to another even though the parents are deprived of the tumors by an operation. I am gratefully indebted to Professor Morgan and Doctor Bridges for helpful suggestions. Conclusions.-1. A non-lethal tumor appeared as a mutation in the lethal tumor strain. 2. The locus of the gene of the new tumor is close to that of the dichaete in the third chromosome. 3. The tumor may occur in any segment of the larva but seems to occur more often in the twelfth and thirteenth segments. 4. The cells of the tumor are rounded or polygonal in shape and show the presence of pigment. 5. Ingrowth of tumor cells into the imaginal discs of the appendages checks the development of the parts. 6. Young tumors were inserted into larvae of normal strains. Five per cent survived the operation, completed metamorphosis and carried the inserted tumor into the adult fly. METALLIC SALTS OF PYRROL, INDOL AND CARBAZOL BY EDWARD C. FRANKLIN DEPARTMENT OF CHEMISTRY, LELAND STANFORD UNIVERSITY Communicated, October 20, 1919 The ammono acids, that is to say, the acids of the ammonia system of acids, bases and salts,1 are derivatives of ammonia in which one or two hydrogen atoms of the ammonia molecule are replaced by negative groups. A number of examples of compounds so related to ammonia are nitramide or nitrosyl amide, NO2NH2; acetamide or acetyl amide, CO CH,CONH2; phthalimide or phthalylimide, CH CO NH; benzene sulfonnitramide or nitrosyl benzenesulfonyl imide, CH,SO2NHNO2; methylnitramine or methyl nitrosyl imide; acetanilide or phenyl acetyl imide; trinitraniline, C6H2(NO2)3NH2; cyanamide, CNNH2; urea, CO(NH2)2 etc., etc. These substances are true acids ranging in acidity from benzenesulfonnitramide, which approaches the ordinary mineral acids in strength, through phthalimide and methyl nitramine, which are well known to possess weak acid properties, to acetamide and urea which are not ordinarily recognized as acids at all. The acid properties of acetamide, urea and other very weak ammono acids, however, show themselves distinctly when in solution in liquid ammonia as has been shown by the writer and his students in earlier papers. The fact that acids too weak to be recognized as such in aqueous solution are still capable of showing acid properties when in solution in liquid ammonia is undoubtedly due to the lower solvolytic2 action of ammonia as compared with water which in turn results from the very slight autoionization of the former solvent. To what extent liquid ammonia is dissociated has not been accurately determined. Since, however, it is a comparatively easy matter to obtain liquid ammonia with a specific conductance less than one-eighth that of water3 it must be considerably less ionized than is the latter solvent. It happens therefore that salts of the weaker ammono acids, which, because of the strong hydrolytic action of water are incapable of exstence in the presence of this solvent, have been easily prepared from liquid ammonia solutions. CH CH and carbazol, C&HA NH CH = CH ammonia hydrogen is replaced by distinctly negative groups. They are therefore to be classed among the ammono acids and as such should react, in liquid ammonia solution, with the more electro positive metals and with their amides, the ammono bases, to form salts. Experiments will be described in a paper to be printed in the Journal of Physical Chemistry showing that pyrrol, indol and carbazol react in liquid ammonia solution with metallic potassium, sodium, calcium and magnesium and with the amides of potassium, sodium, calcium and silver to form the following well defined salts: Sodium pyrrol, CHÂNNа and C1H1NNа. NH ̧. Calcium pyrrol, (CHN)2Ca and (C,H,N)2Ca. 4NH3. Silver pyrrol, C4H1NAg. NH3. Sodium indol, C,H,NNa.xNH3. Calcium indol, (C,H,N) Ca. 4NH3. Magnesium indol, (C.H&N)2Mg. 4NH3. Silver indol, CзH.NAg. NH3. Potassium carbazol, C12H,NK. 2NH, and C12H,NK.NH3. 1 Amer. Chem. J., 28, 1902, (83); 47, 1912, (285); Eighth Int. Cong. App. Chem. 6, 1912, (119) and J. Amer. Chem. Soc., 37, 1915, (2279). 2 Solvolysis is used as a general term to include hydrolysis, ammonolysis, amidolysis, aminolysis, alcoholysis, etc. 3 Franklin and Kraus, J. Amer. Chem. Soc., 27, 1905, (191). GROWTH AND REPRODUCTION IN FOWLS IN THE ABSENCE OF CAROTINOIDS AND THE PHYSIOLOGICAL RELATION OF YELLOW PIGMENTATION TO EGG LAYING BY LEROY S. PALMER DIVISION OF AGRICULTURAL BIOCHEMISTRY, UNIVERSITY OF MINNESOTA Communicated by W. A. Noyes, November 1, 1919 The chemical identification of each of the recognized vitamines as individual substances or chemical groups is greatly to be desired. As the result of certain studies1 which I made on the physiological relation between the yellow carotin and xanthophyll pigments of plants and the yellow lipochromes of animal tissues and fluids, I became impressed with the fact that there seemed to be more than a casual relation between the simultaneous presence of the plant carotinoids and fatsoluble vitamine in butter fat and egg yolk and in the leafy parts of green plants, and the simultaneous absence of carotinoids and fatsoluble vitamine from lard. That phase of my carotinoid studies showing the physiological identity of egg yolk lipochrome with plant xanthophyll suggested that the fowl should be a suitable animal upon which to test the relation of plant carotinoids to growth and reproduction. It was decided to approach the question by attempting to raise a flock of chickens from hatching to maturity on a ration devoid of carotin and xanthophylls. The problem which was presented was therefore mainly one of selecting a ration devoid of yellow plant pigments but which was presumably adequate otherwise for the normal growth of chickens. Three experiments were undertaken. The first was a preliminary experiment, during the winter of 1916-17, to test the efficacy of a ration of white corn, white corn bran, bleached flour, skim-milk and bone meal, beginning with birds weighing about one-half pound each. Several birds reached the period of fecundity on this ration devoid of carotinoids, and the yolks of the eggs laid were remarkably deficient in color. In the second experiment, carried out during the season of 1917-18, 75 White Leghorn chicks were placed on a practically identical carotinoid-free ration immediately after hatching. None of these birds reached maturity, partly, it is believed, because of nutritional difficulties, and partly, it is admitted, because of a number of unfortunate accidents in caring for the birds, which caused the death of the larger share of them. The nutritional difficulties of the second experiment were overcome in the third trial begun in April, 1918, by introducing into the ration both pork liver, which is rich in vitamines,' but devoid of carotinoids, and a roughage of paper pulp. A flock of 50 vigorous, normal White Leghorn chickens were raised from hatching to maturity on this modified white corn and skim-milk ration. The mature birds were free from yellow pigmentation. Not only was normal growth secured on this ration, but the hens in the flock exhibited normal fecundity. Seventeen of the hens whose egg records were kept averaged 52 eggs each during a period of 233 days. Several had considerably higher records. One hen laid 44 eggs during a period of 59 days. Especially interesting was the character of the pigmentation of the egg yolks. The hard-boiled yolks were colorless but the raw yolks had a faintly yellow color. The pigment, however, was neither carotin nor xanthophyll. Acetone readily extracted the coloring matter from the raw yolks but the pigmented fat which could be obtained from this extract failed to give the reduction test with ferric chloride, which I have shown to be characteristic of carotinoids. Attempts to identify the pigment with Barbieri's ovochromine or with bilirubin, were not successful. It was felt that the negative relation between carotinoids and fatsoluble vitamine as exhibited by normal growth and fecundity in chickens on rations devoid of carotinoids could not be regarded as established unless the carotinoid-free eggs should prove fertile and normal chicks be hatched from them. Inasmuch as the cocks and hens of the carotinoid-free flock were kept together throughout the experiment the eggs were presumably fertile. About 90 eggs, in all, were incubated at various times. Forty-one livable chicks were hatched from these eggs. The young chicks appeared normal in every way except for the complete absence of yellow pigmentation from the shanks, beaks, and other skin parts. The newly hatched chicks were immediately placed on a carotinoidfree ration and were cared for as nearly as possible in the same manner as the young chicks of the preceding generation. By the end of three months, however, all had died. Although it is probable that this unfortunate result may be explained on the ground that the chicks were hatched very late in the season and therefore had to combat a period of extreme heat, as well as a very restricted diet during the most precarious period of their growth, nevertheless the question remains open as to whether it is possible to continue the carotinoid-free condition into more than one generation. Physiological relation of pigmentation to egg laying.-Practical poultry men have recognized for several years that a relation exists between the amount of yellow pigment visible in the shanks, ear lobes, beak and vent of hens of the Leghorn, Plymouth Rock, Wyandotte and Rhode Island Red breeds and their previous egg laying activity. Extensive biometric analyses have been made by Blakeslee and Warner' and by these authors with Harris and Kirkpatrick of data collected at the Storrs Agricultural Experiment Station egg laying competitions in order to establish the character of this relation. The results show a positive correlation between pale colored shanks, ear lobes, beak, etc., and a recent more or less heavy egg production. The hypothesis which has been adopted by these investigators to explain the physiological relationship which has been observed between fecundity and pigmentation is that the growth of the egg abstracts the pigment from the body tissues. The idea that the relationship could be explained also on the basis that the egg yolk abstracts fat-soluble pigment from the food, thus precluding its localization in the body tissues, was advanced by Harris, Blakeslee and Warner' in an earlier paper, but was apparently abandoned. The high percentage of fat in the blood of laying hens, as compared with non-laying hens, as shown by Warner and Edmonds,1o and by Riddle and Harris,"1 is believed by the former authors to support the hypothesis that the tissue fat is being transferred to the egg yolk during laying with a consequent subtraction of pigment. The success attained in raising a flock of White Leghorn fowls entirely lacking in pigmentation in both adipose tissue and visible skin parts presented the opportunity for ascertaining the true physiological relation |