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In vivo, in vitro, and computational analysis of dendritic calcium currents in thalamic reticular neurons.

Thalamic reticular (RE) neurons are involved in the genesis of synchronized thalamocortical oscillations, which depend in part on their complex bursting properties. We have investigated the intrinsic properties of RE cells using computational models based on morphological and electrophysiological data. Simulations of a reconstructed RE cells were compared directly with recordings from the same cell to obtain precise values for the passive parameters. In a first series of experiments, the low-threshold calcium current (I(Ts)) was studied via voltage clamp in acutely dissociated RE cells that lack most of their dendrites. Simulations based on a cell with truncated dendrites and Hodgkin-Huxley kinetics reproduced these recordings with a relatively low density of I(Ts). In a second series of experiments, voltage-clamp recordings obtained in intact RE cells in slices showed a higher amplitude and slower kinetics of I(Ts). These properties could be reproduced from the reconstructed cell model assuming higher densities of I(Ts) in distal dendrites. In a third series of experiments, current-clamp recordings were obtained on RE cells in vivo. The marked differences with in vitro recordings could be reconciled by simulating synaptic bombardment in the dendrites of RE cells, but only if they contained high distal densities of I(Ts). In addition, simpler models with as few as three compartments could reproduce the same behavior assuming dendritic I(Ts). These models and experiments show how intrinsic bursting properties of RE cells, as recorded in vivo and in vitro, may be explained by dendritic calcium currents.

Animals↗

Relationship between creatine kinase activity and semen characteristics in subfertile men.

OBJECTIVE: Creatine kinase is an indicator of sperm maturity. We studied whether sperm creatine kinase levels differ between normal healthy donors and subfertile patients and determined the correlation between sperm creatine kinase level and semen quality in subfertile men. MATERIAL AND METHODS: Semen samples were obtained from 76 subfertile and 15 healthy normal donors after 48 to 72 hours of sexual abstinence. Sperm characteristics were assessed with a computer-assisted semen analyzer. Morphology was evaluated by Kruger's strict criteria and World Health Organization methods. The thiobarbituric acid assay was used to measure lipid peroxidation; sperm creatine kinase activity was measured using a commercial kit after detergent extraction (Triton X-100). RESULTS: Creatine kinase levels were significantly higher (P < .001) in subfertile men (median = 0.197 U/10(8) sperm) compared with donors (median = 0.061 U/10(8) sperm). In subfertile men, creatine kinase levels correlated significantly with lipid peroxidation levels (r = .49; P = 0.03) and sperm concentration (r = -.70; P < .001), and with normal sperm forms by Kruger's (r = -.30; P = 0.01) and WHO methods (r = -.32; P < .005). Creatine kinase levels and spermatozoal characteristics did not correlate significantly in donors. Compared with subfertile normospermic men, creatine kinase activity was significantly higher in oligospermic and asthenospermic men (P <.001). CONCLUSIONS: The inverse relationship between creatine kinase level and sperm concentration and morphological forms suggests that creatine kinase levels can be a reliable marker for semen quality in subfertile men. An elevated creatine kinase level and its correlation with lipid peroxidation levels may reflect biochemically immature spermatozoa.

Creatine Kinase↗

Widening of capillary neck and alteration of extracellular matrix ultrastructure in diabetic rat glomerulus as revealed by computer morphometry and improved tissue processing.

Morphological and morphometric studies of glomeruli were carried out in streptozotocin-induced diabetic rats using improved tissue processing and computerized morphometry. Increased mesangial matrix, occupying the enlarged diabetic mesangium, contained an abundance of dark granular material in addition to the microfibrils which were usually found in the control glomeruli. In the diabetic glomeruli, the lamina densa was thick and heterogeneous showing a dense layer both on its epithelial and endothelial aspects, and the lamina rara externa contained more fibrils than in control rats. Detailed estimation of the absolute values of the various compartments of the diabetic glomeruli by using perfusion-fixed materials and a computer-assisted digitizer revealed that the volume and surface area of the mesangium were increased more extensively than those of the capillary; the enlargement of the mesangial-capillary interface area was the most pronounced among the morphometric changes of the diabetic glomeruli; and that the moderate increase in capillary volume was associated with an increased radius. Our quantitative results showed that capillaries in the diabetic glomeruli had an extensively wider neck which may be the first sign of structural damage to the glomerular tuft.

Animals↗

Suppression by isoproterenol of endothelial cell morphology and barrier function changes induced by platelet-activating factor.

Using a model to study vascular permeability on hydrostatically perfused bovine pulmonary artery endothelial cell (EC) monolayers and software to analyze cell morphological parameters automatically in a computer image workstation, we studied the effects of isoproterenol (IPN) on platelet-activating factor (PAF)-induced changes in EC monolayer permeability and cell morphological parameters. Albumin has fortifying effects on endothelial barrier function. As albumin concentration in the perfusate increased (0, 1, 5, 10, 20 mg/ml), EC monolayer hydraulic conductivity (Lp) decreased gradually while Lp of the filter membranes did not change. After treatment of the EC monolayer with PAF 10(-8) mol/liter for 30 min, transmonolayer fluid flow, protein clearance rate, and Lp value increased noticeably. At the same time, cell area decreased and intercellular distance and percentage of intercellular space area in total cell monolayer increased. Pretreatment with 10(-4) mol/liter IPN blocked PAF-induced EC permeability and morphological changes, suggesting that EC contraction and intercellular gap formation are important mechanisms for PAF-induced high vascular permeability. IPN inhibits the effects of PAF via stabilization of EC morphology, protection of intercellular junction, and blockade of intercellular gap formation.

Albumins↗

Segmentation of stained blood cell images measured at high scanning density with high magnification and high numerical aperture optics.

In hematological morphology, it is necessary to resolve and analyze the smallest possible cellular details appearing in the light microscope. A prerequisite for computer-aided analysis of subtle morphological features is measuring the cells at a high scanning density with high magnification and high numerical aperture optics. Contrary to visual observations, the information content in a measured picture can be increased by setting the condensor's numerical aperture (NA) greater than the objective's NA. The complexity and heterogeneity of such cell images necessitate a new segmentation method that conserves the morphological information required in the subsequent image analysis, feature extraction, and cell classification. In our segmentation strategy, characteristic color difference thresholds for each nucleus and cytoplasm are combined with geometric operations, probability functions, and a cell model. All thresholds are repeatedly recalculated during the successive improvements of the image masks. None of the thresholds are fixed. This strategy segments blood cell images containing touching cells and large variations in staining, texture, size, and shape. Biological inconsistencies in the calculated cell masks are eliminated by comparing each mask with the cell model criteria integrated into the entire segmentation process. All 20,000 leukocyte images from 120 smears in our leukemia project were segmented with this method.

Blood Cells↗

Morphometry of normal and teratozoospermic canine sperm heads using an image analyzer: work in progress.

Combining the traditional morphologic evaluation of spermatozoa with computer assisted image analysis adds randomness, objectivity, repeatability and accuracy to morphometric measurements. We collected semen from 10 fertile, normospermic dogs aged 1 to 7 yr and from 3 teratozoospermic breed-matched dogs. Sperm head morphology was examined in Giemsa-stained smears by light microscopy, using a computer-assisted image analyzer and by transmission electron microscopy. We found significant variation in sperm head area, length, width and degree of roundness among normospermic individual dogs, indicating that it would be necessary to examine many more dogs before the size and shape of normal dog spermatozoa could be determined. The normospermic dogs were used as controls for the teratozoospermic cases. Case 1: A 2-yr-old subfertile Cavalier King Charles Spaniel had semen with small and narrow-based sperm heads and a proximal cytoplasmic droplet in most of the spermatozoa. With the image analysis system, sperm heads were shown to be smaller and more oval than in normospermic dogs. The variatons in size and shape were similar in magnitude to those of control dogs. An examined infertile half-brother had similar semen quality. Case 2: A 3-yr-old Petit Basset Griffon Vendeen with 2 unsuccesfull matings exhibited spermatozoa with severe abnormalities. Measured by image analyzer, sperm heads were irregular in shape and very small in area. One of the two littermates examined had semen of the same quality as the case dog. Case 3: A 3-yr-old fertile Golden Retriever had semen with giant sperm heads in about 50% of spermatozoa. Image analyzing results revealed 2 populations of different sized sperm heads. Giant heads consisted of 52.2% of all spermatozoa. The results of the study reported here suggest that the image analysis technique may be useful in evaluating structural changes in sperm morphology, supplementing visual assessment that is used in conventional methods.

Journal Article↗

Fluorine-18 deoxyglucose and false-positive results: a major problem in the diagnostics of oncological patients.

Fluorine-18 deoxyglucose (FDG) is not a very tumour-specific substance, and its accumulation in benign lesions with increased glucose metabolism may give rise to false-positive results and hence cause FDG positron emission tomography (PET) to display relatively low specificity (frequently below 85%). Correct interpretation of FDG PET studies is predicated upon detailed knowledge of morphological abnormalities, and the importance of the correlation of functional and morphological information, as derived from computed tomography or magnetic resonance imaging, is discussed. It is emphasized that image fusion programs cannot substitute for understanding of functional and morphological methods. The reconstruction of PET cross-sections is considered, and it is concluded that an iterative image reconstruction method is to be favoured, given its advantages in reducing image artefacts and improving quantification of radioactivity concentrations. The differentiation of malignant and benign lesions when using FDG PET is then reviewed; false-positive findings may be obtained, for example, in patients with acute inflammatory lesions, chronic pancreatitis, retroperitoneal fibrosis or salivary gland tumours. It is suggested that these problems may be alleviated by means of multitracer studies, e.g. using carbon-11 labelled aminoisobutyric acid for quantification of A-type amino acid transport. Finally, the effects of radiotherapy and chemotherapy on FDG uptake and the problems that accrue from these effects are reviewed. Both radiotherapy and chemotherapy can cause increased FDG uptake, complicating diagnosis and evaluation. Knowledge of the effects of different treatment procedures on regional FDG metabolism is therefore necessary for correct interpretation of the PET data.

Artifacts↗

Neurobiology of the aging brain: morphological alterations at synaptic regions.

Computer-assisted morphometric studies have been carried out on nerve cell terminal regions in rats of different ages. The numerical density (Nv), the average area (S) and the surface density (Sv) of ethanol phosphotungstic acid stained (E-PTA) synaptic junctions were evaluated in cerebellar glomeruli and dentate gyrus supragranular layers. The volume density (Vv), the average volume (V) and the numerical density (Nvm) of synaptic mitochondria were measured in the cerebellar glomeruli of young (3 months), adult (11 months) and old (28 months) animals. We found that during aging Nv and Sv undergo a significant decrease, whereas S is significantly increased. The mitochondrial Vv is unchanged in all the age groups analysed, whereas in old rats V is increased and Nvm decreased, respectively. We interpret our results as supporting that the old CNS retains part of its remodelling activity and is capable of adaptive morphological response at synaptic terminal regions.

Journal Article↗

Direct comparison of computed tomography and magnetic resonance imaging for characterization of posterior left atrial morphology.

OBJECTIVE: Accumulating evidence points to the central importance of the posterior left atrium (PLA) for atrial fibrillation (AF). Catheter ablation intended to cure AF is increasingly practiced; performance and assessment of this procedure is enhanced by accurate imaging of PLA anatomy. Prior reports have suggested that both computed tomographic (CT) and magnetic resonance (MR) imaging techniques provide accurate PLA images. These techniques have never been compared directly. MATERIALS AND METHODS: Twenty patients referred for catheter ablation underwent preoperative imaging using both CT and MR. Each technique was used to create a multidimensional image of the PLA. RESULTS: Within patients, morphologic and dimensional PLA indices, including number of individual pulmonary venoatrial junctions, presence of ostial branches, circumference of each venoatrial junction, venoatrial junction "non-circularity", and distance between ipsilateral superior and inferior venoatrial junctions, were well correlated. CONCLUSIONS: CT and MR-based images of the PLA appear comparable. Technique selection should involve considerations of toxicity, tolerance, and local resources.

Adult↗

Morphology of cerebral lesions in children with congenital hemiplegia. A study with computed tomography.

This study has analysed the results of CT scans of the brains in children with congenital hemiplegia. The material consists of 111 out of a total of 151 children with this cerebral palsy syndrome in a population-based series. We have classified the morphological findings in five groups. The groups are designed to reflect the phase of maturation of the brain when the insult happened. The groups are: 1. Maldevelopment, 2. Periventricular atrophy, 3. Cortical-/subcortical atrophy, 4. Miscellaneous, 5. Normal. In contrast to previous reports we found a high proportion (17%) with maldevelopment. However the dominating morphological pattern was periventricular atrophy, consistent with a hypoxic-ischemic insult to the immature brain, seen in 42%. Cortical and/or subcortical atrophy was found in 12%. Three children (3%) presented with morphological patterns not possible to classify. The group with no pathology according to CT was 26%.

Atrophy↗

Computed tomography in the study of the development of the skull base: 1. Normal morphology.

We examined 11 human fetuses (18 to 30 weeks' gestation) with high resolution CT to study the developmental ossification of the two main component structures of the skull base, the temporal and sphenoid bones. Computed tomography, with its excellent contrast discrimination, allowed clear visualization of the morphology of ossifying centers. Skull base growth during this gestational period was found to be a dynamic event with rapid changes in morphology and interosseous relationships. The growth of the sphenoid body on CT corroborates the studies done by Kodama, who used dehydrated skulls. The temporal bone proceeded in a stepwise fashion; complete encapsulation of the membranous labyrinth occurred before it grew anteroposteriorly to form the apex and mastoid.

Gestational Age↗

Comparative study of morphological and time-frequency ECG descriptors for heartbeat classification.

The prompt and adequate detection of abnormal cardiac conditions by computer-assisted long-term monitoring systems depends greatly on the reliability of the implemented ECG automatic analysis technique, which has to discriminate between different types of heartbeats. In this paper, we present a comparative study of the heartbeat classification abilities of two techniques for extraction of characteristic heartbeat features from the ECG: (i) QRS pattern recognition method for computation of a large collection of morphological QRS descriptors; (ii) Matching Pursuits algorithm for calculation of expansion coefficients, which represent the time-frequency correlation of the heartbeats with extracted learning basic waveforms. The Kth nearest neighbour classification rule has been applied for assessment of the performances of the two ECG feature sets with the MIT-BIH arrhythmia database for QRS classification in five heartbeat types (normal beats, left and right bundle branch blocks, premature ventricular contractions and paced beats), as well as with five learning datasets-one general learning set (GLS, containing 424 heartbeats) and four local sets (GLS+about 0.5, 3, 6, 12 min from the beginning of the ECG recording). The achieved accuracies by the two methods are sufficiently high and do not show significant differences. Although the GLS was selected to comprise almost all types of appearing heartbeat waveforms in each file, the guaranteed accuracy (sensitivity between 90.7% and 99%, specificity between 95.5% and 99.9%) was reasonably improved when including patient-specific local learning set (sensitivity between 94.8% and 99.9%, specificity between 98.6% and 99.9%), with optimal size found to be about 3 min. The repeating waveforms, like normal beats, blocks, paced beats are better classified by the Matching Pursuits time-frequency descriptors, while the wide variety of bizarre premature ventricular contractions are better recognized by the morphological descriptors.

Algorithms↗

Standardization of sampling and staining methods for the morphometric evaluation of sperm heads in the Cynomolgus monkey (Macaca fascicularis) using computer-assisted image analysis.

Automated sperm morphology analysis (ASMA) technology has improved the assessment of sperm morphology, but the results depend on the use of adequate and standardized procedures. In this study the Sperm-Class Analyzer (SCA) ASMA system was used to assess sperm head morphometry in the Cynomolgus monkey and to evaluate the influence of sample size, intraslide variation, and the use of three staining techniques on the accuracy of image processing and sperm head morphometry. Haematoxylin is the staining technique of choice for Cynomolgus spermatozoa, as optimum contrast of sperm heads with the surrounding background allows efficient segmentation, i.e. sperm head boundary detection, making the image analysis process more accurate. The analysis of 100 spermatozoa is recommended since a larger sample size did not result in more accurate sperm head morphometry. There were no differences in either the percentage of correctly binarized sperm heads or sperm head dimensions among samples obtained from different zones of the slides, although differences in stain intensity (grey level) were detected. The measurements made on Haematoxylin, Diff-Quik and Hemacolor-stained slides yielded different values for all of the sperm head parameters under consideration. This result demonstrates that the procedures of fixation and staining significantly affect the dimensions of sperm heads.

Animals↗

Three-dimensional reconstruction of pulmonary veins in patients with atrial fibrillation and controls: morphological characteristics of different veins.

Multidetector computed tomography can be used to evaluate the anatomy of pulmonary veins (PVs) in patients with AF. The study evaluated two groups. Group 1 included 61 patients assessed following PV ablation with ultrasound of RF energy. Group 2 included 15 patients undergoing ablation for AF and 14 control subjects without a history of AF matched for age and sex. The anatomy of the PVs was analyzed in this group prior to the ablation and compared to controls. Computed tomography was used to measure the ostium of the left superior, left inferior, right superior, right inferior PVs, and the left atrial appendage size. In group 1, PV stenosis was seen in 14 (30%) of 46 patients undergoing ablation with RF energy and in none of the 15 patients receiving ablation with ultrasound energy. In group 2, the ostium size was not different between patients with AF and controls. Similarly, the ostium of the PV that appeared to trigger AF was not larger than the ostium of the remaining veins. A "clustering pattern" of PV branches near the right inferior PV ostium was seen in almost every patient, independent of the presence of the arrhythmia. Computed tomography frequently detects PV stenosis following RF ablation. Ultrasound ablation does not appear to result in PV narrowing. Overall, patients with AF do not have larger sizes of PV ostia. Multiple ramifications from the right inferior PV ostium is a common pattern and may represent a protective anatomic variant.

Atrial Fibrillation↗

A Golgi analysis of the primate globus pallidus. II. Quantitative morphology and spatial orientation of dendritic arborizations.

The morphology of pallidal neurons was analyzed quantitatively in Golgi-impregnated brains of men and macaques (Macaca irus). Selected neurons were drawn with a camera lucida and reconstructed from serial sections. Dendritic arborizations were analyzed in three dimensions using a video computer microscope (Yelnik et al., '81). Morphological (topological and metrical) parameters were computed, and the overall geometry of arborizations was studied in three dimensions with the aid of principal component analysis (Yelnik et al., '83). Statistical tests were used in order to compare human with simian large neurons and the lateral with the medial pallidum. All neurons were found to belong to a single neuronal population. Particular neurites may be added randomly onto pallidal dendrites, mainly in the lateral pallidum. Large pallidal neurons are characterized by sparsely branched dendritic arborizations (4 stems, 13 tips) with thick, smooth, and long dendrites (longest dendrite = 1,000 micron, total dendritic length = 7,600 micron). All arborizations are discoidal in shape with mean dimensions of 1,500 X 1,000 X 250 micron. Pallidal discs are always parallel to the lateral border of each pallidal nucleus and thus perpendicular to striatal axons to which they present their greatest extent. They may be traversed by a large number of these axons. The existence of other pallidal neuronal groups, "intermediate" and "local circuit" neurons, identified in their fine morphological features by François et al. ('84), was confirmed quantitatively in the present study.

Animals↗

Use of Ti-coated replicas to investigate the effects on fibroblast shape of surfaces with varying roughness and constant chemical composition.

A two-stage replica technique with a subsequent titanium (Ti)-coating treatment was used to faithfully replicate topographies of polished, acid-etched, machined-like, finely blasted, coarsely blasted, coarsely blasted and acid-etched, and Ti plasma-sprayed Ti surfaces. The replicas were used to study the influence of different rough surface topographies on the response of human fibroblasts in vitro under conditions of constant surface chemistry for all surfaces. The surface topographies of the replicas were characterized using non-contact laser profilometry, scanning electron microscopy (SEM), and stereo-SEM, whereas surface chemistry was examined using X-ray photoelectron spectroscopy. Fibroblasts were trypsinized and plated onto the Ti-coated epoxy-resin replica surfaces for 24 h and observed with SEM. Fluorescein-5-thiosemicarbazide was used to stain the cell components including cell membrane, and the stained cells were optically sectioned using epifluorescent microscopy. The optical sections were computationally reconstructed to obtain three-dimensional images and cell volume and cell thickness determined. The different surface topographies were found to alter cell thickness and cell morphology. However, cell volume as computed from three-dimensional reconstructions was not affected by surface features. The results suggest that cells distort themselves to accommodate to rough surfaces but their volume is not significantly altered.

Cell Size↗

The morphometric analysis and recognition an amyloid plaque in microscope images by computer image processing.

This paper presents an approach of the two-dimensional image processing application in recognition of amyloid plaque in microscope images of the brain tissues. The authors propose to create universal amyloid plaque computer pattern and special multivariate image segmentation techniques based on collected images and statistical information. This recognition image procedure is divided into 3-dimensional statistical colour and morphological shape identifications. The developed computer system will collect and store image data and exchange them by network with other collaborated systems.

Alzheimer Disease↗

Canine fresh and cryopreserved semen evaluation.

There is little doubt that objective assessment of multiple parameters related to the functional and morphological characteristics of spermatozoa, increase the predictability of the fertilizing potential of a semen sample. Conventional microscopic methods for sperm evaluation in combination with the more objective computer-assisted sperm motility and morphology analyzers and flow cytometry, have allowed investigators to obtain precise information about the morphofunctional status of spermatozoa, which already has resulted in a better understanding of sperm biology and of some of the mechanisms involved in sperm cryoinjury. In addition, assays based on in vitro fertilization provide valuable information about the functional ability of spermatozoa when interacting with the oocyte. Although objective methods for dog sperm evaluation are available for many researchers, considerable effort has still to be invested in order to standardize physiological sperm parameters for the results to be interpretable under unified criteria. In the present paper some of the classical and new methods currently used for dog sperm evaluation are reviewed, though not exhaustively, covering some aspects of the sperm cell that are useful in estimating its functional ability.

Acrosome↗