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PubMed1984

[Characteristics of trophoblast cell reproduction in the connective zone of the rat placenta. I. Determination of the degree of ploidy and the number of Barr bodies in interphase nuclei].

The degree of ploidy in the interphase nuclei was determined in the connective zone of the rat's placenta on days 13 and 14 of embryo development. On day 13, the ploidy in the majority of nuclei was 2c or 4c; on day 14, the 4c nuclei were dominating, the share of 8c nuclei increasing. The number of Barr's bodies in each nucleus of the placental connective zone tends to increase with the increase in ploidy level. This is an evidence of a "genuine polyploidy" as a mechanism of the initial polyploidization of the given cell population.

Animals↗
PubMed1983

Prevention of interphase death in rat thymocytes by bisulfite.

The effect of sodium bisulfite, a specific inhibitor of chromatin proteolysis, on radiation damage in rat thymocytes in vitro was examined. Rat thymocytes irradiated with 1 kR X rays in vitro were incubated at 37 degrees C with 10 mM glucose for 4 to 6 hr. During that time development of interphase death as judged by erythrosin B uptake, release of low molecular weight DNA (free DNA), and reduction in cell size was measured. Sodium bisulfite added to the cells at the beginning of incubation exerted a marked preventive effect on radiation damage. The effect was enhanced with increasing concentration of bisulfite from 0.25 to 2 mM. The effect of bisulfite was reversible; i.e., removal of bisulfite from the cells resulted in the reappearance of the radiation damage.

Animals↗
PubMed1983

[Electron microscopic study of the chromonema and chromomeres in mitotic and interphase chromosomes].

Study of ultrathin sections of the Chinese hamster normal cells and cells treated with solutions containing decreasing concentrations of Ca++ and Mg++ ions revealed several discrete levels of DNP-fibres organization in mitotic and interphase chromosomes. In total Ca++ and Mg++ concentrations from 4 mM to 0.3 mM, the chromosomes are formed by filamentous elements or chromonema of 100-nm diameter. During gradual decrease of Ca++ concentration to 0.2-0.1 mM the chromosomes decondensate into discrete chromatin structures--chromomeres. In decondensation, the chromomeres acquire a "rosette-like" structure with an electron dense core and DNP-fibers radiating from it. Levels of DNP compactization in chromosomes are supposed to be as follows: the nucleosomal fibres of 10-nm diameter, nucleomeric fibres of 25-nm diameter and chromonema, i.e. a filamentous structure of 100-nm diameter consisting of chromomeres.

Animals↗
PubMed1984

[Interphase chromatin in lymphocytes in sex differentiation disorders].

Interphase chromatin of peripheral lymphocytes was studied in patients aged 6 to 20 years with Turner and Morris's syndromes by AO labeled fluorometry using the authors' modification of DNP cell thermal denaturation. It was shown that the lymphocyte chromatin melting profiles represent the curves with seven maxima at the following temperatures: 47 degrees, 55 degrees, 65 (+/- 2)degrees, 78 (+/- 1)degrees, 82, 88 (+/- 1)degrees, 92 (+/- 2)degrees C (P less than 0.01). There were no statistically significant differences between clinically and karyotypically different groups either in the parallelism of the melting profiles or in the fluorescence intensity of AO connected with cell DNP. In the male control group, the similar curve was obtained for lymphocyte chromatin in 25 and for normal human spermatozoa in 100% of cases, i.e. deviation specificity was revealed in the lymphocyte melting profiles, from a "classical" normal variant to the so-called male variant with sex differentiation breaks (Turner, Morris and Klinefelter's syndromes). Possible cytogenetic mechanisms of breaks are discussed.

Adolescent↗
PubMed1984

On the fusion of nucleoli in interphase.

By comparing unstimulated and phytohaemagglutinine (PHA)-stimulated human lymphocytes from peripheral blood it was found that the fusion of nucleoli in interphase is not merely a passive phenomenon but is strongly correlated to the metabolic activity of cells. The fusion is independent of the cell cycle (DNA-replication cycle).

Adult↗
PubMed1980

A quantitative scanning electron microscopic study of microvilli in interphase and mitosis of carcinogen-transformed fibroblasts.

SEM revealed different surface characteristics of C3H/10T 1/2 cells upon exposure to carcinogens. Long and short microvilli appeared as cytological markers in three morphologically different cell types of different oncogenic potential following exposure to DMBA. The results present evidence that our recording of microvilli in interphase cells is only negligibly influenced by microvilli-bearing mitotic cells. This report further substantiates that the evaluation of the oncogenic transformation in the C3H/10T 1/2 mouse fibroblast cell system by the estimation of cells bearing long microvilli seems to be a new promising approach utilizing SEM.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1982

The H1 histones and their interphase phosphorylated states in differentiated and undifferentiated cell lines derived from murine teratocarcinomas.

The histone H1 fraction from the murine embryonal carcinoma cell line F9 can be resolved by two-dimensional gel electrophoresis into at least 15 components. Ten of these components incorporate 32P from administered orthophosphate. The digestion of 3H- and 32P-labeled samples with alkaline phosphatase indicates that this cell line has at least five H1 subtypes, three of which have not been observed in the mouse before. Every subtype is phosphorylated to some extent. There are, however, striking differences among them in the degree of phosphorylation in interphase, both in the proportion of molecules present as phosphorylated forms and in the numbers of phosphate groups per molecule. Phosphorylations of two types are discernible; most phosphorylations do not significantly affect the mobility of the protein in sodium dodecyl sulfate-containing gels but one does, probably reflecting a phosphorylation-induced alteration in protein conformation. Four subtypes undergo only the first type of phosphorylation; the other subtype undergoes both types of phosphorylation. The same 5 subtypes are also present in two other independently derived embryonal carcinoma cell lines. In contrast, two differentiated (endoderm-like) cell lines derived from teratocarcinomas lack H1b, and one also lacks H1d. Our results are consistent with the view that the H1 subtypes differ from each other functionally. They also show that neither H1b nor H1d is essential for cell division.

Animals↗
PubMed1981

[Computer classification of the fusion profiles of human lymphocyte interphase chromatin].

AO fluorescent microscopy coupled with thermal denaturation of DNP cells as modified by the authors was used to study the structure of lymphocyte interphase chromatin from 164 normal persons. Analysis of the data processed by means of a Sperry Univac Computer 90/30 has demonstrated that in 40% of the cases, the melting profiles of DNP cells from normal persons represent, irrespective of the sex, a complicated but consistently repeated curve with 6 peaks at certain temperatures, i.e. that different test subjects have a pronounced "similarity" in the characteristics discussed. In the remaining cases, there have been recorded diverse but consistent deviations that correlated with the sex of the test subjects. No peak at a temperature of 85 degrees and its appearance at 82 degrees C were the most frequent deviations seen in the male group. There have been obtained altogether 5 subgroups for females and 7 subgroups for males. Thus an attempt has been made to classify for the first time the melting profiles of DNP cells from normal persons with the use of computer.

Acridine Orange↗
PubMed1993

[Prostate cancers and potential precancerous conditions: DNA cytometric investigations and interphase cytogenetics].

The topic of this investigation was to compare precancerous lesions of the prostate (prostatic intraepithelial neoplasia -PIN- and atypical hyperplasia -AH-) and invasive carcinomas concerning DNA ploidy (image cytometry/ICM) and morphologically feasible chromosomal aberrations (interphase cytogenetics/NISH). The aim was to find clues to formal pathogenesis of prostatic cancer. Prostatic tissue of 76 patients (76 areas with carcinoma, 71 with PIN, and 12 with AH) was examined by means of ICM. In 44 cases of coincidental PIN and carcinoma, the gradings of PIN and carcinoma correlated. C-values, 2,5c-exceeding-rate, and aneuploidy rate turned out to increase in PIN and carcinoma with increasing grading (P < 0.01). In some of these cases NISH was carried out in serial sections by applying centromer-(X,Y,1,7,8,10,17,18) and telomer-(1p) specific DNA probes. The result of this approach was an increase in the number of chromosomal aberrations and chromosomes involved correlating with the grading. Our conclusion is that PIN 1 could be regarded as the precancerous lesion mainly to higher differentiated carcinomas, whereas PIN 2 and 3 should be considered a preneoplastic condition mainly of moderately and low differentiated carcinomas.

Chromosome Aberrations↗
PubMed1994

Enhancement of hybridization efficiency in interphase cytogenetics on paraffin-embedded tissue sections by microwave treatment.

To enhance the efficiency of in situ hybridization in interphase cytogenetics on formalin-fixed and paraffin-embedded tissue sections we studied the effect of microwave heating as an addition to tissue pretreatment. The modified protocol resulted in a reduction of proteolytic digestion time and of effective enzyme concentration. Apart from that, reproducibility was raised and in large sections successful hybridization was more homogenous. We conclude, that the given technical recommendations should be useful for the study of chromosomal aberrations in routinely processed tissues.

Chromosome Aberrations↗
PubMed1995

Clonal analysis of progenitor cells by interphase cytogenetics in patients with acute myeloid leukemia and myelodysplasia.

Interphase cytogenetics was used to investigate the clonal origin of bone marrow (BM) cells, peripheral blood (PB) cells, and in vitro cultured progenitor cells of five patients with acute myeloid leukemia (AML) and myelodysplasia (MDS). A new in situ hybridization (ISH) technique was used to examine the origin of the progenitor cells. Two patients with respectively, trisomy 8 and polyploidy as ISH marker were studied both at presentation and during remission. At presentation, the in vitro cultured clusters of both cases appeared diploid. Therefore, despite the abnormal growth patterns, the cultured progenitors could have been residual normal cells. Alternatively, they could have originated from a preleukemic clone with a normal karyotype. In both cases abnormal BM and/or PB cells (less than 6%) were detected with ISH during remission, indicating partially or completely clonal remissions in these patients. Both patients have relapsed. One patient with trisomy 10 as ISH marker was analyzed during myelodysplastic phase and after progression to AML. On both occasions, abnormally appearing clusters were cultured. However, only part of the clusters carried trisomy 10. The presence of a subclone characterized by trisomy 10 and an abnormally growing (pre)leukemic clone without trisomy 10 may explain this observation. Monosomy 1 and 17 were respectively used as ISH markers in two other AML patients. All in vitro cultured clusters carried the numerical abnormality. Long-term liquid cultures of these leukemias were performed for 10-20 days. In both cases, no residual normal clonogenic cells could be detected. Therefore, the selective growth advantage of normal progenitor cells in long-term marrow cultures could not be demonstrated in these two patients with leukemia. This paper illustrates the usefulness of ISH to study the biology of AML at the clonogenic level during preleukemic phase, active disease, remission, and under in vitro culture conditions. It is a sensitive technique which allows individual analysis of large numbers of small aggregates and single cells in culture.

Adult↗
PubMed1994

[The localization of intermediate filaments in polykaryons in the interphase and mitosis].

Localization of vimentin filaments has been investigated in polykaryons formed by cell fusion in culture of Chinese hamster cells (line Ag17). A network of vimentin filaments was observed around each nucleus of the interphase polykaryons. This compartmentalization of the network persists after beginning of mitoses up to metaphase, both in bipolar and multipolar mitotic cells. The picture of localization of intermediate filaments is most clear in polykaryons with mitotic asynchrony of the nuclei at early stages of mitosis. In anaphase, the central part of the network disappears, and in the polykaryon only peripheral part of the network is observed. The compartmentalization of the network of intermediate filaments is disrupted by such a way. In telophase, bright peripheral fluorescence may by the result of the beginning of network reorganization, and this process will be finished in daughter cells. The role of intermediate filaments in distribution of chromosomes and centrioles between daughter cells during mitosis is discussed.

Animals↗
PubMed1994

[The centriolar complex of heterophasic homokaryons in the interphase and mitosis].

Two patterns of heterophasic centrioles occur in interphase polykaryons obtained by cell fusion: in the form of a united complex, or as groups scattered around the cytoplasm. By the start of mitosis, replication of centrioles is finished. There is no cellular center in polykaryons with mitotic asynchrony of nuclei. In these polykaryons pairs of centrioles are situated near each nucleus. In polykaryons with mitotic asynchrony the spindle forms asynchronously and mainly near condensed chromosomes. At metaphase, the spindles display a complicated multipolar organization. Associated by interpolar microtubules, the spindle poles are not always the nearest to each other and therefore kinetochore microtubules may be directed not to the nearest centrioles. Pairs of centrioles are located at the poles. The problem is discussed of the correlation between the centriole number and polykaryon ploidy, as well as of the role of multipolar mitosis in distribution of centrioles and chromosomes between daughter hybrid cells.

Animals↗
PubMed1995

Interphase cytogenetic analysis of single cell suspensions prepared from previously formalin-fixed and paraffin-embedded tissues.

Fluorescence in situ hybridization (FISH) provides a rapid and accurate method for the detection of chromosomal aneuploidy. We have developed a technique for the use of FISH on single cell suspensions produced from either formalin-fixed or paraffin-embedded tissues. Preparation of such tissues involves sequential rehydration, enzymatic digestion to release single nuclei, and hybridization with a fluorescently labeled chromosome-specific centromeric probe. In a clinical setting formalin-fixed tissue from many tissue types is readily available for additional retrospective study. FISH on formalin-fixed tissues is especially beneficial in follow-up studies of cases involving termination after prenatal diagnosis or patients with a malignant disease where previous routine cytogenetics established the chromosomal aneuploidy. The use of this technique eliminates the biases of cytogenetic analysis due to clonal selection in tissue culture, the low number of cells analyzed, and the restriction to only dividing cell populations. We have demonstrated that this application of interphase cytogenetics to the study of various formalin-fixed tissues is amenable to the detection of chromosomal aneuploidies and has specific advantages over cytogenetic analysis.

Aneuploidy↗
PubMed1995

Correlation of microscopic phenotype with genotype in a formalin-fixed, paraffin-embedded testicular germ cell tumor with universal DNA amplification, comparative genomic hybridization, and interphase cytogenetics.

We present a strategy for the evaluation of numerical copy number changes of DNA segments within a solid tumor genome that allows the correlation of microscopic phenotype with genotype in formalin-fixed, paraffin-embedded tumor material. Cells from a human testicular germ cell tumor and adjacent tissue areas with normal seminiferous tubules were selected separately from microscopically analyzed histological tissue sections, and DNA was extracted from the selected areas. After universal DNA amplification, the amplification products were subjected to comparative genomic hybridization. The results confirmed balanced chromosome copy numbers for the normal tissue area, although the analysis of the tumor tissue area revealed numerous gains and losses of chromosome segments. The comparative genomic hybridization results were used to select DNA probes for interphase cytogenetics on serial sections. We conclude that this technique allows the screening of selected tissue areas for numerical DNA alterations, thus enabling a direct phenotype-genotype comparison.

Adult↗
PubMed1994

[Damage to and interphase death of Ehrlich ascites carcinoma tumor cells at different growth stages during energy starvation and heat shock].

The reaction of the Ehrlich ascite carcinoma cells, being at different phases of their growth, to the energy deprivation (rotenone in glucose-free medium) and heat shock (HS) was investigated. The criteria of this reaction were interphase death (according to Trypan blue staining) and structural changes (appearance of big blebs). It was found that proliferating cells (from log phase), judging from the two criteria, were more sensitive to a separate action of both energy deprivation and HS, than the resting ones (from stationary phase). Under combined actions (energy deprivation plus HS), when cell damage is much accelerated, the difference in their sensitivity was revealed only in relation to structural damages. Under the action of starvation and HS, changes in ATP content in the cells of both ages were similar; the reaction of cells to both the agents, after removing the calcium from medium (by chelator), was not changed. It means that specificity of the reaction of cells being at different growth phases to the damaging agents is not determined by disturbance in their energetic and calcium homeostases. In the proliferating cells, the cytoskeletal protein aggregation under energetic deprivation proceeded faster than that in the resting ones. It is proposed that the reaction of cells being on different growth phases depends on the stability of cytoskeletal proteins and on the content of stress proteins.

Adenosine Triphosphate↗
PubMedDate not supplied

[Functional activity of the nucleolar organizer in the interphase granulocytic nuclei in chronic myeloid leukemia].

Silver staining was used to investigate transcription activity of ribosomal genes of leukemia granulocytes in the interphase in 32 patients with PA-positive chronic myeloid leukemia (CML) and 26 healthy subjects. Visual calculation of nucleolar argentophilic granules (AG) and measurements of AG total area on cytoanalyzer Morphoquant were made. Introduction of a specially devised computer program and combination of silver staining with autoradiography in vitro (3H-thymidine) made it possible to quantify the activity of the ribosome genes in each phase of the mitotic cycle (G0/1, S, G2). Moreover, a significant difference was established in AgNO3 staining capacity of mature granulocyte nuclei in normal condition and in CML.

Autoradiography↗