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At least 613 records · Page 34Linked to original sources
PubMed1994

[Nucleotide sequence of DNA isolated from protein cores of rosette-like structures (elementary chromomeres) of mouse interphase chromosomes].

Data are presented on the primary nucleotide sequence of 15 DNA fragments cloned from protein cores of rosette-like structures of mouse interphase chromosomes. According to data of a computer analysis, these DNA fragments represent protein-noncoding regions of interphase chromosomes, contain nucleotide sequences causing DNA strand bending, and contain a large amount of cis-regulatory transcriptional elements and sites closely related to sites of DNA topoisomerase II cleavage. Five clones demonstrated partial length homology with satellite DNA, short and long interspersed sequences, and extrachromosomal DNA from mouse thymocytes. Six clones showed homology with sites of initiation of replication of viruses, yeast, and mice. Six clones were found to contain consensus sequences for non-homologous recombination in somatic cells and six clones contained consensus sequences of human VTR1.1 loci.

Animals↗
PubMed1995

Interphase fluorescence in situ hybridization identifies chromosomal abnormalities in plasma cells from patients with monoclonal gammopathy of undetermined significance.

Karyotypic studies in patients with monoclonal gammopathy of undetermined significance (MGUS) have been hampered by a low percentage of bone marrow plasma cells (BMPC), which are predominantly nonproliferating. By combining cytomorphology and interphase fluorescence in situ hybridization (FISH) we investigated whether or not chromosomal abnormalities occur in BMPC from patients with MGUS. Studying chromosomes 3, 7, 11, and 18, which we found to be frequently aneuploid by FISH in multiple myeloma (MM), we observed three hybridization signals for one of these chromosomes 3 were most common, occurring in 38.9% of patients, followed by gains of chromosomes 11 (25%), 7 (16.7%), and 18 (5.6%) Among BMPC, the frequency of aneuploid cells was 18.9% +/- 13.9% (mean +/- SD) for chromosome 3, 22.3% +/- 9.2% for chromosome 11, 23.2% +/- 22.0% for chromosome 7, and 6.1% +/- 2.3% for chromosome 18. In five patients, chromosomal abnormalities were shown to be restricted to BMPC expressing cytoplasmic immunoglobulins corresponding to the serum paraprotein. No gain of hybridization signals was observed in normal and reactive plasma cells. In one patient with MGUS, metaphase cytogenetics revealed one abnormal metaphase with 47, XY, +4, and trisomy 4 was also demonstrated in a subpopulation of BMPC by interphase FISH. FISH results from patients with MGUS and newly diagnosed MM at stage IA (n = 14) indicated that aberrations involving > or = 2 chromosomes occurred significantly more often in early stage MM (P < .01). With respect to clinical and laboratory features, MGUS patients with and without chromosomal abnormalities were indistinguishable. Our results indicate that MGUS already has the chromosomal characteristics of a plasma cell malignancy.

Adult↗
PubMed1995

Interphase cells of the centric diatom, Thalassiosira fluviatilis, lack detyrosinated, nontyrosinatable and acetylated tubulin.

Within eukaryotic cells tubulin generally exists in protein families composed of closely related isoforms generated either by differential gene transcription or by posttranscriptional and posttranslational mechanisms. In this study, immunological approaches were used to examine the contribution of posttranslational modifications to tubulin heterogeneity in a centric diatom, Thalassiosira fluviatilis, and to show the spatial distribution of microtubules in these cells during their interphase. After blotting to nitrocellulose, tubulin in cell-free homogenates of T. fluviatilis was recognized by several general tubulin antibodies including one to the tyrosinated isoform, but not by antibodies to detyrosinated, nontyrosinatable nor acetylated tubulins. Immunofluorescent staining of methanol-fixed cells revealed a net-like reticulum of microtubules originating at or near the cell nucleus. For all antibodies, except one (TU-01), results obtained by immunofluorescent experiments corrobated the analysis of blotted tubulins. Furthermore, microtubules exhibited differential staining patterns corresponding to the intensity of antibody reactivity on blots. Antibody to detyrosinated tubulin, as well as TU-01, yielded a spotty pattern of fluorescence on chloroplasts. Microtubules in T. fluviatilis support normal cell function in the absence of detectable amounts of three common posttranslationally modified tubulins, perhaps due to the rigid silica frustule which maintains shape and to the absence of flagella in interphase cells.

Acetylation↗
PubMedDate not supplied

Chromosome and interphase analysis of placental mosaicism in intrauterine growth retardation.

Confined placental mosaicism is found in a higher percentage of cases of unexplained intrauterine growth retardation (IUGR) than in those of normal pregnancies. To test this hypothesis of cytogenetically abnormal placental cells associated with IUGR, we identified patients in whose fetuses IUGR was suspected during the antepartum period by clinical and serial ultrasonography and Doppler examinations and confirmed by immediate neonatal physical examination. At birth placental biopsy samples and cord blood were collected and coded. Similar specimens were obtained from non-IUGR pregnancies during that period to avoid bias in evaluation of results. These specimens were processed for standard cytogenetic studies and fluorescent in situ hybridization (FISH) of interphase cells to detect any chromosomal mosaicism in a double-blind fashion. Results were obtained on 26 IUGR placentas. Of these placentas, 22 were cytogenetically normal on standard karyotype analysis and interphase fluorescent studies. Four placentas were shown to have some chromosomal aneuploidy: one with the same chromosome anomaly in multiple cells and the other with multiple single-cell aneuploidy. Of these placentas, cytogenetic and FISH studies showed one to have a monosomy 21 cell line, two placentas showed mosaicism on karyotyping, which was not confirmed by FISH, and one placenta had a suggested chromosomal instability. All placentas had normal anatomy regardless of the chromosome findings and no chromosome anomalies were seen in any of the infants. Non-IUGR placentas showed no chromosomal-confined mosaicism and all had normal findings on placental pathologic examination.

Aneuploidy↗
PubMed1994

Interphase cytogenetic studies of bladder cancer.

Transitional cell carcinomas of human urinary bladder were studied by interphase fluorescence in situ hybridization (FISH). With current hybridization to isolated nuclei, 26 tumors were investigated and nonrandom +7, -9 and -10 were identified. Monosomy 11, tetraploidies and polyploidies were detected in invasive and poor-differentiated tumors. Hybridization on frozen sections offers another means of analysing surgical samples. FISH to vesical washings can be applied to monitor tumor progression. Hybridizations on paraffine sections and on tissues previously stored in liquid nitrogen allow retrospective studies of the archived materials. Our data suggest that the interphase FISH can become a powerful tool for cytogenetic studies of bladder cancer.

Carcinoma, Transitional Cell↗
PubMed1994

Fluorescence in situ hybridization of interphase nuclei isolated from whole blood of transgenic mice.

Homozygosity in transgenic mice is typically confirmed by the time-consuming practice of breeding suspected homozygotes to non-transgenics and looking for 100% transmittance of the transgene to offspring. We have devised a simple reproducible method for isolating white blood cell nuclei from small quantities of mouse whole blood and have utilized fluorescence in situ hybridization to confirm the presence of one (hemizygous) or two (homozygous) copies of the transgene locus in interphase nuclei fixed to microscope slides. This method should have other applications where in situ hybridization of interphase nuclei is desirable.

Animals↗
PubMed1994

[The quantitative study of the areas of active rDNA location and of the other parameters of the interphase nucleolus detectable by silver staining during the cell differentiation of the rat trophoblast].

Different quantitative parameters of nucleolar silver staining have been studied in the cambial rat trophoblast cells on the 12th, 13th and 14th days of gestation. It has been shown that the number of Ag-positive granules in the nucleoli varied from 10 to 120. The number and the total area of silver stained granules in the nuclei increased progressively in the course of polyploidization, but was not doubled passing to the next ploidy level. Nevertheless, nucleolar area increased proportionally to the ploidy degree. The mean number and the total area of Ag-stained granules as well as the nucleolar area estimated for each ploidy level did not change significantly in the course of placenta development, suggesting an unchanged level of NOR activity at the studied stages of trophoblast cell differentiation. The data obtained on the interphase nucleoli differ from the data of the analysis of the metaphase Ag-NOR at the same period of placenta development, suggesting a diversity in the interphase and metaphase NOR organization. A proportion of cells with different number of nucleoli in the cambial rat trophoblast cells was maintained unchanged in the studied period of the placenta development, the majority (80-90%) of cells contained from 1 to 3 nucleoli. Such a proportion was similar in the cells of different levels of ploidy up to 16c. In this connection the association of NORs is suggested to be in relation with switching from the polyploid mitotic cycle to the endoreduplication leading to polyteny.

Animals↗
PubMed1993

Interphase molecular cytogenetic analysis of epithelial ovarian carcinomas.

Karyotype information on ovarian carcinomas has been limited because the tumors are often difficult to culture and the resultant metaphases can have complex numerical and structural chromosomal anomalies. Fluorescent in situ hybridization is a rapid method of determining centromere copy number in metaphase cells and interphase nuclei. Fluorescent in situ hybridization was used to determine the numerical centromere complement of chromosomes X, 8, 12, and 17 and HER-2/neu gene amplification within interphase nuclei of 25 primary epithelial ovarian carcinomas. Touch preparations of the carcinomas were hybridized with two-color combinations of directly labeled alpha-satellite centromeric chromosome enumeration probes and a directly labeled HER-2/neu probe. Modal centromere copy numbers for each of the four chromosomes were used to determine numerical abnormalities relative to the flow cytometric DNA ploidy level for each tumor. Four cases were found to be normal with respect to the four chromosomes studied. In the remaining 21 cases a relative loss of chromosomes 17 (16 cases) and X (nine cases) and a relative gain of chromosomes 12 (10 cases) and 8 (nine cases) were the most common findings. In addition, the HER-2/neu gene was amplified in two of the 25 tumors. In conclusion, fluorescent in situ hybridization is an excellent method for rapid determination of numerical abnormalities and gene amplification in ovarian carcinomas.

Biomarkers, Tumor↗
PubMed1994

Analysis of interphase cells for the Philadelphia translocation using painting probe made by inter-Alu-polymerase chain reaction from a radiation hybrid.

Fluorescence in situ hybridization (FISH) probe for the identification of the Philadelphia (Ph) translocation [t(9;22) (q34;q11)] in chronic myelogenous leukemia cells was developed by inter-Alu-polymerase chain reaction of DNA from an interspecific somatic cell hybrid containing approximately 5 Mb of human DNA covering the ABL gene region on human chromosome 9q34. This probe was large enough to be effective in identifying the genomic domains yet small enough to resolve them in more than 90% of bone marrow interphase cells. Combination of the probe with a cosmid contig probe for the BCR region of chromosome 22 in two-color FISH reduced the frequency of false-positive identification of the Ph chromosome to less than 1%. The procedure allows detection of as few as 1% Ph+ cells independent of the cycling status or BCR/ABL expression level of cells, and the quantitation of non-Ph chromosome-containing interphase nuclei in the marrow of patients judged 100% Ph+ by standard cytogenetics.

Humans↗
PubMedDate not supplied

Comparative interphase cytogenetics using FISH on human ovarian carcinomas.

Analyses using fluorescence in situ hybridization (FISH) were performed on cytogenetic slides of 25 human ovarian carcinomas. Biotinylated alphoid DNA probes were used for the monocolor and bicolor detection of chromosomes #1, #7, #8, #11, #12, #17 and #20 in interphase nuclei of the target cell material. Specific loss of chromosomes #17 and #20 was the most significant finding in aberrant cell populations of the tumors, as was gain of chromosomes #7, #1, #8 and #11. By the use of probe combinations, the presence of combined gains and losses of several chromosomes within the same cell subpopulations could be shown in a series of tumors, while in others the significant numerical chromosome abnormalities found characterized various different cell populations. In summary, FISH could be shown to be a powerful tool of interphase cytogenetics, provided that its limitations are considered conscientiously.

Adult↗
PubMed1993

[Interphase cytogenetics--a new technique for analysis of acquired genetic changes in tumor cells].

Newer genetic investigations of solid tumours by means of in situ hybridization (ISH) is reviewed. In situ hybridization, ISH, is a relatively new molecular biological method which can be used as a supplement to analysis of chromosome aberrations in tumour cells. Labelled DNA or RNA sequences (probes) are bound to specific sequences on cell chromosomes. After hybridization, the complementary sequences of the probe can be visualized and observed in a microscope. In contrast to classical cytogenetics where the chromosomes are studied in the mitosis, ISH can also be carried out on non-mitotic cells. The term "interphase cytogenetics" refers to this application of ISH, which can demonstrate the distribution of cells with different genotype without previous cell culturing. Interphase cytogenetics has been used for the analysis of several different tumour types where it gives fast and reliable information about specific cytogenetic aberrations in tumour cells. It is to be anticipated that this laboratory technique will be employed increasingly in the diagnosis and follow-up of certain malignancies.

Chromosome Aberrations↗
PubMedDate not supplied

Interphase cytogenetics and flow cytometry analyses of renal tumours.

Fluorescent in situ hybridization (FISH) can be used to determine chromosome changes in human neoplasia. In our study, we have tested the feasibility of FISH to interphase cells of renal carcinoma to evaluate chromosome aneuploidies. We carried out in parallel in situ hybridization and flow cytometric studies in order to evaluate the possible correlation between numerical chromosome abnormalities and ploidies detected by flow cytometry (FCM). The ploidy of chromosomes 7, 11, 17 and 18 was investigated in three cases of this tumour utilizing specific probes. We found evaluable and comparable results in every case of renal carcinoma analyzed for both FISH and DNA FCM analyses and our results indicate that fluorescent in situ hybridization with chromosome-specific repetitive DNA probes can serve as a cytogenetic tool for the detection of numerical specific chromosome abnormalities of interphase nuclei of renal carcinoma.

Aneuploidy↗
PubMedDate not supplied

Detection of numerical chromosomal abnormalities in malignant cells in fine needle aspirates by fluorescence in situ hybridization of interphase cell nuclei with chromosome-specific probes.

The feasibility of detecting numerical chromosomal abnormalities (NCA) in malignant cells on fine needle aspirates (FNA) using the fluorescent in situ hybridization (FISH) technique was tested on clinical specimens from patients with various neoplasms. Directly labeled DNA probes specific for chromosomes 8 and 12 were used for in situ hybridization to interphase cell nuclei. Thirty-nine of 42 samples from various sites were evaluable. Based initially on the Papanicolaou-stained slides, there were 32 malignant and 7 benign samples. Blind analysis (200 cells per sample) showed that all benign samples had a normal number of chromosomes, while 27 of 32 malignant samples showed different NCA composed of 5-95% of the cell population and ranging from 1 to 10 chromosome signals per cell. We conclude that interphase cytogenetic cell analysis of FNA by FISH is (1) feasible and gives superior signals for detection of NCA, and (2) relatively simple, with a turnaround time of less than 24 hours. This method may have diagnostic and prognostic application in the study of the biologic behavior of malignant neoplasms.

Adenocarcinoma↗
PubMed1993

High frequency of monoallelic retinoblastoma gene deletion in B-cell chronic lymphoid leukemia shown by interphase cytogenetics.

Inactivation of the retinoblastoma tumor-suppressor gene (RB-1) has been associated with tumorigenicity in various human malignancies. In chronic lymphoid leukemias of B-cell origin (B-CLL) an involvement of RB-1 has been suggested based on cytogenetic data. We examined RB-1 and its chromosomal locus 13q14 in 35 cases of B-CLL by dual-color in situ hybridization to interphase nuclei and by G-banding analysis of metaphase chromosomes. In one patient (pt) a monosomy 13, and in three other pts deletions involving or encompassing band 13q14 were detected by conventional cytogenetic analysis. In contrast, in situ hybridization to interphase nuclei showed a monoallelic RB-1 deletion in 11 cases (31%). One pt showed a translocation with the breakpoint in 13q1?4 on G-banding, but on in situ hybridization analysis the RB-1 signals were not affected. Our data show that RB-1 deletions can be diagnosed accurately by in situ hybridization on the one-cell level. The frequency of RB-1 deletions detected in this study is significantly higher than previously assumed in B-CLL, and seems to be in the same range as in retinoblastoma.

Alleles↗
PubMed1996

Interphase fluorescence in situ hybridization and reverse transcription polymerase chain reaction as a diagnostic aid for synovial sarcoma.

Identification of the t(X;18)(p11.2;q11.2) that is associated with a high proportion of synovial sarcoma can be a useful diagnostic aid. The translocation results in fusion of the SYT gene on chromosome 18 to either the SSX1 or the SSX2 gene, two homologous genes within Xp11.2. Two-color interphase fluorescence in situ hybridization and reverse transcription polymerase chain reaction were assessed as approaches to identify the rearrangement in well characterized cases. The presence of the translocation, and the specific chromosome X gene disrupted, were inferred from the configuration of signals from chromosome-specific centromere probes, paints, and markers flanking each gene in preparations of interphase nuclei. Rearrangement was found in two cell lines and eight of nine tumor samples, including analysis of five touch imprints. This was consistent with cytogenetic data in four cases and reverse transcription polymerase chain reaction analysis using primers known to amplify both SYT-SSX1 and SYT-SSX2 transcripts. The transcripts were distinguished by restriction with LspI and SmaI. Contrary to previous suggestions, there was no obvious correlation between histological subtype and involvement of the SSX1 or SSX2 gene. These approaches could also be applied to the identification of tumor-free margins and metastatic disease.

Adolescent↗
PubMed1996

Automated counting of in situ hybridization dots in interphase cells of leukemia samples.

Twenty-seven samples (cell cultures prepared for routine cytogenetics) of leukemia patients with known cytogenetic abnormalities were stained by in situ hybridization for interphase cytogenetics with centromere specific probes for chromosome Nos 4, 6, 7, 8, 9, 12, 17, 18, X and Y. The number of hybridization domains per nucleus was quantified using a semi-automated system developed in our laboratory. Results of this automated counting procedure (with and without verification of the counting results by the operator) were compared with conventional cytogenetic data and with visual scoring of the number of hybridization dots. The findings show that the system is capable of analysing 1000 cell nuclei in less than 30 min, including the necessary verification of the results by the operator. Automated counting and visual scoring were in good agreement. Conventional cytogenetics and interphase cytogenetics agreed in only 50% of the cases, confirming other studies showing that conventional cytogenetic results are not always representative for the majority of the cell population.

Adolescent↗
PubMed1996

[Interphase cytogenetics of the breast tumors with fluorescence in situ hybridization (FISH) on cytologic preparation--its practice and clinical applications].

Fluorescence in situ hybridization (FISH) study with the chromosome specific probes is performed in the interphase nuclei of the routinely processed cytologic preparation of the breast tumors. Numerical aberrations on the chromosomes 1, 3, 11 or 17 were detected in more than 80% of the malignant tumors, but not in the benign tumors. Marked heterogeneity of the polysomies is noted in the malignant tumor cells. A few malignant cases revealed monosomy of chromosome 17. No apparent correlation between the numerical abnormalities and the histological features in malignant tumors is identified. These results suggest that the interphase cytogenetics with FISH for the breast tumors may be useful for differential diagnosis of malignancy. The practice and the clinical applications of the FISH study are discussed.

Biopsy, Needle↗
PubMed1996

Interphase fluorescence in situ hybridization in a Turner syndrome variant with IsoXq. A case report.

BACKGROUND: Performing fluorescence in situ hybridization (FISH) on uncultured amniotic fluid cells has been known to produce rapid diagnoses of major chromosomal aberrations. However, if the aberration involves a structural chromosomal anomaly, the FISH result may be not only uninformative but misleading. CASE: FISH with alpha-satellite probes of chromosomes X and Y was performed on the uncultured amniocytes of a prenatal amniotic fluid specimen. Eighty-five percent of the hybridized interphase nuclei displayed two signals when probing with X; no significant hybridization was found when probing with Y. This FISH result was interpreted as normal, disomic for chromosome X. Cytogenetic analysis later, however, disclosed the fetal karyotype to be 46,X,i(Xq). CONCLUSION: Interphase FISH with an alpha-satellite probe (or probe made of repeat centromeric sequences) may be useful in the detection of a numerical anomaly of a chromosome but not of a structural anomaly within the chromosome itself.

Amniotic Fluid↗