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The positive inotropic action of the newer cardiotonic phosphodiesterase inhibitors such as indolidan, milrinone, and imazodan has been previously attributed to selective inhibition of cGMP-inhibitable Type IV (high affinity) cAMP phosphodiesterase activity. However, the subcellular binding site(s) for this class of compounds has not been defined. We have characterized the binding of [3H]LY186126, an analogue of indolidan, in subcellular fractions prepared from rabbit and sheep ventricular myocardium. Binding required magnesium ion and exhibited rapid association and dissociation kinetics. Specific binding (defined by ligand displacement with 5 microM indolidan) to enriched rabbit sarcoplasmic reticulum (SR) membrane vesicles was saturable (Bmax = 714 +/- 77 fmol/mg of protein) and of high affinity (Kd = 6.2 +/- 1.4 nM). Linear and nonlinear analyses of the binding isotherms fit a single-site model. Mixed SR preparations from sheep myocardium exhibited binding characteristics (Bmax = 944 +/- 115 fmol/mg; Kd = 8.5 +/- 2.3 nM) comparable to those of rabbit cardiac SR. Further subfractionation of sheep SR indicated that the binding sites were equally distributed between free (Bmax = 630 fmol/mg; Kd = 4.4 nM) and junctional SR (Bmax = 569 fmol/mg; Kd = 10.9 nM). Specific binding of [3H]LY186126 was also demonstrated in the cytosolic subfraction of rabbit myocardium that contained Type IV phosphodiesterase activity (Peak III from anion exchange chromatography). Competition for [3H] LY186126 binding studied in rabbit SR showed that, of the compounds tested, lixazinone (RS 82856) competed most effectively (IC50 = 0.030 +/- 0.008 nM), followed by indolidan (0.14 +/- 0.05 nM), cGMP (17.8 +/- 2.6 nM), milrinone (39.3 +/- 13.2 nM), and imazodan (192 +/- 73 nM). In contrast, rolipram, which does not inhibit SR-associated Type IV phosphodiesterase activity, was not effective at competing for [3H]LY186126 binding (IC50 greater than 30 microM). These results indicate that [3H]LY186126 has specific binding sites in myocardial subcellular fractions that contain cGMP-inhibitable Type IV (high affinity) cAMP phosphodiesterase activity.

The mean generation time of Haemophilus influenzae in simulated blood cultures is 103 to 107 min. With 0.56 to 0.58 doublings per h, even large inocula of 256 cells per ml reach only 2 X 10(6) cells per ml and produce no visible evidence of growth after 24 h of incubation. Hemin and NAD added to simulated blood cultures triple the rate of growth of H. influenzae, so that even small inocula produce visible turbidity after overnight incubation. With a mean generation time of 36 min, a single cell of H. influenzae in simulated blood culture supplemented with hemin and NAD undergoes 30 doublings in 18 h, producing 2(30) (1.07 X 10(9] cells and visible turbidity.

Studies were made of the synthesis of beta-galactosidase by Escherichia coli in the presence of 5-fluorouracil, streptomycin, and subinhibitory concentrations of chloramphenicol. The preferential inhibition of beta-galactosidase synthesis observed in the presence of the above drugs was found to be caused by catabolite repression.

Studies were made of the synthesis of ß-galactosidase by Escherichia coliin the presence of 5-fluorouracil, streptomycin, and subinhibitory concentrations of chloramphenicol. The preferential inhibition of ß-galactosidase synthesis observed in the presence of the above drugs was found to be caused by catabolite repression.