Structural Plasticity Is a Feature of Rheostat Positions in the Human Na+/Taurocholate Cotransporting Polypeptide (NTCP)

In the Na⁺/taurocholate cotransporting polypeptide (NTCP), the clinically relevant S267F polymorphism occurs at a “rheostat position”. That is, amino acid substitutions at this position (“S267X”) lead to a wide range of functional outcomes. This result was particularly striking because molecular models predicted the S267X side chains are buried, and thus, usually expected to be less tolerant of substitutions. To assess whether structural tolerance to buried substitutions is widespread in NTCP, here we used Rosetta to model all 19 potential substitutions at another 13 buried positions. Again, only subtle changes in the calculated stabilities and structures were predicted. Calculations were experimentally validated for 19 variants at codon 271 (“N271X”). Results showed near wildtype expression and rheostatic modulation of substrate transport, implicating N271 as a rheostat position. Notably, each N271X substitution showed a similar effect on the transport of three different substrates and thus did not alter substrate specificity. This differs from S267X, which altered both transport kinetics and specificity. As both transport and specificity may change during protein evolution, the recognition of such rheostat positions may be important for evolutionary studies. We further propose that the presence of rheostat positions is facilitated by local plasticity within the protein structure. Finally, we note that identifying rheostat positions may advance efforts to predict new biomedically relevant missense variants in NTCP and other membrane transport proteins.     

Phase Equilibria in the System CaF2-AlF3-Na3AlF6 and Part of the System CaF2-AlF3-Na3AlF6-Al2O3

The complete phase-equilibrium diagram of the system CaF₂-AlF₃-Na₃AlF₆ and the subsolidus portion of the system CaF₂-AlF₃-Na₃AlF₆-Al₂O₃ were established from microscopic, powder X-ray diffraction, quench, and DTA data obtained from samples encapsulated in sealed tubes and either reacted in the solid state or melted and recrystallized. The system Na₃AlF₆-CaF₂ contains a simple eutectic with no compound formation or solid solution. The system CaF₂-AlF₃-Na₃AlF₆ contains two ternary compounds, NaCaAlF₆ and NaCaAl₂F₉, which melt incongruently at 735° and 712°C, respectively; NaCaAlF₆ exists in three polymorphic forms with transitions at 610° and 722°C and NaCaAl₂F₉ is body-centered cubic with a₀= 10.765 Å. The three binary and two ternary compounds divide the system into eight compatibility triangles. Along the NaCaAlF₆-Na₃AlF₆ join, 7 mol% NaCaAlF₆ is soluble in α-cryolite at 525° and 42% in β-cryolite at 731°C. The quaternary system Na₃AlF₆-AlF₃-CaF₂-Al₂O₃ contains eight compatibility tetrahedra.     

3D Active Edge Silicon Detector Tests With 120 GeV Muons

3D detectors with electrodes penetrating through the silicon wafer and covering the edges were tested in the SPS beam line X5 at CERN in autumn 2003. Detector parameters including efficiency, signal-to-noise ratio, and edge sensitivity were measured using a silicon telescope as a reference system. The measured sensitive width and the known silicon width were equal within less than 10 mum.     

AN ANALYSIS OF PHASE DISTRIBUTION AND TURBULENCE IN DISPERSED PARTICLE/LIQUID FLOWS

An analysis of dilute, turbulent particle/liquid two-phase flow is presented. The three-dimensional conservation equations that govern turbulent motion in solid/fluid flows are derived using ensemble averaging, and the unknown terms in these equations are constituted to achieve closure. These closure terms include the shear stress due to interparticle collisions, the corresponding terms in the Reynolds stress equation, the force and dissipation due to particle-wall collisions, and the interfacial work due to particle/turbulent eddy interaction. The resultant two-fluid model was then evaluated using a computational fluid dynamic (CFD) solver and the predictions were compared with experimental data. Good agreement was observed for a variety of flow conditions.     

Differential binding of tropane-based photoaffinity ligands on the dopamine transporter

Benztropine and its analogs are tropane ring-containing dopamine uptake inhibitors that produce behavioral effects markedly different from cocaine and other dopamine transporter blockers. We investigated the benztropine binding site on dopamine transporters by covalently attaching a benztropine-based photoaffinity ligand, [125I]N-[n-butyl-4-(4"'-azido-3"'-iodophenyl)]-4', 4"-difluoro-3alpha-(diphenylmethoxy)tropane ([125I]GA II 34), to the protein, followed by proteolytic and immunological peptide mapping. The maps were compared with those obtained for dopamine transporters photoaffinity labeled with a GBR 12935 analog, [125I]1-[2-(diphenylmethoxy)ethyl]-4-[2-(4-azido-3-iodophenyl)ethy l]p iperazine ([125I]DEEP), and a cocaine analog, [125I]3beta-(p-chlorophenyl)tropane-2beta-carboxylic acid, 4'-azido-3'-iodophenylethyl ester ([125I]RTI 82), which have been shown previously to interact with different regions of the primary sequence of the protein. [125I]GA II 34 became incorporated in a membrane-bound, 14 kDa fragment predicted to contain transmembrane domains 1 and 2. This is the same region of the protein that binds [125I]DEEP, whereas the binding site for [125I]RTI 82 occurs closer to the C terminal in a domain containing transmembrane helices 4-7. Thus, although benztropine and cocaine both contain tropane rings, their binding sites are distinct, suggesting that dopamine transport inhibition may occur by different mechanisms. These results support previously derived structure-activity relationships suggesting that benztropine and cocaine analogs bind to different domains on the dopamine transporter. These differing molecular interactions may lead to the distinctive behavioral profiles of these compounds in animal models of drug abuse and indicate promise for the development of benztropine-based molecules for cocaine substitution therapies.     

Prevalence and predictors of sexual dysfunction in long-term survivors of marrow transplantation

PURPOSE: To describe the prevalence of sexual difficulties in men and women after marrow transplantation (MT), and to define medical, demographic, sexual, and psychologic predictors of sexual dysfunction 3 years after MT. PATIENTS AND METHODS: Four hundred seven adult MT patients were assessed pretransplantation. Survivors repeated measures of psychologic and sexual functioning at 1 and 3 years posttransplantation. RESULTS: Data were analyzed from 102 event-free 3-year survivors who defined themselves as sexually active. Men and women did not differ in sexual satisfaction pretransplantation. At 1 and 3 years posttransplantation, women reported significantly more sexual dysfunction than men. Eighty percent of women and 29% of men reported at least one sexual problem by 3 years after MT. No pretransplantation variables were significant predictors of 3-year sexual satisfaction for women. For men, pretransplantation variables of older age, poorer psychologic function, not being married, and lower sexual satisfaction predicted sexual dissatisfaction at 3 years (R2=.28; P < .001). Women who were more dissatisfied 3 years after MT did not receive hormone replacement therapy (HRT) at 1 -year posttransplantation and were less satisfied at 1 year, but not pretransplantation (R2=.35; P < .001). CONCLUSION: Sexual problems are significant in the lives of MT survivors, particularly for women. Although HRT before 1 year posttransplantation improves sexual function, it does not ensure sexual quality of life. Intervention for women is needed to apply hormonal, mechanical, and behavioral methods to prevent sexual difficulties as early after transplantation as possible.