In an EDTA/Hoechst 33258 assay system, a linear increase in fluorescence with increase in cell number between 2 X 10(3) and 1 X 10(5) was obtained if a dye concentration of 800 ng/ml was used. For a given number of cells, the enhancement of fluorescence was found to be greater than that of a theoretically equivalent of DNA. A standard curve for the assay was derived by plotting enhancement of fluorescence against cell number. The effect of storage on the fluorescence of intact monolayers, cellular or commercial DNA, or dye-DNA complexes made it essential that the assay was carried out on fresh samples.

No abstract

A factor that stimulates the proliferation of human umbilical vein endothelial cells has been shown to be present in serum-free medium conditioned by the prior growth of a cell line (1PT) derived from a poorly differentiated bronchial carcinoma. Preliminary characterization of this factor has revealed that it is a heat-labile, acid-stable proteinaceous material, the activity of which is not diminished by treatment with a reducing agent. In its partially purified state it has been shown to be anionic and to be associated with material exhibiting a broad molecular weight range of 35 X 10(3) to 100 X 10(3). It does not bind strongly to heparin-Sepharose and its mitogenic effect on endothelial cells is not potentiated by heparin. These properties suggest that this factor may differ from other previously described tumour-derived endothelial mitogens.

Concentrations of cytosolic androgen receptor, DNA and soluble protein, contents of DHT, and in-vivo uptake of 3H-DHT were measured in rat ventral prostates at 5-day intervals during sexual development. Regarding prostate weight two phases of growth were noted being separated by a period of stagnation from Day 40 to 45. Cytosolic androgen receptor, particle-bound DHT, and uptake of 3H-DHT into the 100,000-g sediment showed a clear pattern: a maximum in the prepubertal animal at age Day 20, a minimum at age Day 30 (4 days after the early pubertal rise of LH, testosterone, and DHT) followed by a second maximum on Day 55 (2 days before the beginning of fertility), and a second minimum in the young mature animal on Day 70. An intermediate peak seen at age Day 37 was not significant. Neither the time-dependent profile of the cytosolic androgen receptor nor the contents and in vivo uptake of DHT were correlated to concentrations of circulating gonadotrophins, growth hormone, and sex-steroids measured during puberty in the same strain of animals. Therefore, the regulating mechanism remains unclear.