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THE POSTERIOR LOBE OF THE PITUITARY GLAND
1099
THE POSTERIOR LOBE OF THE PITUITARY GLAND
with any regularity in animals of an extract of a normal tissue.
by
the
injection
EFFECT ON GASTRIC SECRETION
ITS
RELATIONSHIP TO THE STOMACH AND TO THE BLOOD PICTURE
BY E. C.
DODDS, M.V.O., D.Sc., M.D., F.R.C.P. Lond.
COURTAULD PROFESSOR OF BIOCHEMISTRY
IN
THE
UNIVERSITY
OF LONDON
HILLS, B.Sc., Oxon.* NOBLE, M.D. Toronto †; AND
G. M. R. L.
P. C.
WILLIAMS,
B.Sc. Lond.*
(From the Courtauld Institute of Biochemistry, Middlesex Hospital, London) been shown (Dodds, Noble, and and Noble 2) that it is possible to prepare an extract of the posterior lobe of the pituitary gland which when injected subcutaneously into animals will produce an acute haemorrhagic lesion of the acid-bearing area of the stomach ; and the animals usually recover completely in a period of some ten days. The same effects can be produced in rabbits by the oral administration of the extract. These lesions have been described in previous papers in detail. It has been shown that if the contents of the stomach be rendered alkaline prior to injection, it is impossible to obtain the characteristic reaction. The most significant points concerning the gastric lesion are, firstly, its extremely accurate limitation to the area of the stomach in which oxyntic cells are found, and, secondly, the complete absence of lesions in other parts of the alimentary canal. It has also been demonstrated that similar lesions can be produced in all the laboratory animals testednamely, monkey, cat, rabbit, guinea-pig, rat, and IT has
previously
Smith’; Dodds
mouse.
The
present communication deals with the
exten-
sion of these investigations, and it will be seen that the extract exerts a profound influence upon the blood picture. PRODUCTION OF CHRONIC ULCERS
In view of the acute lesions produced by a single injection, it was decided to study the effects of repeated doses of the extract. A typical experiment was as follows : a rabbit was given 40 c.cm. of standard B.P. pituitrin by stomach-tube every 7 days over a period of 8 weeks. During this time the animals appeared to be in good condition and did not exhibit any unusual symptoms after the initial injection. At autopsy a typical punched-out ulcer was found on the greater curvature which had perforated but was healed by omentum. Chronic ulcers have been produced following a single subcutaneous dose of 75 mg. per kg. of the acetone picric acid extract of the posterior lobe of the pituitary. An ulcer has also been found following four subcutaneous injections of 5 c.cm. of standard B.P. pituitary extract, given every other day. These ulcers are always single and situated in the acidbearing area of the stomach. Their gross appearance is typically that seen in chronic gastric ulcers in man. We believe that this is the first occasion on which chronic gastric ulcers have been produced *
Working with a grant from the Medical Research Council. † Working with the Ellen Mickle Fellowship from the University of Toronto.
An attempt was made to study the mechanism for the production of the lesion. Since it has been shown that the rendering of the stomach contents alkaline is capable of abolishing the typicai iesion, it follows that gastric acidity must in some way be associated with this phenomenon. A technique for studying the reaction of rabbits’ stomachs to histamine was worked out. The following is a very brief account of a typical experiment. The rabbits were put on a milk and bran diet for two weeks to allow the stomach contents to become sufficiently liquid to be washed out. Under sodium amytal anaesthesia the pylorus was ligatured, the most dependent portion of the stomach opened, and an external fistula was established. Through this the contents were removed and the stomach was washed out. In the control series of animals the resting juice was collected for half an hour. This was followed by the subcutaneous injection of 0-25 mg. per kg. of
histamine.
Thirty-minute specimens
were
collected,
titrated for free hydrochloric acid and for total acidity. In the experimental group the above technique was followed except that these animals received 2 c.cm. per kg. of standard B.P. pituitrin intravenously, and then after 30 minutes the histamine injection. The control series showed a typical profuse secretion due to the histamine with the free HCI reaching a peak of 80-130 c.cm. of N/10 HC1 per 100 c.cm. of juice in 90-120 minutes. In the experimental group we have never found any free HCI secreted, even though large doses of histamine have been repeated at the end of 5 hours. This reaction has also been obtained using the acetone picric acid extract administered intravenously. From these and other experiments it would appear that injection of the extract is capable of causing a temporary inhibition of secretion of hydrochloric acid, in such a way that the stomach will not react to the histamine stimulus for a period of some 6 hours. It is interesting to note at this juncture that other previous workers have called attention to the fact that injection of pituitary extract reduced gastric and
were
acidity.34
EFFECTS ON BLOOD
It has been shown5 that very profound changes occur in the blood of animals injected with this extract. Thus, if a rabbit be injected subcutaneously with the preparation, it will be found on the fifth day that a severe anasmia has developed, the blood count being reduced from 6,000,000 down to as low as just over 1,000,000 red cells. The haemoglobin is also reduced but not proportionately to the same extent. On the fifth or sixth day a reticulocytosis appears which may rise to as high as 50 per cent. There is a marked anisocytosis and a definite macrocytosis. If the animals be killed obvious changes are seen in the spleen, such as haemorrhagic infarction, and there is evidence of a stimulation of the blooddestroying system. The intestines are full of bile, and it would appear that the anaemia may possibly be ascribed to an increased blood destruction. If left alone the animals will recover completely. DISCUSSION
impossible at the present time to offer any complete explanation of the various phenomena recorded, but the following remarks may assist in an attempt to estimate the significance of these investigations. In the first instance it must be It is
1100 the substance, or substances, reaction are contained only in the pituitary gland. Exhaustive controls have been made with other tissues of the body, employing not only doses identical to those which prove active in the case of the pituitary, but with doses many times larger. We have, however, been unable to obtain with these any action upon the stomach or blood picture. The following is a list of tissues from which extracts have been prepared : spleen, liver, kidney, testes, muscle, pancreas, suprarenal, brain, stomach, thyroid, and placenta. As pointed out in previous publications, the factor or factors necessary for the production of a gastric lesion and blood picture are contained in the commercial pituitary extract (B.P.) as well as in the pituitary extract made by the acetone-picric 6 acid process by the method of Dodds and DiclLens.6 It seems at first sight that the quantity of material injected is so great as to make it doubtful whether the phenomena described are really exaggerations of a natural process. It must be borne in mind, however, that the B.P. pituitary extract is standardised for the oxytocic and vasopressive principles in units which bear no relation to the, phenomena described. By application of the method of preparation and purification of the vasopressive principle described by Kamm et al.it has been found that the actual weight of an effective dose of the active principle may be reduced to a figure quite compatible with those of other hormones. On the weight basis, therefore, it appears that no theoretical objection could be made to our observations on these
pointed out that responsible for this the posterior lobe of .
grounds. A full
explanation of the connexion between the pituitary gland and the stomach and the blood picture can only be arrived at by a study of the hypophysectomised animal, and this is in activeprogress
CLINICAL
present time in association with Dr. A. S. Parkes and Mr. 1. W. Rowlands. From the clinical aspect, our observations might possibly provide an explanation for a number of For example, in hitherto puzzling phenomena. Simmonds’s disease it is known that there is atrophy of the organs of the reticulo-endothelial system.8 9 Again, in certain cases of basophil adenoma of the anterior lobe of the pituitary gland, ulceration of the stomach 10 and also erythraemia have been recorded." In cases of duodenal ulcer polycythsemia, has also been observed.l2 The occurrence of acute gastric lesions following operations on the base of the brain has been recorded on several occasions in the publications of Harvey Cushing.13 From these investigations it seems justifiable to consider the possibility of a hormonal connexion between the posterior lobe of the pituitary gland, the stomach, and the blood picture.
at the
One of us (E. C. D.) is indebted to the Medical Research Council for a grant towards the defrayment of the cost of material for this investigation. REFERENCES
Dodds, E. C., Noble, R. L., and Smith, E. R.: THE LANCET, 1934, ii., 918. 2. Dodds, E. C., and Noble, R. L.: Jour. Soc. Chem. Ind., 1934, liii. (Chemistry and Industry), 1026. 3. Sollmann, T.: Manual of Pharmacology, London, 1932,
1.
p. 477. 4. Cascao de Anciaes, J. H.: Compt. rend. soc. de biol., 1926, xcv., 313. 5. Dodds, E. C., and Noble, R. L.: Nature, May 11th, 1935. 6. Dodds, E. C., and Dickens, F. : Brit. Jour. Exp. Path., 1924, v., 115. 7. Kamm, O., Aldrich, T. B., Grote, I. W., Rowe, L. W., and Bugbee, E. P. : Jour. Amer. Chem. Soc., 1928, l., 573. 8. Riecher, H. H., and Curtis, A. C.: Jour. Amer. Med. Assoc., 1932, xcix., 110. 9. Calder, R. M.: Bull. Johns Hopkins Hosp., 1932, 1., 87. 10. Swan, W. G. A., and Stephenson, G. E.: THE LANCET, Feb. 16th, 1935, p. 372 11. Coshing, H. : Bull. Johns Hopkins Hosp., 1932, 1., 137. 12. Boyd, W.: Amer. Jour. Med. Sci., 1934, clxxxvii., 589. 13. Cushing, H.: Surg., Gyn., and Obst., 1932, lv., 1.
AND LABORATORY NOTES
PARALYTIC ILEUS CURED WITH PROSTIGMIN
Evacuans
were
given.
This
was
vomited and the dose
repeated at 2 P.M. He vomited at 7.30 P.M. and was complaining of abdominal pain and distension. No result from aperient was obtained, so he was given two vegetable
laxative tablets at 8 P.M. He vomited off and on the whole night; the abdomen remained distended and there was no result (either flatus or A REPORT to the Therapeutic Trials Committee faeces) from the aperient. Hourly foments were applied to abdomen and on April 3rd a turpentine enema produced of the Medical Research Council published a year the a good fagcal result and he was put on to 1 c.cm. pituitrin, ago1 described the use of prostigmin in a series to be followed by 0’5 c.cm. three-hourly. Towards noon of cases of abdominal distension following operation. his distension had increased and he was unable to pass The dose ranged from 1.0 to 4’0 c.cm., but the larger flatus alone or with the help of a flatus tube. Hiccup quantity was used on one occasion only and the usual began and his distension increased. Nothing abnormal be felt per rectum. Towards midnight he was given practice was to givee 1’0 c.cm., repeated if necessary. could another turpentine enema, which gave a good, formed None of the patients mentioned in that report seem result, but the abdomen still remained tensely distended to have been as gravely ill as the one who is the and the skin was stretched and shining, and the patient of the The case is present note. subject interesting looked drawn, haggard, and altogether extremely ill, not only because almost every known remedy (except his pulse and temperature did not appear to although acetylcholine) was previously given, but because the reflect the severity of his abdominal condition. He was dose of prostigmin administered was the large one at 12.40 A.M. on April 4th put on to eserine, gr. 1/100 recommended by Dr. M. B. Walker for the treatment every four hours. He vomited green bile-stained fluid and hiccupped off of myasthenia gravis. and on the whole day. The abdomen at 4.30 P.M. was even A. B., aged 23, was seen on March 30th, 1935, with more distended and no gurgling could be heard with the acute appendicitis. Temp. 101’F; pulse-rate 88. He had stethoscope. A turpentine enema gave a fluid return with had abdominal pain for 48 hours before sending for medical streaks of blood-stained mucus, but no flatus was passed. assistance. The operation revealed a gangrenous appendix, An ox-gall enema was given at 6.20 P.M. with no result, which was removed and a drainage-tube inserted. On followed at 9.30 P.M. by 20 c.cm. of anti-gas gangrene the following day the tube was changed and the patient serum. An hour later the patient was given a spinal was given a turpentine enema, with a very small faecal anaesthetic (Stovaine) with no result. At midnight he result. On April 2nd, at noon, 2 drachms of Cascara was given an enema of milk and molasses with no result. He was still vomiting and had a very bad night; his 1 Carmichael, E. A., Fraser, F. R., McKelvey, D., and temperature on April 5th was 100°, the pulse-rate varying Wilkie, D. P. D.: THE LANCET, 1934, i., 942.
BY W. E.
DAVID, M.C., M.B. Sydney
THE POSTERIOR LOBE OF THE PITUITARY GLAND
with any regularity in animals of an extract of a normal tissue.
by
the
injection
EFFECT ON GASTRIC SECRETION
ITS
RELATIONSHIP TO THE STOMACH AND TO THE BLOOD PICTURE
BY E. C.
DODDS, M.V.O., D.Sc., M.D., F.R.C.P. Lond.
COURTAULD PROFESSOR OF BIOCHEMISTRY
IN
THE
UNIVERSITY
OF LONDON
HILLS, B.Sc., Oxon.* NOBLE, M.D. Toronto †; AND
G. M. R. L.
P. C.
WILLIAMS,
B.Sc. Lond.*
(From the Courtauld Institute of Biochemistry, Middlesex Hospital, London) been shown (Dodds, Noble, and and Noble 2) that it is possible to prepare an extract of the posterior lobe of the pituitary gland which when injected subcutaneously into animals will produce an acute haemorrhagic lesion of the acid-bearing area of the stomach ; and the animals usually recover completely in a period of some ten days. The same effects can be produced in rabbits by the oral administration of the extract. These lesions have been described in previous papers in detail. It has been shown that if the contents of the stomach be rendered alkaline prior to injection, it is impossible to obtain the characteristic reaction. The most significant points concerning the gastric lesion are, firstly, its extremely accurate limitation to the area of the stomach in which oxyntic cells are found, and, secondly, the complete absence of lesions in other parts of the alimentary canal. It has also been demonstrated that similar lesions can be produced in all the laboratory animals testednamely, monkey, cat, rabbit, guinea-pig, rat, and IT has
previously
Smith’; Dodds
mouse.
The
present communication deals with the
exten-
sion of these investigations, and it will be seen that the extract exerts a profound influence upon the blood picture. PRODUCTION OF CHRONIC ULCERS
In view of the acute lesions produced by a single injection, it was decided to study the effects of repeated doses of the extract. A typical experiment was as follows : a rabbit was given 40 c.cm. of standard B.P. pituitrin by stomach-tube every 7 days over a period of 8 weeks. During this time the animals appeared to be in good condition and did not exhibit any unusual symptoms after the initial injection. At autopsy a typical punched-out ulcer was found on the greater curvature which had perforated but was healed by omentum. Chronic ulcers have been produced following a single subcutaneous dose of 75 mg. per kg. of the acetone picric acid extract of the posterior lobe of the pituitary. An ulcer has also been found following four subcutaneous injections of 5 c.cm. of standard B.P. pituitary extract, given every other day. These ulcers are always single and situated in the acidbearing area of the stomach. Their gross appearance is typically that seen in chronic gastric ulcers in man. We believe that this is the first occasion on which chronic gastric ulcers have been produced *
Working with a grant from the Medical Research Council. † Working with the Ellen Mickle Fellowship from the University of Toronto.
An attempt was made to study the mechanism for the production of the lesion. Since it has been shown that the rendering of the stomach contents alkaline is capable of abolishing the typicai iesion, it follows that gastric acidity must in some way be associated with this phenomenon. A technique for studying the reaction of rabbits’ stomachs to histamine was worked out. The following is a very brief account of a typical experiment. The rabbits were put on a milk and bran diet for two weeks to allow the stomach contents to become sufficiently liquid to be washed out. Under sodium amytal anaesthesia the pylorus was ligatured, the most dependent portion of the stomach opened, and an external fistula was established. Through this the contents were removed and the stomach was washed out. In the control series of animals the resting juice was collected for half an hour. This was followed by the subcutaneous injection of 0-25 mg. per kg. of
histamine.
Thirty-minute specimens
were
collected,
titrated for free hydrochloric acid and for total acidity. In the experimental group the above technique was followed except that these animals received 2 c.cm. per kg. of standard B.P. pituitrin intravenously, and then after 30 minutes the histamine injection. The control series showed a typical profuse secretion due to the histamine with the free HCI reaching a peak of 80-130 c.cm. of N/10 HC1 per 100 c.cm. of juice in 90-120 minutes. In the experimental group we have never found any free HCI secreted, even though large doses of histamine have been repeated at the end of 5 hours. This reaction has also been obtained using the acetone picric acid extract administered intravenously. From these and other experiments it would appear that injection of the extract is capable of causing a temporary inhibition of secretion of hydrochloric acid, in such a way that the stomach will not react to the histamine stimulus for a period of some 6 hours. It is interesting to note at this juncture that other previous workers have called attention to the fact that injection of pituitary extract reduced gastric and
were
acidity.34
EFFECTS ON BLOOD
It has been shown5 that very profound changes occur in the blood of animals injected with this extract. Thus, if a rabbit be injected subcutaneously with the preparation, it will be found on the fifth day that a severe anasmia has developed, the blood count being reduced from 6,000,000 down to as low as just over 1,000,000 red cells. The haemoglobin is also reduced but not proportionately to the same extent. On the fifth or sixth day a reticulocytosis appears which may rise to as high as 50 per cent. There is a marked anisocytosis and a definite macrocytosis. If the animals be killed obvious changes are seen in the spleen, such as haemorrhagic infarction, and there is evidence of a stimulation of the blooddestroying system. The intestines are full of bile, and it would appear that the anaemia may possibly be ascribed to an increased blood destruction. If left alone the animals will recover completely. DISCUSSION
impossible at the present time to offer any complete explanation of the various phenomena recorded, but the following remarks may assist in an attempt to estimate the significance of these investigations. In the first instance it must be It is
1100 the substance, or substances, reaction are contained only in the pituitary gland. Exhaustive controls have been made with other tissues of the body, employing not only doses identical to those which prove active in the case of the pituitary, but with doses many times larger. We have, however, been unable to obtain with these any action upon the stomach or blood picture. The following is a list of tissues from which extracts have been prepared : spleen, liver, kidney, testes, muscle, pancreas, suprarenal, brain, stomach, thyroid, and placenta. As pointed out in previous publications, the factor or factors necessary for the production of a gastric lesion and blood picture are contained in the commercial pituitary extract (B.P.) as well as in the pituitary extract made by the acetone-picric 6 acid process by the method of Dodds and DiclLens.6 It seems at first sight that the quantity of material injected is so great as to make it doubtful whether the phenomena described are really exaggerations of a natural process. It must be borne in mind, however, that the B.P. pituitary extract is standardised for the oxytocic and vasopressive principles in units which bear no relation to the, phenomena described. By application of the method of preparation and purification of the vasopressive principle described by Kamm et al.it has been found that the actual weight of an effective dose of the active principle may be reduced to a figure quite compatible with those of other hormones. On the weight basis, therefore, it appears that no theoretical objection could be made to our observations on these
pointed out that responsible for this the posterior lobe of .
grounds. A full
explanation of the connexion between the pituitary gland and the stomach and the blood picture can only be arrived at by a study of the hypophysectomised animal, and this is in activeprogress
CLINICAL
present time in association with Dr. A. S. Parkes and Mr. 1. W. Rowlands. From the clinical aspect, our observations might possibly provide an explanation for a number of For example, in hitherto puzzling phenomena. Simmonds’s disease it is known that there is atrophy of the organs of the reticulo-endothelial system.8 9 Again, in certain cases of basophil adenoma of the anterior lobe of the pituitary gland, ulceration of the stomach 10 and also erythraemia have been recorded." In cases of duodenal ulcer polycythsemia, has also been observed.l2 The occurrence of acute gastric lesions following operations on the base of the brain has been recorded on several occasions in the publications of Harvey Cushing.13 From these investigations it seems justifiable to consider the possibility of a hormonal connexion between the posterior lobe of the pituitary gland, the stomach, and the blood picture.
at the
One of us (E. C. D.) is indebted to the Medical Research Council for a grant towards the defrayment of the cost of material for this investigation. REFERENCES
Dodds, E. C., Noble, R. L., and Smith, E. R.: THE LANCET, 1934, ii., 918. 2. Dodds, E. C., and Noble, R. L.: Jour. Soc. Chem. Ind., 1934, liii. (Chemistry and Industry), 1026. 3. Sollmann, T.: Manual of Pharmacology, London, 1932,
1.
p. 477. 4. Cascao de Anciaes, J. H.: Compt. rend. soc. de biol., 1926, xcv., 313. 5. Dodds, E. C., and Noble, R. L.: Nature, May 11th, 1935. 6. Dodds, E. C., and Dickens, F. : Brit. Jour. Exp. Path., 1924, v., 115. 7. Kamm, O., Aldrich, T. B., Grote, I. W., Rowe, L. W., and Bugbee, E. P. : Jour. Amer. Chem. Soc., 1928, l., 573. 8. Riecher, H. H., and Curtis, A. C.: Jour. Amer. Med. Assoc., 1932, xcix., 110. 9. Calder, R. M.: Bull. Johns Hopkins Hosp., 1932, 1., 87. 10. Swan, W. G. A., and Stephenson, G. E.: THE LANCET, Feb. 16th, 1935, p. 372 11. Coshing, H. : Bull. Johns Hopkins Hosp., 1932, 1., 137. 12. Boyd, W.: Amer. Jour. Med. Sci., 1934, clxxxvii., 589. 13. Cushing, H.: Surg., Gyn., and Obst., 1932, lv., 1.
AND LABORATORY NOTES
PARALYTIC ILEUS CURED WITH PROSTIGMIN
Evacuans
were
given.
This
was
vomited and the dose
repeated at 2 P.M. He vomited at 7.30 P.M. and was complaining of abdominal pain and distension. No result from aperient was obtained, so he was given two vegetable
laxative tablets at 8 P.M. He vomited off and on the whole night; the abdomen remained distended and there was no result (either flatus or A REPORT to the Therapeutic Trials Committee faeces) from the aperient. Hourly foments were applied to abdomen and on April 3rd a turpentine enema produced of the Medical Research Council published a year the a good fagcal result and he was put on to 1 c.cm. pituitrin, ago1 described the use of prostigmin in a series to be followed by 0’5 c.cm. three-hourly. Towards noon of cases of abdominal distension following operation. his distension had increased and he was unable to pass The dose ranged from 1.0 to 4’0 c.cm., but the larger flatus alone or with the help of a flatus tube. Hiccup quantity was used on one occasion only and the usual began and his distension increased. Nothing abnormal be felt per rectum. Towards midnight he was given practice was to givee 1’0 c.cm., repeated if necessary. could another turpentine enema, which gave a good, formed None of the patients mentioned in that report seem result, but the abdomen still remained tensely distended to have been as gravely ill as the one who is the and the skin was stretched and shining, and the patient of the The case is present note. subject interesting looked drawn, haggard, and altogether extremely ill, not only because almost every known remedy (except his pulse and temperature did not appear to although acetylcholine) was previously given, but because the reflect the severity of his abdominal condition. He was dose of prostigmin administered was the large one at 12.40 A.M. on April 4th put on to eserine, gr. 1/100 recommended by Dr. M. B. Walker for the treatment every four hours. He vomited green bile-stained fluid and hiccupped off of myasthenia gravis. and on the whole day. The abdomen at 4.30 P.M. was even A. B., aged 23, was seen on March 30th, 1935, with more distended and no gurgling could be heard with the acute appendicitis. Temp. 101’F; pulse-rate 88. He had stethoscope. A turpentine enema gave a fluid return with had abdominal pain for 48 hours before sending for medical streaks of blood-stained mucus, but no flatus was passed. assistance. The operation revealed a gangrenous appendix, An ox-gall enema was given at 6.20 P.M. with no result, which was removed and a drainage-tube inserted. On followed at 9.30 P.M. by 20 c.cm. of anti-gas gangrene the following day the tube was changed and the patient serum. An hour later the patient was given a spinal was given a turpentine enema, with a very small faecal anaesthetic (Stovaine) with no result. At midnight he result. On April 2nd, at noon, 2 drachms of Cascara was given an enema of milk and molasses with no result. He was still vomiting and had a very bad night; his 1 Carmichael, E. A., Fraser, F. R., McKelvey, D., and temperature on April 5th was 100°, the pulse-rate varying Wilkie, D. P. D.: THE LANCET, 1934, i., 942.
BY W. E.
DAVID, M.C., M.B. Sydney