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PubMed1984

Cytofluorometric nuclear DNA determinations on human corneal endothelial cells.

In order to investigate polyploidization of human corneal endothelium, nuclear DNA cytofluorometry was performed. Touch smears of 31 human corneal endothelium taken from 21 autopsies and 10 donor eyes were fixed with absolute methanol and subsequently stained by the pararosaniline-Feulgen nuclear reaction. The optimum condition for Feulgen hydrolysis of the specimens was determined, and all smears were hydrolysed in 1 N HCl for 5 min at 60 degrees C. After the Feulgen reaction, the pararosaniline-DNA fluorescence was measured with an epi-illumination cytofluorometer (Olympus AH-QRFL). Polyploid nuclei, which are the result of abortive cell division after DNA synthesis, appeared with DNA contents of tetraploid (19 cases) and octaploid (two cases) classes. All other nuclei had DNA quantities within the range of the diploid class. Nuclei containing DNA between diploid and tetraploid amounts were not found. It was concluded that most human corneal endothelial cells are of post mitotic Gl population but that a few are in the generative G0 phase.

Adult↗
PubMed1987

Chromosomal aberrations induced by the restriction endonucleases EcoR I, Pst I, Sal I and Bam HI in CHO cells.

4 widely used cohesive end-producing restriction endonucleases (REs), EcoR I, Pst I, Sal I and Bam HI were tested in CHO cells for their aberration-inducing effects. It was demonstrated that all these REs significantly increased the frequencies of aberrant cells, the aberration frequencies per cell and the aberration frequencies per chromosome. The effects of REs on chromosomal aberrations are similar to ionizing radiation, but more minutes and interchange figures are observed. Polyploid cells are more susceptible to RE treatment, an interesting finding which may be explained by the mechanisms leading to the formation of polyploid cells.

Animals↗
PubMedDate not supplied

The role of blood platelets in nucleoside metabolism: regulation of megakaryocyte development and platelet production.

In higher vertebrates, different types of blood cells develop from common precursors. Mammals are unique in possessing two types of blood cells--erythrocytes and platelets--which lack nuclei. Although platelets display consistent and easily-recognisable morphological and ultrastructural characteristics and show extreme metabolic and functional versatility, they are not true cells, being produced by fragmentation of giant polyploid precursors called megakaryocytes. At present, the physiological mechanisms which regulate megakaryocyte development and platelet production are not well understood. Platelets are actively involved in metabolism of purine derivatives and a significant platelet role in pyrimidine metabolism has also been demonstrated (see previous papers). Here an attempt is made to integrate information about platelet involvement in nucleic acid precursor metabolism with current concepts of haematopoiesis, particularly megakaryocyte development and platelet production. It is concluded (i) that megakaryocytic cells are immediate descendents of haematopoietic stem cells which have become polyploid as a result of genetic damage or metabolic imbalances, (ii) megakaryocytes and platelets are the ultimate regulators of stem cell development because they control the availability of thymidine and (iii) that the production of megakaryocytes and platelets is a physiological safety mechanism which prevents fixation of genetic damage and protects other cells from potentially cytotoxic and genotoxic stimuli.

Animals↗
PubMed1995

Image cytometry determination of ploidy level, proliferative activity, and nuclear size in a series of 314 transitional bladder cell carcinomas.

Image cytometry was carried out on 281 superficial (Ta and T1) and 33 invasive (T2 to T4) bladder cancers. The parameters used to characterize these bladder tumors were: (1) histopathological grading, (2) clinical staging, (3) tumor size, (4) deoxyribonucleic acid (DNA) index (DI), (5) DNA histogram type (DHT), (6) percentage of euploid (diploid plus tetraploid) cells, (7) percentage of polyploid cells (> 5C DNA content), (8) proliferative activity (S phase fraction value), and (9) nuclear area (NA). The proliferative activity of the tumors was not related to either histopathological grade or to clinical stage, but it was related to the DHT parameter, which made it possible to identify diploid, hyperdiploid, triploid, hypertriploid, tetraploid, and polymorphic tumors. The hypertriploid tumors exhibited a significantly lower proliferative activity than the nonhypertriploid ones. Although both the DI and the NA values correlated significantly with histopathological grading, only the NA values correlated significantly with clinical staging. We further observed that some grade III bladder tumors were definitely diploid, whereas some grade I tumors were highly aneuploid. We thus hypothesize that the ploidy level of a given tumor reflects its age directly and its aggressiveness only very indirectly. In our opinion aneuploidy is only an indirect marker of aggressiveness because it reflects the fact that a malignant tumor is old, ie, has been present in a patient over a long period of time and has had ample time to express its malignancy at the clinical level. A significant relationship was accordingly obtained between tumor size and ploidy level with the highest proportion of aneuploid tumors and the highest percentage of polyploid cell nuclei being observed among the largest bladder tumors.

Adult↗
PubMed1986

A flow cytofluorometric study on age-dependent ploidy class changes in mouse hepatocyte nuclei.

To quantitate the age-dependent hyperploidization of mouse hepatic cell nuclei, a cytophotometric study using FACS III was performed. The isolated nuclei from the liver of C57BL/6CrSlc mice at various ages were stained with chromomycin A3. The fraction of polyploid nuclei was increased with age and invariably greater in male than in female animals of corresponding ages. The highest ploidy class observed was 16C, but from the analysis of polyploidization ratio, increase in 4C and 8C nuclei was considered to be the characteristic of the age-related change.

Aging↗
PubMed1983

A quantitative study of nuclear DNA of trophoblastic cells: proliferation kinetics.

The nuclear DNA contents of labeled and unlabeled nuclei of trophoblastic cells of 8 normal pregnancies, 2 missed abortions, and 14 cases of trophoblastic diseases were measured by Feulgen-DNA cytofluorometry combined with autoradiography. Syncytiotroblasts were rarely labeled and were considered to be inactive with regard to cell proliferation. Most of the labeled cells were cytotrophoblasts. The predominantly labeled cells ranged between 2 and 4C in cases of normal pregnancy and missed abortion, and a shift to a higher ploidy was seen in the more grave trophoblastic disease. From the point of cellular proliferation kinetics of trophoblastic cells, it is presumed that the cells in cases of normal pregnancy and type I hydatidiform mole are in the 2C cell cycle and in type II hydatidiform mole and in invasive mole there are heterogeneous cell populations, most of which are in the 2C cell cycle while the remaining are undergoing polyploidization or aneuploidization. In choriocarcinoma, the cells are not in the 2C cell cycle and also are probably undergoing polyploidization or aneuploidization in the hyper-4C area.

Abortion, Missed↗
PubMed1980

Sister-chromatid exchanges and chromosomal aberrations in cultured Chinese hamster cells treated with pesticides positive in microbial reversion assays.

The induction of sister-chromatid exchanges (SCEs), chromosomal aberrations and polyploids was investigated in cultured Chinese hamster cells treated with pesticides or a related compound positive in microbial reversion assays. The chemicals tested were captan, captafol, 1,2-dibromoethane (EDB), 1,2-dibromo-3-chloropropane (DBCP), 5-nitro-1-naphrhonitrile (NNN), p-dimethylaminobenzenediazo sodium sulfonate (DAPA), 2-hydrazinoethanol (HEH), vamidothion, dichlorovos (DDVP), N-nitroso-ethylenethiourea (N-nitroso-ETU), and 2,4-dinitrophenyl thiocyanate (NBT). A significant and dose-dependent increase in the frequency of SCEs and chromosomal aberrations was observed in the cell cultures treated with captan, captafol, EDB, DBCP, HEH, DDVP, vamidothion, DAPA or N-nitroso-ETU or NBT produced a significant increase in the frequency of polyploid cells, whereas the other agents did not. When compared with results from microbial reversion assays, a close correlation was observed between the ability to induce SCEs or chromosomal aberrations and the mutagenic potency in bacteria (r:0.71-0.84).

Aneuploidy↗
PubMed1987

Induction of chromosome aberrations in lymphocytes of mice after subchronic exposure to benzene.

The induction of chromosome aberrations in lymphocytes of mice after subchronic exposure to benzene was investigated. 4 groups of 5 Swiss (ICR) male mice were given orally a solution of benzene every day for 14 days except days 5 and 10. The daily doses were 0, 36.6, 73.2 and 146.4 mg/kg. Mice were sacrificed on day 15, lymphocytes were obtained by perfusion of the spleen and the cells were cultured in RPMI 1640 medium. After 48 h of culture, cells were harvested for cytogenetic analysis. A significant dose-dependent increase in the frequency of cells with chromatid aberrations were found (p less than 0.001). A significant increase in polyploid cells were also observed (p less than or equal to 0.05). This study represents the first report on the induction of chromosome aberrations and polyploid cells in lymphocytes of mice after subchronic exposure to benzene. Such dual activity of benzene suggests that benzene may be responsible for more human health problems than currently estimated.

Animals↗
PubMed1986

Chromosomal changes in a documented case of malignant histiocytosis: significance of polyploidy.

A case of malignant histiocytosis was studied by cytology, cytochemistry, electron microscopy, and cytogenetics. It was shown that the malignant cells expressed a fully differentiated histiocytic pattern with high macrophagic activity. This correlated with the presence of polyploid metaphases. The significance of polyploid cells in the definition of malignant histiocytosis is discussed.

Aged↗
PubMed1990

Cytogenetics of colorectal adenocarcinomas.

The occurrence of nonrandom chromosomal anomalies in colorectal adenocarcinomas could be demonstrated from the cytogenetic study of 100 cases. The most frequent changes are a rearrangement of chromosome 17, leading to the loss of its short arm and a loss of one chromosome 18. Three types of tumors with abnormal karyotypes can be defined. First are the monosomic-type near-diploid tumors (MD), characterized by a monosomy of both 17p and chromosome 18 mostly associated with other recurrent monosomies. In two of three cases, one or several minor derived polyploid subclones are also observed. Second are the monosomic-type polyploid tumors (MP), which have a pattern of chromosome imbalance very similar to that of MD tumors. They derive from MD tumors by endoreduplication followed by complete disappearance of the original MD clone. Third are the trisomic-type tumors (TT), which lose either 17p or chromosome 18 or none, most of the anomalies being gains of entire chromosomes. These TT tumors never undergo endoreduplication. In addition, seven tumors with normal karyotypes were found and may constitute another category (NT). A nonrandom distribution of these tumor types in relation to tumor site was observed, since in the distal colon, TT and NT tumors are underrepresented and endoreduplications are significantly more frequent. The level of chromosomal mutagenesis is two- to threefold higher in MD and MP than in TT tumors. More than 95% of the rearrangements are unbalanced, and most of them result from breakpoints located in juxtacentromeric heterochromatin. A good correlation is found between our results and the available molecular data on allelic losses. The involvement of recessive tumor suppressor genes in colorectal tumorigenesis and the possible relationship between chromosomal imbalances and deviations in metabolic pathways is described.

Adenocarcinoma↗
PubMed1991

Evolution of a near-triploid karyotype in a secondary erythroleukemia.

We report a case of erythroleukemia (EL;FAB M6), preceded by a myelodysplastic phase, in a 50-year-old male 8 years after treatment for Hodgkin's lymphoma. Cytogenetic analysis of bone marrow at time of diagnosis of EL revealed three cell lines: 1) 28 of 53 cells (53%) were hypodiploid, 43,XY,-5,-7,-12; 2) 23 of 53 cells (43%) were near-triploid, stemline 67-69,XY,+2,del(5)(q11.2),+del(5)(q11.2),+6,-7,+8,-9,-11,-12,+15,-16,der (17)t (17;?) (p11.2;?),-18,-20,-20,+22,+r, + mar (relative to a complete triploid cell); 3) 2 of 53 cells (4%) were normal 46,XY. The relative monosomies of 5, 7, and 12 in both abnormal lines suggest that the near-triploid line evolved from the hypodiploid line. A single hypodiploid cell with both del(5) and der(17) chromosomes that appeared identical to those in the near-triploid line suggests that polyploidization occurred after these structural rearrangements. While EL is not characterized by any well-defined structural abnormality, reported cases are frequently hypodiploid, with occasional cases of polyploidization, as in our patient, EL in adults without previous neoplasia or recognized mutagenic exposure has been shown to have loss or deletion of chromosomes 5 and 7, also characteristic of myelodysplastic syndromes and secondary leukemia. Our patient had a relative lack of chromosomes 5 and 7 in both abnormal clones, as well as a del(5)(q11) in the near-triploid line. This case of EL clearly demonstrates the evolution of a complex near-triploid line from a hypodiploid line, with chromosome abnormalities typical of both EL and secondary leukemia.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1993

Karyotypic analysis of CCRF-CEM and drug-resistant cell lines with stable and unstable ploidy.

Karyotypic studies were performed on methotrexate (MTX)- and vincristine (VCR)-resistant cell lines derived from the human T-cell leukemia CCRF-CEM cell line. We noted karyotypic selection and additional chromosome change associated with acquisition of drug-resistance. Furthermore, we observed that both the parental and MTX-resistant sublines, CEM/MTX R1-3, had a tendency to ploidy change. Karyotypic studies of malignant cells have shown that polyploidy is frequently a consequence of a single sporadic event, followed by growth and selection of the polyploid clone [1]. In this study, however, various karyotypic clones were identified with near- and pseudotetraploid karyotypes that appeared to be derived from different near- and pseudodiploid sidelines. Polyploidy was invariably associated with loss of at least two of the four chromosomes 8 whether the pseudodiploid sideline from which it was derived carried one or both chromosomes 8. In contrast, neither polyploid clones nor loss of chromosome 8 was noted in the CEM/VCR R cells.

Chromosome Aberrations↗
PubMed1995

Cytogenetics of malignant gliomas: I. The autosomes with reference to rearrangements.

Autosomal chromosome abnormalities are far from always detectable and, when detected, far from fully consistent in malignant gliomas. In 15 of 41 malignant gliomas, we found autosomal chromosome aberrations ranging from solitary trisomy to a wildly abnormal polyploid complement. The sequence of chromosome events appears to proceed from the normal to the near-diploid state (via structural and numerical changes) to near-tetraploidy (via polyploidization), and finally toward near-triploidy (via chromosome loss and additional rearrangements). Characteristic chromosome changes of trisomy 7 and monosomy 10 were repeatedly found, usually together in the same cell clones. In only one case was trisomy 7 an isolated change. We observed structural rearrangements of chromosomes 7 and 10 which may be of some use in mapping specific genes duplicated or deleted by the whole-chromosome changes of chromosomes 7 and 10. Nonrandom structural changes of other autosomes, including chromosomes 1, 5, and 11, fit with the model of malignant glioma as a process involving multiple genes. An unusual concentration of breakpoints in 12q13, juxtaposing it to at least five other regions, reflects the presence of genetic information in 12q13 important to the development of malignant gliomas.

Adult↗
PubMed1995

A cytogenetic study of malignant fibrous histiocytoma.

We report the results of cytogenetic analysis of malignant fibrous histiocytoma of soft tissue (MFH). Seven of 12 successfully cultured MFHs had complex clonal aberrations, including translocations, deletions, and unidentifiable marker chromosomes. Telomeric associations were observed in five and the double minute phenomenon in four of seven MFHs with abnormal karyotypes. In one case (a storiform-pleomorphic MFH, grade IV) with a complex polyploid karyotype, two clonal ring chromosomes were present, one interpreted as r(19)(p13q13), one unidentified. In two tumors, clonal structural rearrangements of chromosome 1 were seen: del(1)(q21) in a storiform-pleomorphic MFH, grade IV, and add (1)(q21 or q32), t(1;10)(p22;q22) in a myxoid MFH, grade I. The remaining five MFHs had normal karyotypes, but in two of them nonclonal, structural aberrations were found. The modal chromosome number in the studied MFHs varied widely, but the majority of tumors with abnormal karyotypes had polyploid chromosome complements (five of seven cases). Our results confirm many of the previous findings and indicate that double minutes (dmins) may be more frequent in MFH than previously reported.

Adolescent↗
PubMed1993

Cell-cycle and ploidy analysis in bone marrow and liver cells of rats after long-term consumption of irradiated wheat.

Rats were fed for 4 or 90 days either with 70% freshly irradiated wheat (0.25, 0.75 or 2.25 kGy) and 30% complementary feed or with a control diet. None of the parameters examined (food consumption, body weight, haematological analysis, histopathological inspection of thymus, lung, liver, spleen and kidney, DNA analysis of bone marrow cells and nuclei from liver cells by flow cytometry) showed any statistically significant association with the feeding regimen. Minor changes in ploidy of liver cells and cell cycling of bone marrow cells were detectable (wheat-irradiation dose-dependent increase in G2/M-phase bone marrow cells up to 0.6%, decrease of 8C nuclei up to 1.1% in liver cells). From the pattern of alterations observed in our study, radiolytic by-products of wheat irradiation with a spindle poison-like activity can be excluded. Polyploid cells do not accumulate within the 90-day feeding period. The minor effects on cell cycle and ploidy observed are qualitatively comparable with the effects seen after food restriction in animal studies. It is suggested that an altered composition of fatty acids (the components of wheat most sensitive to irradiation) is responsible for these marginal effects. Our findings may explain the earlier findings of Bhaskaram and Sadasivan (American Journal of Clinical Nutrition 1975, 28, 130-135) who reported an increase in the number of polyploid cells in the lymphocytes of malnourished children fed irradiated wheat. The most likely mechanisms for such an effect are adaptive, constitutively regulated processes, similar to those which respond to food restriction. It is concluded that the consumption of irradiated wheat does not, therefore, pose any health risk to humans.

Analysis of Variance↗
PubMed1977

Chromosome studies on bone marrow cells of Chinese hamsters fed a radiosterilized diet.

Metaphase preparations of chromosomes from bone marrow cells of Chinese hamsters were examined for mutagenic effects following the feeding of a radiosterilized diet. No increase in the incidence of structural chromosomal aberrations was observed. As far as numerical aberrrations were concerned, the proportion of cells with polyploidy increased to between 4 to 5 times the control level, irrespective of the moisture content of the diet. This polyploidy effect occurred very early, being detectable within 24 h, if the diet fed had been irradiated with an absorbed dose of 4.5 - 10(6) rad. The incidence of polyploidy remained below 0.5%, however, nor did it rise with higher radiation doses. When the feeding of the irradiated diet was stopped, the proportion of polyploid cells returned to the control level within a maximum of 6 weeks. If the diet was stored (initially) for 6 weeks following irradiation before being fed to the animals no increase in the number of polyploid cells was noted. These results are not interpreted as a mutagenic effect of the irradiated diet.

Animals↗
PubMed1982

Repetitive DNA and polyploidy in selachians.

1. The DNA reassociation kinetics have been studied in 6 selachian species: Raja asterias, Raja montagui, Dasyatis violacea, Torpedo marmorata, Torpedo ocellata and Oxynotus centrina. 2. The results obtained show that the genomes of the two Torpedo, Dasyatis and Oxynotus are polyploid if compared with those of the two rays, though this finds no correspondence in the diploid chromosome number. 3. The phenomena of polyploidization would often be followed by wide chromosome rearrangements and by a progressive divergence of the various repetitive DNA sequences. 4. The existence of polyploidy in almost all the main superorders of living Selachians suggests that this mechanism of genomic evolution may have played an important role in the phylogeny of this class.

Animals↗
PubMed1990

Flow cytometric studies of the survival of cytochalasin-induced polyploidy in Chinese hamster ovary cells.

A method has been developed to estimate the post-irradiation survival of cytochalasin B-induced polyploidization of adherent Chinese hamster ovary cell using the flow cytometer. After exposure to radiation, surviving cells are allowed to become polyploid in the presence of cytochalasin, and are detached using trypsin, fixed by the addition of glutaraldehyde and stained using mithramycin. DNA content distributions are polymodal, and the absolute number of cells per culture in any given ploidy class is estimated by reference to a non-fluorescent bead internal standard, detected using forward scatter. Post-irradiation survival is defined as the ability to reach a given DNA content, and is reduced exponentially with dose. A bioassay to determine optimum cytochalasin concentrations can be derived from the relative size of the 2C (G0/G1) peak in the DNA content distribution. At culture densities greater than about 8 x 10(4) cell/cm2 the relative number of cells reaching at least 16C is reduced, but this inhibition is partially reversible by an increase in the medium glucose concentration, but not by the use of cytochalasin D or dihydro B.

Animals↗