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Computerized 3-D reconstruction of small blood vessels from high voltage electron-micrographs of thick serial cross sections.

Morphological analysis of compartments in terms of volumes and the orientation of the cells and their nuclei within very small mesenteric arteries and arterioles is being carried out. Samples were from simultaneously sham operated and instrumented control and one-clip two kidney Goldblatt sustained (30 days) hypertensive dogs. Collection of specimens was standardized, i.e. sampling location, perfusion fixation, embedding, staining, and sectioning by methods proven to preserve in vivo dimensions. Serial thick cross sections (0.5 mu) were subjected to high voltage electromicrography (1.0 MeV) at 2500x and then linearly enlarged 3x photographically. The glossy micrographs (7500x) were made into montages and the contours of the smooth muscle layer (vascular smooth muscle), the endothelium (E), nuclei (N), and the internal lamina (IL) were digitized on a large GTCO digi-pad with input to a microcomputer (PC-Limited, Turbo) equipped with a 20 MB hard disk and coprocessor. Solid and transparent image reconstructions and computerized dissection of cell compartments were performed. Compartment volumes of vascular smooth muscle, IL, E and N, and individual cell and nuclear volumes, as well as nuclear/cytoplasmic ratios were calculated. Image reconstructions are presented. The emphasis is on methods and these are discussed in terms of useability for determining accurate microvessel morphology in health and disease, with particular emphasis on assessment of hypertrophy and hyperplasia in experimental hypertension, and architectural angiogenesis in general.

Animals↗

Correlation of laser-Doppler-velocity measurements and endothelial cell shape in a stenosed dog aorta.

Laser-Doppler-velocity measurements were carried out in an elastic 1:1 true-to-scale silicone rubber model of a dog aorta with stenosis. The model was constructed from a cast of a severely stenosed dog aorta (71% of its area). The stenosis in the dog aorta was prepared by wrapping a cotton band around the aorta. This band was tightened until the presence of a thrill or a bruit was felt distal to the band. Twelve weeks later the animal was sacrificed and a cast was prepared from the aorta. From this vascular cast, the cross-sectional area was calculated. Endothelial cell geometry and orientation was studied using computerized analysis to determine the cell area and shape index. An elastic silicone rubber model was prepared from the cast to measure the velocity profiles and to estimate the local wall shear stress. Velocity measurements were done at steady and pulsatile flow using a Newtonian aqueous-glycerol solution and a non-Newtonian blood-like fluid. From those velocity measurements the velocity gradients near the wall were determined and the shear stress calculated. The flow distal to the stenosis separates from the wall at physiological conditions. The endothelial cells are smaller and more elongated in the throat; distal to the stenosis they are larger and rounder. The shape index distribution along the stenosed aorta is correlated with the level of wall shear stress. It is shown that even low changes in the wall shear stress have an influence on the orientation of the endothelial cells.

Animals↗

Contact angles of substances used for internal tamponade in retinal detachment surgery.

In order to ascertain the tamponade effect of air and silicone oil we examined the contact angles subtended by ex vivo human retina, Teflon and Perspex to find a suitable experimental material which would mimic the surface properties of the retina at a three-phase interface. Using the captive bubble technique to measure the contact angle, it was found that air subtended a larger contact angle (38.8 degrees) with the retina than did silicone oil (18.2 degrees). On coating the Perspex surface with protein (PCP), it was observed that the surface properties were modified such that PCP subtended contact angles with air (43.0 degrees) and silicone oil (16.4 degrees) similar to those subtended by ex vivo human retina. Using PCP as an experimental material that mimics ex vivo human retina, spherical chambers were employed in order to examine qualitatively and to quantify the arc of contact obtained with air and silicone oil. It was found that air gave a greater arc of contact for the same percentage fill than silicone oil.

Air↗

Radial secretory glia conserved in the postnatal vertebrate brain: a study in the rat.

Secretory glial cells in the roof of the last diencephalic prosomer, ependymocytes and hypendymocytes, form the subcommissural organ. The cells of this complex were labelled immunocytochemically, using an antiserum against their specific secretory products. The study aims at the characterization of this cell type in the rat as an anatomical model situation. Radially oriented secretory glial cells remain after birth behind the posterior commissure in the mesencephalic aqueduct. At about postnatal day 10, the cell bodies descend into the conventional ependyma and at postnatal day 25 they assume a compact, rounded appearance. The secretory product they release is involved in the formation of Reissner's fiber. This differentiation in phenotype is not accompanied by a change of the intermediate filament expression. In the adult rat these cells had been labelled immunopositive for cytokeratins 8 and 18 as well as vimentin but not for glial fibrillary acidic protein. DiI-marking from the third ventricle and from the dorsal surface of the brain shows that the basal processes of ependymocytes and hypendymocytes project to the external and internal glial limiting membrane, respectively, through the commissural fiber bundles. Also the subependymal located hypendymocytes have apical processes with contacts to the cerebrospinal fluid. When this secretory cell population is studied with respect to cyto-architectonical changes during ontogeny the results lead to a new understanding of the subcommissural cells. They are not specialized ependymal cells in a regionally restricted and secondary differentiated ependymal area, but rather descendants of an ontogenetically ancient, specific type of radial glia. Characteristic features for all subcommissural cells are that they: (1) appear very early during ontogeny, (2) are derived from a radial oriented glial cell type, (3) carry at least one kinocilium, (4) possess an original intermediate filament pattern, (5) release a secretory product.

Affinity Labels↗

Formation of field discontinuities and islands in visual cortical maps.

The representations of visual hemifields in the extrastriate areas of various species exhibit field discontinuities and islands. We propose that these violations of retinotopy are a developmental consequence of the elongated shape of the respective cortical areas. To substantiate this claim, we investigated a model of activity-driven map formation. In agreement with observations, this model yields maps with field discontinuities if the cortical areas exceed a threshold elongation. Moreover, within the same model island representations in the periphery and the area centralis can also be understood. A multistability of the solutions in the model gives a very simple explanation for the observed interindividual variability of maps in cats. The model leads to a prediction of the radial dependence of the areal magnification factor near field discontinuities, which could be accessible for a high precision mapping experiment.

Animals↗

Kinematic networks. A distributed model for representing and regularizing motor redundancy.

Motor control in primates relates to a system which is highly redundant from the mechanical point of view--redundancy coming from an imbalance between the set of independently controllable variables and the set of system variables. The consequence is the manifestation of a broad class of ill-posed problems, problems for which it is difficult to identify unique solutions. For example (i) the problem of determining the coordinated patterns of rotation of the arm joints for a planned trajectory of the hand; (ii) the problem of determining the distribution of muscle forces for a desired set of joint torques. Ill-posed problems, in general, require regularization methods which allow to spell acceptable, if not unique, solutions. In the case of the motor system, we propose that the basic regularization mechanism is provided by the potential fields generated by the elastic properties of muscles, according to an organizational principle that we call "Passive Motion Paradigm". The physiological basis of this hypothesis is reviewed and a "Kinematic Network" (K-net) model is proposed that expresses the kinematic transformations and the causal relations implied by elasticity. Moreover, it is shown how K-nets can be obtained from a kinematic "Body Model", in the context of a specific task. Two particularly significant results are: (i) the uniform treatment of closed as well as open kinematic chains, and (ii) the development of a new method for the automatic generation of kinematic equations with arbitrary topology. Moreover, the model is akin to the concept of "motor equivalence" in the sense that it provides families of motor equivalent trajectories parametrized by tunable motor impedances.

Biomechanical Phenomena↗

Automated calculation of gallbladder ejection fraction.

We have developed a computer method which by automatically locating the border of the gallbladder in each image and subtracting a varying location-dependent background corresponding to the gallbladder border overcomes the difficulties associated with quantitating gallbladder contraction. These difficulties are attributable to significant and changing background activity, imprecise manual delineation of the gallbladder, and the changing position and shape of the gallbladder. Validation studies using a gallbladder phantom showed the method to be very accurate. No significant difference (P greater than 0.05) was observed between the expected and calculated ejection fractions. Ten patient studies were analyzed, with a resulting range of gallbladder ejection fractions of 22-79%. Excellent reproducibility was obtained with an average intraobserver coefficient of variation of 3.5%, and no statistically significant difference in interobserver measurements (P greater than 0.05). The regression line for interobserver measurements had a slope of 0.96 +/- 0.08, an intercept of 1.6 +/- 4.0%, and a correlation coefficient of 0.99. From these preliminary results we conclude that this method offers a reliable means of quantitating gallbladder contraction.

Cholecystokinin↗

A geometric model of the rat kidney.

Firstly details are collected concerning the required parameters of a simple linear regression in order to evaluate statistically results of measurements, which can also be present in the form (chii, yi +/- SDi). In this way lines of regression are determined for connections between the kidney weight and the body weight, between the lengths of the proximal tubules (and the proximal convolution) of the three various types of nephron and the kidney weight, between the length of the distal convoluted tubule, likewise the number of glomerula, and the kidney weight and finally between the single nephron filtration rate and the length of the proximal tubule. Starting from a model body weight for the rat of 200 g and considering the percentage of thin segments in the tissue of the renal pyramid, a loop of Henle with a length of 8.1 mm for the thin part and a length of 2.4 mm for the ascending thick limb was calculated for the model nephron from the lengths of the loops of the three types of nephrons. In contrast to former model formulations concerning the collecting duct system, the tree-like branched structure was considered for the first time and a linear approximation to the relation between both the circumference line and the cross section area and the lenggh of the collecting ducts was determined. The geometric model relates only to the tubular system and takes no notice of the blood vessels.

Animals↗

A volume conductor study of compound action potentials of nerves in situ: the forward problem.

A computer model for the stimulation of compound nerve action potentials, based on superposition of volume conducted single nerve fibre potentials, is presented. The model assumes that the intracellular fibre potential, the fibre diameter distribution and the electrical conductivities of different tissues are known. Volume conductor fields are calculated in the spatial frequency domain. The influence of important parameters in the model is evaluated numerically. It is shown that it is necessary to give up the usual assumption of homogeneity and isotropy in the extracellular medium. In the present model parameters are introduced which allow an overall description of the complex morphological and physiological structure of the nerve trunk. Simulation results indicate that the model is a rather promising tool in studying the main properties of compound action potentials which up till now have not been sufficiently well understood.

Action Potentials↗

Effects of certain training parameters on detection of simulated breast cancer.

Forty-two female subjects, constituting 14 groups, palpated a silicone breast model in a lump detection task. Three variables, number of trials (amount of practice), search pattern requirement (restricted vs. unrestricted), and the presence or absence of lumps, were manipulated according to 3 x 2 x 2 design. The effect of all modes of practice on detection performance in posttest 1 was evidenced by an increase in percent of lumps detected and a decrease in detection threshold, but no form of practice was superior to the others. Following posttest 1 a brief search-training procedure was administered to 39 subjects which further lowered detection threshold, increased percent of lumps detected, and increased trial duration in posttest 2. The results indicate the effectiveness of the training procedure for improving detection ability and suggest the need for a more complex model to determine more specifically the effects of the practice models employed.

Adolescent↗

Beyond topographic mapping: towards functional-anatomical imaging with 124-channel EEGs and 3-D MRIs.

A functional-anatomical brain scanner that has a temporal resolution of less than a hundred milliseconds is needed to measure the neural substrate of higher cognitive functions in healthy people and neurological and psychiatric patients. Electrophysiological techniques have the requisite temporal resolution but their potential spatial resolution has been not realized. Here we briefly review progress in increasing the spatial detail of scalp-recorded EEGs and in registering this functional information with anatomical models of a person's brain. We describe methods and systems for 124-channel EEGs and magnetic resonance image (MRI) modeling, and present first results of the integration of equivalent-dipole EEG models of somatosensory stimulation with 3-D MRI brain models.

Brain↗

In vitro studies of intragastric digestion.

Following consumption of a meal, 99% of the large food particles are emptied only after intragastric fragmentation has reduced their diameter to less than 2 mm. An in vitro model was constructed to evaluate some of the factors which may play a role in the process of intragastric digestion. Gastric mixing of food was simulated in a silicone rubber tube (ID 19 mm) placed in a peristaltic pump. Peristaltic waves progressed upwards along the tube at a frequency of 0, 1, or 3/min, reducing the internal diameter of the tube to 5 mm. Cooked chicken liver particles (2-2.8 mm in diameter) were placed in the tube with one of the following: (1) 150 mM NaCl, (2) 150 mM HCl with or without pepsin, or (3) phosphate buffer at pH 7, 5.4, or 2.6 + pepsin. After 30 min, the extent of particle reduction and of solubilization of proteins were determined and expressed as percent of the initial liver weight. The diameter of liver particles was reduced to a greater extent in NaCl than in pH 7 buffer or acid solutions with or without pepsin. In contrast, the amount of proteins solubilized was enhanced two- to threefold by acid pepsin solutions compared to NaCl or pH 7 phosphate. The present in vitro studies suggest that changes in motor and/or secretory activity of the stomach significantly modify intragastric digestion.

Dietary Proteins↗

Topometry of normal intrahepatic bile ducts.

This paper focuses on practical problems which may also prove to be of theoretical importance, by presenting a method of establishing an exact topometry of the intrahepatic bile ducts. Measurements were made on corrosive casts of the intrahepatic bile ducts from 13 human livers. On the basis of the topometric results presented it is possible to construct an adequate model by means of plastic tubes and to use it for the study of bile flow.

Bile Ducts, Intrahepatic↗

Stability of the human spine in neutral postures.

The present study aimed to identify some of the mechanisms affecting spinal compressive load-bearing capacity in neutral postures. Two spinal geometries were employed in the evaluation of the stabilizing mechanisms of the spine in standing neutral postures. Large-displacement finite-element models were used for parametric studies of the effect of load distribution, initial geometry, and pelvic rotation on the compression stability of the spine. The role of muscles in stabilization of the spine was also investigated using a unique muscle model based on kinematic conditions. The model with a realistic load configuration supported the largest compression load. The compressive load-bearing capacity of the passive thoracolumbar spine was found to be significantly enhanced by pelvic rotation and minimal muscular forces. Pelvic rotation and muscle forces were sensitive to the initial positioning of T1 and the spinal curvatures. To sustain the physiological gravity load, the lordotic angle increased as observed in standing postures. These predictions are in good agreement with in vitro and in vivo observations. The load-bearing potential of the ligamentous spine in compression is substantially increased by controlling its deformation modes through minimal exertion of selected muscles and rotation of the pelvis.

Humans↗

Advantages and limitations of two software calipers in quantitative coronary arteriography.

Software calipers allowing the measurement of the distances between pairs of manually defined picture elements in digitized images may be useful tools for a rapid assessment of the morphology of coronary vessels, e.g. for choosing the appropriate balloon or stent sizes before or during cardiac intervention procedures. In this paper we have studied extensively the advantages and limitations of two manual software calipers--one developed for a PC-based cinefilm analysis workstation, the other for the Philips DCI system. Based on analyses of a perspex vessel phantom with 17 sectors of known size filled with different concentrations (50 and 100%) of the contrast agent and acquired at two kV-levels (68 and 92 kV), it was found that the cinefilm approach is characterized by a very small overall (averaged over te data from three observers) systematic overestimation of 0.03 mm, and the DCI system by a systematic underestimation of 0.07 mm; the worst case accuracy value for an individual observer on frames with 100% contrast dye concentration was 0.20 mm for cinefilm, and -0.34 mm for the DCI, respectively. The overall variabilities in the measurements (precision) were almost identical for the two approaches (overall 0.07 and 0.08 mm for the cinefilm and digital approaches, respectively, and worst case for individual observers on the 100% contrast frames, 0.16 and 0.13 mm, respectively. Inverting the images (bright or dark contrast containing structures) of the phantom at 100% contrast concentration and acquired at 62 kV had no significant effect on the results obtained with the cinefilm analysis system (overall accurary -0.12 mm for both situations), whereas it had on the results from the DCI system (overall accuracies -0.29 (dark vessels on bright background) and -0.08 mm (bright vessels on dark background), respectively). Enhancing the digital images on the DCI with unsharp masking techniques did not significantly influence the measurement accuracy and precision. Finally, it was found that woven dacron, polyurethane and polyvinylchloride catheters filled with 100% contrast dye can be measured with an overall accuracy of better than 0.13 mm on the DCI system. On the PC-based system the woven dacron and polyvinylchloride catheters would result in an overall accuracy better than 0.17 mm, and the polyurethane catheter better than 0.30 mm. The evaluation study has made clear that the nylon catheter should not be applied in QCA-studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiac Catheterization↗