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Polyclonal cervical tumors detected by flow cytometry.

In a prospective study, the tumors of 167 patients with invasive squamous cell carcinomas of the uterine cervix were investigated by flow cytometric measurements of the DNA-content. In 37 patients (22%) 2 or more stemlines could be detected, indicating clonal heterogeneity. The proportion of multiple stemlines was equal in stages IB to III but significantly higher in stage IV (p less than 0.01). In 14 of the 37 polyclonal tumors the DNA-value of the second peak was twice the value of the first peak, suggesting possible polyploidization. For those tumors with evidence of polyploidization that had values in the diploid, tetraploid or octaploid region (n = 9) the mean age was 35.6 +/- 11.7 years and for grossly aneuploid tumors (n = 5) the mean age was 73 +/- 12.6 years. The differences of mean age between the two groups differed significantly (p less than 0.001). Flow cytometry proved to be a useful, rapid, and sensitive method for detection of multiple stemlines.

Adult↗

PubMed1986

Correlation between automated DNA ploidy measurements of Hürthle-cell tumors and their histopathologic and clinical features.

The treatment of Hürthle-cell tumors of the thyroid is controversial because of their rarity and the inconsistent histopathologic criteria for their diagnosis. In order to obtain more objective criteria for the management of Hürthle-cell tumors, the nuclear DNA content of cells from 20 cases was measured with the MicroTICAS system and the correlation between the DNA distribution patterns and the clinical and histopathologic findings was evaluated. Three main DNA patterns were found: euploid, polyploid and aneuploid. The euploid or polyploid Hürthle-cell tumors came from patients who did not develop distant metastases or recurrence whereas the aneuploid variants came from patients who died of their disease and/or developed distant metastases and recurrence. Various correlation analyses were performed between DNA ploidy and age, sex, size of tumor, growth pattern, pleomorphism, invasion and metastases. Our data suggests that an aneuploid DNA pattern or one with a large percentage of aneuploid nuclei with DNA content exceeding 5N may predict eventual metastases or recurrence from Hürthle-cell tumor.

Adenoma↗

PubMed1986

[Cell kinetics and histological types of gastric cancer as studied by DNA-RNA cytofluorometry after acridine orange fluorescence staining].

In order to clarify the relation between histological classification and biological behavior of individual tumor cells, we quantitatively analyzed the cell kinetics of 56 advanced gastric cancers using DNA-RNA cytofluorometry after acridine orange fluorescence staining and compared the results with histological findings. The gastric cancers examined could be classified into two main groups based on the ploidy patterns constituting the cell population kinetics. In group I, tumor cells were found to be chiefly composed of diploid cells with many S-phase cells even at their advanced growth stages, and their morphological characteristics were shown to correspond to undifferentiated carcinoma (according to Nakamura & Sugano). In group II, tumor cells were composed of various classes of polyploid cells as well as of diploid cells with an increased number of S-phase cells, and the extent of polyploidization appeared to progress with the growth of the tumor. The morphological characteristics of this group tended to correspond to differentiated adenocarcinoma. It is, therefore, concluded that both the histological structures and cell morphology have close relationships with cell kinetics in the human gastric cancers.

Acridine Orange↗

PubMed1979

[Regenerative reactions of myocardiocyte nuclei in ischemic heart disease].

The ploidy of myocardiocytes nuclei in the left ventricle was determined stereometrically and microspectrophotometrically in 20 cases of ischemic heart disease (IHD) and 9 control cases. An increase in the number of nuclei and the degree of their ploidy and their correlation with the myocardium mass were established. The number of polyploid nuclei was particularly high in the periphery of postinfarction scars. These data indirectly indicate the participation of polyploidization and amitosis of myocardiocytes nuclei in the development of myocardial regenerative hypertrophy.

Adult↗

PubMed1979

[Studies on phenotype, development, and viability of human spontaneous abortuses with acrocentric trisomies and polyploidies: with reference to the relationship of the viability to the origin of extrachromosomes (author's transl)].

The specimens to be presented consist of 65 trisomics and 37 polyploids which were obtained from a cytogenetic survey of human spontaneous abortion with various chromosome banding techniques. The phenotype of the 102 abortuses was analysed macroscopically before and after fixation of the tissues. The developmental age was estimated with the crown-rump measurement and from stage of Streeter's horizon on embryos and fetuses, or from the size of the placenta when an embryo was lost. The degree of phenotypic abnormality and the developmental age were used on determining the viability of abortuses. This viability was examined in each type of chromosomal anomlies. Three of 4 13-trisomics and all of 4 21-trisomics showed normal phenotype and advanced development, while most of 14-trisomics, 15-trisomics and 22-trisomics had the various severe phenotypic anomalies such as blighted ovum and disorganized embryo, and less developed. These findings suggest that abortuses with the chromosomal anomalies which can be found in the newborns longer survive than those with other anomalies. In other words, these anomalies have a better viability among abortuses. Eleven trisomics and 11 polyploids were ascertained for the origin of extra chromosomes by sequential QFQ- and RFA-banding. Of 7 abortuses with trisomy 21 studied, 4 originated at the maternal meiotic non-disjunction and none was of paternal origin. On the other hand, several published studies have shown that the paternal origin is not uncommon in Down children. Therefore, as far as trisomy 21 is concerned, it is concluded that the trisomy 21 of paternal origin has much better survival rate than that of maternal origin. In triploidy, the present study could not define the relationship between the viability and the origin, since 9 of 10 abortuses studied and all of 4 triploid newborns so far reported are of paternal origin.

Abnormalities, Multiple↗

PubMed1984

[Feulgen-DNA cytofluorometry of the liver cell carcinomas].

Twenty-four autopsy cases of liver cell carcinomas were investigated by Feulgen-DNA cytofluorometry (NIKON SPM-RF 1-D) on paraffin-embedded tissue materials. The methods employed for tissue preparation and cytofluorometry were previously reported by us. The ploidy pattern based on cytofluorometry of the liver cell carcinomas exhibited wide DNA content distributions, in which many S phase cells having intermediate DNA content values ranging from diploid level to various, higher ploidy levels were included. From the comparative studies on both the histopathological findings and the ploidy pattern of the liver cell carcinomas, it was indicated that the peak value of the DNA content distribution was shifted from the diploid to the tetraploid level with the increasing appearance of the polyploid cells, and also that the degree of polyploidization appeared to correspond to the extent of the histological malignancy.

Adult↗

PubMed1980

Endomitosis in grasshoppers. I. Nuclear morphology and synthesis of DNA and RNA in the endopolyploid cells of the inner parietal layer of the testicular follicle.

Changes in chromosome number (n-16n) due to somatic polyploidization were observed in the inner parietal layer cells (IPLCs) of the testicular follicle in 30 grasshopper species. A distribution into a geometrical series originating from the diploid value was established for the chromosome number of most of the nuclei of the IPLCs. However, nuclei with chromosome numbers of n, 3n, 5n, 7n were also found. POLYPLOId cells showed morphological patterns indicative of classical endomitosis. Endometaphase was most frequently encountered in the IPLCs. At this stage, the chromosomes were sufficiently condensed so that they were individually identifiable and their numbers could be counted. A Feulgen stain-positive fibrillar halo around the central part of chromosomes produced a lampbrush-like appearance. It was shown that the presence of many endometaphases in the IPLCs was not correlated with intense chromosome replication. 3H-thymidine incorporation data as well as cytophotometric DNA measurements did not demonstrate any appreciable DNA synthesis in the IPLCs. However, these cells were found to incorporate intensely 3H-uridine, especially in the endometaphase stage. Based on these data a reconsideration of the morphological patterns of endomitosis is suggested. The chromosomes of endopolyploid cells are considered as suitable models for studies of the functional organization of actively transcribed, individually visible chromosomes.

Animals↗

PubMed1983

[Quantity of DNA, sex chromatin bodies and nucleoli in the nuclei of the muscle cells of normal and hypertrophied human atria].

Nuclei of myocytes of normal and hypertrophied human heart atrium were studied on squash preparations. Judging by the sex chromatin body pattern, it is the polyploidization that is responsible for the increased DNA contents in these nuclei. The cytophotometric DNA measurements demonstrated the up to 93% occurrence of polyploid nuclei even in myocytes of normal atrium. Myocytes of hypertrophied atrium contained nuclei of higher ploidy degrees. The total area of nucleoli per nucleus was shown to be proportional to a degree of ploidy.

Adult↗

PubMed1980

[Comparative histo- and cytospectrophotometric study of the DNA content in the nuclei of lung cancer cells].

Histo- and cytospectrophotometric examinations of the lung carcinoma (26 observations, operation material) established that such features as hetero- and polyploidy, a considerable increase in the average content of DNA and "index of DNA accumulation" progressed as long as the degree of tumour differentiation decreased. Differences between poorly differentiated forms of squamous cell carcinoma and adenocarcinoma were found (the latter had more octoploid and other polyploid cells, higher values of the "index of DNA accumulation" and a significantly higher average DNA content in the nuclei than in squamous cell carcinoma) as well as between small- and large-cell variants of undifferentiated cancer (the latter had more octoploid and other polyploid cells, higher "index of DNA accumulation" and significantly higher average DNA content in the nuclei than in undifferentiated small-cell carcinoma). The cytospectrophotometric method for DNA determination may be used as an auxilliary test for the determination of the histological form, the degree of differentiation of lung carcinoma prognosis, and choice of the method of treatment.

Adenocarcinoma↗

PubMed1984

[Impulse cytophotometry and catamnesis in breast neoplasms].

CEll suspensions from 62 cancers of breast were investigated by flow cytophotometry after staining with pepsin-Ethidium bromide. In all patients a radical operation was made, followed by radiotherapy. The surviving patients were observed for 5 years. From the DNA-histograms 48% of the tumours were classified as diploid (or peridiploid), 40% as polyploid and 11% as aneuploid. Concerning ploidy a rank with worsening prognosis was found from diploid over polyploid to aneuploid cancers. In this sense bad prognosis means: dead or relapse during 5 years, histologically undifferentiated tumours, great primary tumour, metastasis in lymph nodes. In diploid tumours bad prognosis is indicated by a higher part of S- and G2-phase nuclei in the flow histograms. Most of the findings, but not all, are proved statistically. Now as ever, histological differentiation is still the most easily statement for prognosis.

Aneuploidy↗

PubMed1978

[Correlation between the number of centrioles and ploidy of mouse liver hepatocytes].

Using the electron microscope it was shown that in interphase hepatocytes with ploidies equal to 2n, 2n.2, 4n, 4n.2 and 8n, the number of centrioles per cell exactly corresponded to the ploidy of the cell. Both in mononuclear and binuclear cells all the centrioles are accumulated in one complex in which each pair of centrioles forms a diplosome. In binuclear cells, the complex of diplosomes is situated at equal distances from each nucleus, thus making the cell centre. The involvement of the supernumerous centrioles in polyploid metaphase cells was detected for the regenerating liver of old mice. It was found that each mitotic pole had at least four centrioles. In the pole, a pair of centrioles forms diplosomes tightly connected to each other. It is suggested that the initially tetraploid cell might divide in this manner. In addition, a question is discussed on how the existence of centrioles can be associated with the mechanism of polyploidization.

Animals↗

PubMed1980

[Flow cytometric studies of human brain tumors. Part I: Human malignant brain tumors (author's transl)].

This report concerns the distribution of the DNA content in cells obtained from seven nonneoplastic, human brain tissue specimens and from eight different kinds of malignant human brain tumors of 30 patients. Analysis was carried out by flow cytometry suing suspensions of single separated cells stained with propidium iodide as DNA-intercalating fluorochrome. Normal, non-stimulated lymphocytes served as diploid controls. An average of 91% of the cells from non-neoplastic brain tissue was diploid (2C) and these cells were presumable in G0 or G1 stage of the cell cycle. In specimens from malignant brain tumors the proportion of diploid cells in G0 or G1 stage varied between 35 and 89%. In these specimens polyploid (including triploid, tetraploid and hypertetraploid) cells were frequently observed. From the distribution of their DNA content the 14 specimens of glioblastomas were classified into 3 types. Specific patterns were noted in type III-b which was characterized by two peaks having the same heights. The cerebral metastasis displayed the greatest variations in ploidy and these tumors, on basis of their DNA distribution were divided into 3 types. As shown in the present study the cytofluorometric analysis of malignant brain tumors revealed characteristic features: 1) multiple DNA peaks, 2) wide ploid variations ranging from 2C to 8C, 3) variability of DNA content patterns and distribution, 4) relatively large portions of cells in S and G2 + M phase of the cell cycle, 5) presence of heteroploid and polyploid cells. Because flow cytometry has distinct advantages over other methods in permitting rapid analysis of large populations of cells with a high degree of reproducibility, results obtained with this technique may be of significant clinical importance.

Brain Neoplasms↗

PubMed1980

Combined autoradiographic and cytophotometric study of proliferating cell populations in human gliomas with description of a logarithmic screen method of cytophotometry.

Cell populations of 19 human brain tumors were studied by the combination of autoradiographic and cytophotometric methods. In some tumors there were up to 20% polyploid cells, but in the majority of cases the number of labeled polyploid cells was minimal. In 17 cases, labeled cells were within the range 2c to 4c or 1c to 3c: i.e., mainly near-diploid or hypodiploid cells took part in the proliferation process. In two cases (a malignant astrocytoma and an oligodendroglioma) there were two classes of proliferating cells, diploid and tetraploid. In some cases, large populations of S and G2 cells were found, which were considered to be arrested within the cycle.

Astrocytoma↗

PubMed1981

[Biorhythm changes in DNA synthesis in hepatocytes exposed to organophosphates and strontium-89].

The authors irradiated albino rats with I microc/gr strontium89 with previously modelled liver injury by FOS (Agria 1050). A temporary inhibition of the methodical activity was established as well as aberrant mitoses and strongly manifested polyploidity of the hepatic cells. Those phenomena faded quickly (within 8 days), with the exception of polyploidity, which as a sign of physiological regeneration the authors admit to be adaptive reaction directed to the restoration of the functional level of the liver on account of the hepatocytes preserved. Prior to the onset of tissue regeneration, the cellular one should be terminated, due to which, the mytotic activity was highly inhibited on the 3rd day of the irradiation, the DNA-synthesis cycle being lengthened and mainly on account of tgi, i.e. during the reparative biosynthesis the cell is incapable/of developing in the mitotic phase. After the presynthetic period is over the hepatocytes rapidly overcome the inhibition, tgi is considerably shortened and from presynthesis they almost immediately enter the period of mitosis. That requires an additional protein synthesis, manifested by the changes in serum protein. S and G1 proved to be most sensitive to DNA-synthesis blocking. The presynthetic period is essentially lengthened due to lesions of the cellular structures, synthesizing substances, necessary for entering S - i.e. the hepatocytes pass T, equal to the inhibition between G1 and S, its duration being proportional to the degree of the lesion of the organ.

Animals↗

PubMed1980

[Analysis of the depression of the mitotic activity of the corneal epithelium in white rats under chronic pyrogenal stress].

It was established previously that stable depression of cell division in corneal epithelium under chronic stress is not related to variations in the index of labeled nucleus (ILN). The paper demonstrates that five-day long pyrogenal injection did not alter the daily ILN: it was 67% in control rats and 69.7% in the experimental group. Cytophotometric logical sections showed that chronic stress led to a significant increase in the number of tetraploid nuclei (from 3.2 to 7.7%). No material changes were observed in the content of nuclei belonging to other classes. No polyploidization or G(2) growth evidence were recorded either. The absence of progressive rise in tetraploid nuclei and of more pronounced polyploidization in the presence of long-term depression of the cell cycle and stable ILN is explained by circadian variations in the mitotic activity reaction to stress. During chronic stress the depression of the cell cycle observed in the day and evening time was not recorded in the morning hours at the height of the mitotic activity. This circadian shunt is conducive to maintaining modal ploidity of corneal nuclei.

Animals↗

PubMed1995

[The DNA and protein content of atrial cardiomyocytes and the size and ultrastructure of the cardiomyocytes in children with congenital heart defects].

Right atrium myocytes were obtained during open heart surgery from 4 patients with atrial septal defect, and from 1 patient with Fallot's tetrad, aged from 3.5 to 6 years. DNA and total protein contents were measured consecutively in isolated cells using two-wave-length scanning cytophotometry following double staining after Feulgen and with naphthol yellow S. All the biopsies contained polyploid myocytes: the number of 4c cells varied from 18 to 41%, that of 8c cells from 0 to 4%. The average values of cells area and protein content for tetraploid cells were 1.5 times higher that for diploid ones. In all the cases normal and little changed myocytes were dominating (69-91%). The highest number of cells with moderate and severe degenerative changes was registered in the cases of maximum degree of cell hypertrophy and polyploidization.

Cardiomegaly↗

PubMed1995

[Genetic structure and origin of the tetraploid toad Bufo danatensis Pisanetz, 1978 (Amphibia, Bufonidae) of Central Asia. Differentiation of geographic forms and genetic relationship between diploid and tetraploid species].

A hierarchical system, consisting of several levels of genetic divergence, is formed by distribution of genetic distances between diploid taxa of the green toad species complex (subspecies Bufo viridis viridis--B. v. turanensis and species B. viridis--B. sp.) and taxa representing western (B. viridis s.l.) and eastern (B. raddei) Pale-arctic regions. Formally, genetic differentiation of the tetraploid toad B. danatensis from the parapatric diploid species reaches the species level. Genetic diversity of the tetraploid species is more than twofold higher than the total diversity of two modern diploid species. This suggests that B. danatensis was formed with the participation of other unknown species in the past. The polyploid species B. danatensis is assumed to have originated by hybridization of two species groups that arose approximately 2.6 million years ago in Mediterranean and Central Asian regions of the western Palearctic. The genetic heterogeneity of geographical populations of the polyploid species indicates its polyphiletic origin. The heterogeneity range suggests that formation of the tetraploid species continued for a very long period (about 1 million years).

Animals↗

PubMed1993

Effects of the recombinant hematopoietic growth factors interleukin-3, interleukin-6, stem cell factor, and leukemia inhibitory factor on the megakaryocytic differentiation of CD34+ cells.

Using a liquid culture system and human CD34+ marrow cells, we examined the effects of recombinant interleukin (IL)-3, IL-6, stem cell factor (SCF), and leukemia inhibitory factor (LIF) on megakaryocyte (MK) growth, endoreplication, and maturation. MK proliferation, ploidy distribution, and volume were studied by flow cytometry. IL-3 was the only cytokine that, alone, induced a marked increase in MK proliferation. At a high CD34+ cell concentration, addition of IL-6, SCF, and LIF to IL-3--containing medium increased the number of MK (approximately 20%). At a low CD34+ cell concentration, IL-3 alone was a less potent inducer of MK growth, but IL-6, SCF, and their combination had a marked effect, increasing the number of MK by a factor 1.7, 2.9, and 4.4, respectively. These differences may be related to the endogenous release of cytokines in the culture. The effects of these cytokines were subsequently tested on a more mature type of MK progenitor (CD34+ cells isolated after 6 days of incubation in liquid culture). IL-3 remained the most potent cytokine, but IL-6 or SCF alone also increased MK number in comparison to unstimulated cultures. The ploidy distribution of MKs grown with IL-3 was not markedly changed by the addition of the other cytokines, with the exception of SCF, which induced a significant increase in the mean ploidy. However, in all cultures, glycoprotein (GP)IIIa+ 2N and 4N cells were present in large but variable numbers (35% to 75%). The number of these low-ploidy MKs directly correlated with MK proliferation. Therefore, we subsequently explored the absolute number of polyploid MK produced in culture. SCF, IL-6, or their combination, in association with IL-3, increased the number of polyploid MK up to fourfold. In addition, they improved the maturation of MK grown in the presence of IL-3, leading to the synthesis of demarcation membranes and platelet shedding. A similar effect of growth factors on the maturation of day 6 CD34+ cells was observed. We conclude that IL-6 and SCF have a broad range of activities on megakaryocytopoiesis, acting both on the early and late stages. However, the proliferative properties of these cytokines largely predominate in our cultures. Therefore, in the absence of a specific MK regulator, this study further extends the need for a combination of growth factors to maximize megakaryocytopoiesis.

Adult↗