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Biomedical subjects

M Bauchinger

Publications and source records attributed to M Bauchinger.

At least 55 records · Page 3Linked to original sources
PubMed1996

Detection of centromeres in vinblastine- and radiation-induced micronuclei of human lymphocytes using FISH with an alpha satellite pancentromeric DNA probe.

Fluorescence in situ hybridisation (FISH) with a human alphoid satellite pancentromeric DNA probe was used to detect centromeres in micronuclei of human lymphocytes induced by gamma irradiation and by Vinblastine sulfate. In a cytokinesis-block micro-nucleus assay a dose-dependent increase of micronuclei was detected for both agents. 72-89% of vinblastine-induced micronuclei, but only 7-48% of radiation-induced micronuclei showed centromere-positive fluorescence signals. Vinblastine treatment frequencies of centromere-negative micronuclei did not increase compared to control values, nor did frequencies of centromere-positive micronuclei in irradiated lymphocytes. Since FISH with an alpha satellite DNA probe allows the direct detection of centromeric DNA sequences the spindle damaging or clastogenic effectiveness of a compound can be easily and reliably examined in a cytokinesis-block micronucleus assay in human lymphocytes.

Adult↗
PubMed1996

Combined immunophenotyping and FISH with sex chromosome-specific DNA probes for the detection of chimerism in epidermal Langerhans cells after sex-mismatched bone marrow transplantation.

Langerhans cells (LC) of the skin represent bone marrow-derived dendritic antigen-presenting cells and are therefore important in pathophysiological processes such as rejection, graft-versus-host disease, and graft-versus-leukemia-reaction after bone marrow transplantation (BMT). For understanding of these diseases, the evaluation of the chimeric status of LC following BMT is of great interest. To analyze the sex chromosome constitution of LC in the skin, we established a modified and refined technique of combined immunophenotyping and fluorescence in situ hybridization (FISH) and investigated frozen sections of skin biopsies from nine patients after allogeneic sex-mismatched BMT and of two healthy donors for control. LC were specifically labeled using a fluorescent CD1 a antibody and hybridized simultaneously with X and Y chromosome-specific DNA probes. The results of this practical application on nine leukemia patients show the appearance of donor-type LC and the persistence of host-type LC at various times (36 up to 1395 days) after sex-mismatched BMT. Complete chimerism of LC could not be detected in any case. The frequency of recipient-specific LC ranged from 7% to 92% and showed no correlation with time postgrafting. We conclude from our results of 1461 analyzed LC that combined immunophenotyping and interphase cytogenetic analysis by FISH is the method of choice for the assessment of chimerism in a particular cell type after sex-mismatched BMT. Its practical application on other tissues affected by BMT-related pathophysiological processes reveals further knowledge of the time-dependent course of chimeric patterns after BMT.

Acute Disease↗
PubMed1996

Combined cytogenetic and molecular genetic analyses of fifty-nine untreated human prostate carcinomas.

G-banding analyses and molecular genetic investigations (fluorescence in situ hybridization (FISH) and loss of heterozygosity (LOH) studies) were performed in 59 tumor and nontumorous samples of human prostate carcinoma. Clonal chromosome aberrations were detected in 16 tumors of which nine were poorly differentiated (G3) and 11 in an advanced stage (pT3). Six cases showed numerical chromosome aberrations. The most common numerical aberrations were trisomy 7 and loss of the Y chromosome each present in three tumors. Clonal structural aberrations were detected in 12 tumors. Deletions could be observed in two cases affecting chromosome 6q23 and in two cases affecting chromosomal region 16q. A structural variant of the pericentromeric heterochromatin of chromosome 9 became apparent in six cases. The Y chromosome was involved in clonal translocations in two cases, additionally an inversion occurred on chromosome 19 in one case. All clonal chromosomal changes were found exclusively in the tumor sample. For an analysis of the pericentromeric heterochromatin of chromosome 9, FISH using a chromosome 9-specific sat III DNA probe was carried out on metaphase preparations of tumor and nontumorous tissues of two cases showing var(9)(qh). The FISH data suggest a deletion in the pericentromeric heterochromatin. Loss of heterozygosity studies on chromosomal regions 10q and 16q were carried out because both chromosomes were frequently affected by nonclonal structural aberrations. Loss of heterozygosity could be verified in 11 cases.

Aged↗
PubMed1996

Genotoxicity of 4-chloro-o-toluidine in Salmonella typhimurium, human lymphocytes and V79 cells.

In the absence of a metabolizing system (S9 mix) 4-chloro-o-toluidine (4-COT) was found to be ineffective in a combination of assays for gene mutations in Salmonella typhimurium, for chromosome aberrations and sister chromatide exchanges in human lymphocytes, and for the induction of spindle disturbances in V79 Chinese hamster cells. In the presence of S9, 4-COT was also ineffective in producing structural or numerical changes in mammalian cells, but the yields of 4-COT induced revertants in S. typhimurium strains TA 100 and TA 98 were about 2-fold higher than those in controls.

Animals↗
PubMed1996

Quantification of FISH-painted chromosome aberrations after domestic radon exposure.

Chromosome painting (target chromosomes 1, 4, 12) was performed in peripheral lymphocytes from 25 occupants of nine houses with indoor radon concentrations of 210-3000 Bqm-3. Compared to a control group, the mean frequency of symmetrical translocations of the radon group was slightly but not significantly (p < 0.10) increased. A similar tendency became apparent for a comparison of two groups of subjects with cumulative radon exposures above and below 2800 Bqm-3 y. It is concluded that FISH-based measurements of stable symmetrical translocations should reflect the cumulative radon exposure to haematopoietic compartments such as the red bone marrow rather than to mature blood lymphocytes. Since, however, radon-derived bone marrow doses are low and control frequencies of translocations are very high (about 10-fold higher than the value for conventionally scored dicentrics), the observed relative increase (1.5-fold) of the translocation frequency in blood lymphocytes is too small to discriminate chronic radon exposure from background.

Adolescent↗
PubMed1996

Chromosome translocations in thyroid tissues from Belarussian children exposed to radioiodine from the Chernobyl accident, measured by FISH-painting.

Chromosome painting of chromosomes 1, 4 and 12 was performed on metaphase preparations of cultured thyroid cells to analyse the frequency of radiation-induced stable chromosome translocations in papillary thyroid carcinomas from 40 Belarussian children exposed to radioiodine from the Chernobyl accident, and from 31 reference case. As expected, we found the highest translocation frequencies in secondary thyroid tumours after radiotherapy, but there were also high frequencies in tumour tissues as well as in non-tumourous tissues from childhood papillary carcinoma samples from Belarus. Among the Belarussian tumours the cases from the Gomel region exhibited the highest frequency of translocations and five cases lie within the range of frequencies observed in secondary thyroid tumours after radiotherapy. The findings support the assumption that radiation was the principal cause of the tumours in Belarus, but they indicate also that only a minority of the Belarus cases, which have developed papillary carcinomas, were exposed to very high doses of radioiodine.

Adolescent↗
PubMed1996

Chromosome analysis by fluorescence in situ hybridization: further indications for a non-DNA-proportional involvement of single chromosomes in radiation-induced structural aberrations.

The frequencies of symmetrical complete and incomplete translocations and dicentrics induced in human lymphocytes after in vitro irradiation with 3Gy X-rays were analysed by the use of fluorescence in situ hybridization (FISH). Single whole chromosome painting (WCP) probes, specific for chromosomes 1-4, 6-10, 12, 14 and X were hybridized separately. A human pancentromeric DNA-probe was used simultaneously for unequivocal centromere detection. For both aberration types, symmetrical translocations and dicentrics, a significant deviation from a DNA-proportional distribution was found. In general, chromosomes with a higher DNA content (chromosomes 1-3, 6 and 7) were less frequently involved in the formation of symmetrical translocations and dicentrics than expected according to their DNA-content, whereas smaller chromosomes were more frequently involved. The only exception was chromosome 4, exhibiting the highest translocation frequency of all chromosomes analysed. Ratios of the yields of symmetrical translocations to the yields of dicentrics varied between 0.9 and 1.8 for the single chromosomes. The present results substantiate our previous data obtained with identical chromosomes but examined in four different triple combinations.

Chromosome Aberrations↗
PubMed1995

Quantification of low-level radiation exposure by conventional chromosome aberration analysis.

Chromosome dosimetry, in its conventional form largely based on scoring of dicentrics and ring chromosomes in human blood lymphocytes, is the most widely distributed and reliable biological technique in radiological protection to estimate individual whole-body doses of about 100 mGy of low-LET radiation. Attempts to detect and quantify effects even of lower acute doses or protracted and chronic exposures have been repeatedly performed and the results revealed inherent limitations of this approach. Most relevant items, such as extrapolating from high-dose to low-dose effects, the influence of background frequency of dicentrics on the dose estimates, dose accumulation and concomitant temporal decline of the yields of unstable dicentrics or the statistical analyses of the data and their implications for quantifying low-level radiation exposure will be discussed in this report.

Chromosome Aberrations↗
PubMed1995

Automated metaphase finding: an assessment of the efficiency of the METAFER2 system in a routine mutagenicity assay.

The efficiency of the automated metaphase finding system METAFER2 is assessed in a routine mutagenicity assay using an aneuploid rat liver cell line treated with various promutagens. Data sets generated by automated and manual selection of metaphases are compared. It is demonstrated that METAFER2 routinely allows an efficient automatic identification of metaphases not only in lymphocyte preparations, but also in preparations from mammalian cell lines with varying chromosome numbers. Although larger slide areas are required for automated compared to manual metaphase scanning, the automatic system is faster by a factor of about 5. The interactive visual elimination of metaphases of insufficient quality is an easy and fast procedure. METAFER2 allows an unbiased selection of metaphases irrespective of their appearance as homogeneously stained first or harlequin-stained second division cells. Random selection of metaphases is neither influenced by various structural chromosome changes nor by increased frequencies of sister-chromatid exchanges.

Aneuploidy↗
PubMed1995

Effect of activated oxygen species in human lymphocytes.

The cytogenetic effectiveness of activated oxygen species (AOS) generated by the superoxide forming xanthine-xanthine oxidase (X/XO) system was studied in human lymphocyte cultures. The observed chromosome damage was exclusively of the chromatid type. In the experiments a clear dependence of aberration induction on XO concentration and exposure time could be demonstrated. While using anti-AOS agents, the H2O2 antagonist catalase and the hydroxyl radical scavenger formate reduced X/XO induced chromosome damage whereas superoxide dismutase (SOD) did not. In the presence of SOD, aberration frequency was even enhanced. The results indicate that the chromosome damage is caused indirectly via H2O2 formation from spontaneous dismutation of superoxide, whereas H2O2 might be reduced intracellularly giving rise to the highly reactive hydroxyl radical. This effect might be enhanced by SOD, possibly by raising the intracellular amount of easily membrane passing H2O2. Thus, referring to chromosome aberrations, SOD, which is generally reported to protect from AOS, is capable of increasing oxygen mediated biological damage. This observation might be explained by the involvement of DNA associated transition metal, like iron or copper ions, in reducing H2O2. DNA bound copper ions, thought to be necessary for maintenance of DNA quaternary structure, might represent a generator complex for the hydroxyl radical by reduction of X/XO derived hydrogen peroxide. This might cause 'site specific damage' to the DNA which is subsequently converted into chromatid-type aberration by S-dependent misreplication and/or misrepair. This is different to the formation of radiation induced chromosome aberrations which arise by an S-phase independent mechanism.

Antioxidants↗
PubMed1995

Combined FISH painting and harlequin staining for cell cycle-controlled chromosome analysis in human lymphocytes.

We present a simple method that allows scoring of FISH-painted chromosomes exclusively in the first division human lymphocytes. It consists of a combination of FISH with chromosome-specific libraries and a differential sister chromatid staining after BrdU treatment of lymphocyte cultures. The method allows a precise quantification of induced chromosome damage for human biodosimetry.

Cell Cycle↗
PubMed1995

Chromosome painting in highly irradiated Chernobyl victims: a follow-up study to evaluate the stability of symmetrical translocations and the influence of clonal aberrations for retrospective dose estimation.

Follow-up fluorescence in situ hybridization (FISH) measurements of symmetrical translocations were performed in peripheral blood lymphocytes from 12 highly irradiated victims of the Chernobyl nuclear power plant accident biannually, between September 1991 and July 1994, to investigate the persistence of these aberration type with time post-exposure. Translocations were determined using biotin-labelled painting DNA probes for human chromosomes 1, 4 and 12 and a digoxigenin-labelled alpha-satellite pancentromeric DNA probe. In 11 of 12 cases the translocation frequencies remained fairly constant during the observation period, which allows to generate comparable dose estimates on the various sampling times. In one case (no. 9) the existence of a cell clone containing the consistent chromosome rearrangement t(1;13) (q25;q14) was identified using FISH in rehybridized slides with a digoxigenin-labelled painting DNA probe for chromosome 13 and a separate G-banding analysis. To obtain reliable dose estimates, total translocation frequency has to be corrected for the high contribution (16.5-23.5%) of this clonal translocation.

Accidents↗
PubMed1995

A proposed system for scoring structural aberrations detected by chromosome painting.

The advent of chromosome painting has brought the realization that structural aberrations can be far more complicated than previously imagined. Various investigators have devised their own nomenclature systems to deal with this difficulty, with the result that the terminology has become inconsistent and confusing. Recently, an international group of cytogeneticists experienced in chromosome painting gathered to address this issue. Results of the meeting are presented in this report, which provides a nomenclature system capable of describing chromosome aberrations that occur between painted and unpainted chromosomes, as well as aberrations involving only painted chromosomes. The nomenclature is flexible enough to describe accurately even the extensively rearranged chromosomes. As a consequence of this flexibility, the scheme upon which the nomenclature is based differs substantially from other systems of aberration classification. We call this system the Protocol for Aberration Identification and Nomenclature Terminology (PAINT).

Chromosome Aberrations↗
PubMed1995

Cytogenetic research after accidental radiation exposure.

Currently, chromosome aberration analysis in peripheral lymphocytes is the most sensitive method to estimate individual doses in accidental radiation exposures. The assessment of dose is particularly reliable in cases with acute, uniform, whole-body exposure or after irradiation of large parts of the body. However, the scenarios of most radiation accidents largely result in partial-body exposures or a non-uniform dose distribution. This complicates dose estimation especially in cases with protracted or fractionated exposures. Problems exist also for the dose reconstruction of radiation exposures occurring a long time before sampling. To overcome these problems, the Qdr method or the "contaminated Poisson" method can be used to determine meaningful dose estimates from data based on conventional scoring of dicentrics. Scoring of so-called stable translocations by the newly developed technique of chromosome painting should be particularly useful for estimating doses of past exposures or of dose accumulation. After incorporation of radionuclides with largely localized depositions in certain organs or tissues, realistic individual dose estimates cannot be achieved. Exemplified by incidents involving larger groups of the population such as in Chernobyl and Goiania and by single cases with serious overexposures, chromosome dosimetry is evaluated in the present article.

Brazil↗
PubMed1994

Dicentric and translocation analysis for retrospective dose estimation in humans exposed to ionising radiation during the Chernobyl nuclear power plant accident.

Chromosome analyses were carried out in peripheral blood lymphocytes obtained between September 1991 and March 1992 from 15 persons exposed to ionising radiation during the Chernobyl nuclear power plant accident. At present, all are being treated for symptoms of the delayed stage of the cutaneous radiation syndrome. Biological dose-equivalent estimates were determined, either by measuring the frequency of dicentric and ring chromosomes in first division unstable cells from conventional preparations (Qdr method), or by measuring the frequency of stable translocations using two-colour fluorescence in situ hybridisation (FISH) with composite whole chromosome-specific DNA libraries for human chromosomes 1, 4 and 12 (chromosome painting) and a degenerate alpha-satellite pancentromeric DNA probe. With both methods fairly comparable individual estimates between 1.1 and 5.8 Gy were obtained for 12 of 15 individuals. Three individuals exhibited no elevated aberration frequencies. Perspectives and limitations of chromosome painting for dose reconstruction of past radiation exposures are discussed.

Cells, Cultured↗
PubMed1994

Chromosome aberrations in peripheral lymphocytes from occupants of houses with elevated indoor radon concentrations.

Chromosome analyses were performed in blood lymphocytes of 25 subjects continuously living in houses with indoor radon (222Rn) concentrations exceeding 4-60-fold the German average of 50 Bqm-3. The mean frequency of cells containing dicentrics + ring chromosomes (1.3 +/- 0.3/1000 cells) and the incidence of dicentrics + ring chromosomes per cell (1.5 +/- 0.4 x 10(-3)) were significantly increased compared to the control levels (0.54 +/- 0.11 x 10(-3) for both endpoints). Taking into account the individual radiation history over the last 10 years prior to blood sampling and the life time of peripheral lymphocytes, weighted cumulative radon exposures at the time of blood sampling between 700 and 6300 Bqm-3a were derived. Although individual exposures could not be inferred from the aberration rates, a tendency for an exposure-effect relationship became apparent for two groups of subjects with a mean weighted cumulative radon exposure above and below 1800 Bqm-3a.

Adolescent↗