A method for the estimation of urinary testosterone

1. A method has been developed for the estimation of testosterone in human urine by using acid hydrolysis followed by a quantitative form of a modified Girard reaction that separates a ;conjugated-ketone' fraction from a urine extract; this is followed by column chromatography on alumina and paper chromatography. 2. Comparison of methods of estimation of testosterone in the final fraction shows that estimation by gas-liquid chromatography is more reproducible than by colorimetric methods applied to the same eluates from the paper chromatogram. 3. The mean recovery of testosterone by gas-liquid chromatography is 79.5%, and this method appears to be specific for testosterone. 4. The procedure is relatively rapid. Six determinations can be performed by one worker in 2 days. 5. Results of determinations on human urine are briefly presented. In general, they are similar to earlier estimates, but the maximal values are lower.     

Adhesion of phospholipid vesicles to Chinese hamster fibroblasts: Role of cell surface proteins

The adhesion of artificially generated lipid membrane vesicles to Chinese hamster V79 fibroblasts in suspension was used as a model system for studying membrane interactions. Below their gel-liquid crystalline phase transition temperature, vesicles comprised of dipalmitoyl lecithin (DPL) or dimyristoyl lecithin (DML) absorbed to the surfaces of EDTA- dissociated cells. These adherent vesicles could not be removed by repeated washings of the treated cells but could be released into the medium by treatment with trypsin. EM autoradiographic studies of cells treated with[(3)H]DML or [(3)H]DPL vesicles showed that most of the radioactive lipids were confined to the cell periphery. Scanning electron microscopy and fluorescence microscopy further confirmed the presence of adherent vesicles at the cell surface. Adhesion of DML or DPL vesicles to EDTA-dissociated cells modified the lactoperoxidase-catalyzed iodination pattern of the cell surface proteins; the inhibition of labeling of two proteins with an approximately 60,000- dalton mol wt was particularly evident. Incubation of cells wit h (3)H-lipid vesicles followed by sodium dodecyl sulfate (SDS)- polyacrylamide gel electrophoresis showed that some of the (3)H-lipid migrated preferentially with these approximately 60,000-mol wt proteins. Studies of the temperature dependence of vesicle uptake and subsequent release by trypsin showed that DML or DPL vesicle adhesion to EDTA- dissociated cells increased with decreasing temperatures. In contrast, cells trypsinized before incubation with vesicles showed practically no temperature dependence of vesicle uptake. These results suggest two pathways for adhesion of lipid vesicles to the cell surface-a temperature-sensitive one involving cell surface proteins, and a temperature-independent one. These findings are discussed in terms of current models for cell-cell interactions.