Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria

Apoptotic cell death involves a ritual series of morphological changes, presumably reflecting a conserved molecular pathway. We now report that the nuclear events typical of apoptosis can be reproduced in "apoptotic" Xenopus egg extracts. In this cell-free system, nuclear assembly and protein import are initially normal; after 2-4 hr, however, a process of nuclear destruction ensues involving chromatin condensation and the shrinkage and fragmentation of the nuclei. This apoptotic process, which also occurs in nuclei added exogenously, is blocked by the addition of baculovirus-expressed Bcl-2 protein. To block the disintegration of nuclei that are added later, Bcl-2 must be present during this latent period. "Apoptosis" in these extracts requires a dense organelle fraction enriched in mitochondria. The cell-free system described here provides a novel tool for understanding intracellular events in apoptosis and the inhibitory function of Bcl-2.     

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

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

Autoradiographic assessment of blood flow heterogeneity in the hamster heart

OBJECTIVE: Provide regional flow measurement in the hearts of small mammals using a new, higher-resolution technique based on the deposition of a molecular marker. METHODS: We determined the instantaneous extraction and retention of the "molecular microsphere" radiolabeled desmethylimipramine in retrogradely perfused hamster hearts. In a separate series of experiments, autoradiography was used to measure regional myocardial deposition densities in hamster hearts of about 0.5 g with spatial area resolution of 16 x 16 microns. RESULTS: Radiolabeled desmethylimipramine is almost 100% extracted during a single transcapillary passage and is retained in the tissue for many minutes. Autoradiographic images demonstrated a spatial flow heterogeneity with standard deviations of 31 +/- 4% of the mean flow (N = 5) in 16 x 16 x 20-micronm3 voxels. This is equivalent to the projections made using fractal relationships from cruder observations obtained with microspheres in the hearts of baboons, sheep, and rabbits. CONCLUSIONS: Autoradiography using a molecular deposition marker provides quantitative information on myocardial flow heterogeneities with resolution at the size of cardiac myocytes. Because the regions resolved are smaller than the volume of regions supplied by single arterioles, the results must slightly exaggerate the true heterogeneity of regional flows.     

US-guided core breast biopsy: use and cost-effectiveness

OBJECTIVES: We sought to determine, using serial echocardiography, the hydrodynamic mechanisms involved in the occurrence of hemolysis after mitral valve repair. BACKGROUND: Recently, fluid dynamic simulation models have identified distinct patterns of mitral regurgitant flow disturbances in patients with mitral prosthetic hemolysis that were associated with high shear stress and may therefore produce clinical hemolysis. Rapid acceleration, fragmentation, and collision jets were associated with high shear stress and hemolysis whereas slow deceleration and free jets were not. METHODS: We reviewed serial echocardiographic studies of 13 consecutive patients with hemolytic anemia after mitral valve repair who were referred for mitral reoperation between January 1985 and December 1996 (group 1). Thirteen patients undergoing reoperation for mitral regurgitation after mitral valve repair but without hemolysis served as controls (group 2). RESULTS: The mitral regurgitant jet was central in origin in 12 group 1 patients and 9 group 2 patients (Fisher exact test, p= 0.3). The other patients had para-ring regurgitation. Group 1 patients had collision (n=11), rapid acceleration (n=2) or fragmentation (n=1) jets whereas group 2 patients had slow deceleration (n=11) or free jets (n=2) (Fisher exact test, p < 0.0001). One patient with hemolysis had both collision and rapid acceleration jets. The "culprit" jet could be identified on the postbypass transesophageal echocardiography (TEE) study in only 1 patient at the time of initial mitral repair. Twelve group 1 patients underwent reoperation, with subsequent resolution of hemolysis in all patients. At reoperation, the initial repair was found to be intact in 8 (67%) patients. CONCLUSION: Distinct patterns of flow disturbance associated with high shear stress were identified by color Doppler imaging in patients with hemolysis after mitral valve repair. The majority (92%) of these color flow disturbances were not present during intraoperative postbypass TEE study after initial mitral repair and subsequently developed in the early postoperative period.     

Hypothermic coagulopathy in trauma: effect of varying levels of hypothermia on enzyme speed, platelet function, and fibrinolytic activity

Comparative study of protein metabolism in neurons of layers III and V of the sensorimotor cortex was carried out in two groups of Wistar rats, which differed in learning results: "bad" (60% of population) and "good" learners (40%). It was found out that the associative neurons (layer III) were most sensitive to cognitive load. In "bad" learners, an increase in nuclear and cytoplasmic dimensions and rise in protein concentration and content took place in these neurons, while in the efferent neurons (layer V) the protein content increased only in the cytoplasm. In "good" learners, the cognitive load led to a decrease in all the cytochemical parameters in neurons of the layer III while in the neurons of the layer V the content and concentration of proteins increased both in nuclei and cytoplasm. It is suggested that the character of protein metabolism changes produced by information load can be considered as a reflection of individual peculiarities of cognitive activity, and the extent of cytochemical changes as a reflection of complicity of a cognitive task.     

Modelling the three-dimensional elastic constants of parallel-fibred and lamellar bone

The complex hierarchical structure of lamellar bone makes understanding structure–mechanical function relations, very difficult. We approach the problem by first using the relatively simple structure of parallel-fibred bone to construct a mathematical model for calculating Young's moduli in three-dimensions. Parallel-fibred bone is composed essentially of arrays of mineralized collagen fibrils, which are also the basic structural motif of the individual lamellae of lamellar bone. Parallel-fibred bone structure has orthotropic symmetry. As the sizes and shapes of crystals in bone are not well known, the model is also used to compare the cases of platelet-, ribbon- and sheet-reinforced composites. The far more complicated rotated plywood structure of lamellar bone results in the loss of the orthotropic symmetry of individual lamellae. The mathematical model used circumvents this problem by sub-dividing the lamellar unit into a thin lamella, thick lamella, transition zone between them, and the recently observed back-flip lamella. Each of these is regarded as having orthotropic symmetry. After the calculation of their Young's moduli they are rotated in space in accordance with the rotated plywood model, and then the segments are combined to present the overall modulus values in three-dimensions. The calculated trends compare well with the trends in microhardness values measured for circumferential lamellar bone. Microhardness values are, as yet, the only measurements available for direct comparison. Although the model is not directly applicable to osteonal bone, which is composed of many hollow cylinders of lamellar bone, the range of calculated modulus values and the trends observed for off-axis calculations, compare well with measured values. © 1998 Chapman & Hall     

Optical computing: introduction by the feature editors

This feature issue of Applied Optics: Information Processing contains 19 papers on Optical Computing. Many of these papers are expanded versions of presentations given at the Optical Society of America's Sixth Topical Meeting on Optical Computing held in Salt Lake City, Utah, in March 1995. This introduction provides a brief historical account of the series of optical computing meetings and a brief review of the papers contained in this special issue.     

Photorefractive processing for large adaptive phased arrays

An adaptive null-steering phased-array optical processor that utilizes a photorefractive crystal to time integrate the adaptive weights and null out correlated jammers is described. This is a beam-steering processor in which the temporal waveform of the desired signal is known but the look direction is not. The processor computes the angle(s) of arrival of the desired signal and steers the array to look in that direction while rotating the nulls of the antenna pattern toward any narrow-band jammers that may be present. We have experimentally demonstrated a simplified version of this adaptive phased-array-radar processor that nulls out the narrow-band jammers by using feedback-correlation detection. In this processor it is assumed that we know a priori only that the signal is broadband and the jammers are narrow band. These are examples of a class of optical processors that use the angular selectivity of volume holograms to form the nulls and look directions in an adaptive phased-array-radar pattern and thereby to harness the computational abilities of three-dimensional parallelism in the volume of photorefractive crystals. The development of this processing in volume holographic system has led to a new algorithm for phased-array-radar processing that uses fewer tapped-delay lines than does the classic time-domain beam former. The optical implementation of the new algorithm has the further advantage of utilization of a single photorefractive crystal to implement as many as a million adaptive weights, allowing the radar system to scale to large size with no increase in processing hardware.     

A Bayesian network approach to assess and predict software quality using activity-based quality models

ContextSoftware quality is a complex concept. Therefore, assessing and predicting it is still challenging in practice as well as in research. Activity-based quality models break down this complex concept into concrete definitions, more precisely facts about the system, process, and environment as well as their impact on activities performed on and with the system. However, these models lack an operationalisation that would allow them to be used in assessment and prediction of quality. Bayesian networks have been shown to be a viable means for this task incorporating variables with uncertainty.ObjectiveThe qualitative knowledge contained in activity-based quality models are an abundant basis for building Bayesian networks for quality assessment. This paper describes a four-step approach for deriving systematically a Bayesian network from an assessment goal and a quality model.MethodThe four steps of the approach are explained in detail and with running examples. Furthermore, an initial evaluation is performed, in which data from NASA projects and an open source system is obtained. The approach is applied to this data and its applicability is analysed.ResultsThe approach is applicable to the data from the NASA projects and the open source system. However, the predictive results vary depending on the availability and quality of the data, especially the underlying general distributions.ConclusionThe approach is viable in a realistic context but needs further investigation in case studies in order to analyse its predictive validity.