Part 15

+--------+----------+-----------+---------------------------- | | |Treatment | Date. | Weight.| Food. |(2 per cent| Symptoms. | | | caffein). | ---------+--------+----------+-----------+---------------------------- |_Grams._| _Milk_ | _cc._ | | | _(cc)._ | | Apr. 28 | | 300 | | Apr. 29 | 1,280 | 300 | | Apr. 30 | 1,290 | 300 | | May 1 | 1,315 | 300 | | May 2 | 1,330 | 300 | | May 3 | | 300 | | May 4 | 1,360 | 300 | | May 5 | 1,365 | 300 | | | |_Meat_ | | | |_(grams)._| | | | | | May 6 | 1,395 | 60 | | May 7 | 1,365 | 80 | | May 8 | 1,340 | 180 | | May 9 | 1,380 | 180 | | May 10 | 1,400 | 180 | | May 11 | 1,425 | 180 | 14.5 |No symptoms. May 12 | 1,470 | 180 | None. | May 13 | 1,485 | 180 | None. | May 14 | 1,510 | 115 | 14.5 | Do. May 15 | 1,500 | 115 | 14.5 | Do. May 16 | 1,485 | 115 | 14.5 |Passed worms. May 17 | 1,480 | 115 | 14.5 | May 18 | 1,485 | 115 | 19.5 |Feces soft and black; | | | | almost diarrhea. May 19 | 1,495 | 115 | 19.5 | May 20 | 1,500 | 115 | 19.5 |Scratches her eyes and | | | | chases her tail. May 21 | 1,500 | 115 | 17.0 | May 22 | | 115 | 17.0 | May 23 | 1,470 | 115 | 17.0 | May 24 | 1,465 | 115 | 20.0 | May 25 | 1,450 | 115 | 20.0 |Feces soft and black. May 26 | 1,450 | 115 | 23.0 |Diarrhea and worms. May 27 | 1,355 | 115 | 23.0 |Refused to eat all food. May 28 | 1,270 | 115 | 23.0 |Threw up worms, stiff, and | | | | has skin over both eyes. May 29 | | | |Found dead. ---------+--------+----------+-----------+----------------------------

Highest amount of caffein given, 362 mg per kilo. No autopsy.

Examination of the results obtained in the experiments of series C shows that young and growing dogs tolerate large amounts of caffein. In four subjects of this series, Nos. 1, 2, 3, and 6, no effect was observed when moderately large amounts (160 to 200 mg per kilo of caffein) were fed. Symptoms were noticed only when these amounts of caffein were increased from 50 to 60 per cent. The other two dogs, Nos. 4 and 5, of this series were less resistant, however, to caffein, as 0.16 gram of the drug per kilo induced well-marked symptoms. Since these were fed meat, while Nos. 1, 2, and 3 received milk, the difference in toxicity may be due to the diet employed, but No. 6, which likewise received a meat diet, failed to show the effects of caffein when 200 mg per kilo were fed. On the other hand, it should be noticed that No. 1 died after receiving 360 mg per kilo, No. 2 survived a dose of 334 mg, while No. 3 died after a dose of 322 mg per kilo of caffein. The fatal doses for Nos. 4, 5, and 6 were 287, 335, and 300 mg per kilo, respectively. Although the differences are too small to justify any definite conclusion regarding the effect of a milk diet or of a meat diet on the toxicity of caffein, the results nevertheless suggest a reasonable possibility that caffein is more toxic to young dogs when on an exclusively meat diet than when fed milk. It is perfectly evident, however, that the resistance to caffein in either case is very great, almost twice that of adult subjects. As shown in series A and B, 125 to 175 mg per kilo proved fatal to all but two animals in these experiments, while symptoms of toxicity appeared after much smaller doses. In other respects the behavior of young dogs toward caffein was the same as that of the adult. In neither case was cumulation nor tolerance observed under the conditions of these experiments. The findings at autopsy were likewise similar, as gastro-enteritis was the chief lesion observed on macroscopic examination. It might be mentioned, however, in this connection, that the symptoms of caffein intoxication in young dogs often presented marked differences from those observed in those of more advanced age. The resemblance of the effects of caffein in young puppies and in rabbits was very striking. In both, the tonic with clonic convulsions were observed after a sufficient quantity of caffein was administered. In the dogs which were fully grown a large dose of caffein was usually followed by tonic convulsions and almost instantaneous death.

Moderately large amounts of caffein fed daily to puppies for several days--in some cases as long as 10 days--induced mild symptoms only. No cumulative effect was observed in any of the experiments of series C. There seems to be tolerance of certain functions toward caffein, but no general tolerance of the body could be obtained in these experiments. Caffein is apparently less toxic for adult dogs on high than on low protein diet. In young and growing dogs caffein is somewhat less toxic when milk, rather than meat, forms the exclusive diet. Some pathological conditions apparently increase the toxicity of caffein also in dogs. The symptoms of caffein intoxication observed in young dogs are in some respects different from those in full grown and older animals, and resemble those noticed in rabbits.

DISCUSSION OF RESULTS.

It was pointed out at some length in the introduction that the toxicity of some drugs may not be the same for all forms of life. This observation was also made by some investigators who experimented with caffein on different species of animals. Thus Maurel(55) stated that caffein is twice as toxic for the frog as for the rabbit when administered by mouth. Fröhner's(26) experiments, on the other hand, made on domestic animals, failed to show great differences in the toxicity of caffein. According to this observer, horses seem to be more susceptible than cattle, goats, and swine, the minimum toxic dose being the same for all of these, while the resistance of the dog to caffein is about midway between that of the horse and the other animals mentioned. It may be remarked, however, that Fröhner made only 13 experiments. That these data are inadequate for the formation of any conclusions as to the toxicity of caffein is evident since the most striking effect of caffein observed in the work herein reported was the comparatively wide range of variation in the resistance of individuals of the same species to this drug. This was found to be the case even when the conditions of experimentation were approximately uniform, and was observed whatever the mode of administration of the drug employed. The toxicity for different individuals also varied in acute as well as in chronic intoxication. It is for this reason that the number of tests employed were often quite large, for no conclusions of any value could be drawn without averaging the results of a sufficiently large number of experiments. Furthermore, it is to be borne in mind that the action of a drug may differ according to the mode of its introduction into the body and that different species of animals may vary in this regard. This is especially true of some substances when given by mouth, the range in toxicity for certain species of animals being much greater when thus administered than when injected subcutaneously or intravenously.

Maurel's(56) investigations are of interest in this connection, as his work embraces a systematic study of the toxicity of a large number of substances in the rabbit, pigeon, and frog when given by mouth, subcutaneously, intravenously, or when injected into the muscles. According to this investigator the range of variation of the toxicity of a substance is widest when given by mouth. Potassium sulphocyanid, for example, is about 2.5 times as toxic for the frog as for the rabbit when given by mouth. Quinin hydrobromid is three times as toxic for the frog as for the pigeon, while for the rabbit it is twice as toxic as for the pigeon. When given by hypodermic injection the toxic dose per kilo weight is practically the same for all three species. The difference of resistance according to the mode of administration is even more marked for spartein sulphate. When given by mouth the toxicity for the rabbit is six times as great as for the frog, but when injected subcutaneously the toxic dose is about the same for the rabbit and for the frog. The relation of the mode of administration to toxicity is further shown in the following substances: For the rabbit the minimum fatal dose per kilo of quinin hydrobromid is 1.5 grams administered by mouth, 0.5 gram when injected subcutaneously, and 0.07 gram by the intravenous path, while strychnin sulphate is twice as toxic administered intravenously as subcutaneously, and six times as toxic as when administered by mouth. The mode of introduction, however, does not always affect the toxicity of a substance. This is made evident by the action of strychnin on frogs in which, according to Maurel(56), the toxic dose is the same whether given by mouth or injected into the subcutaneous tissues. This appears to hold true also for other animals as demonstrated by the experiments of Hatcher(35) on the cat, in which he observed that strychnin is as readily absorbed from a full stomach as from the subcutaneous tissues. These findings are extremely interesting, especially in view of Maurel's(57) work on the subject, according to which he finds that a substance is much less toxic when given by mouth than when administered by hypodermic injection or intravenously. That this generalization does not admit, however, of universal application is made evident by the work of various experimenters. Claude Bernard(10) observed that curara is as poisonous for the pigeon when given by mouth as when injected subcutaneously, while Zalesky(86) found that samandarin is more toxic for frogs when introduced into the stomach than by injection into the lymph sacs. Our experiments with caffein likewise show that Maurel's generalization does not always hold good, since it was found in experiments with gray rabbits that the minimum fatal dose is but moderately greater by mouth than by the subcutaneous path.

Equally interesting is the observation of the writer, that in the guinea pig the difference in the toxicity between the subcutaneous and intraperitoneal injections is very slight, while in the cat the toxicity of caffein is the same whether given by mouth or injected into the subcutaneous tissues, and is markedly less when injected into the peritoneal cavity. The experiments on dogs show considerable variation of effective dose when given by mouth, but the interesting observation was made that the toxic dose by mouth may be smaller in some cases than the average dose by subcutaneous injection. If the resistance to caffein by subcutaneous injection of the different species of animals experimented upon in the present research be compared, it will be noticed that the gray rabbit or Belgian hare, which is more resistant than the other varieties employed, stands more caffein in proportion to the weight of the body than the other animals.

Although the minimum fatal dose was found to be somewhat larger for the guinea pig than for the gray rabbit when caffein was injected intraperitoneally, it was on the contrary smaller by other paths of introduction, and approximated quite closely the minimum fatal dose for rabbits of the other varieties. Cats as well as dogs were found to be distinctly less resistant to caffein than the herbivora.

There are a number of factors far more important than zoological differences which influence the toxicity of caffein. Some of these are age, season, and pathologic conditions. As these factors have already been dwelt upon in their appropriate places, further discussion might seem unnecessary, but owing to their importance in determining the action of a drug, emphasis is desirable. Especially is this the case with pathological conditions in relation to toxicity. While no positive proof of diminished resistance to caffein in pathological conditions was obtained by subjecting the suggestion to experimental test, it was observed in these experiments on rabbits that death occurred in some individuals after small doses which are usually not even toxic. The findings at autopsy indicate the presence of pathological conditions. The same was observed in some experiments on cats and dogs. It is extremely probable, therefore, that disease modifies the reaction of the organism to caffein as well as to other drugs.(78)

That the resistance to drugs may vary according to the age of the subject has been maintained by a number of pharmacologists. According to Guinard,(30) young dogs, rabbits, and guinea pigs are very susceptible to morphin, resembling children in this regard.[E] The minimum fatal dose for these animals is about one-third less than for the adult. This is not true, however, for the young of other species. Cats under 15 days of age tolerate twice the toxic dose of morphin for the adult cat. Young beeves and goats are likewise more resistant to this alkaloid than adults. On the other hand, according to Livon,(54) young guinea pigs are more sensitive to alkaloids than adults. The toxicity of caffein, as shown in the present investigation, was found to be less in the young than in the adult. In dogs the young subjects are in some instances almost twice as resistant as adults. The difference was found to be less in cats and rabbits than in dogs, but it was quite marked.

[E] A case of accidental poisoning reported recently by Wichura (Münich. Méd. Woch., 1911, No. 30, p. 1618) throws some doubt on the accepted view that the susceptibility of young children to morphin is greater than that of adults. Wichura also found that the therapeutic doses of codein preparations ordinarily recommended for children in pleuritic cough are not effective in this condition.

The effect of season on the toxicity of drugs has been discussed in the section on the experiments on guinea pigs, which were more resistant to caffein in the fall than in February and March. The effect of season seems to vary with the animal, but it may also differ with the substance employed. In Noe's(65) studies on this subject cantharidin was found to be more toxic for the hedgehog in November than in July. The effect of season was different for morphin, as it was observed that the resistance of the hedgehog was greater at the end of the summer than earlier in the season.

The relation of diet to toxicity of drugs has been studied by Hunt.(39) His experiments indicate that this is an important factor in the resistance to acetonitril. The studies here reported on the effect of diet on toxicity of caffein in rabbits were confined to experiments with oats and carrots and do not show any modification of the resistance to caffein. The question of diet in chronic intoxication in dogs, however, suggests that in these animals diet may affect the toxicity of caffein, although the data on this subject are far from satisfactory. There is nevertheless sufficient evidence to suggest that a high protein diet for the adult dog tends to greater resistance of the animal to caffein and similarly the growing dog tolerates larger quantities of caffein on a milk diet than on a diet of meat.

This brings us to a consideration of the behavior of caffein in chronic intoxication. Although in both rabbits and dogs absence of cumulation was evident, in other respects decided differences in the resistance to caffein were observed. While the rabbit tolerates more than twice the single dose of caffein per kilo for the dog, the result is quite different in repeated dosage of the drug, the rabbit succumbing to continued administration of much smaller doses of the drug than the dog. This is probably due to lesions of the gastro-intestinal canal caused by caffein which occasions loss of appetite much more readily in the rabbit than in the dog. The abundant energy reserve in the dog makes it possible for this animal to stand inanition much longer than the rabbit and other herbivora. The difference in the behavior of the rabbit and dog toward caffein is interesting as showing complete reversal of resistance in acute and chronic intoxication. From the statement in the introduction it is evident that the size of the single toxic or lethal dose of a substance is in no wise an index of the active degree of its toxicity. The experiments with caffein here reported furnish additional evidence that this is true, at least for the rabbit.

GENERAL SUMMARY AND CONCLUSIONS.

The toxicity of caffein in the rabbit varies with the mode of its administration, being least when given by mouth and greatest by intravenous administration. The toxicity is from 15 to 20 per cent greater by subcutaneous injections than by mouth, but is about half of that when injected into the peritoneal cavity. No difference was observed in the toxicity of caffein whether administered into gluteal or into the lumbar muscles. When introduced by this route the toxicity was found to be less by one-third than when it is injected into the peritoneal cavity, but is about 30 per cent more toxic than the subcutaneous injections. White or black rabbits were found to be less resistant to caffein than gray rabbits.

The resistance of the guinea pig to caffein, as of the rabbit, is greatest when given by mouth. The minimum fatal dose is less by intraperitoneal injections, but greater than by subcutaneous injections, thus differing from the rabbit in this regard. The adult cat is less resistant than the guinea pig or rabbit to caffein. The minimum lethal dose by mouth is the same as by subcutaneous, and is less than by intraperitoneal, injection. The minimum fatal dose for dogs was found to be the same by mouth as by subcutaneous injection and is almost the same as for the cat. The toxicity of caffein varies in the guinea pig according to season of the year.

Age is likewise a factor in the toxicity of caffein, young animals being more resistant than the full-grown and older animals; this was shown in experiments on rabbits, cats, and dogs. The symptoms of caffein poisoning also were different in puppies and in full-grown dogs. Different diets, such as carrots and oats, did not influence the resistance of rabbits and guinea pigs to caffein. Low protein diet tends to decrease resistance to caffein in dogs. Young growing dogs are less resistant to caffein on a meat than on a milk diet. Caffein is not cumulative in the rabbit or dog, even if administered for a considerable length of time. Some degree of tolerance may be induced in the rabbit under certain conditions, but not in dogs under the conditions of the experiments made in this investigation. The possibility, however, that dogs may acquire tolerance for caffein is not excluded. Although the rabbit tolerates a much larger single dose of caffein than the dog, it was found, in experiments on chronic intoxication that the rabbit is less resistant to caffein than the dog. The toxicity of caffein is probably increased under pathological conditions, since comparatively smaller doses were fatal to rabbits, cats, and dogs, when marked lesions not due to caffein were found at autopsy. Glycosuria was observed in rabbits, guinea pigs, and cats when caffein was given in sufficient amounts.

TABLE 18.--_Acute caffein intoxication: Table showing average minimum toxic and minimum fatal doses for adult animals._

Table headings:

SC: Subcutaneously. BM: By mouth. IP: Intraperitoneally. IM: Intramuscular. IV: Intravenous.

+------+------------------------------------------------- |Effect| Dose per kilo (grams) Animal. |of +---------+-----------+-----------+-------+------- |dose. | SC | BM | IP | IM | IV -------------+------+---------+-----------+-----------+-------+------- Rabbit, gray |{Toxic| 0.15| 0.325| 0.100| 0.13| 0.05 | | | | -0.125 | -0.15| |{Fatal| .30| .350| .150| .20| 0.10 | | | | | | -.16 Rabbit, white|{Toxic| | | | | or black |{Fatal| .20| .290| | | | | | | | | Guinea pig |{Toxic|0.15- .16| .150| .200| | |{Fatal| .20- .24|0.280- .300| .240- .250| | | | | | | | Cat |{Toxic| .12- .14| .125| .125- .150| | |{Fatal| .15| .150| .180- .200| | | | | | | | Dog |{Toxic| | .100- .120| | | |{Fatal| .15- .16| .140- .150| | | -------------+------+---------+-----------+-----------+-------+-------

NOTE.--The doses given in this table are approximate.

CAFFEIN BIBLIOGRAPHY.

1. ABDERHALDEN and BRAHM. Zts. Physiol. Chem., 1909, =62=: 133.

2. ALBERS. Deut. Klin., 1852, =5=: 577.

3. AMAT. Diss. Paris, 1889.

4. AMORY. Boston Med. Surg. J., 1868, =1=: 261.

5. VON ANREP. Arch. Ges. Phys., 1880, =21=: 185.

6. AUBERT. Pflüger's Arch., 1872, =5=: 589.

7. AUER and MELTZER. J. Pharm. Exper. Ther., 1911, =2=: 402.

8. BALDI. Terapia Moderna, 1891, =5=: 617.

9. BENNETT. Brit. Med. J., 1874, =2=: 674.

10. BERNARD, CLAUDE. Leçons sur les substances toxiques, Paris, 1857, p. 292.

11. BINZ. Arch. Exper. Path. Pharm., 1878, =9=: 31.

12. ---- Ibid., 1891, =28=: 197.

13. BRILL. Diss. Marburg, 1861.

14. BUCHHEIM and EISENMENGER. Beitr. Anat. Physiol., 1870, =5=: 115.

15. BUNGE and SCHMIEDEBERG. Arch. Exper. Path. Pharm., 1876, =6=: 233.

16. CHITTENDEN. The Nutrition of Man, 1907.

17. COGSWELL. Lancet (London), 1852, =2=: 488.

18. DANILEWSKI. Arch. Exper. Path. Pharm., 1894, =35=: 105.

19. DRZEWINA. Compt. rend. Soc. biol., 1911, =70=: 772.

20. DUMAS and PELLETIER. Ann. chim. phys., 1823, =24=: 182.

21. EDMUNDS. J. Amer. Med. Assoc., 1907, =48=: 1744.

22. FILEHNE. Arch. Phys., 1886, p. 72.

23. FLEISCHER and LOEB. Archives of Internal Medicine, 1909, =3=: 78.

24. FOCKE. Arch. Pharm., 1903, =241=: 678.

25. FRERICHS. Handwörterbuch Phys., 1846, =3=: 721.

26. FRÖHNER. Monats. prakt. Tierh., 1892, =3=: 529.

27. GENTILHOMME. Bull. Soc. Med. Reims, 1867, =5=: 93.

28. GOUREWITCH. Arch. Exper. Path. Pharm., 1907, =57=: 314.

29. GUINARD. Compt. rend. Soc. biol., 1900, =2= (2d ser.): 727.

30. ----. Compt. rend., 1893, =113=: 520.

31. ----. La Morphine et l'apomorphine, Paris, 1903.

32. GUNN. Arch. Int. Pharm. Ther., 1909, =19=: 319.

33. HALE. U. S. Public Health and Marine-Hospital Service. Hyg. Lab. Bul. 53, p. 43.

34. HARRINGTON. Amer. J. Phys., 1898, =1=: 385.

35. HATCHER. J. Amer. Med. Assoc., 1910, =60=: 746.

36. HENNEGUY. Diss. Montpellier, 1875.

37. HOFMEISTER. Arch. Exper. Path. Pharm., 1894, =33=: 198.

38. HOPPE. Écho Méd. Neuchâtel, 1858.

39. HUNT. U. S. Public Health and Marine-Hospital Service. Hyg. Lab. Bul. 69, p. 51.

40. ----. Ibid., Bul. 33.

41. IGERSHEIMER and STAMI. Arch. Exper. Path. Pharm., 1909, =61=: 18.

42. JACOBI and GOLOWINSKI. Arch. Exper. Path. Pharm., Supplement Bd. 1908, p. 286.

43. JOBST. Ann. Pharm., 1838, =25=: 63.