Activation-induced subcellular redistribution of Gs alpha.

We have examined the subcellular distribution of alpha s, the alpha subunit of the heterotrimeric G protein Gs, by using immunofluorescence microscopy. In transiently transfected HEK293 cells, wild-type alpha s localizes to the plasma membrane. However, a mutationally activated alpha s (alpha sR201C) is diffusely distributed throughout the cytoplasm. Similarly, cholera toxin activation of alpha s causes it to redistribute from the plasma membrane to cytoplasm in stably transfected cells. In HEK293 cells stably transfected with alpha s and the beta 2-adrenergic receptor (beta-AR), stimulation of the beta-AR by the agonist isoproterenol also causes a translocation of alpha s from the plasma membrane to cytoplasm. Replacing the agonist with antagonist allows alpha s to return to the plasma membrane, demonstrating the reversibility of alpha s translocation. Receptor-activated alpha s does not colocalize with internalized beta-AR at endosomes. Incubation of cells in hypertonic sucrose to inhibit clathrin-coated pit-mediated endocytosis of agonist-activated beta-AR failed to block agonist-stimulated redistribution of alpha s. These findings demonstrate that activated alpha s reversibly undergoes a translocation from the plasma membrane to cytoplasm and begin to address the relationship between regulated trafficking of a seven-transmembrane receptor and its cognate G protein.     

Studies of cyclic AMP action using mutant tissue culture cells

Summary S49 mouse lymphoma cell mutants, each with a specific defect in its ability to generate or respond to cyclic AMP, have been isolated. Analysis of the properties of these cells has begun to provide information on complex and significant biologic problems related to the cyclic AMP system. Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture Association, New Orleans, Louisiana, June 6–9, 1977. The work was supported in part by National Science Foundation Grant BMS 75-06764 and National Institutes of Health Grants GM 16496 and GM 00001. P.C. is the recipient of National Institutes of Health Research Career Development Award GM 00308. P. A. I. is an Established Investigator of the American Heart Association.     

Studies of cyclic AMP action using mutant tissue culture cells.

S49 mouse lymphoma cell mutants, each with a specific defect in its ability to generate or respond to cyclic AMP, have been isolated. Analysis of the properties of these cells has begun to provide information on complex and significant biologic problems related to the cyclic AMP system.     

Activated-sludge process dynamics with continuous total organic carbon and oxygen uptake measurements

The dynamic responses of a bench-scale activated-sludge process to step changes and square-wave inputs in the feed flow and concentration were measured. Instrumentation permitted the continuous measurement of the oxygen uptake rate and dissolved organic carbon responses. Notable were the sensitivity of the oxygen uptake rate to process changes and the reliability of the dynamic oxygen electrode method. The responses were found to be greatly influenced by the organic loading, FS₀/XV, which was incorporated into a load-dependent kinetics model. Simulations showd good agreement with experiment in the case of the square-wave disturbances. Because of the changing and complex nature of the activated sludge it was necessary to reestimate the parameter set for each run.     

Brucellosis in a Veterans' Hospital, 1963

In Canada, about 100 sporadically occurring cases of brucellosis are reported yearly. Three patients were admitted to one Montreal hospital in the first seven months of 1963; all were employed in or around a packing plant. One had pain and electrocardiographic changes suggestive of Brucella myocarditis; he recovered promptly. Symptoms of neurasthenia and anxiety are common and were observed in two of these three cases. Two had positive blood cultures; the third showed conclusive agglutination to Br. abortus (1:10,240). In some patients with frequent previous exposures to infection the agglutination response may be equivocal and difficult to interpret. All three patients responded well to tetracycline; streptomycin and corticosteroids also have a role in the therapy of some cases. There may also be some place for combined antibiotic and vaccine therapy. The program of control of the disease in cattle in Canada is aimed at eradication by 1967.