Thymic alterations in feline GM1 gangliosidosis

GM1 gangliosidosis is an inherited metabolic disease characterized by progressive neurological deterioration with premature death seen in children and numerous animals, including cats. We have observed that thymuses from affected cats greater than seven months of age (GM1 mutant cats) show marked thymic reduction compared to age-matched normal cats. The studies reported here were done to describe alterations in the thymus prior to (less then 90 days of age) and during the development of mild (90 to 210 days of age) to severe (greater than 210 days of age) progressive neurologic disease and to explore the pathogenesis of the thymic abnormality. Although histologic examination of the thymus from GM1 affected cats less than 210 days of age showed no significant differences from age-matched control cats, thymuses from GM1 mutant cats greater than 210 days of age were significantly reduced in size (approximately 3-fold). Histologic sections of lymph nodes, adrenal glands, and spleens from GM1 gangliosidosis-affected cats showed no significant differences. Flow cytometric analyses showed a marked decrease in the percentage of immature CD4+CD8+ thymocytes (p < 0.001) and significantly increased CD4-CD8+ cells (p < 0.01) in GM1 mutant cats greater than 210 days of age when compared to normal age matched cats. Co-labelling with CD4, CD8, and CD5 indicated an increase in the percentage of GM1 mutant cat thymocytes at this age which were CD5high, suggesting the presence of more mature cells. Cytometric analyses of subpopulations of peripheral lymphocytes indicated an increase in CD4-CD8+ cells (p < 0.05) with concurrent decreases in CD4+CD8- and CD4-CD8- cells (which were not significant). Similar analyses of thymocyte and lymphocyte subpopulations from cats < 210 days of age showed no significant differences between GM1 mutant and normal cells. GM1 mutant cats at all ages had increased surface binding of Cholera toxin B on thymocytes, indicating increased surface GM1 ganglioside expression. Increases were highly significant in GM1 mutant cats greater than 210 days of age. In situ labelling for apoptosis was increased in GM1 mutant cats between 90 to 200 days of age when thymic masses were within normal limits. In GM1 mutant cats over 200 days of age, decreased labelling was observed when thymic mass was reduced and the CD4+CD8+ subpopulation, known to be very susceptible to apoptosis, was significantly decreased. These data describe premature thymic involution in feline GM1 gangliosidosis and suggest that increased surface GM1 gangliosides alters thymocyte development in these cats.     

The role of estrogen and progesterone in the regulation of reproductive behaviour in female ring doves(Streptopelia risoria) under long vs. short photoperiods

The role of estrogen and progesterone in the regulation of reproductive behavior was studied in female ring doves (Streptopelia risoria) under long vs. short photoperiods. The doves held under long (16 hours/day) or short (8 hours/day) photoperiods were treated for 15 days with saline, estrogen, or progesterone, alone or in combination. 7 days after the start of hormone therapy, the doves were placed with reproductively active males for 4 hours/day for 9 days. No difference was seen in egg-laying, courtship or nest-building by control birds under the 2 photo regimes. Ovarian follicles remained small in all doves receiving hormone treatment. Oviducts of doves receiving progesterone alone remained small in the short-photoperiod group but at long photoperiods the oviducts became enlarged. Estrogen stimulated oviduct growth at both photoperiods wheras the combined hormone treatment resulted in greater oviduct development than estrogen alone (p less than .02). Female soliciting and nest-building activity remained low in progesterone-treated doves under short photoperiod but increased rapidly under long photoperiod. Doves treated with estrogen and estrogen plus progesterone performed considerable soliciting and nest-building. No marked tendency was seen for doves under long-photoperiod conditions to be more active in nest-oriented behavior. Copulatory behavior by the female occurred infrequently in all hormone-treated doves.     

Exercise tolerance at 4 and 6 ATA

Seven normal male subjects performed 5-min bicycle exercise ranging from 50-100% maximum oxygen uptake at 4 ATA and three were also studied at 6 ATA. At all pressures, the subjects breathed 0.2 ATA O2 plus nitrogen. All subjects were able to perform maximum work at all pressures. No pressure-dependent variations in heart rate, O2 uptake, or CO2 output were noted. At both 4 and 6 ATA, ventilation was decreased at exercise levels greater than 80% maximum O2 uptake. The magnitude of the decrease was not great, however, and signified only minor CO2 retention. In some instances exercise ventilation closely approached the 15-S maximum breathing capacity and these subjects noted severe dyspnea, possibly due to dynamic compression of large airways. In three subjects, respiratory frequency was measured as well as minute ventilation; this relationship did not change with depth. Subjects performing heavy exercise at 6 ATA noted disturbances of consciousness, presumably due to N2 narcosis.     

Statistical methods for the analysis of tumor multiplicity data

The fit of tumor multiplicity data from 93 mouse skin, lung, and liver carcinogenicity experiments to Poisson, negative binomial, and normal distributions was studied. The data were fitted well by the negative binomial distribution. This distribution has two parameters, the mean tumor multiplicity and an exponent determined by the interanimal homogeneity of tumor response. The value of the latter parameter was related to animal strain and the target tissue studied in the carcinogenicity experiments. The null distribution of the two-sample likelihood ratio test based on the negative binomial with common exponent model for tumor multiplicity data was shown by simulation studies to be approximately chi 2 with 1 d.f. Simulation also indicated that the likelihood ratio test has sufficiently better performance when the negative binomial model is valid to make its use more attractive than the more commonly used Wilcoxon test or Student t test. Charts for estimating the number of animals per group that are required to detect specified differences in tumor multiplicities are provided for several commonly used assays.