Biomedical subjects

L M Cruz-Orive

Publications and source records attributed to L M Cruz-Orive.

At least 37 records · Page 2Linked to original sources

PubMed1994

Estimation of fetal volume by magnetic resonance imaging and stereology.

The current methods to monitor fetal growth in utero are based on ultrasound image measurements which, lacking a proper sampling methodology, may be biased to unknown degrees. The Cavalieri method of stereology guarantees the accurate estimation of the volume of an arbitrary object from a few systematic sections. Non-invasive scanning methods, and magnetic resonance imaging (MRI) in particular, are valuable tools to provide the necessary sections, and therefore offer interesting possibilities for unbiased quantification. This paper describes how to estimate fetal volume in utero with a coefficient of error of less than 5% in less than 5 min, from three or four properly sampled MRI scans. MRI was chosen because it does not use ionizing radiations on the one hand, and it offers a good image quality on the other. The impact of potential sources of bias such as fetal motion, chemical shift and partial voluming artefacts is discussed. The methods are illustrated on four subjects monitored between weeks 28 and 40 of gestation.

Adult↗

PubMed1994

Fluctuations of visual field interpretation related to cluster and scotoma analysis by one interpreter.

A total of 189 numerical printouts of the visual fields of 189 eyes of individuals either suspected of or verified as having glaucoma and displaying varying degrees of glaucomatous visual field damage were analyzed by one expert interpreter. The amount of clusters (or scotomas) was counted and delineated in each visual field under both a more stringent and a more liberal criterion. The interpretations of the visual fields differed significantly in both groups, as did the subjective certainty the interpreter allotted to his assessment. In the group of certain decisions, only 144 defects were assessed as opposed to 364 when a more stringent rather than a more liberal criterion was adopted. In uncertain cases, a reverse trend was observed, since only 11 as opposed to 324 defects were assessed when a more liberal rather than a more stringent criterion was adopted. Both trends may be explained by an increase in sensitivity at the expense of specificity (i.e., a change in the receiver-operating characteristic of the expert when the latter is pressed to change the criterion in a subjective cluster or scotoma assessment task). The ambiguity of the task and the difference in the criterion adopted by the interpreter significantly influences the assessment of normality and pathology, in particular its extent and spatial distribution.

Adult↗

PubMed1993

Vertical LM sectioning and parallel CT scanning designs for stereology: application to human lung.

Practical, unbiased stereological methods are described to estimate lung volume and external surface area, and total volume and surface area of relatively large and anisotropic structures (bronchi and arteries) inside the lung. The volume of each of five lung strata was estimated first by fluid displacement and then by computed tomography (CT) using Cavalieri's method; the reliability of CT was assessed through a calibration procedure, and image thresholding criteria for an accurate volume estimation using CT were established. The parallel, perfectly registered CT section images were also used to estimate the external surface area of each stratum by the spatial grid method. Unbiased estimation of internal surface areas in lung is a long-standing problem: since the structures are large and essentially void, large sections are needed; to facilitate identification, thin sections have to be used for light microscopy, and since such structures are anisotropic, the sections should be vertical. A practical stereological design is demonstrated here on an infant lung, which fulfils all these requirements. This study illustrates the potential of using unbiased stereology to characterize infant pulmonary hypoplasia.

Female↗

PubMed1993

Unbiased volume estimation with coaxial sections: an application to the human bladder.

A problem of current medical interest is how to estimate the volume of an object from coaxial sections. Such images can be produced in ultrasound scanning, when the planar scan sections all emanate from a common axis. Two unbiased volume estimators for this sampling regime, previously published without derivation, are derived and presented here in some detail. The estimators are based on an ancient theorem of Pappus of Alexandria (c. AD 320). One of the estimators was used to estimate the volume of urine in the bladder of six human volunteers. The case example was attractive because the corresponding volumes could be voided and directly measured after the scanning session, thus confirming the unbiasedness of the estimator used. Sample size guidelines are also given.

Adult↗

PubMed1993

Unbiased estimation of human body composition by the Cavalieri method using magnetic resonance imaging.

The classical methods for estimating the volume of human body compartments in vivo (e.g. skin-fold thickness for fat, radioisotope counting for different compartments, etc.) are generally indirect and rely on essentially empirical relationships--hence they are biased to unknown degrees. The advent of modern non-invasive scanning techniques, such as X-ray computed tomography (CT) and magnetic resonance imaging (MRI) is now widening the scope of volume quantification, especially in combination with stereological methods. Apart from its superior soft tissue contrast, MRI enjoys the distinct advantage of not using ionizing radiations. By a proper landmarking and control of the scanner couch, an adult male volunteer was scanned exhaustively into parallel systematic MR 'sections'. Four compartments were defined, namely bone, muscle, organs and fat (which included the skin), and their corresponding volumes were easily and efficiently estimated by the Cavalieri method: the total section area of a compartment times the section interval estimates the volume of the compartment without bias. Formulae and nomograms are given to predict the errors and to optimize the design. To estimate an individual's muscle volume with a 5% coefficient of error, 10 sections and less than 10 min point counting (to estimate the relevant section areas) are required. Bone and fat require about twice as much work. To estimate the mean muscle volume of a population with the same error contribution, from a random sample of six subjects, the workload per subject can be divided by square root of 6, namely 4 min per subject. For a given number of sections planimetry would be as accurate but far more time consuming than point counting.

Body Composition↗

PubMed1993

Spatial distribution of curve length: concept and estimation.

The length of a curvilinear feature, such as a dendrite tree of a neuron, can, in principle, be estimated by the recent, non-invasive method of total vertical projections (TVPs). Curve length is a measure of size, but it reports nothing about curve shape. The shape of a tree-like structure can be described to some extent by the distribution of branch length in properly defined regions of three-dimensional (3-D) space. A definition of curve length distribution in three dimensions is proposed and implemented here on a human neuron. The relevant 3-D regions overlap after projection, and therefore the TVPs method cannot be used directly to estimate the corresponding feature lengths. However, using the ANALYZE software system running on a SUN SPARC workstation, dendrite subsets sitting in predefined regions of space were rendered in different colours and measured separately by the TVPs method using a cycloid test system. In combination with non-invasive image acquisition and processing techniques, the length distribution concept is likely to be useful in the metrical analysis of either microscopic or macroscopic arborizations in a wide variety of contexts, including living cells and organisms.

Dendrites↗

PubMed1993

Morphology and stereology of the female canine urethra correlated with the urethral pressure profile.

Perfusion-fixed urethrae of five sexually intact nulliparous female beagles in anestrus, between 2 and 3 years of age, were morphologically and morphometrically investigated. Absolute and relative volumes of different tissue components were stereologically estimated per urethral quarter. A discrete smooth muscle sphincter could neither be identified in the bladder neck nor in the urethra. However, the circularly arranged smooth muscle fibers in the proximal half of the urethra were regarded as being capable of sphincteric function. The connective tissue, composed of collagenous and longitudinally orientated numerous elastic fibers, accounted for the most predominant constituent. It comprised 77.3 to 78.2% of the total volume in the proximal three urethral quarters. The vascular plexus was most prominently developed in the proximal two urethral quarters, and its relative and absolute volumes significantly exceeded those of the third quarter. The discovered direct connections between small arteries and the sinusoids of the vascular plexus emphasize its importance as a major contributory component to urinary continence. The striated musculus urethralis was essentially confined to the distal half of the urethra and most strongly developed in the fourth quarter, where it surrounded the urethra transversely at its ventral and lateral aspects. The caudal muscle fibers continued dorsally to the vagina. A transitional epithelium bounded the urethral lumen. Its superficial cell layer progressively flattened towards the external urethral orifice, changing into a stratified squamous lining as also seen in the vestibulum. Recorded pressures, or pressure changes, could neither be correlated with volumes nor with morphology of particular tissue elements in the different urethral quarters.

Animals↗

PubMed1993

Evaluation of numerical displays of visual field data by human observers--the definition of scotomas and clusters by human interpreters.

Numerical printouts of 23 visual field examinations performed with glaucoma program G1 were analyzed by 3 perimetrists. The task consisted of defining the existence of clusters or scotomas. Considerable discrepancy with regard to the existence or the size of clusters and/or scotomas was found among the three perimetrists. The median difference in number of clusters as compared for each pair of interpreters was found to differ from zero at a significance level of 5% (Wilcoxon's signed-rank nonparametric test). These findings suggest that the interpretation of a visual field status by human observers will always remain subjective and to a large extent contradictory. This situation calls for the implementation of interpretative computer software to supplement the currently available algorithms.

Adult↗

PubMed1993

The efficacy of cw Nd:YAG laser trabeculoplasty.

The long-term (mean follow up, 51 weeks) efficacy of cw Nd:YAG laser trabeculoplasty in reducing intraocular pressure (IOP) was studied in 106 eyes of 75 patients. Fifty patients (73 eyes) of this group had primary open-angle glaucoma; 11 (17), mixed glaucoma; 10 (12), pseudoexfoliation syndrome; and 4 (4), secondary glaucoma of varied etiology. Fifty laser impacts were delivered over 180 degrees of the circumference of the chamber angle with an Nd:YAG laser working in the cw mode (spot diameter, 100 microns; exposure duration, from 200 to 300 milliseconds; constant power level, 9 W, resulting in 1.8 to 2.7 J/pulse and a total energy dose/session of 90 to 135 J). In a few cases, an exposure duration of 400 milliseconds was used, corresponding to a pulse energy of 3.6 J/pulse and a total energy dose/session of 180 J. Success was defined as a reduction in IOP to 22 mm Hg or less without any further laser or surgical intervention. The cumulative success rate was 97.2% at 1 month, 93.3% at 3 months, 83.3% at 6 months, 78.7% at 12 months, and 71.5% at 24 months. Thus, the efficacy of cw Nd:YAG laser trabeculoplasty is comparable to that reported for trabeculoplasty performed using the argon laser (ALT), the Nd:YAG laser working in the free-running mode, and the diode laser. Biomicroscopy failed to reveal any serious inflammatory reactions persisting longer than 1 week. We conclude that cw Nd:YAG laser trabeculoplasty is a safe and effective alternative to ALT.

Adult↗

PubMed1992

Estimating neuron dendritic length in 3D from total vertical projections and from vertical slices.

The recently developed method of total vertical projections is illustrated to estimate the total dendritic length of a human Substantia Nigra neuron. Next, the length of the different orders of dendritic branches, and the mean segment length for each order--commonly regarded as important parameters in neuron physiology--are also estimated. Finally, it is shown how to estimate the mean dendritic length in a population of neurons from vertical slices of arbitrary and unknown thickness. Being unbiased and highly efficient, the proposed methods offer interesting alternatives to current procedures used for the metric analysis of neuron arborizations.

Dendritic Cells↗

PubMed1991

The application of total vertical projections for the unbiased estimation of the length of blood vessels and other structures by magnetic resonance imaging.

A new stereological method has recently been developed to estimate the total length of a bounded curve in 3D from a sample of projections about a vertical axis. Unlike other methods based on serial section reconstructions, the new method is unbiased (i.e., it has zero systematic error). A basic requirement, not difficult to fulfill in many cases, is that the masking of one structure by another is not appreciable. The application of the new method to real curvilinear structures using a clinical magnetic resonance (MR) imager is illustrated. The first structure measured was a twisted water-filled glass tube of known length. The accuracy of the method was assessed: With six vertical projections, the tube length was measured to within 2% of the true value. The second example was a living bonsai tree, and the third was a clinical application of MR angiography. The possibility of applying the method to other scientific disciplines, for example, the monitoring of plant root growth, is discussed.

Blood Vessels↗

PubMed1990

Increase in hepatocyte and nuclear volume and decrease in the population of binucleated cells in preneoplastic foci of rat liver: a stereological study using the nucleator method.

Gamma-glutamyltranspeptidase-positive hepatocyte foci were produced in female rats given a single dose of diethylnitrosamine neonatally after birth and, after weaning, a diet containing phenobarbitone for 30 wk. The nucleator method, a new stereological approach, provided an efficient, unbiased estimate of mean cell volume in focal lesions and extrafocal areas. It also provided an unbiased sample of cells to estimate hepatocyte nuclear volume and the percentage of binucleated cells. The results showed an increase in the mean volume of mononucleated cells--from 4,700 micron3 in extrafocal areas to 12,700 micron2 in foci--and of binucleated cells--from 6,900 micron3 to 25,000 micron3. This demonstrated the hypertrophic effect of the carcinogenic treatment in focal lesions. A striking reduction in the proportion of binucleated cells was also observed in the preneoplastic lesions. Nuclear volume measurements from mononucleated and binucleated hepatocytes were used to assess ploidy. An apparent increase in nuclear ploidy, with no change in cellular ploidy, was noted in focal tissue when compared with nonfocal tissue. This appeared to be caused by an increase in mononucleated tetraploid cells and a reduction in binucleated cells with two diploid nuclei, indicating an altered mitotic mechanism in focal lesions. The significance of these changes in cell volume, apparent ploidy levels and binuclearity in preneoplastic foci is discussed in relation to the hepatocarcinogenic process.

Animals↗

PubMed1990

Ultrastructural changes in chemically induced preneoplastic focal lesions in the rat liver: a stereological study.

Ultrastructural changes were investigated and quantified, using a stereological approach, in early gamma-glutamyltranspeptidase (GGT)-positive focal lesions, induced in the rat liver by treatment with a single initiating dose of diethylnitrosamine (DENA) followed by promotion with phenobarbitone (PB) for 30 weeks. Within the extra-hepatocyte environment of focal tissue, the mean volume occupied by Ito cells was markedly decreased, whilst that occupied by endothelial and Kupffer cells was increased, when compared to uninvolved tissue from the same rat livers. The bile canaliculi were dilated, but no significant differences in the mean volume occupied by the sinusoidal and Disse spaces were noted. In focal hepatocytes there was a striking overproduction of lipid droplets and proliferation of smooth endoplasmic reticulum (sER). Whorls of concentrically arranged, parallel ER membranes were found only in the hepatocytes of preneoplastic foci, in association with the proliferated sER, and never in the surrounding, uninvolved tissue. The increase in mean volume of the sER, lipid droplet and cytoplasmic matrix compartments, together with the appearance of whorls, were the major contributing factors to the marked hypertrophy seen in focal hepatocytes. The mean volume of the rough endoplasmic reticulum, mitochondrial, lysosomal, peroxisomal and nuclear compartments per hepatocyte also increased, but contributed to a lesser extent to the cellular hypertrophy. It is speculated that whorls may be structural adaptations, resulting from a possible alteration in the normal feedback control of cholesterol synthesis, for the production of sterols and the biogenesis of sER in eosinophilic-type focal cells. The significance of changes observed in focal tissue, and the high biological variation noted between foci, is discussed in relation to the hepatocarcinogenic process.

Animals↗

PubMed1990

Estimating the total number of lymphatic valves in infant lungs with the fractionator.

The fractionator is illustrated by means of a biomedical example involving the estimation of the number of lymphatic valves in lungs of infants who had died from sudden infant death syndrome (SIDS) and other known causes. The method is unbiased irrespective of tissue deformations and it does not require external information such as section thickness. An upper bound of the coefficient of error of the estimate of the number of valves within one lung was 6.5%, despite the fact that the number of valves counted per lung at the last stage ranged between 11 and 37 only. The upper bound includes the biological variation of the number of valves among infant lungs. Some theoretical remarks are also made on the efficiency of the fractionator. It is suggested, for instance, that the initial sampling stages cause more impact on the precision of the final estimator than the subsequent stages, and that an optimal arrangement of fragments submitted to systematic sampling should have the smallest fragments at the ends, with fragment contents increasing smoothly toward the middle of the series.

Female↗

PubMed1990

Assessment of particle retention and clearance in the intrapulmonary conducting airways of hamster lungs with the fractionator.

A modified version of the fractionator was used to estimate the total number of polystyrene microspheres retained in the airways of hamster lungs at two different time points after inhalation. A systematic three-stage subsampling procedure with known sampling fractions was adopted. First, each lung was cut into slices, from which primary disectors were sampled systematically with a known sampling fraction. From each primary disector, smaller sub-disectors were subsampled, and the corresponding sampling fraction was estimated by point counting. Finally, a few particles were counted at the microscopic level in the sub-disectors, and the final estimate of total particle number (which is unbiased irrespective of any tissue deformations) was easily computed as a product of the counted number times the reciprocal of the successive sampling fractions. The error variance of each estimate was assessed from the data using a new estimator. An average of 6% of the deposited particles were retained on the epithelial surface of the intrapulmonary conducting airways shortly after the inhalation, from which at least one-third was already phagocytosed by macrophages. After 24 h, an average of 87% of the particles retained shortly after the inhalation had been cleared. The proportion of particles ingested by macrophages had increased to at least 87% in three out of four animals studied.

Animals↗

PubMed1990

Recent stereological methods for cell biology: a brief survey.

With the advent of many new tools over the last five years, stereology has become simpler, assumption-free, and more efficient but, at the same time, new terms and concepts have proliferated, which risk overwhelming potential users. The present review is intended to meet the urgent need for a structured classification and evaluation of the newest stereological methods. Being fairly comprehensive, the exposition is necessarily succinct: the reader is referred to selected references for the necessary details and examples.

Animals↗

PubMed1989

Image analysis of electron micrographs relating to mineralization in calcifying cartilage: theoretical considerations.

Biological mineralization kis a cell-mediated process which is believed to be triggered by a "nucleating agent." Various matrix structures, such as matrix vesicles, collagen fibrils and macromolecules, have been claimed to be the source of this substance, since these components have been found by transmission electron microscopy (TEM) of thin sections to be associated with early mineral crystals. Systematic image analysis of the relationships revealed in electron micrographs between specific matrix components and early mineral deposits has shown that unequivocal image interpretation is not possible. This is due principally to the problems posed by overprojection and truncation phenomena, since the structures being analyzed lie within the same dimensional range as thin section thickness. Various examples are illustrated and discussed. The site at which mineral crystals are initially laid down thus cannot be identified with any matrix structure using thin section TEM. Possible technical approaches to resolve this problem of image analysis are discussed.

Animals↗

PubMed1988

Application of the Cavalieri principle and vertical sections method to lung: estimation of volume and pleural surface area.

A practical methodology is proposed for the stereological analysis of lung and other organs using recently developed unbiased procedures. This study concentrates on the unbiased estimation of lung volume using Cavalieri's principle compared with the fluid displacement method and measurement of pleural surface area using vertical sections. Furthermore, the proposed design, in addition to the sampling of extensive slices for the initial steps, also allows sampling of vertical sections for light and electron-microscopical stereology. The procedures are described in detail by reference to biological data from the right lungs of four rabbits. We found excellent agreement between estimates of lung volume measured with Cavalieri's principle and those measured by fluid displacement. Pertinent details of the statistical analysis of the sources of variation (namely biological, systematic sectioning and point-counting variation) are given in an appendix.

Animals↗