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Phenomenon of karyolysis in cytophotomorphometric examinations

Phenomenon of karyolysis in cytophotomorphometric examinations

Annales Academiae Medicae Stetinensis Suppl 43: 1-87

Studies were performed on hepatocytes of ten rabbits' livers. The one-year-old animals were sacrificed by a blow in the suboccipital region. Under aseptic conditions each liver was divided into 9 parts. One of them was examined at once (zero time) the remaining parts were placed in sterile vessels lined with gauze moistened in Ringer's solution, which created an environment of 100% humidity. Autolysis was being conducted for 8 hours at temperature 37 degrees C, carrying out the investigations with hourly intervals. The cells for studying were obtained by preparing contact smears that stemmed from a fresh cross-section of the liver segment. The smears were fixed in formalin vapour 40% proceeding for 2 hours at temperature 21 degrees C, and subsequently in 96% ethanol for 0.5 hour. Feulgen's reaction was run in accordance with the routine principles by using the basic fuschin NI 42510 of Fischer Firm (USA). The morpho-photometric measurements of nuclei were taken by means of computed device "Morphoquant" of Carl Zeiss Firm, Jena, 1982. The following measurements of the cell nuclei were accomplished, namely: A. morphometric--surface, circumferential length, bulged surface, ratio of diameters, unstained surface; B. photometric--medium extinction, circumferential extinction, total extinction. The results of studies were documented by 95 diagrams. On the basis of measurements taken, it has been established that, during the 7 hour-long period of autolysis, nucleus of hepatocyte is subjected to dynamic transformations, whereas the plotted function does not follow a linear course. During the observation the circumference and the surface resemble a polynomial with three extremes (Fig. 12, 21, 39); similarly the mean extinction and circumference extinction (attention is attracted by its severe course as compared to surface and circumference) (Fig. 57, 66). At the time of experiment the differentiation of nuclei population undergoes changes in the direction of its uniformity which concerns, to greater or lesser degree, all the investigated features (compare figures of histograms). The isolated group of tetraploidal nuclei is characterised by occupying another place. It is defined by 3 variables in spatial arrangement (Fig. 76, 80). The analysis of correlation allowed for determination of the dynamic connections, between the studied variables, which seem to be specific for a given experimental time (Tab. 1, 2). On the ground of the performed studies 7 final conclusions have been formulated: 1. Autolysis of the nuclei of hepatocytes is non-linear phenomenon; (its course resembles multinominal graph 6 degrees with three extremes in 1, 4 and 6th hour. 2. Photomorphometric parameters allow for determining the status of the nucleus of hepatocyte in the course of autolysis. 3. The fourth hour of autolysis is a characteristic point after which no response of nucleus to deteriorating oxygen condition was observed. 4. In postmortem autolysis of the nuclei of hepatocytes pyknosis is found to appear, there is no phenomenon of karyorrhexis. 5. After a 7-hour-long experiment, Feulgen's diffusional reaction starts in cytoplasm. 6. A group of tetraploid nuclei is being isolated in the course of the autolysis of hepatocytes. 7. In the course of experiment there appeared a small quantitative loss of nuclear DNA.

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Accession: 046973586

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PMID: 9857529

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