Anticoccidial drugs were evaluated for activity and for the development of resistance in a model of Toxoplasma gondii growing in human fibroblast cultures. Of 13 anticoccidial drugs tested, 9 had selective antitoxoplasma activity (50% inhibitory concentration, in micrograms per milliliter): decoquinate (0.005), arprinocid-N-oxide (0.015), robenidine (0.03), the aryl triazine CP-25,415 (0.2), toltrazuril (0.4), clopidol (1), dinitolmide (Zoalene; Dow) (10), and the carboxylic acid ionophores monensin (0.001) and salinomycin (0.04). Glycarbylamide, amprolium, nicarbazin, and the 6-(p-bromophenoxy)-7-chloro analog of halofuginone (Stenorol; Roussel-UCLAF) (CP-63,567) were toxic for the fibroblasts. Since Eimeria tenella has a similar drug susceptibility profile, anticoccidial drugs can be viewed as a potential source of new antitoxoplasma therapies. The development of resistance has limited the usefulness of most of these drugs as anticoccidial agents; in coccidia, resistance to all except the ionophores occurs readily in vivo. We explored the development of resistance in T. gondii by attempting to select mutants in vitro from parasites mutagenized with ethylnitrosourea. Mutants that had 20-to 50-fold-reduced susceptibility to decoquinate, arprinocid-N-oxide, and CP-25,415 were obtained. Ionophore-resistant T. gondii mutants were also selected in vitro; however, there was only a twofold difference in susceptibility between these mutants and the wild type. For three drugs (clopidol, robenidine, and toltrazuril), we were unable to select resistant mutants. For experimental anticoccidial drugs, there is currently no in vitro method for assessing the risk of development of resistance in Eimeria species. Our results suggest that T. gondii may offer a useful surrogate for this assessment.

The extra-ovarian contribution to progesterone levels in the circulation has been measured in ovariectomized ewes in which pregnancy was maintained by treatment with medroxyprogesterone acetate. Since adrenal production of progesterone is low this procedure allows the determination of placental secretion. Placental production of progesterone has been shown to rise initially between 50 and 70 days of gestation, with a second phase of increase between 90 and 120 days of gestation. Both periods of increased secretion of progesterone were reflected in rises in progesterone production by placental explants in organ culture and in the activity of 3 beta-hydroxysteroid dehydrogenase in the tissue. The activity of 3 beta-hydroxysteroid dehydrogenase exceeded the rate of progesterone production by approximately 30-fold, and it appears unlikely, therefore, that this enzyme is rate-limiting in the synthesis of progesterone. Surgical reduction of the number of cotyledons led to compensation by an increase in weight of individual cotyledons with no significant increase in specific 3 beta-hydroxysteroid dehydrogenase activity or rate of production of progesterone in organ culture. It was concluded that the rise in placental production of progesterone in the middle period of gestation is unlikely to result either from an increase in 3 beta-hydroxysteroid dehydrogenase activity or from exposure to factors stimulating placental growth.

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A technique has been developed for the study of placetal steroid synethesis in tissue culture. Explants of sheep placentas recovered in the last third of gestation produced progesterone and converted labelled androstenedione into unconjugated and conjugated oestrogens. Cortisol added to the medium stimulated 17 alpha-hydroxylase and aromatizing enzyme activities. The results are discussed in relation to the proposal that the rise in secretion of fetal cortisol before parturition has a direct effect on the activation of placental enzymes catalysing the conversion of progesterone to eostrogens.

Epithelial cells and stromal cells of the rabbit endometrium were separated by successive enzymic digestion of the uterine mucosa. Isolated cell types were obtained in high yield, with good viability, and were maintained in monolayer cultures for up to 2 weeks. Epithelial cells in monolayers appeared as polygonal cells, displayed contact inhibition, and showed the presence of microvilli on the cell surface, with many desmosomes. Stromal cells grew rapidly to confluence, displayed overgrowth, and had a fibroblastic appearance with an absence of junctional complexes between cells. Indirect immunofluorescence showed uteroglobin on the surface of epithelial but not of stromal cells, and only epithelial cells secreted uteroglobin into the medium. These results confirm the identity of the cells and provide biochemical evidence for the epithelial cellular origin of uteroglobin. The method allows the culture of separate endometrial cell types, which retain their morphology and differentiated function in vitro.