Enhancing Effect of Corticotrophin*

ENHANCING E F F E C T O F CORTICOTROPHIN* UPON

E X O P H T H A L M O S PRODUCED BY T H Y R O T R O P H I C H O R M O N E

J O A Q U I N BARRAQUER, M.D.,

A N D J O S É M.

CAÎÏADELL,

M.D.

Barcelona, Spain

Some fishes respond with an increase in the protrusion of the ocular globes as a result of different hormonal stimuli. Albert2 pro­ duced exophthalmos in Fundulus heteroclitus by means of intracelomic injections of an­ terior pituitary extracts. Dobyns and Steelman5 showed the possibility of obtaining the same effect by injecting preparations of pi­ tuitary gland without thyrotrophic action. Pickford9 on the contrary, observed that there is a narrow relation between exophthalmos-producing and thyroid-stimulating ef­ fects of thyrotrophic preparations. The same phenomenon was observed in the treatment with tri-iodothyronine and dessicated thy­ roid.7 On the other hand, Matty and coworkers 8 succeeded in producing exophthal­ mos by injecting androgens in two species of teleost fishes (Sparisoma squalidum and Scarus croicensis) and by the injection of L-thyroxine and tri-iodothyronine in these two species and also in the Bathystoma aurolineatum. In Spain it is impossible to obtain Fundu­ lus heteroclitus or any of the species of fishes with which the above-mentioned authors have worked. In a series of previous tests we in­ vestigated the possible exophthalmic action of the anterior pituitary extracts in the fol­ lowing fishes: Barbus fluviatilis, Ciprinus carpio, Leuciscus erythrophtalmus, Fundulus hispanicus, Carassius auratus, Perca fluviatillis, Tinea tinea, Gobitis fossilis, Gobius fiuviatilis and Ameiurus nebulosus. All these, except the last, under diverse experimental conditions can develop exophthalmos follow­ ing injection of thyrotrophin. Since we were able to obtain Carassius auratus (var. japonicus) in unlimited quantities we used this

species for almost all our investigations. Among the many types of gold fish, the Carassius auratus var. japonicus, subvar. bicaudatus show a larger sensitivity and uniformity of response. This subvariety has a more globulous body and a flatter head. From this fish procèdes by genetic selection the Carassius macrophthalmos in which enor­ mous protrusion of the ocular globes is sometimes a constitutionally inherited char­ acteristic (fig. 1). We have presented numerous clinical and experimental arguments in previous papers 4 ' 5 confirming that the exophthalmos character­ istic of Graves' disease and other related forms of ophthalmopathy (edematous exoph­ thalmos, malignant exaphthalmos, infiltrative ophthalmopathy, exophthalmic ophthalmoplegia, and so forth) are secondary to a hyperf unction of anterior pituitary. Accord­ ing to Aterman 3 the adrenal cortex also par­ ticipates in the pathogenesis of these condi­ tions. Experimenting with golden hamsters we have confirmed that the presence of the adrenal glands is absolutely indispensable to the provoking of exophthalmos by injections of thyrotrophin and that previous treatment with corticotrophin or adrenal steroids rein­ forces the exophthalmic action of the thyro­ trophic hormone.5 The series of experiments that follow were planned to study the effect of cortico­ trophin on the exophthalmos produced in Carassius auratus (var. japonicus, subvar. bicaudatus) by intracelomic injections of thy­ rotrophin and its influence on the eye pro­ trusion obtained by injection of thyrotrophin directly into the retrobulbar tissues. EXPERIMENTAL METHODS

PART I

* From the Department of Endocrinology, Uni­ versity of Barcelona School of Medicine, and the Institute Barraquer.

Groups of 10 Carassius, about two years of age and uniform in size (9-10 cm. long)

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JOAQUlN BARRAQUER AND JOSÉ M. CANADELL

Fig. 1 (Barraquer and Cafiadell). Telescopic-eyed gold fish (Carassius macrophthalmus) with genetic constitutional exophthalmos and a common gold fish (Carassius auratus).

were injected intracelomically at intervals of eight hours with thyrotrophic extract, the doses varying from 0.25 to 25 Heyl-Laqueur units. Other similar groups were injected with corticotrophin, chorionic gonadotrophin, growth hormone, human serum, and normal saline solution. The treatment was followed for a space of five consecutive days, examin­ ing the aspect of the fishes and measuring the protrusion of ocular globes with a caliper with a Vernier scale before giving the in­ jections. PART II

Other groups of gold fish were treated with the same doses of thyrotrophin adding to each injection two international units of corticotrophin. Control groups were similarly treated with corticotrophin and chorionic gonadotrophin, growth hormone extract, hu­ man serum and normal saline solution. PART III

In this experiment the exophthalmic ef­ fects of the retrobulbar injections of thy­ rotrophin were studied. The material was injected only once across the conjunctival sac of both eyes with an insulin syringe and a needle of small caliber. The dose was 0.05 cc. (5.0 u. Heyl-Laqueur). Control fishes

were injected with the same volume of cor­ ticotrophin, corionic gonadotrophin, growth hormone and normal saline solution. The re­ sulting ocular protrusion was determined immediately after the injections and at regu­ lar intervals of eight hours. PART IV

The fishes were previously treated intra­ celomically with a dose of 5.0 u. of corticotrophin-zinc and afterward exophthalmos was brought about in the same manner as in the fishes of the previous experiment. RESULTS

The intracelomic injections of thyrotro­ phin at doses of 10 u. Heyl-Laqueur (total quantity injected 150 u.) produced a variable exophthalmic effect. Only in two fishes of 10 injected was a small augmentation of the ocular protrusion and the intercorneal dis­ tance increased 1.5 mm. and 1.8 mm., re­ spectively. Administering 25 u. by injection (total dose 375 u.) exophthalmos was ob­ served in seven fishes, four of which were perfectly evident on simple inspection and without the measuring of intercorneal dis­ tance being necessary (fig. 2). The increase in the intercorneal distance was 3.8 mm. (±1.15 mm.). Larger doses of 25 u. were

CORTICOTROPHIN AND EXOPHTHALMOS

80S

Fig. 2 (Barraquer and Cafiadell). Exophthalmos produced with systemic treatment with thyrotrophin. (At right) control fish.

ill tolerated and smaller doses of 10 u. had no effect. Exophthalmos was not observed in any of the fishes of the control groups injected with corticotrophin, growth hor­ mone, chorionic gonadotrophin, human serum, or normal saline solution. In the animals with positive results, the exophthalmos began to appear on the third day of the experiment, became more evident in the two following days, and disappeared completely at the end of two or three days after ceasing to inject thyrotrophin. We observed that the injection of corti­ cotrophin reinforced the exophthalmic effect of the thyrotrophin without increasing the intensity of the maximum result. Injections of 10 u. of thyrotrophin, with a very small exophthalmic effect (see above), produced exophthalmos in six fishes with increase of the intercorneal distance of 2.9 mm. ( ± 0 . 6 mm.) when it was injected with corti­ cotrophin. Injections of 25 u. of thyrotrophin caused exophthalmos in nine fishes. The evolution of exophthalmos followed the same course as in the animals injected only with thyrotrophin without complementary treat­ ment with corticotrophin. Control groups in­ jected with thyrotrophin and chorionic gon­ adotrophin, growth hormone, human serum or normal saline solution developed exoph­

thalmos of the same kind as that of the fishes treated only with thyrotrophic hor­ mone. These products are devoid of the enhancing effect of corticotrophin. The retrobulbar injections of a small quan­ tity of normal saline produced a transitory increase of the ocular protrusion, could ob­ tain an augmentation in the intercorneal dis­ tance of five mm. or more with perfect tol­ erance by the fishes. The exophthalmos persisted for a space of some four hours, then diminished progressively and disappeared in the course of the following eight hours. In­ jection of gonadotrophic hormone, cortico­ trophin, growth hormone or human serum produced the same phenomenon but, possibly because of the proteic composition of these fluids, the exophthalmos persisted for about 24 hours. On the contrary the exophthalmos produced by injection of thyrotrophin is always more persistent and after 24 hours the increase of the intercorneal dis­ tance is still 80 percent ( ± six percent) of the initial value and after 48 hours it is 12 percent ( ± seven percent). In the fishes treated previously with corticotrophin-zinc one observes that the increase of the ocular protrusion produced by thyro­ trophin is much more persistent. After 24 hours the increase in the intercorneal dis-

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JOAQUÎN BARRAQUER AND JOSÉ M. CANADELL

Fig. 3 (Barraquer and Canadell). Ocular protrusion provoked by retrobulbar injections of thyrotrophin 24 hours before. First fish on the left was previously treated with long-acting corticotrophin ; the fish in the middle did not receive corticotrophin ; on the right is the control fish in which exophthalmos was produced by injecting human serum. Note the persistence of ocular protrusion of both fishes treated with thyrotrophic hormone and the greater degree of exophthalmos in the fish that received cortico­ trophin. tance still equals 82 percent ( ± three per­ cent) of the initial value and after 48 hours it is 58 percent ( ± five percent) and after 72 hours it is 22 percent ( ± 10.5 percent) (fig. 3 ) . DISCUSSION

T h e difficulties of producing exophthalmos in some fishes by the injection of anterior pituitary extracts or near pure thyrotrophic preparations suggest the existence of an exophthalmic factor distinct from the thyro­ trophic hormone. 1 · 6 ' 7 Nevertheless even though it is a trifle risky to apply the re­ sults of our experiments directly as a basic explanation of the pathogenesis of the exoph­ thalmos of Graves' disease, they confirm our hypothesis 5 that this phenomenon probably develops by the effect of a combination of several endocrine factors. T h e action of thyrotrophic hormone is the essential desencadenating factor; the corticotrophin and the activity of the adrenal cortex have a potentializing effect possibly increasing the sensibility of the retrobulbar tissues to the exophthalmogenous action of thyrotrophin. The experiences of Langford 8 and of Matty, et al.9 in that they showed the possibility of producing exophthalmos in fishes by thyroid hormones seem to oppose the thyrotrophic

hypothesis, since the administration of thyroxin produces an inhibiting effect on the pit­ uitary secretion of thyrotrophin. However, in previous experiments 5 we observed that large doses of thyroid hormone increase the thyro­ trophic exophthalmos, while weak doses of the same are capable of reducing it. These results—apparently paradoxal— could be explained by the toxic action of the thyroxin, which is capable of stimulating the anterior pituitary-adrenal system, 10 increases the corticotrophic and corticoid secretion and indirectly creates experimental conditions analogous to those obtained by injecting thyrotrophin and corticotrophin. SUMMARY

Gold fish (Carassius auratus, var. japonicus, subvar. bicaudatus) respond to the administration of thyrotrophic hormone with exophthalmos. Eye protrusion increases with simultaneous injections of corticotrophin. Retrobulbar injections of normal saline, hu­ man serum or pituitary hormones produce a transient exophthalmos, which is very much more persistent when thyrotrophin is in­ jected. Previous treatment of fishes with long-acting corticotrophin prolongs the du­ ration of the exophthalmos caused by the

CORTICOTROPHIN AND EXOPHTHALMOS retrobulbar injections of thyrotrophin ; this effect is not observed when other fluids are injected. Instituto Barraquer. ACKNOWLEDGMENTS

We are indebted to the representative in Spain of the N. V. Organon Laboratories (Oss, Holland)

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for their generous supply of thyrotrophin, corticotrophin and gonadotrophic preparations used in these experiments. To. Dr. S. Pages Maruny for giving us growth hormone and thyrotrophic hor­ mone. To Dr. Hernân Mïnguez of Valencia (Spain), who sent us a large quantity of different species of fishes from Lake La Albufera (Va­ lencia). To Mr. A. Jonch, Director of the Zoo and Aquarium of Barcelona, for allowing us to see a large number of fishes from the aquarium.

REFERENCES

1. Aird, R. B.: Ann. Inter. Med., 15:564, 1941. 2. Albert, A.: Endocrinology, 40:303, 1947. 3. Aterman, K.: Acta Endocrinol., supl. XX, vol. 15, 1954. 4. Barraquer, J., and Cafiadell, J. M. : Publ. Mon. Instituto Barraquer, 1957, p. 4. 5. Cafiadell, J. M., and Barraquer, J.: Rev. Infor. Méd. Terap., 31:305, 1956. 6. Dobyns, B. M., and Steelman, S. L. : Endocrinology, 52:705, 1953. 7. Friedgood, H. B.: Bull. Johns Hopkins Hosp., 54:48, 1934. 8. Langford, H. G.: Endocrinology, 60:390, 1957. 9. Matty, A. J., Menzel, D., and Bardach, ]. E.: J. Endocrinol., 17:314, 1958. 10. Pickford, G. E.: Endocrinology, 55:589, 1954. 11. Swann, H. G.: Proc. Soc. Exper. Biol. & Med., 40:520, 1939.

NEW TEST CHARTS FOR T H E MEASUREMENT OF VISUAL ACUITY AT FAR AND NEAR DISTANCES* L O U I S E L.

SLOAN,

PH.D.

Baltimore, Maryland Measurements of far and near acuity are needed not only to establish eligibility for blind pensions, tax reductions, and other special benefits, or for admission to sight sav­ ing and other special schools, but also to de­ termine fitness for many different job assign­ ments in industry and in the military serv­ ices. The ophthalmologist uses measurements of acuity primarily as an aid in determining the error of refraction and in diagnosing and following the progress of ocular disease. Be­ cause of certain inadequacies in presently available test charts, the Committee on Optics and Visual Physiology of the A.M.A. ap­ pointed a subcommittee to draw u p specifica* From the Wilmer Ophthalmological Institute of The Johns Hopkins University and Hospital. The development of these charts was supported by Grant B-810 from the National Institute of Neuro­ logical Diseases and Blindness, Public Health Serv­ ice, Bethesda, Maryland.

tions for new charts for the testing of far and near visual acuities. This paper describes the special features of charts developed at the W i l m e r Institute and approved by this sub­ committee. t C H O I C E OF OPTOTYPES

T h e optotypes recommended for testing lit­ erate subjects are a specially selected 1,2 set of 10 capital letters, namely Z N H R V K D C O S. Because these include vertical, hori­ zontal, oblique and curved contours they are well suited for use in subjective tests for de­ termination of the error of refraction. T h e design of the letters follows the Snellen prin­ ciple in that the over-all height and width are five times the width of the strokes. T h e let­ ters are of the familiar Gothic form and t The members of this subcommittee are Gerald Fonda, M.D., Chairman, James E. Lebensohn, M.D., Kenneth Ogle, Ph.D. and Louise L. Sloan, Ph.D.