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Real-Time Four-dimensional Imaging of the Heart with Multi-Detector Row CT.

An interactive four-dimensional (4D) visualizing system for the heart was developed by the authors. The system realizes high-resolution three-dimensional (3D) imaging with temporal resolution in a beating heart by using eight or more data sets reconstructed from multi-detector row computed tomography (MDCT) with a retrospective electrocardiograph-gated reconstruction algorithm. The motion of heart walls, papillary muscles, septa, and valves can now be observed in 4D multiplanar reformations (MPRs), as with sonography, while coronary arteries, coronary sinuses, and cardiac veins can be analyzed during the optimal phase in 4D volume-rendering images, as with angiography. All parameters such as window width, window level, field of view, panning, tilt, thresholds, opacity, color, and segmentation function are completely interactive in 4D imaging. Two longitudinal views and one latitudinal view of a heart can be simultaneously visualized in the three relative 4D MPR views. These newly developed capabilities in viewing both 3D volume and temporal resolution data, functional data, and even multiphase data with registration add considerable diagnostic potential. The advent of this real-time 4D visualizing system has enhanced the capabilities of MDCT.

Algorithms↗

Real-time four-dimensional imaging of the heart with multi-detector row CT.

An interactive four-dimensional (4D) visualizing system for the heart was developed by the authors. The system realizes high-resolution three-dimensional (3D) imaging with temporal resolution in a beating heart by using eight or more data sets reconstructed from multi-detector row computed tomography (MDCT) with a retrospective electrocardiograph-gated reconstruction algorithm. The motion of heart walls, papillary muscles, septa, and valves can now be observed in 4D multiplanar reformations (MPRs), as with sonography, while coronary arteries, coronary sinuses, and cardiac veins can be analyzed during the optimal phase in 4D volume-rendering images, as with angiography. All parameters such as window width, window level, field of view, panning, tilt, thresholds, opacity, color, and segmentation function are completely interactive in 4D imaging. Two longitudinal views and one latitudinal view of a heart can be simultaneously visualized in the three relative 4D MPR views. These newly developed capabilities in viewing both 3D volume and temporal resolution data, functional data, and even multiphase data with registration add considerable diagnostic potential. The advent of this real-time 4D visualizing system has enhanced the capabilities of MDCT.

Echocardiography, Four-Dimensional↗

Age-dependent responses to nerve injury-induced mechanical allodynia.

BACKGROUND: Developmental differences in responses to acute and chronic nerve injury have received minimal attention. This study examines developmental differences in behavioral responses to a proximal (closer to the spinal cord) (L5 and L6 spinal nerve root ligation) or to a more distal (closer to peripheral innervation) (partial sciatic nerve ligation) nerve injury in rats paralleling the infant to young adult human. METHODS: Withdrawal thresholds to von Frey filament testing in the hind paw were determined before and various times after either spinal nerve root ligation or partial sciatic nerve ligation in rats aged 2, 4, and 16 weeks. Control rats of these ages were observed serially without surgery. Times for withdrawal thresholds to mechanical stimuli to return to 80% of that of the hind paw in the control animals were compared among the different ages in the two models. RESULTS: Baseline withdrawal thresholds in younger rats were lower (P < 0.05). In the 2-week-old animals, distal injury partial sciatic nerve ligation did not cause a reduction in withdrawal threshold from baseline. This was different from the spinal nerve root ligation group and the older animals in the partial sciatic nerve ligation group. However, when compared with age-matched control animals, both nerve injuries resulted in reduced withdrawal thresholds (P < 0.05). The resolution of hypersensitivity to mechanical stimulation, as measured by return of threshold to 80% of controls, occurred more quickly in 2-week-old than in 4- and 16-week-old animals in both injury models (P < 0.05). CONCLUSION: These data suggest that resolution of sensitization to A-fiber input occurs more rapidly in young animals. In addition, distal injury has less of a sensitizing effect on A-fiber input than proximal injury in the younger animals. The authors speculate that neuroimmune responses, especially at the site of injury, are developmentally regulated and less likely to produce chronic pain when injury occurs at a young age.

Aging↗

Polymorphisms of octopine dehydrogenase (Odh) in mollusks and implications for the neutralism-selectionism hypothesis.

Octopine dehydrogenase (Odh) was examined in several species of bivalves and gastropods and complemented with bibliographic data, to assess the controversy between neutralism and selectionism in explaining the maintenance of genetic variation in natural populations. This debate was the center of the molecular evolution and population genetic research in the 1970s and 1980s, but waned thereafter, without resolution. Although DNA data have been produced, implications are not understood. We examined the polymorphims of Odh in several species of bivalves and gastropods, and the kinetic properties (apparent Km) of the different isozymes in the scallop Euvola ziczac that indicates an apparent case of overdominance of the heterozygous individuals. The question "which of the two hypothesis is correct" has shifted with time to "how much influence did each factor have in the maintenance of genetic variation".

Amino Acid Oxidoreductases↗

Estimating unobserved reflection intensities in Laue diffraction by the maximum-entropy method.

In protein crystallography, the use of low-resolution reflections is important in defining the molecular mask and polypeptide backbone. However, in Laue data collection, the loss of low-resolution reflection data (>2dmin) can be as high as 40-50%, even after the deconvolution of multiples. To estimate the reflection intensities that are not recorded in data collection, a new method is presented based on maximizing the entropy of the Patterson function subject to the constraints imposed by the intensities of the observed reflections. The method has been tested with Laue diffraction data from hen egg-white lysozyme. All unobserved reflections within 5 A resolution were estimated, and their inclusion in the electron-density-map calculation significantly improved the connectivity. This method could also be applied to improve the completeness of monochromatic data.

Animals↗

Dynamics and phylogenetic implications of MtDNA control region sequences in New World Jays (Aves: Corvidae).

To study the evolution of mtDNA and the intergeneric relationships of New World Jays (Aves: Corvidae), we sequenced the entire mitochondrial DNA control region (CR) from 21 species representing all genera of New World jays, an Old World jay, crows, and a magpie. Using maximum likelihood methods, we found that both the transition/transversion ratio (kappa) and among site rate variation (alpha) were higher in flanking domains I and II than in the conserved central domain and that the frequency of indels was highest in domain II. Estimates of kappa and alpha were much more influenced by the density of taxon sampling than by alternative optimal tree topologies. We implemented a successive approximation method incorporating these parameters into phylogenetic analysis. In addition we compared our study in detail to a previous study using cytochrome b and morphology to examine the effect of taxon sampling, evolutionary rates of genes, and combined data on tree resolution. We found that the particular weighting scheme used had no effect on tree topology and little effect on tree robustness. Taxon sampling had a significant effect on tree robustness but little effect on the topology of the best tree. The CR data set differed nonsignificantly from the tree derived from the cytochrome b/morphological data set primarily in the placement of the genus Gymnorhinus, which is near the base of the CR tree. However, contrary to conventional taxonomy, the CR data set suggested that blue and black jays (Cyanocorax sensu lato) might be paraphyletic and that the brown jay Psilorhinus (=Cyanocorax) morio is the sister group to magpie jays (Calocitta), a phylogenetic hypothesis that is likely as parsimonious with regard to nonmolecular characters as monophyly of Cyanocorax. The CR tree also suggests that the common ancestor of NWJs was likely a cooperative breeder. Consistent with recent systematic theory, our data suggest that DNA sequences with high substitution rates such as the CR may nonetheless be useful in reconstructing relatively deep phylogenetic nodes in avian groups.

Animals↗

Multimodal integration of high-resolution EEG and functional magnetic resonance imaging data: a simulation study.

Previous simulation studies have stressed the importance of the use of fMRI priors in the estimation of cortical current density. However, no systematic variations of signal-to-noise ratio (SNR) and number of electrodes were explicitly taken into account in the estimation process. In this simulation study we considered the utility of including information as estimated from fMRI. This was done by using as the dependent variable both the correlation coefficient and the relative error between the imposed and the estimated waveforms at the level of cortical region of interests (ROI). A realistic head and cortical surface model was used. Factors used in the simulations were the different values of SNR of the scalp-generated data, the different inverse operators used to estimated the cortical source activity, the strengths of the fMRI priors in the fMRI-based inverse operators, and the number of scalp electrodes used in the analysis. Analysis of variance results suggested that all the considered factors significantly afflict the correlation and the relative error between the estimated and the simulated cortical activity. For the ROIs analyzed with simulated fMRI hot spots, it was observed that the best estimation of cortical source currents was performed with the inverse operators that used fMRI information. When the ROIs analyzed do not present fMRI hot spots, both standard (i.e., minimum norm) and fMRI-based inverse operators returned statistically equivalent correlation and relative error values.

Analysis of Variance↗

Direct determination of layer packing for a phospholipid solid solution at 0.32-nm resolution.

Electron diffraction intensity data were collected from a 2:3 binary solid solution of two homologous phosphatidylethanolamines (1,2-dimyristoyl-sn-glycerophosphoethanolamine and 1,2-dipalmitoyl-sn-glycerophosphoethanolamine) epitaxially oriented by cocrystallization with naphthalene. The layer packing was determined directly by predicting the value of 12 of the 17 phases from sigma 1- and sigma 2-triplet invariants in space group P1. A reverse Fourier transform of the resulting potential maps provides estimates for three other phases and the two remaining ones were found by generating maps for the 2(2) = 4 possible phase combinations and then testing the smoothness of the potential profile of the hydrocarbon chain packing. The same phase solution can be found by translating a molecular model (based on the known x-ray crystal structure of a shorter homologue) past the unit cell origin. The solid solution is found to retain a stable polar group packing while the statistical occupancy of two terminal-chain carbons is expressed by a reduced potential profile at the nonpolar interface at the bilayer center.

Crystallography, X-Ray↗

Resolution improvement of X-ray diffraction data of crystals of a vesicular stomatitis virus nucleocapsid protein oligomer complexed with RNA.

Vesicular stomatitis virus (VSV) is a non-segmented negative-stranded RNA virus. The nucleocapsid (N) protein of VSV is found tightly associated with the viral genomic RNA and this complex serves as the template for transcription and replication. A method for the soluble expression of the N protein in Escherichia coli has previously been reported. An N protein-RNA oligomer was isolated from this system, the stoichiometry of which was determined to be ten molecules of the N protein bound to approximately 90 nucleotides of RNA. Here, the crystallization of this protein-nucleic acid complex is presented. The crystals belong to space group P2(1)2(1)2, with unit-cell parameters a = 165.65, b = 235.35, c = 75.71 A and a diffraction limit of 6 A. Self-rotation function analysis has shown the oligomer to have tenfold rotational symmetry. In a search to identify heavy-atom derivatives, uranyl acetate was discovered to stabilize the crystals, giving them an increase in diffraction limits to beyond 2.9 A. Based on the internal symmetry of the oligomer, the size of the oligomer determined previously by negative-stained electron microscopy, the space-group symmetry and packing considerations, the packing arrangement in the crystal has been determined.

Crystallization↗

Crystallographic refinement of trichosanthin at 2.6A resolution.

The molecule model of trichosanthin has been rebuilt by using the electron density map improved by the solvent flatten and in accordance with the primary structure put forward by Collins. The crystallographic refinement of two trichosanthin molecules (3828 nonhydrogen atoms) in an asymmetric unit has been carried out by means of the restrain least-square procedure and diffraction data to a resolution of 2.6 A. The results are: an R factor 0.223 and the r.m.s. deviation of the bond length = 0.023 A. The new molecular model is in good agreement with the electron density map calculated with the coefficient 2Fo-Fc.

Crystallography↗

A bound on the energy resolution required for quantitative SPECT.

Scattered radiation is one of several physical perturbations that limit the accuracy of quantitative measurements in single-photon emission computed tomography (SPECT). Improvement in detector energy resolution leads to a reduction of scatter counts and a corresponding improvement in the quantitative accuracy of the SPECT measurement. In this study, simulated SPECT projections of a simple myocardial perfusion phantom were used to investigate the effect of detector energy resolution on the data. The phantom consists of a spherical shell of radionuclide within a 15 cm radius water-filled cylinder. Each projection contains on the order of 3 x 10(5) counts. The results demonstrate that a full-width, half-maximum energy resolution of 3-4 keV is sufficient to render the error due to scatter insignificant compared to the uncertainty due to photon statistics in this case. Further simulations verify that because smaller objects produce less scatter, they can be imaged accurately with degraded energy resolution. These results are useful when designing prototype systems that utilize solid-state detectors and low-noise electronics to achieve improved energy resolution.

Biometry↗

Structural insights into the beta-mannosidase from T. reesei obtained by synchrotron small-angle X-ray solution scattering enhanced by X-ray crystallography.

A molecular envelope of the beta-mannosidase from Trichoderma reesei has been obtained by combined use of solution small-angle X-ray scattering (SAXS) and protein crystallography. Crystallographic data at 4 A resolution have been used to enhance informational content of the SAXS data and to obtain an independent, more detailed protein shape. The phased molecular replacement technique using a low resolution SAXS model, building, and refinement of a free atom model has been employed successfully. The SAXS and crystallographic free atom models exhibit a similar globular form and were used to assess available crystallographic models of glycosyl hydrolases. The structure of the beta-galactosidase, a member of a family 2, clan GHA glycosyl hydrolases, shows an excellent fit to the experimental molecular envelope and distance distribution function of the beta-mannosidase, indicating gross similarities in their three-dimensional structures. The secondary structure of beta-mannosidase quantified by circular dichroism measurements is in a good agreement with that of beta-galactosidase. We show that a comparison of distance distribution functions in combination with 1D and 2D sequence alignment techniques was able to restrict the number of possible structurally homologous proteins. The method could be applied as a general method in structural genomics and related fields once protein solution scattering data are available.

Circular Dichroism↗

The fold of human aquaporin 1.

The fold of human aquaporin 1 is determined from cryo-electron microscopic data at 4.5 A resolution. The monomeric structure consists of two transmembrane triple helices arranged around a pseudo-2-fold axis connected by a long flexible extracellular loop. Each triplet contains between its second and third helix a functional loop containing the highly conserved fingerprint NPA motif. These functional loops are assumed to fold inwards between the two triplets, thereby forming the heart of the water channel. The helix topology was determined from the directionality pattern of each of the six transmembrane helices with respect to the membrane, together with constraints defined by the sequence and atomic force microscopy data. The directionality of the helices was determined by collecting the best-fitting orientations resulting from a search through the three-dimensional experimental map for a large number of alpha-helical fragments. Tests on cryo-electron crystallographic bacteriorhodopsin data suggest that our method is generally applicable to determine the topology of helical proteins for which only medium-resolution electron microscopy data are available.

Aquaporin 1↗

Temporal gap resolution in listeners with high-frequency sensorineural hearing loss.

Temporal gap resolution was measured in five normal-hearing listeners and five cochlear-impaired listeners, whose sensitivity losses were restricted to the frequency regions above 1000 Hz. The stimuli included a broadband noise and three octave band noises centered at 0.5, 1.0, and 4.0 kHz. Results for the normal-hearing subjects agree with previous findings and reveal that gap resolution improves progressively with an increase in signal frequency. Gap resolution in the impaired listeners was significantly poorer than normal for all signals including those that stimulated frequency regions with normal pure-tone sensitivity. Smallest gap thresholds for the impaired listeners were observed with the broadband signal at high levels. This result agrees with data from other experiments and confirms the importance of high-frequency signal audibility in gap detection. The octave band data reveal that resolution deficits can be quite large within restricted frequency regions, even those with minimal sensitivity loss.

Audiometry, Pure-Tone↗

Immunocytochemistry of the amphibian embryo--from overview to ultrastructure.

Amphibian embryos are standard research objects to study pattern formation and morphogenesis. Due to their external development and robust nature, experimental manipulations such as microinjections or transplantations can be easily performed. However, most immunocytochemical approaches addressing the specific localization of proteins are hampered by the fragility of the large and yolky embryonic cells which render high resolution staining difficult. Immunocytochemical data are therefore often restricted to either overall patterns in whole embryo preparations or to immunofluorescent localization with limited resolution on sections. High resolution or ultrastructural protein localization data are rare and can be achieved only with time consuming procedures. Here, a comparative study of immunocytochemical methods suitable for light and electron microscopy using different kinds of plastic resins is presented. Three main approaches are described: preembedding staining of whole embryos, postembedding staining of ultrathin sections and preembedding staining of vibratome sections. All the procedures are designed to study protein expression in early amphibian embryos en gros as well as en detail and the described techniques are suitable to combine two or three levels of resolution on the very same biological specimen. Examples are presented and advantages and disadvantages of the different protocols are discussed.

Animals↗

Crystal structure study of Opsanus tau parvalbumin by multiwavelength anomalous diffraction.

The crystal structure of a small calcium-binding protein, the parvalbumin IIIf from Opsanus tau in which Tb was substituted for Ca, has been analysed by multiwavelength anomalous diffraction. Data at a resolution of 2.3 A were collected at three wavelengths near the L3 absorption edge of Tb (1.645-1.650 A), using the synchrotron radiation emitted by a storage ring and a multiwire proportional counter. The phases of the reflections were determined from this single derivative, without native data. Prior to any refinement, the resulting electron density map shows a good agreement with the model of the homologous carp parvalbumin in regions of identical amino-acid sequence.

Animals↗