We report the discovery of a new potent allosteric effector of sickle cell hemoglobin, GBT440 (), that increases the affinity of hemoglobin for oxygen and consequently inhibits its polymerization when subjected to hypoxic conditions. Unlike earlier allosteric activators that bind covalently to hemoglobin in a 2:1 stoichiometry, binds with a 1:1 stoichiometry. Compound is orally bioavailable and partitions highly and favorably into the red blood cell with a RBC/plasma ratio of ∼150. This partitioning onto the target protein is anticipated to allow therapeutic concentrations to be achieved in the red blood cell at low plasma concentrations. GBT440 () is in Phase 3 clinical trials for the treatment of sickle cell disease (NCT03036813).

Compliance with oral self-administered allopurinol (daily medication) and prednisone (intermittent medication) as well as compliance with monthly scheduled clinic appointments, were examined in 108 patients with newly diagnosed hematologic malignancy. Baseline levels of compliance (control group) were compared to results obtained after implementation of three intervention packages, whose aim was to increase compliance. The packages included combinations of education, home psychologic support and restructuring, and training in pill taking. A 24-hour profile of the two drugs and their metabolites was first determined. Serum samples were then obtained monthly over 6 months and analyzed for presence of the drugs. Control patients were fully compliant with allopurinol only 16.8% of the time. This rate increased significantly (44% to 48% of the time) for those who received any one of the intervention programs. With respect to prednisone, control patients were compliant 26.8% of the time, with no real improvement after interventions. Finally, self reports overestimated compliance by a factor of two when compared to drug analysis. The results indicated that full compliance with oral medications was remarkably low among our patients who have treatable and in some cases curable hematologic malignancy. However, compliance can be significantly improved by the use of various intervention packages.

We report a new technique for assessing the amount of information extracted from the icon that follows a briefly presented picture. The problem of how to measure such information was formulated in terms of how much physical exposure of a picture an icon is worth. Consider two types of stimulus presentations, each with a base duration of d ms. The first is a d-ms picture followed by an icon, and the second is a d + a-ms picture not followed by an icon. How large does a have to be so that equivalent amounts of information are extracted in the two cases? To answer this question, we showed people pictures and later tested their memory for the pictures. We found that the physical exposure duration needed to reach a particular level of performance was approximately 100 ms longer when an icon was not permitted versus when the icon was permitted. This value was independent of the base duration and the luminance of the picture. Moreover, the same value was obtained using three different kinds of memory test and four different sets of pictures. We conclude that an icon is worth approximately 100 ms of additional physical exposure duration. A reasonable explanation for this robust equivalence between icon and stimulus is that the same encoding processes are responsible for extracting information from the icon and from the physical stimulus. Therefore, any variable that affects these encoding processes must affect extraction of information from the icon and the physical stimulus in an identical manner. This prediction was confirmed for one such variable, picture luminance.

By combining the tools of site-directed mutagenesis and [3H]ouabain binding, the functional role of glutamic acid 327 in the fourth transmembrane domain of the sheep alpha 1 isoform of Na+,K(+)-ATPase was examined with respect to its interactions with ouabain, Na+,K+,Mg2+, and inorganic phosphate. Using site-directed mutagenesis, this glutamic acid was substituted with alanine, aspartic acid, glutamine, and leucine. The mutant proteins were constructed in a sheep alpha 1 protein background such that [3H]ouabain binding could be utilized as a highly specific probe of the exogenous protein expressed in NIH 3T3 cells. Na+ competition of [3H]ouabain binding to the mutant forms of Na+,K(+)-ATPase revealed only slight alterations in their affinities for Na+ and in their abilities to undergo Na(+)-induced conformational changes which inhibit ouabain binding. In contrast, K+ competition of [3H]ouabain binding to all four mutant forms of Na+,K(+)-ATPase displayed severely altered interactions between these proteins and K+. Interestingly, [3H]ouabain binding to the mutant E327Q was not inhibited by the presence of K+. This mutant was previously reported to be functionally able to support cation transport with a 5-fold reduced K0.5 for K(+)-dependent ATPase activity (Jewell-Motz, E. A., and Lingrel, J.B. (1993) Biochemistry 32, 13523-13530; Vilsen, B. (1993) Biochemistry 32, 13340-13349). Thus, it appears that this glutamic acid in the fourth transmembrane domain may be important for stabilizing a K(+)-induced conformation within the catalytic cycle of Na+,K(+)-ATPase that is not rate-limiting in the overall ATPase cycle but that displays a greatly reduced affinity for ouabain.

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