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[Diagnostic advances in chromosomal analysis].

It is now known that chromosome disorders form a major category of genetic disease, accounting for a large proportion of all reproductive wastage, congenital malformations, and mental retardation, as well as playing an important role in the pathogenesis of malignancy. A variety of new techniques can be used to identify the chromosomal location of genes directly that promises to revolutionize the field of chromosomal analysis.

Chromosome Aberrations↗

Chromosome analysis of brain tumors in childhood.

We performed a chromosome analysis of 26 pediatric brain tumors, including 20 primitive neuroectodermal tumors (PNETs). 5 astrocytomas, and 1 immature teratoma. Specimens were treated with collagenase, placed in overnight or short-term cultures, and harvested for chromosome analysis. Numerical and/or structural abnormalities were noted in 14 of the 20 PNETs and 4 of the 5 astrocytomas. In 13 PNETs, so-called medulloblastoma in the cerebellum, an i(17q) was the most frequent structural abnormality, accounting for 30% (4/13). Double minute chromosomes (dmin) were observed in one tumor. Near-diploidy was demonstrated in three of these PNETs, hyperdiploidy in three, and near-tetraploidy in three. We could not find any correlation of these cytogenetic findings with the prognosis. In the remaining seven PNETs other than medulloblastoma, the karyotypes of five PNETs demonstrated a variety of numerical and structural abnormalities. As to the astrocytomas, losses of chromosomes 7 and 9 with dmin were observed in two, and structural abnormalities of chromosomes 1 and 17 were also observed in two tumors. In our limited cases, however, we could not find the same chromosome abnormalities that are well known in adult astrocytomas. A congenital immature teratoma showed hyperdiploidy with increased numbers of chromosomes 3, 6, and 12. We conclude that i(17q) is an important chromosome abnormality in medulloblastomas, and that the oncogenesis of pediatric astrocytomas might be different cytogenetically from that of adult astrocytomas.

Adolescent↗

The clinical usefulness of chromosome analysis by flow cytometry.

Historical and clinical aspects of chromosome analysis by flow cytometric methods are reviewed. A new method of preparing small samples (1.0 mL of blood) of peripheral lymphocytes for flow karyotype analysis using phytohemagglutinin and interleukin-2 is presented in detail. Figures of flow karyotypes, partial banded karyotypes, and idiograms of patients with inv(8), rec dup(8), rob t(14;21), and t(1;22) are presented, as well as examples of univariate and bivariate flow histograms from other researchers' published works. The clinical utility of these techniques is explored, with specific reference to recent work in chromosome polymorphisms and cultured amniocyte lines. We conclude that, although flow karyotyping is not a replacement for classical banded chromosome analysis, used as an adjunct, this new technique has some clinical usefulness relating to its capability to resolve small differences in chromosomal DNA content.

Animals↗

How necessary is a chromosomal analysis in growth-retarded girls?

In growth-retarded girls, shortness may be the only clinical sign of Turner syndrome. Therefore recommendations have been published that a chromosomal analysis should be carried out in every girl showing growth retardation. The present study was carried out to examine whether the head circumference of small girls can serve as a criterion and hence limit the need for expensive chromosomal analysis. The head circumferences of 45 girls with Turner syndrome were compared with those of 41 growth-retarded girls. It could be shown that the heads of the patients with Turner syndrome had a normal circumference, whereas the heads of the control group were significantly smaller. In view of this result, the risk of overlooking Turner syndrome in diagnosing growth retardation without a chromosomal analysis could be estimated to be 1:9000. The conclusion is drawn that a chromosomal analysis is not necessary in girls whose growth retardation is combined with a small head circumference and who do not show other malformations.

Chromosome Aberrations↗

Biodosimetry after accidental radiation exposure by conventional chromosome analysis and FISH.

A 137Cs source was stolen from a radioactive waste depository in Estonia on 21 October 1994 and kept in a private house for 4 weeks. This resulted in the death of one person, acute radiation injuries to four people and exposure of several other people to lower doses of radiation. Analysis of chromosomal aberrations in peripheral blood lymphocytes was used in the assessment of radiation exposure of 18 people involved in the accident. Dose estimation assessment based on the frequencies of dicentric chromosomes was performed both by the standard method and by considering possible dose protraction and non-uniform exposure. Considerable differences in dose estimates were obtained depending on the approach used, ranging from about 1 Gy to almost 3 Gy in the patients most heavily exposed. In view of the deterministic health effects observed in some of the subjects, it was concluded that the dose estimates involving information on dose protraction were more realistic than those obtained by comparison with the standard high dose-rate calibration curve. Chromosome painting analyses using fluorescence in situ hybridization, with a probe cocktail for chromosomes 1, 2 and 4 and centromere detection, were performed in parallel. Good agreement on dicentric chromosome frequencies was observed between the conventional and painting analyses. The frequencies of complete translocations were comparable with the frequencies of dicentric chromosomes. In addition to the complete translocations, a pronounced increase in the frequency of incomplete translocations was observed. Dose estimates performed on the basis of FISH translocation frequencies were consistent with the dicentric analysis.

Adolescent↗

Chromosome analysis in hematologic disorders. The leukemias.

For two decades, cytogenetic studies have been used to rule in (or out) the Philadelphia (Ph1) chromosome associated with chronic myeloid leukemia. Beyond this single purpose, chromosome studies have generally not been utilized in or applied to the practice of hematology-oncology. This report presents male and female patients, teens to 70s in age, with representative hematologic disorders, in whom the cytogenetic findings were useful clinically. These cases illustrate the following principles: (1) hematologic disorders can be characterized by chromosome analysis; (2) chromosome findings help in the diagnosis, prognosis, and treatment of blood diseases; (3) blood and bone marrow samples can be processed routinely for cytogenetic analysis; (4) these samples can be transported long distances from clinic to laboratory; and (5) the contemporary practice of hematology and oncology requires chromosome analysis for fuller evaluation and understanding of hematologic conditions.

Adult↗

Clinical performance of a system for semiautomated chromosome analysis.

Until recently equipment for automated chromosome analysis has not been used for routine purposes in clinical cytogenetic laboratories. During a 3 1/2-year period the chromosome laboratory of Rigshospitalet has tested the Magiscan chromosome system under routine conditions and performed the first evaluation of its clinical performance. The system consists of an image processor with a light pen for manual interaction connected to a hard-copy printer and a microscope with a TV camera and a motorized scanning stage for eight slides. Automated metaphase finding takes place without operator assistance. An operator is involved in the analysis after the metaphases are located. Using two of these complete systems, we have performed a total of 4,691 chromosome analyses comprising a count of 10 metaphases, of which three were "eyeball" karyotyped and one was "machine" karyotyped. Presently, two-thirds of our prenatal analyses (amniotic-cell cultures) are carried out with these two machines. A third Magiscan system without scanning stage is used as a "karyotyping-only" system to produce hard-copy karyograms in those cases in which metaphases are manually located and counted in the microscope. Since the end of 1984, 4,773 additional machine karyograms have been produced with this system. With a complete system, a prenatal analysis can be carried out in an average of 35 min. The average time for a machine karyotype is 7 min. Since 1984 the productivity of the laboratory has increased 17%-20% without enlarging the staff.

Female↗

Diagnosis of pleural effusions by chromosome analysis.

Cytogenetic changes, namely numerical (aneuploidy) and morphologic chromosome abnormalities including markers, clearly indicate the presence of malignant cells. To compare the utility of chromosome and cytogenetic analysis, the idiopathic pleural effusions of 60 patients were subjected to a double-blind analysis. A fluid was considered "positive" for malignancy if there was a marker chromosome, or if 10 percent of metaphases were hyperdiploid. Of the 33 neoplastic effusion, 30 (91 percent) were diagnosed "positive" by chromosome analysis whereas only 21 (64 percent) were classified "positive" or "suggestive positive" by cytology. All 27 benign effusions were correctly diagnosed by cytology; however, two were labelled "positive" by chromosome analysis. Combining the results of cytologic and chromosome analysis did not increase the number of "positive" diagnoses.

Chromosome Aberrations↗

Image quality in digital chromosome analysis systems.

This paper reports on an investigation into the differences in image quality of different components used in a digital image processing system for chromosome analysis. As chromosome aberrations are important tools in the cloning of genes, it is important to know if the introduction of computerized analysis systems increases the risk of missing small aberrations. In this investigation the number of visible bands on a number of chromosomes has been used as a measure of quality. The images compared are microscope ocular images, photographs from a microscope built-in camera, digital images from a high and from a standard resolution camera, presented both on screen and print-out on paper. The main conclusions are that: (1) the view in the microscope ocular gives the best resolution, (2) there are risks of losing vital information using the digital image processing system for chromosome analysis, and (3) this risk is significantly reduced when using a high resolution camera.

Chromosome Banding↗

Automation of routine clinical chromosome analysis. II. Metaphase finding.

Metaphase finding is an essential activity in chromosome analysis, and there is much to gain from its automation. This paper describes software for automatic metaphase finding developed for use as part of a routine clinical chromosome analysis system, principally for samples from blood and amniotic fluid. Since the metaphase finding and analysis programs were intended to be used widely in clinical laboratories, cost and portability were important design features. The metaphase finder has been implemented on a moderately priced, general-purpose image analyzer (Magiscan 2), which controls a standard research microscope with motorized stage and focus. Metaphases are detected using fast gray-level processing on whole fields of view, followed by binary processing to produce a figure of merit for each detected object. Clinical experience has shown that this ability to rank detected objects on the basis of their suitability for analysis is a critical feature in determining the usefulness of an automatic metaphase finder.

Amniotic Fluid↗

A simple reproducible method for prometaphase chromosome analysis.

A method is described for the analysis of chromosomes in prophase and early metaphase. It involves culturing the lymphocytes in medium RPMI-1640, supplemented with 10% autologous plasma instead of fetal bovine serum. Living cells are treated with actinomycin D and colcemid for 1 h prior to harvest and harvested early at 65 h of incubation, using a hypotonic solution formulated by Ohnuki (1968). The method has been tested on several hundred clinical samples on a routine basis. On average, 30% of the dividing cells were in prometaphase.

Azure Stains↗

Chromosomal analysis in multiple myeloma: cytogenetic evidence of two different diseases.

We report the cytogenetic results obtained in 81 multiple myeloma (MM) patients with abnormal karyotypes. Most karyotypes were complex with numerical and structural abnormalities but the analysis of chromosomal abnormalities allowed identification of two cytogenetic patterns depending on the chromosome number: a first hyperdiploid pattern (54%) with recurrent trisomies 3, 5, 7, 9, 11, 15 and 19 and a second pattern (46%) showing either pseudodiploid, hypodiploid or near-tetraploid karyotypes. Structural abnormalities were present in all but five hyperdiploid karyotypes, and frequently involved lymphoid breakpoints: immunoglobulin gene regions (36 cases) or chromosome 11q13 region (21 cases). Numerous other structural aberrations were detected; the most frequent involved chromosome 1 and chromosome 13. Structural abnormalities were significantly more frequent in the second hypodiploid group. When analyzing the results obtained in the 60 patients studied at the time of diagnosis, a prognostic correlation was found between the cytogenetic pattern and overall survival: hyperdiploid patients had a longer survival than patients belonging to the pseudo/hypo/near-tetraploid group (median survival 36.8 vs 18.2 months, P < 0.04). These results suggest that MM could correspond to two closely related diseases.

Adult↗

Clastogenic effects of known and suspect spindle poisons studied by chromosome analysis in mouse bone marrow cells.

The present study was performed within the project 'Genomic Mutations' (sponsored by the Commission of the European Communities) in order to gather all possible experimental information on 10 chemicals selected on the basis of their possible capacity to induce aneuploidy. An analysis of chromosomal aberrations was carried out in bone marrow cells of mice with the first five chemicals: colchicine (COL), econazole (EZ), chloralhydrate (CH), hydroquinone (HQ) and diazepam (DIAZ). The experiments were performed in parallel to micronucleus tests with the objective to distinguish a positive micronucleus response due to chromosomal breakage from that obtained by lagging chromosomes. The results of the micronucleus tests will be reported elsewhere. COL, CH, EZ and DIAZ showed no clastogenic effects in mouse bone marrow cells after single intraperitoneal injection. Polyploid cells were significantly more frequent after COL treatment. HQ showed a dose-dependent induction of chromosomal aberrations at 6 and 24 h after treatment. After 24 h, cells with multiple aberrations up to complete chromosome fragmentation were frequently observed. They indicate that a small fraction of the cell population, probably related to a specific stage of the cell cycle, was particularly sensitive to HQ. A sex difference in clastogenic response to HQ was not observed. It is concluded that of the five chemicals tested only HQ was clastogenic in mouse bone marrow cells under the present experimental conditions.

Aneuploidy↗

Canalization at two extra scutellar bristles: artificial selection, chromosome analysis, and effect of temperature.

Two isofemale lines (P1 and P2) from a wild-type population of Drosophila melanogaster were selected for an invariant phenotype, two symmetrical and anterior scutellar extra bristles. Both P1 and P2 lines responded quickly to direct selection for two extra bristles, but although this phenotype was canalized in the P2 line, it was not in the P1 line. This lack of canalization at two extra bristles, measured by probit width, is the first reported in the literature. Analysis of chromosomal contribution showed the P1 decanalized phenotype to be due to chromosome 2. Synthetic chromosomal combinations were tested at four development temperatures (17 degrees, 21 degrees, 24 degrees, and 30 degrees C), and no correlation was observed between canalization at two extra bristles measured by probit width and minimum phenotypic change due to different temperatures. In this test, development at the highest temperatures was accompanied by an increased tendency to return to the wild phenotype in the canalized phenotypes only, suggesting that this drastic change could be accounted for by the action of the wild-type canalizing system. Decanalized genotypes, or those canalizing at phenotypes other than the wild type, could be explained by mutations which, to a greater or lesser degree, alter the normal genetic system of canalization.

Animals↗

Chromosome analysis in the management of patients with non-Hodgkin's lymphoma.

Analysis of chromosome abnormalities in non-Hodgkin's lymphoma has stimulated a basic inquiry into the mechanisms of gene deregulation in human tumors. A growing literature has also correlated cytogenetic and molecular genetic features of lymphoma with patient prognosis and other clinical features of the disease. The goal of this report is to review those genetic abnormalities which are currently useful in the clinical management of patients with lymphoma, and to provide a conceptual framework for this growing literature.

Chromosome Aberrations↗

[History of modern chromosomal analysis. Differential staining of plant chromosomes].

Differential staining methods found extensive use in karyotype studies of many plant and animal species and provide for reliable identification of all chromosomes of the organism. Below we describe the most widespread methods and history of their advent. In addition, we discuss specific structure of the chromosomes and possible mechanisms responsible for differential segmentation.

Chromosomes, Plant↗

Human sperm chromosome analysis--study on human sperm chromosome mutagenesis induced by carbon disulfide.

The aim of this study was to investigate the effect CS2 of on human sperm chromosomal aberration. The human sperm/hamster egg fusion technique was used to analyze 203 human sperm chromosome complement from 9 healthy volunteers. The incidence of numerical aberration was 1.0% and that of structural chromosome aberration was 5.9% and total abnormalities was 6.9%. Structural aberrations consisted of breaks, deletions, centric rings, fragments, and chromatid exchange. The results from high concentration group (10 mumol.L-1 CS2) showed that the incidence of chromosomal aberration rate was significantly higher than that of the control group. The results indicate that high concentration of CS2 might directly cause mutagenesis of the germ cell.

Adult↗

Chromosome analysis in couples with recurrent abortions.

Chromosome analysis using a G-Banding technique was performed in 35 couples (70 individuals) with a history of two or more spontaneous abortions of unknown cause. Among these individuals, 5 (7%) showed balanced translocations, all of whom were female. The outcome of 31 pregnancies of 10 balanced translocation carriers was as follows: Ten of the offspring had normal phenotypes (32%), 5 (16%) were born with chromosomal abnormalities and 16 (52%) were spontaneously aborted.

Abortion, Habitual↗