clotting, power is concerned the lymph always goes hand in hand with (Preliminary communication.)

Transcription

1 ON THE EFFECT OF PEPTONE ON THE CLOTTING OF BLOOD AND LYMPH. BY L. E. SHORE, M.B., Demonstrator of Physiology in the University of Cambnidge. (Preliminary communication.) MOST of the facts recorded in this short preliminary paper were observed in the course of an investigation of the fate of peptone in the lymphatic system. The action of peptone on the clotting power I deemed of sufficient importance to deserve a special investigation, but I shall do no more here than record some of the effects observed, without attempting to give an explanation of the facts, until I have made more experiments. Schmidt-Miillheim' discovered in 1880 that when *3 to *6 gr. of peptone per kilogrammie of body weight is injected into the blood of the dog, blood drawn from the body at any time during the next hour does not clot spontaneously. Fano0 showed that to produce this effect, the peptone must be injected rapidly in the course of a few minutes, and that no effect is produced if the same amount or even more is introduced slowly and gradually. He also observed that after rapid injection into the blood, the lymph that can be collected from the thoracic duct also does not clot. In cases where the peptone injection was without influence on the blood (6O/o of the cases) the lymph obtained during the same time is also unaffected. He lays great stress on the fact that as far as clotting, power is concerned the lymph always goes hand in hand with the blood, and deduces therefrom a proof that the lymph is completely dependent on the blood and produced therefrom by a physical diffusion. In direct opposition to this stand several observations of my own. I record an experiment showing that slow injection of peptone may lead to blood which clots normally, lymph which does not clot. For 1 Schmidt-Miillheim. Du Bois-Reymonds Arch. Physiol. Abth Fano. Du Bois Reymond's Arch. Physiol. Abth

2 562 L. E. SHORE. the sake of brevity the full protocol of the experiment will not be given. Dog. 8 5 kilos. Lymph from thoracic duct clots normally. 40/0 peptone solution flowing constantly into jugular vein at average rate of 5-6 c.c. per 10 minutes. The lymph of the first 10 minutes after the beginning of the injection clotted in minutes, all the other portions collected in each successive 10 minutes, were fluid 4 houirs later, and with the exception of the 4th and 5th portions, which showed a small weak clot at the bottom of the vessel, were still fluid the next day, 16 hours later. The injection was continued 2 hours 20 minutes. Total amount of peptone injected 3-16 gr. or *37 gr. per kilo. For 1 hour 20 minutes after the injection was discontinued the lymph collected in each successive 10 minutes remained fluid. The experiment was theni stopped. Blood was drawn from the crural artery five times during the course of the injection. 1. Drawn 27 min. after begin. of inject. Small weak clot at bottom of vessel after 2 his 2.,, 42,, Weak clot in 30 minutes. 3.,, 55,, Clotted in 10 minutes. 4.,, 72,, Clotted in 5 minutes. 5.,, 135,, Clotted in 5 minutes. The more usual effect of the slow injection of peptone into the blood is to lead to a period in which the lymph does not clot, followed by a period when it clots quicker and firmer than before. The following experiment illustrates this. Dog. 5 5 kilos. Lymph from thoracic duct clots in 10 minutes. 4o/,, peptone solution running constantly into jugular vein at average rate of 5-2 c.c. per 10 minutes for 2 hours 30 minutes. The lymph flowing out in each 10 minutes was collected. The second portion clotted in 20 minutes. The third and fourth portions were fluid 5 hours later. The fifth clotted in 15 milnutes and the succeeding portions quicker, till after 1 hour 30 nminutes from the beginning of the injection the lymph clotted in a few seconds after it dropped from the cannula. This lasted till 20 minutes after the discontinuation of the injection. The lymph then gradually exhibited its normal clotting power, and 1 hour later clotted as it did before the injection. The period of loss of clotting power is generally a short one, as in this case; it may be only represented by diminished clotting power, the lymph clotting after 2 or 3 hours. In these cases a small network of fibrin filaments is formed only on the sides of the vessel, and a portion remains fluid and can be poured out. The period of rapid

3 PEPTONTE ANYD CLOTTINAG. 563 clotting power is much longer. There is often difficulty in collecting the lymph, owing to clotting in the cannula itself. The clot formed is a very firm gelatinous one. When observed for as long as 3 days, it is noticed that it does not contract. This is in nmarked contrast to the clot of normal lymph. When peptone is injected into the bile duct it passes largely into the thoracic duct. The same effect on the clotting power of the lymph is observed. The period of loss of clotting power follows some 20 minutes after the conmmencement of the injection, and if the injection solution is weak (1 0/O) lasts 40 or 50 minutes, and is followed by the period of rapid clotting, which lasts till the end of the experiment. The injection of a strong solution (5 0/a) has the effect of cutting short the period of loss of or diminished clotting power, and of hastening and intensifying the period of rapid clotting power. As it is known that some 2 0/0 of bile salts added to the blood out of the body prevents clotting I made an experiment to see if the absorption of bile alone would produce the result. No diminution of the clotting power was observed. When peptone is injected into a lymphatic vessel, the lymph obtained from the thoracic duct does not clot. Periods of rapid clotting also occur, but I have not made as yet a sufficient number of experiments to enable me to state definitely the conditions of their occurrence. I can corroborate Fano's observation that after rapid injection of peptone into the blood the lymph does not clot. In all my cases the blood remained, as usual, fluid. A much smaller amount of peptone injected into the blood suffices to prevent clotting of the lymph, so that even with a rapid injection, the same condition as is obtained witb a slow injection can be brought about; viz. clotting blood, and non-clotting lymph. If blood is drawn immediately after the injection of only a small amount of peptone its clotting power is affected. I injected into the jugtular of a 8-6 kilo dog *2 gr. peptone (t023 gr. per kilo.) in i min. Blood was drawn from the crural artery 1, 2, and 5 minutes later; the last of these portions formed a weak clot, in the other two it was not until after the lapse of 4 hours that a few filaments of fibrin were formed on the walls of the vessels. The lymph collected for 1l hours after the injection, remained fluid. It is well known that peptone added to blood out of the body does not diminish its clotting power. I found that if as much as.50/0 is added clotting is prevented. The same thing is observed with lymph. This result is of very little physiological

4 .564 L. E. SHORE. importance as the addition of many salts to the same extent produces a like result. In my work on the fate of peptone, I had repeatedly noticed, that in cases of injection into the blood or into the bile duct peptone could be detected in the lymph of the rapid clotting stage, but not in that of the preceding non-clotting stage. It was quite possible however that in the latter too small a quantity to be detected might be present. I determined then to see the effect of the addition of mere traces of peptone to the lymph out of the body. The first experiment was as follows.- Lymph from thoracic duct quite clear, no blood. Clots rather slowly minutes. 10/0 peptone in normal salt solution. Lymph received into peptone solution as follows and gradually mixed by gentle shaking. I c.c. peptone solution + 2 c.c. Lymph I c.c.,, + 3 c.c.,, all these clotted rapidly. 2 c.c.,, + 2 c.c.,, 10 minutes. 05 c.c.,, c.c. 0-2 c.c.,, c.c.,, fluid. 24 hours. In the last portion, 3 hours later some fine flocculi could be seen, and 2 hours later still these had formed a faint loose network, the next day the contents of the tube were however fluid with a few fibrils attached to the walls. The amount of peptone in this lymph was /0. So striking a result as this I did not obtain again. With such traces of peptone in subsequent experiments I could only obtain a retardation of the clotting, the clot, however, being always small and weak. When rather mnore than this small amount of peptone is added the clotting was as a rule quicker and firmer than normal. The difficulty in obtaining constant results depends mainly on the variation in the lymph. When a trace of blood is present the effect of the peptone is less marked. Differences are also obtained with clear lymph and relatively opaque lymph. This is of great importance as in a fasting animal the opacity depends largely on the number of leucocytes. Clotting can be induced in non-clotting lymph by dilution and passage of CO,, or by dilution alone (1 to 2 of water) in the same way as with peptone plasma. I have made some observations on the proteids of the non-clotting lymph but I can make no definite statement at present. My experiments show that the clotting power of blood and lymph do not vary hand in hand. The formation of lymph

5 PEPTONVE AND CLOTTING. is not due to a mere physical diffusion from the blood vessels. When peptone is injected into the blood the flow of lymph from the thoracic duct is greatly increased. The percentage of proteids in it is also consid6rably increased. H eidenhain holds that the formation of lymph is due to a true secretory activity of the endothelial cells. With rapid injection of peptone into the blood I have observed that, early in the course of the experimeint, lymph that was flowing clear and colourless from the thoracic duct, becomes cloudy and white, and is followed by lymph containing at first a trace only of blood, but gradually becoming redder and redder. The white cloudiness is chiefly due to leucocytes, not to suspended fat, the subsequent red colour to red corpuscles, not to dissolved haemoglobin. Neumeister' has observed that after death from large doses of peptone, there is hyperaemia, and effusion of blood, in the mesentery and abdominal organs. The endothelial cells of the blood capillaries are so profoutndly affected by peptone injection that in casting out water and proteids with the peptone, they allow not only colourless corpuscles but red corpuscles also to escape. Other substances besides peptone, such as extract of leech (Haycraft) and extract of muscle of crayfish (Heidenhain) which act in the same way on the clotting power of blood, act similarly on the endothelial cells, and produce similar results in the flow of lymph. A continuation of my work is in the direction of learning what proteids are removed from the blood by the secretion, and to learn more of the function of the endothelial cells, "die lebende Oberfliche des Blutes,2" by the actioni of other substances on them. 1 Neumeis t er. Zeitschr. f. Biologie. xxiv Alex. S chmidt. Intern. Med. Congress. Berlin,