State dependent dissociation of HERG channel inhibitors.

BACKGROUND AND PURPOSE: Inhibition of HERG channels prolongs the ventricular action potential and the QT interval with the risk of torsade de pointes arrhythmias and sudden cardiac death. Many drugs induce greater inhibition of HERG channels when the cell membrane is depolarized frequently. The dependence of inhibition on the pulsing rate may yield different IC(50) values at different frequencies and thus affect the quantification of HERG channel block. We systematically compared the kinetics of HERG channel inhibition and recovery from block by 8 blockers at different frequencies. EXPERIMENTAL APPROACH: HERG channels were expressed heterologously in Xenopus oocytes and currents were measured with the two-electrode voltage clamp technique. KEY RESULTS: Frequency-dependent block was observed for amiodarone, cisapride, droperidol and haloperidol (group 1) whereas bepridil, domperidone, E-4031 and terfenadine (group 2) induced similar pulse-dependent block at all frequencies. With the group 1 compounds, HERG channels recovered from block in the presence of drug (recovery being voltage-dependent). No substantial recovery from block was observed with the second group of compounds. Washing out of bepridil, domperidone, E-4031 and terfenadine was substantially augmented by frequent pulsing. Mutation D540K in the HERG channel (which exhibits reopening at negative voltages) facilitated recovery from block by these compounds at -140 mV. CONCLUSION AND IMPLICATIONS: Drug molecules dissociate at different rates from open and closed HERG channels ('use-dependent' dissociation). Our data suggest that apparently 'trapped' drugs (group 2) dissociated from the open channel state whereas group 1 compounds dissociated from open and resting states.

First gamma-ray measurements of fusion alpha particles in JET trace tritium experiments.

Gamma-ray spectra from nuclear reactions between fusion-born alpha (alpha) particles and Be impurities were measured for the first time in deuterium-tritium plasmas in the Joint European Torus. The time dependence of the measured spectra allowed the determination of the density evolution of fast alpha particles. Correlation between the decay time of the gamma-ray emission and the plasma parameters in different plasma scenarios was established. Results are consistent with classical slowing down of the alpha particles in discharges with high plasma currents and monotonic q-profiles. In low plasma current discharges and in the discharges with large on-axis current holes (extreme reversal central magnetic shear), the gamma-ray emission decay times are shorter than the classical slowing down times, indicating an alpha-particle confinement degradation in such discharges in line with theoretical predictions.

Fluorescence in situ hybridization vs. epsilometer test for detection of clarithromycin-susceptible and clarithromycin-resistant Helicobacter pylori strains in gastric biopsies from children.

AIM: To compare the results of culture and epsilometer test with fluorescence in situ hybridization for the detection of Helicobacter pylori and the presence of clarithromycin-susceptible and clarithromycin-resistant strains in antral biopsies from children. METHODS: Antral biopsies from 149 unselected children were investigated prospectively; 15 had previously received anti-H. pylori therapy. H. pylori status was defined by histology, rapid urease test and 13C-urea breath test. Fluorescence in situ hybridization was applied on fresh tissue with probes specific for the clarithromycin-susceptible wild type and three clarithromycin-resistant mutants. Susceptibility to clarithromycin was tested by epsilometer test in two laboratories. RESULTS: Culture and fluorescence in situ hybridization gave negative results in all 66 H. pylori-negative children (specificity, 100%). Of 83 infected children, cultures were successful in 75 (90%), epsilometer test in 71 (86%) and fluorescence in situ hybridization in 77 (93%). Eleven children (13%) showed discrepant results between the applied methods, indicating mixed infection. Clarithromycin-resistant isolates were identified in 16 of 73 previously untreated children. CONCLUSIONS: Primary resistance to clarithromycin is common (22%) in H. pylori isolates from children living in Germany. Fluorescence in situ hybridization is an excellent, fast method for the detection of H. pylori and clarithromycin-resistant mutants in gastric biopsies. Multiple biopsies identify mixed infections, indicating that clarithromycin-resistant and clarithromycin- susceptible strains are not evenly distributed within the stomach.

Regulation of expression of terminal oxidases in Paracoccus denitrificans.

In order to study the induction of terminal oxidases in Paracoccus denitrificans, their promoters were fused to the lacZ reporter gene and analysed in the wild-type strain, in an FnrP-negative mutant, in a cytochrome bc1-negative mutant, and in six single or double oxidase-negative mutant strains. The strains were grown under aerobic, semi-aerobic, and denitrifying conditions. The oxygen-sensing transcriptional-regulatory protein FnrP negatively regulated the activity of the qox promoter, which controls expression of the ba3-type quinol oxidase, while it positively regulated the activity of the cco promoter, which controls expression of the cbb3-type cytochrome c oxidase. The ctaDII and ctaC promoters, which control the expression of the aa3-type cytochrome c oxidase subunits I and II, respectively, were not regulated by FnrP. The activities of the latter two promoters, however, did decrease with decreasing oxygen concentrations in the growth medium, suggesting that an additional oxygen-sensing mechanism exists that regulates transcription of ctaDII and ctaC. Apparently, the intracellular oxygen concentration (as sensed by FnrP) was not the only signal to which the oxidase promoters responded. At given extracellular oxygen status, both the qox and the cco promoters responded to mutations in terminal oxidase genes, whereas the ctaDII and ctaC promoters did not. The change of electron distribution through the respiratory network, resulting from elimination of one or more oxidase genes, may have changed intracellular signals that affect the activities of the qox and cco promoters. On the other hand, the re-routing of electron distribution in the respiratory mutants hardly affected the oxygen consumption rate as compared to that of the wild-type. This suggests that the mutants adapted their respiratory network in such a way that they were able to consume oxygen at a rate similar to that of the wild-type strain.

Empirical error-confidence curves for neural network and Gaussian classifiers.

"Error-Confidence" measures the probability that the proportion of errors made by a classifier will be within epsilon of EB, the optimal (Bayes) error. Probably Almost Bayes (PAB) theory attempts to quantify how this confidence increases with the number of training samples. We investigate the relationship empirically by comparing average error versus number of training patterns (m) for linear and neural network classifiers. On Gaussian problems, the resulting EC curves demonstrate that the PAB bounds are extremely conservative. Asymptotic statistics predicts a linear relationship between the logarithms of the average error and the number of training patterns. For low Bayes error rates we found excellent agreement between the prediction and the linear discriminant performance. At higher Bayes error rates we still found a linear relationship, but with a shallower slope than the predicted-1. When the underlying true model is a three-layer network, the EC curves show a greater dependence on classifier capacity, and the linear predictions no longer seem to hold.

Do Gabor functions provide appropriate descriptions of visual cortical receptive fields?

Several recent theoretical models for human spatial vision posit that cortical receptive fields act to minimize simultaneously the product of the standard deviation of the sensitivities to position (delta chi) and to spatial frequency (delta omega) in accord with the uncertainty principle from Fourier analysis. The receptive-field functions resulting from this approach--one-dimensional or two-dimensional Gabor elementary functions--have been shown by others to fit measured receptive fields adequately in some species. However, only complex-valued Gabor functions minimize this product, and these cannot be fitted to single-cell receptive fields. We point out that the derivations of others have an implied metric or measure of positional and spatial-frequency uncertainties and that there is an infinitely large class of such metrics, many of which yield other receptive-field functions that are quite plausible biologically. We review neurophysiological measurements of others and analyze psychophysical masking data and find that in many cases receptive-field functions other than Gabor functions fit better. We conclude that there are insufficient theoretical demonstrations and experimental data to favor Gabor functions over any of a number of other plausible receptive-field functions.

Temporal impulse responses from flicker sensitivities.

We derive visual temporal impulse response functions from the large-field sinusoidal flicker sensitivity measurements made at several luminances by Kelly [J. Opt. Soc. Am. 51, 422 (1961)]. To this end we recognize response function causality and use Kramers-Krönig relations to reconstruct the (unmeasured) temporal phase spectrum and hence also the impulse response functions. At high luminances our derived functions are triphasic, while at low luminances they are biphasic. These functions are model independent, rely on no free parameters, and are consistent with certain theoretical model-dependent functions as well as psychophysical measurements by others.

Receptive-field symmetry probed using converging gratings.

Contrast-sensitivity functions (CSF's) for converging and diverging gratings were obtained under voluntary fixation at several field widths and under retinal stabilization at one field width. In general, these types of gratings had similar CSF's at all temporal frequencies tested, a result that can be explained by plausible spatiotemporal receptive-field models possessing spatial antisymmetry but not by models possessing pure (even) spatial symmetry. Phase condition at the central line in converging and diverging gratings affected grating detectability at high spatial frequencies, as expected.