Characterization of the bioactive form and molecular determinants of recognition of cyclic enkephalin peptides at the delta-opioid receptor

An extensive and systematic search strategy to determine the conformational profile of 12 cyclic disulfide-bridged opioid peptides with varying affinities at the delta receptor has been carried out to identify the structure that is recognized by the delta receptor for each analogue. The methods and procedures used here for the conformational search have already been validated for [D-Pen2,D-Pen5] enkephalin (DPDPE), one member of this family. Use of these methods led to a low-energy solution conformation of DPDPE in excellent agreement with all the geometric properties deduced from its solution nmr spectra. Each of the analogue was subjected to the same procedure, involving a combination of molecular dynamics simulations at high and low temperature. The study was repeated in two environmental conditions, an apolar environment, simulated by using a distance-dependent dielectric constant, and a polar environment by embedding the peptides in a high constant dielectric (epsilon = 80). An automated comparison of the different conformers based on their backbone rms and average distance between the key aromatic moieties was followed by graphic analysis using maximum structural overlap. The cross-comparison of the conformations for each analogue revealed a unique conformer that may be recognized by the delta receptor for each high-affinity analogue that permitted maintaining the critical elements required for recognition in a simple spatial orientation, while maximizing similarity in other regions.     

Design of a bistatic dual-Doppler radar for retrieving vector windsusing one transmitter and a remote low-gain passive receiver

A bistatic dual-Doppler network consisting of an existing single transmitting pencil-beam weather radar and a remotely located, nontransmitting, passive bistatic receiver with a low-gain antenna was constructed and tested during 1993. High-quality dual Doppler vector winds were retrieved from this system. The windfields were compared with those collected with a traditional two-transmitter dual-Doppler system. Several interesting engineering challenges relating to frequency and timing synchronization were resolved in order to retrieve successfully Doppler velocities at a remote bistatic site. Frequency synchronization was achieved by using extremely stable local oscillators, linked by both Global Positioning Satellite (GPS) signals and transmitter sidelobe coupling. Both methods provided the necessary one part in 10⁹ coherence necessary for calculating accurate Doppler velocities. Timing/gating synchronization with submicrosecond accuracy was achieved by using local oscillators at each site linked with GPS and sidelobe coupling. The successful testing of this system demonstrates that inexpensive and practical bistatic multiple-Doppler networks can be deployed. These systems can provide three-dimensional vector winds for a number of purposes in research, aviation, media, weather prediction, education, meteorological modeling and severe weather detection     

Trp82 and Tyr332 are involved in two quaternary ammonium binding domains of human butyrylcholinesterase as revealed by photoaffinity labeling with [3H]DDF

Purified butyrylcholinesterase (BuChE) was photolabeled by [3H]-p-N, N-dimethylamino benzene diazonium ([3H]DDF) to identify the quaternary ammonium binding sites on this protein [Ehret-Sabatier, L. , Schalk, I., Goeldner, M., and Hirth, C. (1992) Eur. J. Biochem. 203, 475-481]. The covalent photoincorporation occurs with a stoichiometry of one mole of probe per mole of inactivated site and could be fully prevented by several cholinergic inhibitors such as tacrine or tetramethylammonium. After complete deglycosylation of the enzyme using N-glycosidase F, the alkylated protein was trypsinolyzed and the digests were analyzed by HPLC coupled to ES-MS. A direct comparison of tryptic fragments from labeled and unlabeled BuChE allowed us to identify the tryptic peptide Tyr61-Lys103 as carrying the probe. Purification of the labeled peptides by anion-exchange chromatography gave a major radioactive peak which was further fractionated by reversed-phase HPLC leading to three, well-resolved, radioactive peaks. Microsequencing revealed that two of these peaks contained an overlapping sequence starting at Tyr61, while the third peak contained a sequence extending from Thr315. Radioactive signals could be unambiguously attributed to positions corresponding to residues Trp82 and Tyr332. This labeling study establishes the existence of two different binding domains for quaternary ammonium in BuChE and exemplifies additional cation/pi interactions in cholinergic proteins. This work strongly supports the existence of a peripheral anionic site in BuChE, implying residue Tyr332 as a key element.     

A methodology for modeling the distributions of medical images and their stochastic properties

The probabilistic distribution properties of a set of medical images are studied. It is shown that the generalized Gaussian function provides a good approximation to the distribution of AP chest radiographs. Based on this result and a goodness-of-fit test, a generalized Gaussian autoregressive model (GGAR) is proposed. Its properties and limitations are also discussed. It is expected that the GGAR model will be useful in describing the stochastic characteristics of some classes of medical images and in image data compression and other applications.     

Image staining and differential diagnosis of ultrasound scans based on the Mahalanobis distance

The covariance matrices associated with each state of health or disease from a previous study are used as the basis of an image staining display technique for aid in quantitative differential diagnosis. A state of health or disease is chosen by the clinician: this selects the covariance matrix from the data base. A region of interest (ROI) is then scrolled through an abdominal B-scan. For each position of the ROI a point in the four-dimensional feature space is calculated. A natural measure of the distance of this point from the center of mass (multivariate mean) of the disease class is calculated in terms of the covariance matrix of this class; this measure is the Mahalanobis distance. The confidence level for acceptance or rejection of the hypothesized disease class is obtained from the probability distribution of this distance, the T(2) probability law. This confidence level is color coded and used as a color stain that overlays the original scan at that position. The variability of the calculated features is studied as a function of ROI size, or the spatial resolution of the color coded image, and it is found that for an ROI in the neighborhood of 4 cm(2) most of the variability due to the finite number of independent samples (speckles) is averaged out, leaving the "noise floor" associated with inter- and intra-patient variability. ROIs on the order of 1 cm(2) may result with technical advances in B-scan resolution. A small number of points on organ boundaries are entered by the user, to fit with arcs of ellipses to be used to switch between organ (liver and kidney) data bases as the ROI encounters the boundary. By selecting in turn various state-of-health or state-of-disease databases, such images of confidence levels may be used for quantitative differential diagnosis. The method is not limited to ultrasound, being applicable in principle to features obtained from any modality or multimodality combination.     

Construction and evaluation of a three-dimensional structure of cytochrome P450choP enzyme (CYP105C1)

The purpose of this work was to develop and carefully evaluateimproved strategies for constructing reliable 3-D models ofP450 isozymes. To this end, a unique combination of steps forbuilding and evaluating a model structure was used to builda homology model of the P450choP isozyme, based on knowledgeof the X-ray structures of P450cam, P450terp, P450BM-3 and P450eryF.Specifically, the reliability of this model was examined bysystematic comparisons of its conformational, energetic, environmentaland packing properties and those of the four reference proteinswith corresponding properties from the database of proteinswith known structures. The results showed that the examinedproperties of this model structure are well within the criteriaestablished for reliable structures and are of nearly as goodquality as those of the reference proteins. In addition, theresult from a 120 ps unconstrained MD simulation of the modelwith structural waters provided evidence that the model is stableat room temperature. This 3-D model can now be reliably usedfor explicit characterization of substrate and inhibitor complexes.Most importantly, although it is envisioned that building modelsfor mammalian P450s will be even more challenging, the stepsdescribed here should be very useful in future constructionof 3-D models of mammalian P450 isozymes.     

ECG changes after paroxetine. 3 case reports

Selective serotonin reuptake inhibitors (SSRI) are known to exhibit far less cardiac side effects than tricyclic antidepressants. We report three cases in which the SSRI, paroxetine, induced clinically relevant ECG changes in patients with high risk profile. In two cases an increase in QTc time was observed, severe bradycardia was also observed twice.     

Subsurface exploration and transient pressure testing from a deep tunnel in fractured and karstified limestones (Lötschberg Base Tunnel,Switzerland)

This paper summarizes the results from a subsurface exploration campaign carried out in fractured and karstified limestones and marls of the Lötschberg Base Tunnel in the Swiss Alps. The exploration program in the 3.2 km long Doldenhorn Section consisted of 50 cored and 13 destructive horizontal predrillings of 250–300 m length each (in total 14,853 m of predrillings). These boreholes were used to locate possible tunnel inflows and characterize their hydraulic properties. A new efficient hydrogeological test protocol has been developed and was systematically and successfully applied. The test protocol consists of short constant rate outflow and recovery tests in open boreholes. The on-site test analysis allowed reliable predictions of the hydrogeological conditions ahead of the advancing tunnel face in brief period of time, leading to decisions regarding further reconnaissance drillings, tunnel driving or sealing measures. Detailed analysis of the transient pressure responses of the hydrogeological tests carried out after tunnel completion led to important new findings regarding flow model identification (type and orientation of conductive fractures), aquifer parameters and boundary effects. Even though the hydrogeological tests had to be carried out within a short time frame (a few hours) they allowed characterization of large scale aquifer properties relevant for long term tunnel inflow behavior.     

Derivation of surface soil moisture from ENVISAT ASAR wide swath and image mode data in agricultural areas

Water and energy fluxes at the interface between the land surface and atmosphere are strongly dependent on surface soil moisture content, which is highly variable in space and time. It has been shown in numerous studies that microwave remote sensing can provide spatially distributed patterns of surface soil moisture. In order to use remote-sensing-derived soil moisture information for practical applications as, for example, flood forecasting and water balance modeling in mesoscale areas, frequent large-area coverage is a prerequisite. New sensor generations such as ENVISAT Advanced Synthetic Aperture Radar (ASAR) or RADARSAT allow for image acquisitions in different imaging modes and geometries. Imaging modes with the capability of large-area coverage, such as the Wide Swath Mode of ENVISAT ASAR, are of special interest for practical applications in this context. This paper presents a semiempirical soil moisture inversion scheme for ENVISAT ASAR data. Different land cover types as well as mixed-image pixels are taken into account in the soil moisture retrieval process. The inversion results are validated against in situ measurements, and a sensitivity analysis of the model is conducted.     

Estimating the uncertainty in the estimated mean area under the ROC curve of a classifier

This article considers the problem of binary classification and its assessment in a distribution-free approach. We estimate the area under the ROC curve (a more general performance metric than the error rate) of a classifier using a bootstrap-based estimator. We then use the method of the influence function to estimate the uncertainty of that estimate from the very same bootstrap samples. Monte Carlo trials show that small-sample estimates can be obtained with little bias.