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Biomedical subjects

I T Young

Publications and source records attributed to I T Young.

35 records · Page 2Linked to original sources

Quantitative analysis of radiation-induced changes in sperm morphology.

When developing spermatogenic cells are exposed to radiation, chemical carcinogens or mutagens, the transformation in the morphology of the mature sperm can be used to determine the severity of the exposure. In this study five groups of mice with three mice per group received testicular doses of X irradiation at dosage levels ranging from 0 rad to 120 rad. A random sample of 100 mature sperm per mouse was analyzed five weeks later for the quantitative morphologic transformation as a function of dosage level. The cells were stained with gallocyanin chrome alum (GCA) so that only the DNA in the sperm head was visible. The ACUity quantitative microscopy system at Lawrence Livermore National Laboratory was used to scan the sperm at a sampling density of 16 points per linear micrometer and with 256 brightness levels per point. The contour of each cell was extracted using conventional thresholding techniques on the high-contrast images. For each contour a variety of shape features was then computed to characterize the morphology of that cell. Using the control group and the distribution of their shape features to establish the variability of a normal sperm population, the 95% limits on normal morphology were established. Using only four shape features, a doubling dose of approximately 39 rad was determined. That is, at 39 rad exposure the percentage of abnormal cells was twice that occurring in the control population. This compared to a doubling dose of approximately 70 rad obtained from a concurrent visual procedure.

Animals↗

An immunocytochemical approach to cell kinetics automation.

Antibodies specific for 5-bromodeoxyuridine (BrdUrd) can be used to measure labeling indices in an automated system by image analysis. The antibody, used with an indirect immunoperoxidase technique, will detect de novo DNA synthesis subsequent to growing the cells for various time intervals in 5-bromodeoxyuridine-containing medium. Asynchronously growing CHO cells were pulsed with 3H-5-bromodeoxyuridine, fixed, denatured and then stained with anti-bromouridine antiserum. Peroxidase-coupled goat anti-rabbit IgG was used as the secondary antibody, and slides were stained with diaminobenzidine. Cells which are positive display a reticular pattern indicative of replicating chromatin. "Labeling indices" were generated by scanning the nuclei by TV image analysis. The percentage of labeled cells by the immunocytochemical technique correlates well with that found by autoradiography. Some of the applications of this automated method include cell kinetics and analysis of S-phase by pattern recognition technique.

Animals↗

Transformation of nuclear morphology during cellular maturation.

Using in vivo labeling of mouse leukocytes with tritiated thymidine, cells in the neutrophilic series were studied to determine the change in their nuclear shape as a function of maturation level. Several morphologic shape parameters including perimeter and bending energy were used to quantify the distribution of the nuclear morphology in a given age cohort. The change in these distributions as a function of calender age level was determined. The two parameters named above were used to test the possibility of inferring the age from the quantitative morphology.

Animals↗

Loss of division potential in vitro: aging or differentiation?

We have examined the hypothesis that diploid cells grown in vitro age, and propose that only proliferative potential and not life-span is telescoped. We suggest that explanted or transplanted diploid cells are driven to divide by the process of subculturing in vitro or in vivo and, in response to this pressure, also complete their differentiation and become refractory to further mitotic stimulation. We conclude that differentiation rather than "mortality" distinguishes diploid from transformed cells and that the former may not age in vitro, but are lost because culture methods are selective for cycling cells.

Cell Cycle↗

The automated classification of mitotic phase for human chromosome spreads.

A system has been developed that automatically recognizes the mitotic phase of human chromosome spreads for karyotyping. Suitable spreads are classified into one of five subphases of mitosis. Classification is performed on the basis of summed chromosome length and most probable chromosome width. Classification requires 100-500 msec. A television camera scans the spread through microscope optics; computer and special purpose electronics process the video signals to generate run length histograms. The histograms are used to determine mitotic phase. Unbanded spreads, 133, were classified with a 4.5% error rate. One hundred banded spreads were classified with a 15% error rate.

Autoanalysis↗

Proof without prejudice: use of the Kolmogorov-Smirnov test for the analysis of histograms from flow systems and other sources.

In this paper the Kolmogorov-Smirnov statistical test for the analysis of histograms is presented. The test is discussed for both the two-sample case (comparing fn1(X) to fn2 (X)) and the one-sample case (comparing fn1 (X) to f(X)). Presentation of the specific algorithmic steps involved is done through development of an example where the data are from an experiment discussed elsewhere in this issue. It is shown that the two histograms examined come from two different parent populations at the 99.9% confidence level.

Immunoglobulin D↗

A technique for multiple-cell chromosome karyotyping.

A new approach to a system for chromosome karyotyping is presented. The system assembles the information about chromosomes from several cells at a time, thereby filtering out noise due to variations in the slide preparations. The system makes i possible to use metaphase spreads which are incomplete due to missing chromosomes, touching and overlapping chromosomes and stain particles. The system gives a precise description of the chromosome complement in terms of distribution function parameters, with the uncertainty of the parameters specificed. The system is adaptive with respect to the initial reference parameter set so that both recognition of normal chromosomes, in spite of the variation displayed among individuals, and identification of aberrant chromosomes are possible. The precise chromosome descriptors can be used to detect differences between the tested individual and various references, in order to find chromosomal abnormalities.

Autoanalysis↗

Automatic detection and localization of sister chromatid exchanges.

Sister chromatids of human metaphase chromsomes from cells which have replicated twice in medium containing 5-bromodeoxyuridine exhibit unequal fluorescence when stained with the dye 33258 Hoechst. Sister chromatid exchanges occurring in these chromosomes are apparent as interchanges of brightly and dully fluorescing chromatids. A technique for detecting such exchanges by computer analysis of chromsome images has been developed and found to campare favorably with manual methods. The exchanges have been localized in the context of quinacrine banding patterns.

Autoanalysis↗

A measurement technique for cell adhesiveness.

Determination of the percentage of cells in clumps on a stained smear of human peripheral blood porvided a useful, accurate technique for measuring cell adhesiveness. Smears of human peripheral blood drawn with EDTA were prepared on a blood slide centrifuge, stained, and examined under a light microscope. Statistical analysis showed that the method resulted in a Poisson distribution of particles on the slide, where a particle was considered to be a simple cell, or two or more cells which appeared to be touching, Analysis of the distributions of erythrocytes and leukocytes showed that clumps were formed before the cells were deposited on the slide. When adhesiveness of erythrocytes or leukocytes was increased by incubation with antiserum to the corresponding cell type, the percentage of that cell type in clumps increased proportionately, Preliminary results using the method showed that normal human donors had similar to 1% of their erythrocytes and 1-5% of their leukocytes in clumps. In chronic myelocytic leukemia, as many as 60% of the leukocytes were in clumps.

Animals↗

3D base: a geometrical data base system for the analysis and visualisation of 3D-shapes obtained from parallel serial sections including three different geometrical representations.

In this paper we discuss a geometrical data base that includes three different geometrical representations of one and the same reconstructed 3D shape: the contour-pile, the voxel enumeration, and the triangulation of a surface. The data base is tailored for 3D shapes obtained from plan-parallel serial sections. It is explained how this geometrical data base is useful with the different processing approaches of a 3D shape, such as analysis and visualisation. Methods of conversion between the geometrical representations are discussed. Examples of the operation of the data base as it is embedded in a data base management system are given by illustrations of retrieval of geometrical information.

Algorithms↗

An analysis technique for biological shape-II.

A discrete function representing the curvature of closed contours has been derived. The function is easily computable and has properties desirable for use in shape analysis. Two curvature measures (as well as the traditional P2A) have then been applied to a set of shapes extracted from leukocyte nuclei in order to evaluate their effectiveness as numeric descriptors of shape complexity. Bending energy, (BEN), was shown to be, on the average, the most sensitive indicator of complex shape, normalized mean absolute curvature (NMAC) was next, and P2A was last. Bending energy could also have important biolgical significance due to its possible relation to developmental processes. Further studies on bending energy will be necessary in order to substantiate this possibility. Many of the discrepancies between P2A, NMAC, BEN, and the study subjects are explainable by the lack of "syntactic", or form, information used by the computer features. If an observer really classifies an object by decomposing it into simpler pieces, then a more sophisticated shape analysis procedure must be used. Such a procedure will be described in a sequel to this paper.

Cell Nucleus↗

An analysis technique for biological shape-III.

The motivation for a syntactic theory of shape was a belief that computer shape analysis could mimic human recognition procedures. The results described here suggest that heuristic techniques can be used to decompose and analyze complex objects. The derived decomposition algorithm gives quite good results when compared with human lobe determinations. In addition, it seems to follow closely the actual procedures used by observers in classifying shapes. The list structure output of the program clearly describes each shape element and greatly facilitates further analysis. Extensions of this work should be directed toward the development of more specific "rules" for shape decomposition, and more complete orientation information in the output list structure.

Computers↗