Effects found in a one-year oral toxicity study in Sprague-Dawley rats of a novel 5-HT1A receptor partial agonist for the treatment of anxiety in humans.

A potent and selective serotonin (5-HT1A) partial agonist with potential as a human anxiolytic drug was given in oral doses of 0, 5, 15, or 50 mg/kg/day by gavage to Sprague-Dawley rats for 6 or 12 mo. Some animals were allowed 1 mo to recover after each treatment period. The 10-fold increase in dose resulted in a 20-fold increase in drug plasma concentration due to saturable first-pass metabolism. This resulted in disproportionately higher concentrations and greater bioavailability of the 15- and 50-mg/kg/day regimens. Drug exposure was associated with decreased spontaneous activity in the 15- and 50-mg/kg rats. The activity of these rats returned to normal during the recovery period. There were significant (p < 0.05) decreases in mean body weights during the study for 50-mg/kg males, with improvement during the recovery periods. No biologically significant effects were noted in clinical laboratory parameters. Based on organ weight increases and histopathological evaluation, drug-related effects after 6 and 12 mo of treatment were in the pituitary (both sexes) and all treated female reproductive organs. In general, these effects persisted into periods of recovery, except for pituitary hyperplasia, which was not apparent following recovery after treatment for 6 mo. After treatment for 12 mo and the following recovery, there were significant increases in adrenal weights in the 15- and 50-mg/kg/day males with no morphological correlate. There was increased pituitary hyperplasia that persisted through the recovery period in all treated groups in both sexes, but there was no increase in pituitary neoplasms. In treated females, there was also morphologic evidence of persistent diestrus (estrogenic effect) evidenced by endometrial squamous metaplasia, increased corpora lutea, vaginal mucification, and decreased uterine size. The clinical and pathological changes seen with these 2 regimens were considered exaggerated pharmacological effects of the drug on serotonin receptor-rich organs.

Veno-occlusive disease of the liver in captive cheetah.

Liver tissues from 126 captive cheetah were evaluated by light microscopy and histochemistry; eight animals were evaluated by electron microscopy. The main hepatic lesion, a vascular lesion resembling veno-occlusive disease (VOD) of the liver and characterized by subendothelial fibrosis and proliferation of smooth muscle-like cells in the central veins, was seen in 60% of the sexually mature cheetah. Although this hepatic vascular lesion was seen in cheetah as young as 1 year of age, the most severe lesions, usually associated with liver failure, were found in cheetah between the ages of 6 and 11. There was no sex predisposition, and in approximately 40% of the VOD cases, liver disease was not suspected clinically or at necropsy. VOD was found in other felidae, especially in the snow leopard. High levels of vitamin A in livers, as well as in diets of the cheetah, could be a contributing factor in the development of VOD in some groups of cheetah.

Dietary estrogens--a probable cause of infertility and liver disease in captive cheetahs.

The cheetah in the wild is "racing towards extinction" mostly due to habitat destruction. Its survival will probably depend on accelerated captive breeding. At this time, however, reproductive failure and liver disease threaten the future of the captive cheetah population. Histopathological evaluation of more than 100 cheetah livers identified venocclusive disease as the main hepatic lesion responsible for liver disease in this species. Analysis of the commercial feline diet by high-performance liquid chromatography and gas-liquid chromatography-mass spectrometry revealed large amounts of two phytoestrogens identified as daidzein and genistein. These compounds were found to be derived from a soybean product that was a component of the cheetah diet, and their concentrations both ranged from 18 to 35 micrograms/g diet. The adult cheetah consequently consumes approximately 50 mg/day of these weak estrogens. When extracts of the diet were tested for estrogenicity using a bioassay, a dose-related increase in uterine weight was observed. In 4 cheetahs studied, withdrawal of this feline diet by substitution with a chicken diet resulted in an improvement in conventional liver function tests and a normalization in the appearance of hepatic mitochondria. We conclude that the relatively high concentrations of phytoestrogens from soybean protein present in the commercial diet fed to captive cheetahs in North American zoos may be one of the major factors in the decline of fertility and in the etiology of liver disease in this species. The survival of the captive cheetah population could depend upon a simple change of diet by excluding exogenous estrogen.

Extrahepatic biliary carcinoma in sloth bears.

Extrahepatic biliary carcinoma was found in each of 5 adult sloth bears that died between 1970 and 1984 while on exhibit in Ohio zoos. The tumor was characterized by numerous mucin-producing neoplastic glands scattered throughout abundant fibrous stroma. The cause of the tumor was not determined.

Pathophysiology of excessive iron storage in mynah birds.

Hepatic iron overload was diagnosed in young and adult Rothschild's mynah birds. By light microscopic evaluation, it was determined that there was progressive accumulation of iron pigment with age in the hepatocytes. The Kupffer's cells of the liver, the reticuloendothelial cells of the spleen, and the phagocytic cells in the small intestine were negative for Prussian blue (iron) pigments. These observations suggested that diet had only a minor influence on the iron distribution in the mynah birds. This was confirmed by comparing iron distribution in the liver and spleen of mynah birds with that in other exotic birds receiving the same diet as the mynah birds. The syndrome of excessive iron overload in the mynah birds shared most of the important histopathologic characteristics with idiopathic (hereditary) hemochromatosis in human beings.

Autoimmune lymphocytic thyroiditis in dogs.

Primary diseases of the thyroid gland, especially lymphocytic thyroiditis and idiopathic follicular atrophy, were the most common lesions associated with clinical hypothyroidism in pet dogs. Lymphocytic thyroiditis resembled naturally occurring lymphocytic thyroiditis in the Obese-strain of White Leghorn chickens and Hashimoto's thyroiditis in man. The morphology of the thyroid lesion and frequent occurrence of circulating thyroglobulin autoantibodies suggested that lymphocytic thyroiditis was immune-mediated in pet dogs. Thyroid lesions similar to naturally occurring autoimmune thyroiditis in experimental dogs were induced by a local thyroidal graft-versus-host reaction. The lesions observed in the thyroid lobe which was not injected with immunocompetent cells appeared to develop from the formation of thyroid antibodies in the gland by migrating host lymphocytes. Autoimmune lymphocytic thyroiditis occurred secondary to an unrelated immune response occurring in target tissue.

Lymphocytic thyroiditis in dogs; induction with a local graft-versus-host reaction.

Fifty million allogeneic canine lymphocytes were injected in the left lobe of the thyroid gland of 10 dogs. The left lobe of the gland was surgically removed at 1 and 4 weeks after injection. The intrathyroidal injection of immunocompetent cells resulted in local graft vs host reaction within the thyroid gland. Multifocal infiltration of lymphocytes, plasma cells, and macrophages associated with scattered, isolated, or small groups of degenerated follicular cells were present in the follicular wall and colloid on day 7. Thyroidal lesions at 1 month after injection were characterized by extensive destruction of follicular cells, disseminated foci of mononuclear cell infiltration, and electrondense deposits between follicular cells and the basement membrane. The contralateral (noninjected) lobe of the thyroid gland at 4 weeks after injection was infiltrated by lymphocytes, macrophages, and many plasma cells. The inflammatory lesions were associated with multifocal areas of degeneration of follicular cells and focal electron-dense thickenings of the basement membrane. The noninjected lobe, following hemithyroidectomy at 1 week after injection of the thyroid gland undergoing graft-vs-host reaction, either had a focal infiltration of polymorphonuclear leukocytes, lymphocytes, macrophages, and mast cells or had no significant lesions.

Induced lymphocytic thyroiditis in dogs: effect of intrathyroidal injection of thyroid autoantibodies.

Multifocal infiltrations of lymphocytes and macrophages were present in the thyroid glands of dogs 1 month after an intrathyroidal injection (0.5 ml) of canine serum containing thyroglobulin autoantibodies. The development of thyroiditis was associated with a gradual increase in the incorporation of tritiated thymidine by the peripheral blood mononuclear cells. There was no alteration in circulating thyroid hormone values or thyroglobulin autoantibody titer for 1 month. Control dogs given a similar volume of canine serum without thyroglobulin antibodies did not have an inflammatory reaction in the thyroid gland, and the phytomitogen responses of lymphocytes did not change, compared with base-line values. An intrathyroidal injection of canine thyroglobulin autoantibodies induced lesions similar to naturally occurring lymphocytic thyroiditis in dogs, indicating that thyroid autoantibodies have an important role in the pathogenesis of immune-mediated thyroiditis.

Histologic and ultrastructural evaluation of thyroid lesions associated with hypothyroidism in dogs.

Thyroid lesions in 16 pet dogs with hypothyroidism were evaluated by light and electron microscopy. Lymphocytic thyroiditis, found in seven dogs, was characterized by diffuse infiltration of the thyroid gland by lymphocytes, plasma cells and macrophages with formation of some lymphoid nodules and destruction of follicles, progressing to replacement of most of the thyroid by fibrous connective tissue. The basement membrane around follicles was thick and had electron-dense deposits. The morphology of the thyroid lesions and the presence of circulating thyroglobulin autoantibodies suggested that lymphocytic thyroiditis was immune-mediated. Idiopathic follicular atrophy, found in nine dogs, was characterized by loss of thyroid parenchyma and replacement by adipose connective tissue. Degeneration of individual follicular cells was present in the early stage, with exfoliation into the colloid and interfollicular area. Most of the thyroid gland consisted of adipose connective tissue with either interspersed small follicles or individual follicular cells that had dilated rough endoplasmic reticulum, large Golgi apparatus, and intracytoplasmic microfollicles in the advanced stage. Follicular atrophy was a degenerative lesion of follicular cells of unknown cause, not associated with inflammatory destruction in the thyroid gland.

Biochemical and immunological investigations on hypothyroidism in dogs.

Circulating antibody titers (1:20 to 1:2560) against thyroglobulin were demonstrated in 48% of pet dogs with hypothyroidism by the chromic chloride passive hemagglutination test. Four of six dogs with acanthosis nigricans (1:20) and one of six male dogs with hyperestrogenism (1:40) had low titers of antibody against thyroglobulin whereas clinically normal pet dogs and dogs with other selected endocrinopathies (hypoadrenocorticism, cortisol-excess, diabetes mellitus) or obesity were consistently negative. Circulating immune complexes evaluated by the mastocytoma cell-assay were present in the sera of 20% of pet dogs with hypothyroidism but were absent in clinically normal dogs. Although variations in dose significantly altered the quantitative response of the thyroid gland to thyrotropin the qualitative pattern of response was similar for T3 but not T4 in clinically normal laboratory beagles. The peak increases for serum triiodothyronine and thyroxine were observed either at eight (0.1 and 0.2 I.U bTSH/5 lbs) or 12 (1.0 I.U. bTYSH/5 lbs) hours postthyrotropin. Dogs with naturally occurring hypothyroidism had a decreased serum T3 and T4 at baseline and eight hours postthyrotropin (1.0 I.U. bTSH/5 lbs) compared to clinically normal pet dogs, laboratory beagles and dogs with other clinical endocrinopathies. The consistent lack of a significant increase of serum T3 and T4 in response to thyrotropin was necessary for the separation of certain hypothyroid from euthyroid pet dogs in which the baseline level of thyroid hormones were equivocal.