which some- the disintegration of leucocytes, and I will conclude what I times follows the injection of nucleo-albumen. It was first have to say on the subject of blood coagulation by a few and more recently confirmed by general remarks on that question. observed by It was Schmidt who originally taught the importance of the is not coagulated by the injection that if the blood Wright, destruction of leucocytes as the main cause of coagulation. it undergoes clotting when shed with great slowness. hypothesis to explain this is that the nucleo-albumen injected These corpuscles are exceedingly prone to undergo changes consists of two parts, one the nuclein, which produces the when exposed to altered conditions ; but a considerable change clotting, and the other is a peptone-like substance which has come over our notions on the subject since that view was hinders clotting. The negative phase is explained by sup- first promulgated. Rauschenbach, one of Schmidt’s pupils, posing that the "peptone" moiety is split off in the circula- recognising that numbers of corpuscles do not disintegrate, tion and produces there the well-known effects of the intra- spoke of a leucocytes which are acted upon and disintegrated " vascular injection of peptone. This is supported by the by the plasma when the blood is shed, and j3 leucocytes which observation that in these animals "peptone" is found remain unaltered. Some very remarkable experiments pubin the urine, and further supported by the experiments of lished within the last few months by Dr. Sherrington,7 who Pekelharing, who demonstrated the existence of peptone in has succeeded in keeping leucocytes alive outside the body the blood under similar circumstances. for several weeks in oxalate plasma, have been very strongly In a previous paper4urged that it was not proven that against the idea that they are specially prone to disintegrate. the"peptone "actually originates from the foreign proteid The word "disintegration" does not, however, mean cominjected, because the nucleo-albumens fail entirely to give plete destruction and disappearance of the corpuscles in any of the peptone or proteose reactions. I think it much question. It signifies merely retrograde changes that end in more probable that the peptone originates from the disintethe death of th% corpuscles. Haycraft regards these changes gration of leucocytes. Peptonuria ordinarily occurs in sup- as being due to mechanical stimulation of living and naked purative diseases where there is a great deal of disintegration protoplasm by foreign solid matter, for if the fluid after being of leucocytes, and I thought one might obtain some light on 5 the subject by the injection of non-proteid materials, such as Verhandlungen der Physiologischen Gesellschaft, Berlin, 1891-92,
primary result leucolysis or destruction of the corpuscles, especially of the multi-nucleated ones. The
an immediate
curare or
white
ever,
one
Wooldridge,
Wright’s
No. 16.
4 Journal of
Physiology, 1892, p. 820.
7
6 Chemisehes Centralblatt, 1892, ii, 82. International Physiological Congress, Liége, 1892.
642 shed is surrounded by a fluid-like oil coagulation is prevented. A CASE OF ACROMEGALY. By watching the corpuscles under the microscope he has shown that they become flattened and irregular ; they then lose their granules, or these retire to one part BY J. MACKIE WHYTE, M.A., M.B. EDIN., M.R.C.S. ENG., PATHOLOGIST TO THE DUNDEE ROYAL INFIRMARY. Such changes are of the cell, leaving the rest clear. better marked in the coarsely granular corpuscles than in the finely granular ones. He regards the shedding out of the THE subject of the following notes first came under the fibrin ferment which accompanies these changes rather as a observation of Dr. Alexander Camphell and myself in process of metabolism than of disintegration. I am inclined to adhere, however, to the old word disintegration, as what September, 1890, on account of ingrowth of nails on both goes on is something different from what occurs in normal her great toes. My description was taken down at that time, llletaboJic processes. As I pointed out some years ago, the but, as it has recently been verified and slightly added to, I disappearance of the granules from the cells inevitably shall speak of her as she is now, merely premising that no suggests a comparison between the process and secretion, in alteration of any importance has occurred during the past which the shedding out of a ferment by secreting cells is two She is a domestic servant, single, aged twentyaccompanied by the disappearance of the granules which seven,years. and good health, being strong and fit for her enjoyed indicate the existence of its zymogen, or mother substance. throws no light on the, This idea has since been verified bv the discoverv of the work till 1886. Her family history The mother is alive and healthy, of slight build, and case. of fibrin the ferment, and so strongly has zymogen medium size. The father died of paralysis five years resemblance to secreting cells been recognised that some below the have gone so far as to speak of the coarsely granular or ago, aged seventy. The maternal grandfather lived to be eighty of age ; the maternal grandmother died of "dropsy," eosinophile cells as unicellular glands having a difEerent years aged fifty. The paternal grandparents died in old age. A origin and function from the neutrophile or finely granular sisterandtwo brothers are alive and healthy ; the youngest cells which appear to be the principal phagocytes. brother died of typhoid fever twelve years ago. From her Hankin has arrived at the conclusion that the alexines or history syphilis and alcoholism can be absolutely protective proteids which confer bacteria-killing power on personal her surroundings have always been favourable blood serum originate from the eosinophile cells, and this excluded, and to health. Fig. 1, engraved from a photograph taken in 1884, bears upon our present question because of the similarity, or it may be identity, between fibrin ferment and the alexines. FIG. 1. There is, however, evidence that cells which are not eosinoin fibrin The cells crusferment. phile may yield explosive tacean blood are hyaline, not eosinophile. (Hardy.) According to Sherrington, too, the eosinophile cells are not fibrino-plastic. I think it would be premature to lay down any definite rule as to which group of corpuscles fibrin ferment comes from ; and further, seeing that substances which produce coagulation can be obtained from cellsas diverse as those in the thymus, brain, liver, testis and kidney, it seems very probable that both kinds of leucocytes in the blood may give rise to fibrin ferment ; they both consist of protoplasm, and the main proteid constituent of all protoplasm is nucleo-albumen. During these lectures I have from time to time stepped aside from my subject proper in order to point out certain practical hearings which I considered to be of importance. I wish that I had had time, and still more the necessary experience, to have done so more fully. If, however, any grains of usefulness are scattered through what I have said, there is no doubt that others will pick them up and work out their practical side. .!!n connexion with blood coagulation and thrombosis I need not stop to point out the immense importance of a correct knowledge of this subject to the pathologist, and I will content myself with two remarks only on its practical aspects. The first of these is to point out the great resemblance between the phenomena observed in the animals in the experiments described and those cases of pulmonary thrombosis which are occasionally seen in the human subject. One can hardly doubt that here we have to deal with poisoning by the same material. Professor Gerald Yeo was witnessing some of our experiments a few days since, and he described to me a Engraved from a photograph taken in 1884 before onset of illness.: It was one of case he had had in King’s College Hospital.9 amputation of the breast for cancer, and the wound was shows her to have been then rather slim in build, with a’ perfectly aseptic ; but ten days after the operation, without bright expression, thin lips and other features, and small, any warning, the patient died suddenly with great dyspnoea. well-shaped hands. About the time of onset of her present Examination of the brain revealed no lesion, but at the illness, namely in 1886, she was subject to certain depressing necropsy a firm fibrinous clot was drawn like a tree with its emotions. In that year she observed that she became more easily branches from the pulmonary artery. The dyspncea in this fatigued, and in the course of a few months great physical and similar cases points to the cause of death in the lungs changes occurred. Menstruation, which had begun at fourteen themselves rather than in the respiratory centre; this and been normal hitherto, became scanty and in six months difference is easily explicable on the ground that the dose of finally ceased. Her body became stouter, so that with her poison enters the circulation more slowly than in our experi- ordinary clothing breathing was greatly impeded. To get up ments where it is injected. The second practical point from the kneeling posture she had sometimes to roll over on which I wish to emphasise is a word of warning. When I her side. Her face changed to such an extent that friends have seen animals die in a few seconds, as though by a who had not seen her for ()JllC time failed to recognise her: lightning flash, after a dose of five cubic centimetres of this The size of her gloves increased aendily from No. 63to No. 8, poison, which could not have contained more than a few and her shoemaker wastroubled with the difficulty of procenti;rammes’of‘ the material, I feel very loth to urge that curing "tops," remarking with somewhat ungallant candour any practical use of such substances should be made in the that he had never known a woman to have so large a foot. human subject, either for the stoppage of hemorrhage or The actual increase in the size is given from No. 4 to No. 7. for the production of a clot in aneurysms. The cure would Profuse perspirations, violent headaches occipital and frontal, in such instances be worse than the disease. and a feeling of stiffness as if she was encased in starch were amongst her troubles. The various changes seem 8 Centralblatt fur Bacteriologie und Parasitenkunde, Bd. xii., Nos. 22 to have been coincident, although the muscular weakness and 23. first attracted her own attention. In spite of her physical dis9 Many similar cases will be found in Sir J. Fayrer’s Clinical and in India. Observations ability she managed to keep her places," being possessed Pathological
.