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X-ray solution scattering studies on vinblastine-induced polymers of microtubule protein: structural characterisation and effects of temperature.

We report here on X-ray solution scattering and electron microscopy studies of microtubule protein in the presence of the antimitotic drug, vinblastine. In buffer conditions used for microtubule assembly, vinblastine caused the formation of coil-like structures. The coils appeared to be made up of two protofilaments. Details of the structure and behaviour of coils in solution were obtained from interpretation of their solution scattering patterns. Upon increasing temperature from 4 to 37 degrees C the pitch of the coils increased from 25.92 to 26.96 nm. However, little change was observed in their mean diameters (38.46 and 38.45 nm, respectively). Increasing the temperature also favoured increased formation and/or elongation of the coils. The effect of temperature on the pitch was fully reversible. Vinblastine-induced assembly of pure tubulin also showed the formation of coils. However, these coils appeared to consist of only one protofilament. Their mean diameters (38.35 nm) were similar to those of the coils formed from microtubule protein.

Animals

[Three-dimensional display of 99mTc-MIBI myocardial scintigraphy].

One of 99mTc-hexakis, 99mTc-methoxyisobutyl isonitrile (MIBI), has been demonstrated to have a myocardial uptake proportional to regional coronary blood flow. In this study, 99mTc-MIBI myocardial scintigraphy were performed for 16 patients with ischemic heart disease. After injection of 740 MBq of 99mTc-MIBI, 64 projection images were collected during a 360-degree rotation. Three-dimensional (3D) display of the left ventricle was reconstructed with depth-shading method from 99mTc-MIBI SPECT images, which were reconstructed by filtered back projection method. In 9 of the patients, left ventricular cineangiography were performed as diagnostic gold standard. Four physicians blinded to patients' clinical informations interpreted 3D images and SPECT images on separate occasions. Diagnosis of hypoperfusion by 3D displays agreed with those of SPECT in 92.9% (104/112 segments), and disagreed in 7.1% (8 segments). Sensitivity and specificity of 3D images were 87.0 and 93.9%, which were not statistically different (p less than 0.05) from that of SPECT images (91.3, 97.0%). Receiver operating characteristic (ROC) analysis revealed nearly identical curves for the two. Although 3D display had nearly identical diagnostic ability with SPECT, observers reported that 3D images were easier to diagnose than SPECT images. An advantage of the 3D display is that the display gives a more realistic impression of the left ventricle to an observer than tomography or planar imaging. Another advantage is that 3D display can reduce the amount of data storage compared with that of SPECT. In conclusion, 3D images may be useful for diagnosis of hypoperfusion of left ventricle.

Adult

The Staphylococcus aureus alpha-toxin channel complex and the effect of Ca2+ ions on its interaction with lipid layers.

Using the techniques of two-dimensional crystallization on supported lipid bilayers together with computer image processing, two distinct two-dimensional crystal types of staphylococcal alpha-toxin complex are formed depending on the presence or absence of Ca2+ ions. Without Ca2+, these are hexagonally packed (in A, a = b = 89.5 +/- 2.5 A; theta = 119.7 degrees) With Ca2+ present, rectangular crystal packing is seen (in A, a = 114.8 +/- 1.6 A, b = 140.2 +/- 0.7 A; theta = 89.1 degrees). A third, banded crystal type is also seen which is interpreted as a side-to-side packing of regular tubules. We use these tubular crystals for cross-correlation searches with top and side-on views of the complex from single particle reconstructions, and with the repeating units from the two-dimensional crystal types. The results lead us to propose a model in which the different two-dimensional crystal types are formed as a result of alpha-toxin hexamers packing in different orientations. In the hexagonal crystals the hexamers lie end-on with a 6-fold axis in projection. On the addition of Ca2+, the hexamers reorient to lie tilted with respect to the support, thus giving rise to a rectangular projection.

Bacterial Toxins

Image processing in digital chest radiography: effect on diagnostic efficacy.

The usefulness of digital image processing of chest radiographs was evaluated in a clinical study. In 54 patients, chest radiographs in the posteroanterior projection were obtained by both 14 inch digital image intensifier equipment and the conventional screen-film technique. The digital radiographs (512 x 512 image format) viewed on a 625 line monitor were processed in three different ways: (1) standard display; (2) digital edge enhancement for the standard display; and (3) inverse intensity display. The radiographs were interpreted independently by three radiologists. The diagnoses were confirmed by CT, follow-up radiographs and clinical records. Chest abnormalities of the films analyzed included 21 primary lung tumors, 44 pulmonary nodules, 16 cases with mediastinal disease and 17 cases with pneumonia/atelectasis. Interstitial lung disease, pleural plaques, and pulmonary emphysema were found in 30, 18 and 19 cases, respectively. The sensitivity of conventional radiography when averaged overall findings was better than that of the digital techniques (P less than 0.001). The differences in diagnostic accuracy measured by sensitivity and specificity between the three digital display modes were small. Standard image display showed better sensitivity for pulmonary nodules (0.74 vs 0.66; P less than 0.05) but poorer specificity for pulmonary emphysema (0.85 vs. 0.93; P less than 0.05) compared with inverse intensity display. We conclude that when using 512 x 512 image format, the routine use of digital edge enhancement and tone reversal at digital chest radiographs is not warranted.

Adolescent

Gd-DTPA-enhanced 3D MR imaging of cervical degenerative disk disease: initial experience.

PURPOSE: To assess whether a single enhanced T1-weighted gradient echo volume sequence, with the appropriate reformatted images, could be equivalent to a more conventional 2D set of MR sequences for the evaluation of cervical extradural degenerative disk disease (bony canal and foraminal stenosis; disk herniation). MATERIALS AND METHODS: Sixty-one patients evaluated for extradural degenerative disease by MR were imaged with a "standard" MR examination (Sagittal T1-weighted spin echo, axial low flip angle gradient echo), were then given 0.1 mmol/kg Gd-DTPA intravenously, and reimaged with either a 3D FLASH (fast low angle shot), TR 40/TE 7/1 excitation), 40 degree flip angle, acquired as 64, 2-mm sagittal partitions, or a 3D turbo FLASH (MP RAGE-magnetization prepared rapid acquisition gradient echo) (10/4/1), 10 degree flip angle acquired as 128, 2-mm coronal partitions. The volume sequences were reconstructed in the axial plane, and right and left 45 degree oblique coronal planes. The two sets of examinations (standard vs volume) were prospectively interpreted by two neuroradiologists for quality of examination, and location, type, and severity of extradural degenerative disease in a random, blinded, independent fashion. RESULTS: There was no significant difference between the standard examination and the 3D MP RAGE for central extradural disease. The 3D FLASH examination was significantly worse than the standard examination in identification of central extradural disease, with an average of 21 herniations not identified, or underestimated in size. Neither the 3D FLASH, nor the 3D MP RAGE examinations showed any significant improvement compared to the routine 2D examination for the location and severity of foraminal disease. CONCLUSION: If extradural degenerative disk disease is being evaluated, then a single enhanced 3D T1-weighted imaging sequence taking 6 minutes can be equivalent to a routine set of mixed 2D spin echo and low flip angle gradient echo sequences.

Cervical Vertebrae

Single photon emission computed tomographic teboroxime imaging with a preprocessing masking technique.

We previously reported that single-head SPECT imaging with teboroxime is feasible. However, excessive hepatic uptake in some patients may interfere with image interpretation. This study examined the feasibility of improving image quality by use of a preprocessing masking technique to subtract hepatic activity. A band of 10 pixels in width adjacent to the inferior cardiac silhouette was marked on the raw planar images, and then SPECT reconstruction was done with the Butterworth filter with a frequency cutoff of 0.3 cycles/cm and the power of 10. The stress and rest images were compared before and after masking in 10 patients who underwent SPECT teboroxime imaging during adenosine-induced coronary hyperemia (140 micrograms/kg/min for 6 minutes). SPECT imaging with a single-head detector was performed with the use of a 180-degree anterior arc (from the 45-degree left posterior oblique projection to the 45-degree right anterior oblique projection); 32 images at 8 seconds per stop were obtained (total imaging time = 6.8 minutes). All images were considered subjectively better after the masking technique was used, especially for assessment of inferior wall perfusion pattern. The maximum count in any pixel was in the hepatic region of interest before masking and in the cardiac region of interest after masking (303 +/- 110 counts vs 166 +/- 55 counts; p < 0.001). The difference was especially pronounced in the images that were obtained when patients were at rest (366 +/- 102 counts vs 184 +/- 64 counts; p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine

Reconstruction of the electrocardiogram during heart surgery.

Electrocardiograms (ECG) recorded during arrhythmia surgery are used for identification of arrhythmias of different morphology. However, the interpretation of an intraoperative ECG is difficult because some leads cannot be recorded and the signals of the remaining leads often differ from those of a preoperative recording because of the sternotomy. Therefore, a method for reconstruction of a complete intraoperative ECG, which resembles a preoperatively recorded ECG, was studied in 24 patients undergoing heart surgery. The reconstruction method involves calculating coefficients for a transformation matrix, using a preoperative ECG recording and a first intraoperative ECG recording. Once this matrix has been established, further intraoperative recordings can be transformed into an ECG which strongly resembles a preoperative ECG. The correlation between reconstructed intraoperative leads and the corresponding preoperative leads was high in the leads Vx and Vy (median correlation coefficient 0.98 and 0.97) and slightly smaller in lead Vz (0.94). Further studies will prove if the method can be useful in arrhythmia surgery.

Aged

A data adaptive reprojection technique for MR angiography.

Inability to detect vessel overlap and vascular loops can compromise the interpretation of magnetic resonance angiograms. A data-adaptive ray tracing (DART) technique was developed to produce the appropriate variations in signal intensity at points of vessel overlap in order to simulate the standard angiographic representation of vessels. In this technique a threshold is utilized to identify vessels in the image slices composing a 3D angiographic data set. A mask, which defines regions slightly larger than the vessel boundaries, is obtained by blurring the vessel information surviving the initial threshold. This mask is converted to binary form prior to multiplication by the original angiographic data set. Following application of an additional threshold to the masked data, line integrals through the regions defined by the mask are performed to obtain an angiographic signal proportional to the integrated vessel signal as in conventional angiography. This integrated reprojection is then uniquely combined with a maximum intensity pixel (MIP) reprojection to produce the final DART image. The application of the DART technique to 2D time-of-flight and 3D phase-contrast angiograms successfully enabled the identification of over-lapping vessels and vascular loops. DART was also found to produce less vessel narrowing than the MIP technique.

Algorithms

Quantitative histological analysis of the human sinoatrial node during growth and aging.

BACKGROUND: Fibrosis or fatty infiltration of the human sinoatrial (SA) node is generally believed to represent replacement of the SA nodal cells by connective tissue. Quantitative analysis, however, has not been performed precisely to validate the interpretation of such histological changes. METHODS AND RESULTS: The actual volume of the SA node and its components were calculated according to the sum of the pixel number representing the colors of SA nodal cells and connective tissue in serial sections using a digital color image analyzer. Average volume occupied by the total SA nodal cells in adolescents and adults (n = 7) was 3.55 +/- 0.45 mm3, which was 2.4 times greater than that in infants (n = 3). The rate of increase was smaller than that of the total SA node (4.2 times, 16.68 +/- 2.56 mm3 in adolescents and adults). The considerable discrepancy in the growth ratio between the SA nodal cells and the total SA node resulted from an increase in the volume of connective tissue (7.4 times). In the elderly (n = 9), the volume of total SA node and SA nodal cells actually decreased (13.10 +/- 1.85 mm3 and 2.18 +/- 0.44 mm3), whereas that of fibrous connective tissue remained unchanged. Constant DNA ploidy patterns of SA nodal cells determined by cytofluorometry indicated that SA nodal cells never synthesize DNA during growth. CONCLUSIONS: Until adulthood, the actual volume of SA nodal cells does not decrease, although the increase in volume ratio of the interstitial tissue to the total SA node has merely given a false impression of involution of SA nodal cells. Atrophy of SA nodal cells, however, occurs during aging together with reduction of the SA node and/or infiltration of fatty tissue.

Adolescent

Polyaxonal amacrine cells of rabbit retina: PA2, PA3, and PA4 cells. Light and electron microscopic studies with a functional interpretation.

Polyaxonal (PA) amacrine cells are a new class of amacrine cell bearing one to six branching, axon-like processes that emerge from the cell body or dendritic trees within 50 microns of the cell body. These slender processes of uniform caliber branch at right angles and in many respects closely resemble the axons of Golgi type II cells found elsewhere in the brain. Of the four types of polyaxonal amacrine cell that we have recognized in rabbit retina, two have been described previously in brief communications. One of these, the PA1 amacrine cell with its interstitially displaced cell body, located in the inner plexiform layer (IPL), has been analyzed extensively in two preceding reports. This paper concerns PA2, PA3, and PA4 amacrine cells. Type 2 polyaxonal (PA2) amacrine cells, identified in Golgi preparations of whole-mounted rabbit retinas, have smaller cell bodies (9-14 microns) than the other three types and these are always displaced to the ganglion cell layer (GCL) or the inner border of the inner plexiform layer (IPL). The dendritic fields of PA2 cells are also smaller than those of other PA amacrine cells, and most of their sparse dendritic branching is narrowly stratified at the border of strata (S) 4 and 5. Some members of this more heterogeneous amacrine cell "type" are bistratified, however, and more highly branched with terminal branches rising to end in S1. PA2 amacrine cells bear a scattering of small dendritic spines and may also exhibit complex dendritic appendages arising at the ends of terminal branches in proximal regions of the dendritic tree. PA2 cells emit one to three axons from the proximal dendritic tree, and about half of the cells bear a single axon. Type 3 polyaxonal (PA3) amacrine cells resemble PA1 cells in the large size of their cells bodies (11-16 microns) and dendritic fields, but differ from the latter in placement of cell bodies, which is in the GCL, and dendritic and axonal stratification, which is multistratified, ranging from S4 to S1, with a concentration in S3 or S4 and a variable contribution to S1. PA3 cells differ from PA1 cells in several other respects, including dendritic branching which occurs at higher frequency and is biased toward temporal retina, and in characteristic bristling dendritic spines, clustered in the intermediate regions of the dendritic tree, that are longer, more variable in appearance and more tightly clustered than the small, uniform spines of PA1 cells that are clustered on proximal dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Ocular

Superimposing two-dimensional gels to study genetic variation in malaria parasites.

Two-dimensional polyacrylamide gel electrophoresis is a valuable tool for studying genetic variation in the human malaria parasite, Plasmodium falciparum. It involves examining the position of protein spots in gel produced from different isolates. Some spots have been seen to vary, while others have had a constant position in all isolates so far examined. These invariant spots provide a reference frame to compare variations in other spots. This paper discusses the usefulness of digital image handling, warping and superimposition in a personal computer environment. Rather than produce a fully automatic interpretation system, we show how the computer may be used as a tool for manipulating gel images, although interpretation of the gels' features remains with the human expert. Autoradiographs are scanned on a desktop scanner, and the images in digital form can be displayed on a monitor attached to a personal computer. The coordinates of the invariant spots on each of several gels are identified by the user. Each of the gels is then warped so that the invariant spots of all the gels coincide as closely as possible. The variable spots are then examined. We have used both affine warping transformations, which match the invariant spots as closely as possible, and thin plate spline transformations, which match them exactly. Colour superimposition proved a useful way of examining the gels.

Animals

Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0).

This paper describes a computer-aided tutorial for biological stereology. Stereology, a type of quantitative morphology, includes a collection of statistical methods that quantify the structural compartments that can be viewed in sections with light and electron microscopy. These methods provide volume, surface, length, shape, and number data, and help define the quantitative relationships among the structural compartments of biological hierarchies. Hierarchies, which connect structural data ranging in size from molecules to organs, serve as a central core to which the data of biological databases can be linked. The tutorial focuses on two objectives. It provides the user primarily interested in using quantitative morphology databases with background information, and offers a set of state-of-the-art tools to researchers wishing to use these methods in the laboratory. The main topics of the tutorial include: introduction to quantitative morphology, symbols/terms, data types, sampling, hierarchies, data interpretation, and utilities. The tutorial runs under the MS-DOS operating system and requires at least an IBM PC AT (or compatible), a color monitor (EGA, VGA), 540 KB of RAM, and 3 MB of hard disk space.

Animals

Oral diagnostic reporting and synchronized image filing using magneto-optical disks.

An experimental radiologic reporting system has been developed and tested. The rewritable and compact magneto-optical disk (MOD) is applied to storing medical images with oral diagnostic reports of these radiologic images. The disk is 5.25 inches in diameter, has 600 MB memory capacity, is erasable, light and compact. Advantages are simultaneous recording of radiologic images and their oral reports by radiologists, and application to circulation of media inside the hospital as well as to filing of medical images. The MOD has a multimedia function of communication and filing. When medical images are taken and stored, oral interpretation by radiologists can be simultaneously added. Physicians can get information of the images and their reports by oral speech at the same time in front of computer workstation. Furthermore, integration of a voice recognition capability is now being undertaken.

Image Processing, Computer-Assisted

New approach to interpretation of technetium-99m pyrophosphate scintigraphy in detection of acute myocardial infarction: clinical assessment of diagnostic accuracy.

A modified classification for interpreting technetium-99m pyrophosphate scintigrams defines the 2+ diffuse pattern of tracer uptake as equlvocal rather than positive for acute myocardial infarction. Results of scintigraphy using this classification were compared with results of standard diagnostic tests for myocardial infarction in 235 patients admitted to a coronary care unit with acute chest pain. Of 81 patients with acute transmural infarction by standard clinical, electrocardiographic and serum enzyme criteria, 76 had a positive, 5 an equivocal and none a negative scintigram. Of 18 with acute nontransmural infarction by standard criteria, 7 had a positive, 9 an equivocal and 2 a negative scintigram. This it was uncommon for a patient with acute myocardial infarction, transmural or nontransmural, to have a definitely negative technetium-99m pyrophosphate study. Ten patients had equivocal evidence of infarction by standard criteria. Of the remaining 126 patients with no evidence of acute myocardial infarction by standard criteria, 87 had a negative, 35 an equivocal and 4 a definitely positive scintigram. Thus the definitely positive scintigraphic pattern was relatively highly specific for acute myocardial infarction. If the 2+ pattern had been considered positive, the specificity of the technique would have been greatly decreased. Computer processing strengthened observer certainty of the visual impression but changed the scintigraphic evaluation in only eight cases. Thus, use of an equivocal pattern renders technetium-99m pyrophosphate imaging both an extremely sensitive and specific method for detecting acute myocardial infarction.

Acute Disease

Initial results for automated computational modeling of patient-specific electromagnetic hyperthermia.

Developments in finite-difference time-domain (FD-TD) computational modeling of Maxwell's equations, super-computer technology, and computed tomography (CT) imagery open the possibility of accurate numerical simulation of electromagnetic (EM) wave interactions with specific, complex, biological tissue structures. One application of this technology is in the area of treatment planning for EM hyperthermia. In this paper, we report the first highly automated CT image segmentation and interpolation scheme applied to model patient-specific EM hyperthermia. This novel system is based on sophisticated tools from the artificial intelligence, computer vision, and computer graphics disciplines. It permits CT-based patient-specific hyperthermia models to be constructed without tedious manual contouring on digitizing pads or CRT screens. The system permits in principle near real-time assistance in hyperthermia treatment planning. We apply this system to interpret actual patient CT data, reconstructing a 3-D model of the human thigh from a collection of 29 serial CT images at 10 mm intervals. Then, using FD-TD, we obtain 2-D and 3-D models of EM hyperthermia of this thigh due to a waveguide applicator. We find that different results are obtained from the 2-D and 3-D models, and conclude that full 3-D tissue models are required for future clinical usage.

Computer Simulation

Subperiosteal resorption: effect of full-frame image compression of hand radiographs on diagnostic accuracy.

Image compression is essential to handle a large volume of digital images, including computed tomographic, magnetic resonance, computed radiographic, and digitized images in a digital radiology operation. Developed during the past few years, full-frame bit allocation performed with the cosine transform technique has been proved to be an excellent irreversible image compression method. This article describes the effect, on the accuracy of diagnosis of subperiosteal resorption, of using the hardware compression module to produce hand radiographs. Receiver operating characteristic analysis of the interpretation of 71 radiographs by five observers demonstrated that there is no statistically significant difference in diagnostic accuracy between the original radiographs and compressed and reconstructed images obtained with a compression ratio as high as 20:1.

Bone Resorption

Short fixation-shock freezing and freeze-drying versus chemical fixation and dehydration: computer assisted image analysis of morphological variables and immunogold labeling density on pituitary secretory granules.

We have performed a computer assisted image analysis evaluation of the effects of two preparation protocols on morphological variables and immunolabeling density of secretory granules using rat adenohypophysis as a model. Glutaraldehyde (GA) fixation for 2 hr and chemical dehydration was compared with short GA fixation (15 min) followed by cryofixation and freeze-drying (CF-FD). The 2 hr GA fixed specimens showed spherical nuclei and secretory granules regardless of their size, contrasting with the more irregularly shaped nuclei and secretory granules after short GA-CF-FD. In the latter group of specimens a correlation could be found between smaller nuclear areas and more irregular shapes. The differences in morphological variables between the two preparation protocols might be due to a better protein stabilization and a reduced collapse of macromolecules after the 2 hr GA fixation, strengthened by the chemical dehydration. The specific immunolabeling with antiserum to growth hormone was much greater but more varied after short time GA-CF-FD than after long GA fixation. Epon surface topography over the granule area also differed: smooth after short time GA-CF-FD and furrowed after long GA fixation. Our results, demonstrating important differences in morphological parameters and immunolabeling density between two common preparation protocols, seem critical for more reliable interpretation in quantitative immunoelectron microscopy. We also emphasize the need for computer assisted image analysis and measurements in immunoelectron microscopy to ensure objective evaluations.

Animals

Evaluation of PACS in ultrasonography.

We review our experience with a picture archiving and communication system to replace film in the ultrasound section of a clinical radiology department. The system includes three ultrasound units connected by a fiberoptic network via acquisition nodes to a central data management system, workstation, and optical jukebox. The system handles 80% of sonographic studies in the department. Image production, interpretation, storage, and retrieval are evaluated. Despite limitations, a picture archiving and communication system can be integrated into a functioning ultrasound section of an active radiology department with minimal disruption and promising results.

Computer Systems