Inactivation of cloned Na channels expressed in Xenopus oocytes

This study investigates the inactivation properties of Na channels expressed in Xenopus oocytes from two rat IIA Na channel cDNA clones differing by a single amino acid residue. Although the two cDNAs encode Na channels with substantially different activation properties (Auld, V. J., A. L. Goldin, D. S. Krafte, J. Marshall, J. M. Dunn, W. A. Catterall, H. A. Lester, N. Davidson, and R. J. Dunn. 1988. Neuron. 1:449-461), their inactivation properties resemble each other strongly but differ markedly from channels induced by poly(A+) rat brain RNA. Rat IIA currents inactivate more slowly, recover from inactivation more slowly, and display a steady-state voltage dependence that is shifted to more positive potentials. The macroscopic inactivation process for poly(A+) Na channels is defined by a single exponential time course; that for rat IIA channels displays two exponential components. At the single-channel level these differences in inactivation occur because rat IIA channels reopen several times during a depolarizing pulse; poly(A+) channels do not. Repetitive stimulation (greater than 1 Hz) produces a marked decrement in the rat IIA peak current and changes the waveform of the currents. When low molecular weight RNA is coinjected with rat IIA RNA, these inactivation properties are restored to those that characterize poly(A+) channels. Slow inactivation is similar for rat IIA and poly(A+) channels, however. The data suggest that activation and inactivation involve at least partially distinct regions of the channel protein

Evidence for the involvement of more than one mRNA species in controlling the inactivation process of rat and rabbit brain Na channels expressed in Xenopus oocytes

The properties of rat and rabbit brain sodium (Na) channels expressed in Xenopus oocytes following either unfractionated or high-molecular- weight mRNA injections were compared to assess the relative contribution of different size messages to channel function. RNA was size-fractionated on a sucrose gradient and a high-molecular-weight fraction (7–10 kilobase) encoding the α-subunit gave rise to functional voltage-dependent Na channels in the oocyte membrane. Single- channel conductance, mean open time, and time to first opening were all similar to the values for channels following injection of unfractionated RNA. In contrast, inactivation properties were markedly different; Na currents from high-molecular-weight RNA inactivated with a several-fold smaller macroscopic inactivation rate and showed a steady-state voltage dependence that was shifted in the depolarizing direction by at least 10 mV relative to that for unfractionated RNA. Single-channel recording revealed that the kinetic difference arose from a greater probability for high-molecular-weight RNA induced channels to reopen during a depolarizing voltage step. Pooling all gradient fractions and injecting this RNA into oocytes led to the appearance of Na channels with inactivation properties indistinguishable from those following injection of unfractionated RNA. These results suggest that mRNA species not present in the high- molecular-weight fraction can influence the inactivation process of rat brain Na channels expressed in Xenopus oocytes. This mRNA may encode β-subunits or other proteins that are involved in posttranslational processing of voltage-dependent Na channels

Evidence for the involvement of more than one mRNA species in controlling the inactivation process of rat and rabbit brain Na channels expressed in Xenopus oocytes

The properties of rat and rabbit brain sodium (Na) channels expressed in Xenopus oocytes following either unfractionated or high-molecular- weight mRNA injections were compared to assess the relative contribution of different size messages to channel function. RNA was size-fractionated on a sucrose gradient and a high-molecular-weight fraction (7–10 kilobase) encoding the α-subunit gave rise to functional voltage-dependent Na channels in the oocyte membrane. Single- channel conductance, mean open time, and time to first opening were all similar to the values for channels following injection of unfractionated RNA. In contrast, inactivation properties were markedly different; Na currents from high-molecular-weight RNA inactivated with a several-fold smaller macroscopic inactivation rate and showed a steady-state voltage dependence that was shifted in the depolarizing direction by at least 10 mV relative to that for unfractionated RNA. Single-channel recording revealed that the kinetic difference arose from a greater probability for high-molecular-weight RNA induced channels to reopen during a depolarizing voltage step. Pooling all gradient fractions and injecting this RNA into oocytes led to the appearance of Na channels with inactivation properties indistinguishable from those following injection of unfractionated RNA. These results suggest that mRNA species not present in the high- molecular-weight fraction can influence the inactivation process of rat brain Na channels expressed in Xenopus oocytes. This mRNA may encode β-subunits or other proteins that are involved in posttranslational processing of voltage-dependent Na channels

Association of Systemic Inflammation With Retinal Vascular Caliber in Patients With AIDS.

PurposeTo evaluate relationships among retinal vascular caliber and biomarkers of systemic inflammation in patients with AIDS.MethodsA total of 454 participants with AIDS had retinal vascular caliber (central retinal artery equivalent and central retinal vein equivalent) determined from enrollment retinal photographs by reading center graders masked to clinical and biomarker information. Cryopreserved plasma specimens were assayed for inflammatory biomarkers, including C-reactive protein (CRP), IL-6, interferon-γ inducible protein (IP)-10, kynurenine/tryptophan (KT) ratio, and intestinal fatty acid binding protein (I-FABP).ResultsIn the simple linear regression of retinal vascular caliber on plasma biomarkers, elevated CRP, IL-6, and IP-10 were associated with retinal venular dilation, and elevated KT ratio with retinal arteriolar narrowing. In the multiple linear regression, including baseline characteristics and plasma biomarkers, AMD was associated with dilation of retinal arterioles (mean difference: 9.1 μm; 95% confidence interval [CI] 5.2, 12.9; P < 0.001) and venules (mean difference, 10.9 μm; 95% CI, 5.3, 16.6; P < 0.001), as was black race (P < 0.001). Hyperlipidemia was associated with retinal venular narrowing (mean difference, -7.5 μm; 95% CI, -13.7, -1.2; P = 0.02); cardiovascular disease with arteriolar narrowing (mean difference, -5.2 μm; 95% CI, -10.3, -0.1; P = 0.05); age with arteriolar narrowing (slope, -0.26 μm/year; 95% CI, -0.46, -0.06; P = 0.009); and IL-6 with venular dilation (slope, 5.3 μm/standard deviation log10[plasma IL-6 concentration]; 95% CI, 2.7, 8.0; P < 0.001).ConclusionsThese data suggest that retinal vascular caliber is associated with age, race, AMD, hyperlipidemia, cardiovascular disease, and selected biomarkers of systemic inflammation

Kidney and pancreas transplantation

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106710/1/j.1600-6135.2004.00399.x.pd

Telescope Alignment From Sparsely Sampled Wavefront Measurements Over Pupil Subapertures

Alignment of two-element telescopes is a classic problem. During recent integration and test of the Space Interferometry Mission s (SIM s) Astrometric Beam Combiner (ABC), the innovators were faced with aligning two such telescope subsystems in the presence of a further complication: only two small subapertures in each telescope s pupil were accessible for measuring the wavefront with a Fizeau interferometer. This meant that the familiar aberrations that might be interpreted to infer system misalignments could be viewed only over small sub-regions of the pupil, making them hard to recognize. Further, there was no contiguous surface of the pupil connecting these two subapertures, so relative phase piston information was lost; the underlying full-aperture aberrations therefore had an additional degree of ambiguity. The solution presented here is to recognize that, in the absence of phase piston, the Zygo measurements primarily provide phase tilt in the subaperture windows of interest. Because these windows are small and situated far from the center of the (inaccessible) unobscured full aperture, any aberrations that are higher-order than tilt will be extremely high-order on the full aperture, and so not necessary or helpful to the alignment. Knowledge of the telescope s optical prescription allows straightforward evaluation of sensitivities (subap mode strength per unit full-aperture aberration), and these can be used in a predictive matrix approach to move with assurance to an aligned state. The technique is novel in every operational way compared to the standard approach of alignment based on full-aperture aberrations or searching for best rms wavefront. This approach is closely grounded in the observable quantities most appropriate to the problem. It is also more intuitive than inverting full phase maps (or subaperture Zernike spectra) with a ray-tracing program, which must certainly work in principle, but in practice met with limited success. Even if such classical alignment techniques became practical, the techniques reported here form a reassuringly transparent and intuitive check on the course of the alignment with very little computational effort

Together, yet still not equal? Sex integration in equestrian sport

Sex segregation is a core organising principle of most modern sports and is a key element in the marginalisation and subordination of girls and women in sport and beyond. In this article I explore the only Olympic-level sport which is not organised around sex segregation – equestrian sport – in order to consider the implications of sex integration for female participants. I draw on a study conducted on elite riders that found that although sex integration in equestrian sport does not lead to female participants being excluded from high-level competition, men continue to perform disproportionately well. This suggests that although sex integration may be an important step towards breaking down gender hierarchies in sport, without accompanying wider changes in gender norms and expectations, sex integration alone will not be enough to achieve greater gender equality in equestrian sport

Incoherent Pair Tunneling as a Probe of the Cuprate Pseudogap

We argue that incoherent pair tunneling in a cuprate superconductor junction with an optimally doped superconducting and an underdoped normal lead can be used to detect the presence of pairing correlations in the pseudogap phase of the underdoped lead. We estimate that the junction characteristics most suitable for studying the pair tunneling current are close to recently manufactured cuprate tunneling devices.Comment: ReVTeX 3.1; 4 pages, 2 EPS figures (included

maxdLoad2 and maxdBrowse: standards-compliant tools for microarray experimental annotation, data management and dissemination

BACKGROUND: maxdLoad2 is a relational database schema and Java(® )application for microarray experimental annotation and storage. It is compliant with all standards for microarray meta-data capture; including the specification of what data should be recorded, extensive use of standard ontologies and support for data exchange formats. The output from maxdLoad2 is of a form acceptable for submission to the ArrayExpress microarray repository at the European Bioinformatics Institute. maxdBrowse is a PHP web-application that makes contents of maxdLoad2 databases accessible via web-browser, the command-line and web-service environments. It thus acts as both a dissemination and data-mining tool. RESULTS: maxdLoad2 presents an easy-to-use interface to an underlying relational database and provides a full complement of facilities for browsing, searching and editing. There is a tree-based visualization of data connectivity and the ability to explore the links between any pair of data elements, irrespective of how many intermediate links lie between them. Its principle novel features are: • the flexibility of the meta-data that can be captured, • the tools provided for importing data from spreadsheets and other tabular representations, • the tools provided for the automatic creation of structured documents, • the ability to browse and access the data via web and web-services interfaces. Within maxdLoad2 it is very straightforward to customise the meta-data that is being captured or change the definitions of the meta-data. These meta-data definitions are stored within the database itself allowing client software to connect properly to a modified database without having to be specially configured. The meta-data definitions (configuration file) can also be centralized allowing changes made in response to revisions of standards or terminologies to be propagated to clients without user intervention. maxdBrowse is hosted on a web-server and presents multiple interfaces to the contents of maxd databases. maxdBrowse emulates many of the browse and search features available in the maxdLoad2 application via a web-browser. This allows users who are not familiar with maxdLoad2 to browse and export microarray data from the database for their own analysis. The same browse and search features are also available via command-line and SOAP server interfaces. This both enables scripting of data export for use embedded in data repositories and analysis environments, and allows access to the maxd databases via web-service architectures. CONCLUSION: maxdLoad2 and maxdBrowse are portable and compatible with all common operating systems and major database servers. They provide a powerful, flexible package for annotation of microarray experiments and a convenient dissemination environment. They are available for download and open sourced under the Artistic License