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LIPOTROPIC EFFECTS LIVER EXTRACT, BI2 AND CHOLINEleased from the pigment glands, some of it reacts with the reactive groups in the mea! and is "bound." Under such circumstances the gossypol is detoxified. For the combination products described in this pulblication the gossypol is rendered more toxic by a process which maintains it in solution. A more thorough study of the reactions of gossypol with amino acids, proteins, and carbohydrates, and of the effect of storage and heat on such products would undoubtedly clarify the nature of this combination product and probably throw light on the reaction that takes place when cottonseed or cottonseed meats are cooked in order to detoxify the gossypol.Summary.(1) Gossypol was combined with proteins, amino acids, and carbohydrate rnaterials by a procedure involving mixing of the g o s s p l and other combining substances in alkaline solutions and su'bsequent lyophilization of the neutralized frozen solutions.( 2 ) Crystalline gossypol when added to the aquarium bath had no visible effect upon goldfish. The water-soluble combination products of gosrypol were toxic to fish in concentrations as small as one part of gossypol product to a hundred thousand parts of water.(3) Separated pigment glands were toxic to fish but were significantly less toxic than the water-soluble combination products of gossypol.I t has been reported that crude liver extract exerts a lipotropic effect and will prevent fibrosis of the liver when administered to rats receiving a high fat diet( 1). The liptropic effect of the liver extract did not seem to be due to choline content. In fact, other supplements which did not give protection against dietary induced liver injury had a higher content of choline than was present in liver extract ( 2 ) . In further investigations a vitamin BIZ concentrate, prepared from liver, also exerted a definite effect in preventing accumulation of fat in the liver of rats(3). In the present study the relationship between dosage of liver extract and lipotropic effect was studied. Observations were also made on the possible liptropic effect of combinations of choline, inositol and folk acid, which are present in small amounts in liver extract and vitamin BIZ concentrate. Crystalline vitamin Bla alone and in Combination with small amounts of choline was also observed for lip0 tropic activity . Methods.Male Sprague-Dawley rats weighing between 120 and 150 g were used. They were fed the high-fat diet (51% lard) and the control diet (6% lard) recently described( 1). These diets were fed for the periods listed in the tables. When the animals were on the synthetic diet, injections of the supplements were begun, ~1 supplements were administered subcutaneously except in one experiment with liver extract as noted in Tzble I. At the end of lobe of the liver and stained with hematoxylin and eosin or with Sudan 111 for fat. The remainder of the liver was analyzed for total *The authors wish to thank Eli Lilly and co. for the liver extract, Merck and Co. for the crystalline vit. B,, and...

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