Circadian complexes: Circadian rhythms under common gene control

Summary Circadian rhythms for food and water consumption were measured in five inbred strains of mice under a photoperiod of 16 h light and 8 h dark (16:8 LD), and under constant light (LL).Significant strain differences were observed which indicate that a common gene difference, or set of differences inMus musculus influences both the phase angle () associating the rhythms with the light-dark cycle, and the periods (_LL) of circadian rhythms for food and water consumption. The biological clock mechanism influenced by this genetic variance is common to both food and water circadian rhythms, and differs among the five inbred strains. A positive genetic correlation was observed between the phase angle () and the period (_LL) of each rhythm. This observation can be understood in terms of a functional relationship between phase and period proposed by Pittendrigh and Daan (1976b) for the entrainment of a circadian oscillator by a light-dark cycle in nocturnal rodents.These results suggest that circadian rhythms for food and water consumption in mice are regulated by a common physiological mechanism, and would respond to natural selection as a single circadian complex under common gene control.     

The distribution of keratin type intermediate filaments in human breast cancer. An immunohistological study

Antibodies to different intermediate filament proteins can be used to distinguish cells of epithelial, mesenchymal, muscle, glial and neuronal origin. Antibodies to prekeratin which characterize cells of epithelial origin, and antibodies to vimentin which recognize cells of mesenchymal origin have been used to study twenty cases of breast carcinoma (sixteen infiltrating ductal carcinomas and four infiltrating intraductal carcinomas), two cases of cystic breast disease, two fibroadenomas and one case of benign cystosarcoma phylloides. The prekeratin and vimentin were detected using specific antibodies to these proteins by immunofluorescence microscopy using alcohol fixed paraffin-embedded tissues. In eighteen out of the twenty carcinomas the tumor cells were strongly and specifically stained by antibodies to prekeratin. DIfferent tumors gave different patterns of prekeratin staining. In contrast, when the same specimens were tested with the vimentin antibody, the tumor cells were unstained, and instead only the usual strong staining to fibroblasts and blood vessels in the stroma was observed. In cystic breast disease, fibroadenomas, and benign cystosarcoma phylloides, cells of epithelial origin were strongly stained by the prekeratin but not by the vimentin antibody.     

Direct isolation in cell-free medium of a spiroplasma fromHaemaphysalis leporispalustris (Acari: Ixodidae) in Maryland

A spiroplasma isolate, was obtained from rabbit ticks (Haemaphysalis leporispalustris) taken from cottontail rabbits in Maryland by inoculation of tick suspensions into SP-4 medium. The isolate was indistinguishable from an experimental vertebrate pathogen (suckling mouse cataract agent spiroplasma) when tested with other plant and tick spiroplasmas in growth inhibition, deformation, and metabolism inhibition tests. The isolated organism had a pathogenic profile for suckling rats and embryonated chicken eggs that differed significantly from that of other suckling mouse cataract agent strains. This is the first report of a direct spiroplasma isolation from ticks in cell-free medium, and confirms the specific association of spiroplasmas of the suckling mouse cataract agent serogroup with rabbit ticks.     

Large-scale rotating filter perfusion system for high-density growth of mammalian suspension cultures

Summary A system has been developed for growth and maintenance of mammalian cells in suspension culture at high density. In principle, the maintenance of constant levels of required nutrients coupled with the removal of toxic cell byproducts can support much higher suspension cell densities than may be obtained in conventional spinners. The system consisted of 4- or 40-liter reaction vessels equipped with a vertically supported rotating cylindrical filter. Agitation was provided by the magnetically driven, rotating filter. Fresh medium was supplied at a rate of 10 to 20 ml/h per 10⁹ cells and the expended medium free of cells was withdrawn through the rotating filter. Both pH and dissolved O₂ and CO₂ were monitored and regulated. Walker 256 carcinosarcoma cells have been grown in these reactors to densities 10-to 30-fold greater than that obtained in Bellco spinners. In addition to high cell densities, the yield of cells per liter of medium used was 2- to 3-fold that obtained in the conventional systems. Both 4-and 40-liter versions of this reactor have been operated without the use of antibiotics. The 40-liter reactor also has been modified for chemostat operation. In a single run, for example, the Walker cell density was maintained between 6 and 10×10⁶ cells/ml with a total yield of 8.7×10¹¹ cells from 360 liters of medium.