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Biomedical subjects

M J Potel

Publications and source records attributed to M J Potel.

17 recordsLinked to original sources

Relative magnitude and asynchrony of directional components of contraction in sedated dog left ventricle.

The purpose of this study was to quantitate the temporal relationships and the extent and speed of shortening in segments of myocardium responsive to contraction in circumferential, longitudinal, and oblique fiber groups. Measurements were made in five sedated dogs (morphine, diazepam) with and without alterations in preload and afterload (nitroprusside, phenylephrine). The measurement interval was the phase of rapid contraction, determined by differentiation of the segment length vs. time. In the control state, percentage segment shortening was greater in circumferential than in longitudinal [15.2 +/- 0.24 (SE) vs. 10.5 +/- 0.80%; P = 0.0020] and in the subepicardial oblique than in the subendocardial oblique fiber directions (16.6 +/- 0.65 vs. 9.7 +/- 0.36%; P = 0.0010). Shortening was proportional to both maximum speed and duration of shortening (r = 0.735 +/- 0.015 and 0.757 +/- 0.017, respectively). Duration of shortening was significantly longer in circumferential than in longitudinal (mean difference 39.3 +/- 6.6 ms; P = 0.0039) and in subepicardial oblique than in subendocardial oblique directions (mean difference 27.7 +/- 5.5 ms; P = 0.0072). Velocities of up to 3.0 segment lengths/s were attained in response to nitroprusside. These data reveal the local anisotropy and asynchrony of contraction in the myocardium; however, they also support the concept of the myocardium as a functional continuum. The dominance of circumferential over longitudinal and subepicardial over subendocardial oblique contractile components indicates their relative contributions to the constriction of the midmyocardial shell.

Animals

Structural analysis of polymers of sickle cell hemoglobin. I. Sickle hemoglobin fibers.

The structure of fibers of deoxyhemoglobin S has been under investigation for several years and a number of different models have been proposed for the arrangement of molecules within the particles. We have used reconstruction and modeling techniques in our analysis of these structures. Several new approaches have been employed in this analysis in order to provide improved estimates of the co-ordinates, pairing, and polarity of the hemoglobin S molecules. Fibers have a variable pitch and, in order to minimize distortions in the reconstructed density maps associated with these variations in pitch, we have developed an iterative procedure to measure the instantaneous pitch and have modified the reconstruction algorithm to incorporate the measured values. This procedure improves the accuracy with which the hemoglobin S molecules can be located in the density maps. Furthermore, the determination of the instantaneous pitch allows us to measure directly the rotation of the individual hemoglobin molecules. These measurements are in excellent agreement with the values predicted using a random angular walk model (as originally proposed for F-actin) to describe the variable pitch. The reconstructions confirm that the fiber consists of 14 strands of hemoglobin S arranged in a hexagonally shaped cross-section. We have determined the pairing of the molecules to form double strands directly from the density maps by identifying the molecules that have intermolecular distances that conform to those of double strands in the Wishner-Love crystal. The seven double strands identified in this manner are consistent with the strand pairings proposed by Dykes et al. (1979) rather than the alternate pairings proposed by Rosen & Magdoff-Fairchild (1985). In addition, we have for the first time determined the polarity of the double strands directly from the reconstruction data. This was achieved using a procedure that amounts to essentially "dissecting" individual double strands from the reconstructed density maps so that their density distribution could be examined independently of the neighboring double strands. Knowledge of the relative polarities of the double strands is essential for determining the intermolecular interactions that stabilize the fiber.

Crystallography

Structural analysis of polymers of sickle cell hemoglobin. II. Sickle hemoglobin macrofibers.

Sickle cell hemoglobin macrofibers are an important intermediate in the low pH crystallization pathway of deoxygenated hemoglobin S that link the fiber to the crystal. Macrofibers are a class of helical particles differing primarily in their diameters but are related by a common packing of their constituent subunits. We have performed three-dimensional reconstructions of three types of macrofibers. These reconstructions show that macrofibers are composed of rows of Wishner-Love double strands in an arrangement similar to that in the crystal. We have measured the orientation and co-ordinates of double strands in macrofibers using cross-correlation techniques. In this approach, the electron density projections of double strands calculated from the known high-resolution crystal structure are compared with regions along the length of the particles in which the distinct pattern of double strands in c-axis projection may be observed. Contrary to assertions by Makinen & Sigountos (1984), our results unambigously demonstrate that adjacent rows of double strands in macrofibers are oriented in an antiparallel manner, as in the Wishner-Love crystal. Adjacent rows of antiparallel double strands are displaced along the helical axis relative to their co-ordinates in the crystal. Electron density models of macrofibers based on the crystallographic structure of the sickle hemoglobin double strand are in good agreement with the projections of macrofibers observed in electron micrographs. We have studied the structure of a closely related crystallization intermediate, the sickle hemoglobin paracrystal. The arrangement of double strands in paracrystals is similar to that in Wishner-Love crystals, except that they are displaced along the a-axis of the crystal. Measurements of the double strand co-ordinates reveal that the distribution of strand positions is bimodal. These results further establish the close structural relationship between macrofibers and paracrystals as intermediates in the crystallization of deoxygenated sickle hemoglobin.

Computer Simulation

Structural analysis of polymers of sickle cell hemoglobin. III. Fibers within fascicles.

We have examined the structure of hemoglobin S fibers, which are associated into large bundles, or fascicles. Electron micrographs of embedded and cross-sectioned fascicles provide an end-on view of the component fibers. The cross-sectional images are rotationally blurred as a result of the twist of the fiber within the finite thickness of the section. We have applied restoration techniques to recover a deblurred image of the fiber. The first step in this procedure involved correlation averaging images of cross-sections of individual fibers in order to improve the signal-to-noise ratio. The rotationally blurred image was then geometrically transformed to polar co-ordinates. In this space, the rotational blur is transformed into a linear blur. The linearly blurred image is the convolution of the unblurred image and a point spread function that can be closely approximated by a square pulse. Deconvolution in Fourier space, followed by remapping to Cartesian co-ordinates, produced a deblurred image of the original micrograph. The deblurred images indicate that the fiber is comprised of 14 strands of hemoglobin S. This result provides confirmation of the fiber structure determined using helical reconstruction techniques and indicates that the association of fibers into ordered arrays does not alter their molecular structure.

Hemoglobin, Sickle

The three-dimensional structure of sickle hemoglobin macrofibers.

The crystallization of deoxygenated sickle cell hemoglobin (HbS) at pH less than 6.5 is preceded by the formation of HbS macrofibers. In stirred solutions, macrofibers appear several hours after the formation of fibers and subsequently aggregate and crystallize. Like fibers, macrofibers are long helical structures consisting of alternating narrow and wide regions. Macrofibers have a pitch of 10,000 A and a diameter averaging 500 A. Here we present a three-dimensional structural analysis of HbS macrofibers. Image reconstructions of these particles show that they consist of 5 antiparallel rows with 8, 11, 12, 11, and 8 double strands per row. Double strands in the outer two rows are asymmetrically disposed resulting in "notches" in opposite corners of the particle cross section. The notches produce characteristic features in electron micrographs of macrofibers. Fourier-filtered images of macrofibers have been produced which reveal their structure in considerable detail. Computed Fourier transforms of macrofibers exhibit spacings of 64 A, 185 A, and 53 A similar to those in the Wishner-Love crystal. A cross-correlation analysis between filtered macrofiber images and electron density projections of Wishner-Love double strands establishes that double strands in adjacent rows are antiparallel. This analysis also shows that the macrofiber rows are displaced axially by differing amounts depending on their distance from the helical axis. Using this data, we have computed a three-dimensional electron density model of the macrofiber based on atomic hemoglobin coordinates. This model reproduces in detail the many features revealed in Fourier-filtered macrofiber images.

Computer Simulation

The restoration of electron micrographs blurred by drift and rotation.

We have investigated the restoration of electron micrographs exhibiting blurring due to drift and rotation. Blurring due to drift arises in micrographs taken of a specimen which is moving relative to the image plane. A related problem is that of rotational blurring which arises in micrographs of thin sections of helical particles viewed in cross section. The twist of the particle within the finite thickness of the section causes the image to appear rotationally blurred about the helical axis. Restoration algorithms were evaluated by applying them to the restoration of blurred model images degraded by additive Gaussian noise. Model images were also used to investigate how an incorrect estimate of the point spread function describing the blur would effect the restoration. Images were, if necessary, geometrically transformed to a space in which the point spread function of the blur can be considered as linear and space invariant as, under these conditions, the restoration algorithms are greatly simplified. In the case of the rotationally blurred images this procedure was accomplished by transforming the image to polar coordinates. The restoration techniques were successfully applied to blurred micrographs of bacteriophage T4 and crystals of catalase. The quality of the restoration was judged by comparisons of the restored images to undegraded images. Application to micrographs of rotationally blurred cross sections of helical macrofibers of sickle hemoglobin resulted in a reduction in the amount of rotational blurring.

Algorithms

Macrofiber structure and the dynamics of sickle cell hemoglobin crystallization.

Fibers of deoxyhemoglobin S undergo spontaneous crystallization by a mechanism involving a variety of intermediate structures. These intermediate structures, in common with the fiber and crystal, consist of Wishner-Love double strands of hemoglobin S molecules arranged in different configurations. The structure of one of the key intermediates linking the fiber and crystal, called a macrofiber, has been studied by a variety of analytical procedures. The results of the analysis indicate that the intermediates involved in the fiber to crystal transition have many common structural features. Fourier analysis of electron micrographs of macrofibers confirms that they are composed of Wishner-Love double strands of hemoglobin molecules. Electron micrographs of macrofiber cross-sections reveal that the arrangement of the double strands in macrofibers resembles that seen in micrographs of the a axis projection of the crystal. This orientation provides an end-on view of the double strands which appear as paired dumb-bell-like masses. The structural detail becomes progressively less distinct towards the edge of the particle due to twisting of the double strands about the particle axis. Serial sections of macrofibers confirm that these particles do indeed rotate about their axes. The twist of the particle is right handed and its average pitch is 10,000 A. The effect of rotation on the appearance of macrofiber cross-sections 300 to 400 A thick can be simulated by a 15 degrees rotation of an a axis crystal projection. The relative polarity of the double strands in macrofibers and crystals can be determined easily by direct inspection of the micrographs. In both macrofibers and crystals they are in an anti-parallel array. On the basis of these observations we conclude that crystallization of macrofibers involves untwisting and alignment of the double strands.

Crystallization

Left ventricular wall motion: its dynamic transmural characteristics.

Cardiac wall motion has been studied extensively. It is usually determined by indirect two-dimensional measurements for the true three-dimensional (3D) motion with its specific speed and direction. Errors are also introduced by using internally fixed reference systems and by the inability to identify precise points on the heart wall during the cardiac cycle. Because of these limitations, the endocardial and epicardial wall motion and their relationship are still unclear. This study was designed to assess endocardial and epicardial wall motion by measuring the direction and speed of implanted markers in an externally fixed 3D coordinate system. Fifty-seven pairs of endocardial and epicardial metallic markers were placed at anterior, lateral, posterior, basal, and apical regions of the left ventricles of 14 normal mongrel dogs. Biplane cineradiographs were performed at 50 frames/sec, and the 3D motions of the markers were analyzed using a specially designed computer system. It was found that the speeds, directions, displacements, and phases of the movements of corresponding endocardial and epicardial points were highly correlated. The correlation coefficients were 0.77 to 0.95 for the mean directions, 0.61 to 0.96 for the mean speeds, and 0.59 to 0.96 for the mean displacements at various regions of the heart, and the periodic movements of the endocardium and epicardium were always in phase. The mean epicardial speeds and displacements are fixed proportions (approximately 70%) of the mean endocardial speeds and displacements despite the differences in absolute values between regions in the same dog and the same regions in different dogs. The correlation coefficients for endocardial and epicardial instantaneous speeds, directions, and velocities ranged from 0.68 to 0.83, 0.81 to 0.88, and 0.77 to 0.86, respectively, for different regions of the heart. The correlation coefficients were significant for both the mean values and the instantaneous values. Thus, when only fixed epicardial points are accessible for wall motion measurements in clinical situations, it is possible to infer the endocardial motion from the epicardial motion.

Animals

Quantitative analysis of cyclic AMP waves mediating aggregation in Dictyostelium discoideum.

We have previously reported the detection of cAMP waves within monolayers of aggregating Dictyostelium discoideum cells (K. J. Tomchik and P.N. Devreotes, 1981, Science 212, 443-446). The computer-assisted analysis presented here of the fluorographic images of the cAMP waves reveals (1) all the waves have a consistent width and height; (2) cAMP concentrations within centers of concentric aggregation territories oscillate periodically while at spiral centers the concentration builds up to a plateau value within 2 mm; (3) cells within the region of intersection of two oppositely directed cAMP waves are stimulated to produce more cAMP than those responding to a single wave; (4) cells start to move when the cAMP level begins to increase and cease movement when the peak cAMP concentration reaches the cell.

Computers

Schooling behavior of tadpoles: a potential indicator of ototoxicity.

Fish and tadpoles in schools use hair cells of their lateral line system to assess their position in relation to neighbors. This suggests that pharmaceutical agents that damage hair cells in the mammalian inner ear may also alter geometry in fish and tadpole schools. We used a computer-based image analysis system to examine the effect of the ototoxic aminoglycoside antibiotic, streptomycin, on school geometry for tadpoles of the African clawed frog Xenopus laevis. Tadpoles exposed to streptomycin in the surrounding water show a general tendency toward clumping, and an increase in the distance over which they orient parallel to neighbors, compared to controls. These behavioral responses appear in 18 min or less, and are evident in some tadpoles exposed to concentrations as low as 5 micrograms/ml. Results suggest that analysis of spatial relations in tadpole schools could serve as a method for rapidly detecting ototoxic potential of agents suspected of damaging hair cells.

Animals

Preaggregative cell motion in Dictyostelium.

The motions of a large number (495) of preaggregative D. discoideum NC-4 cells in sparse fields are recorded on time-lapse film and analysed using a specially constructed computer graphics system. All films are produced under a standard set of conditions, so that the range of cell behaviours under given conditions can be characterized. The mean velocity of pre-aggregative D. discoideum NC-4 is 7.19 micrometers/min. The mean velocity time course has a significant early peak at about 3 h. The distribution of mean velocities is fairly broad with a long high velocity tail. A modified random walk model using the parameters diffusion constant and persistence time describes well the changes in cell direction with time. Persistence can be described as an exponentially distributed 'memory' of movement direction, with a mean of 4.89 min. High velocity cells never have long persistence times, and persistence time shows no relationship with age. A nearest neighbour model of cell spacing shows that cells are randomly (Poisson) distributed at low densities. Measurements of cell contacts are compared to a simple model of contact frequency based on the kinetic theory of gases to show that cells at low densities have an affinity for making collisions. The length of contact durations is indicative of some mechanical adhesion between cells, and cells in contact move significantly though not dramatically slower. A cross-correlation analysis shows that the various parameters of motion are significantly interrelated in numerous ways. Finally mutants and strains related to D. discoideum NC-4 exhibit a number of new behaviours, suggesting that motion is a distinctive characteristic of cell type.

Agglutination

Methods for evaluating cardiac wall motion in three dimensions using bifurcation points of the coronary arterial tree.

An accurate three-dimensional (3D) representation of heart wall motion would be an important means of evaluating cardiac function. To accomplish this, we have developed an interactive computer graphics system designed to enter the time-dependent 3D positions of bifurcations of the coronary arterial tree. These bifurcations are precise markers of the epicardial surface, and their motions accurately represent the motion of the underlying heart wall. We demonstrate techniques for calculating local wall motion, including displacement and velocity, for determining a time-dependent center-of-contraction point towards which the epicardium tends to move and for tracking the mechanical contraction wave using cross-correlation methods. We have applied these techniques to study seven patients with normal left ventriculograms and coronary arteriograms. We have found these methods to be generally applicable and to provide information not obtainable without 3D analysis.

Cineangiography

Three-dimensional left ventricular wall motion in man. Coordinate systems for representing wall movement direction.

We have studied the three-dimensional (3D) motion of left ventricular (LV) epicardial points by tracking one to three dozen coronary artery bifurcations in eleven human subjects. Wall motion was analyzed using several different coordinate systems: (1) cylindrical centered about the LV long axis, (2) spherical with origin at the LV center-of-gravity (COG), and (3) spherical with origin at the LV center-of-contraction (COC), the best-fit 3D point toward which the wall moves. The coordinate systems were studied both fixed and moving with time. Three-dimensional motions were decomposed into three directional components, with high radial (in and out) percentages being regarded as the figure-of-merit of a given coordinate system. Average percentage radial motions were fixed cylindrical 16%, fixed spherical COG 35%, fixed spherical COC 47%, moving cylindrical 17%, moving spherical COG 30%, moving spherical COC 91%. Spherical systems were generally better than cylindrical systems, with the COC representing a better origin than the COG. Moving systems were appreciably better than fixed only for the COC model, indicating that the COC, which traverses up and down the LV midline, moves significantly while the other systems are more stationary. At each instant in time, almost all (91%) of the 3D motion of the entire heart wall is directed toward a single moving 3D point, the COC. Thus, there exists in principle a near-perfect 3D heart wall motion model. Approximately 25% of 3D wall motion is unseen in conventional monoplane views. Also, any model that represents 3D wall motion only along fixed straight 3D lines (eg, end-diastole to end-systole) necessarily ignores 27% of the true 3D heart wall motion.

Adult

Clinical applications of combined cerebral angiograms and brain CT scans.

This paper illustrates the clinical value of a computer technique for superimposition of two different kinds of imaging procedures. The initial application was to overlay cerebral angiograms onto computed tomograms of the brain. Case material from three patients with intracerebral abnormalities is presented. The sum of information from the combined studies exceeds that from either study alone and has value for both diagnosis and treatment of head lesions.

Brain