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Biomedical subjects

M Bauchinger

Publications and source records attributed to M Bauchinger.

At least 37 records · Page 2Linked to original sources
PubMed1998

Molecular genetic characterization of the Philadelphia chromosome detected in reactor personnel highly exposed to radiation from the Chernobyl accident.

Clonal del(22q) chromosome aberrations were coincidentally observed in highly exposed reactor personnel of the Chernobyl power plant accident in the course of retrospective biological dosimetry. These aberrant chromosomes were detected in PHA-stimulated cultures from peripheral blood after FPG staining and revealed a morphology similar to a Philadelphia chromosome. A rearrangement of the BCR gene on 22q11 could be confirmed in unstimulated peripheral blood by RFLP analysis from three of four del(22q) carrying cases. FISH analysis of the del(22q) carrying cases with BCR- and ABL-specific DNA probes additionally exhibited a BCR-ABL fusion in 5.2 to 9% of cells in unstimulated blood. Breakpoints within the BCR gene could be located either in the M-bcr or the m-bcr region and thus, a specific breakpoint region could not be detected in these four patients. Since typical clinical leukemic symptoms associated with the translocation (9;22)(q24;q11) could not be observed in these highly irradiated subjects (1.1 to 5.8 Gy), the role of this particular aberration in the development of a radiation-induced leukemia remains obscure.

Chromosome Deletion↗
PubMed1998

Genetic heterogeneity in a prostatic carcinoma and associated prostatic intraepithelial neoplasia as demonstrated by combined use of laser-microdissection, degenerate oligonucleotide primed PCR and comparative genomic hybridization.

We combined laser-assisted microdissection from H&E-stained paraffin sections, degenerated oligonucleotide-primed polymerase chain reaction (DOP-PCR), and comparative genomic hybridization (CGH) to analyse chromosomal imbalances in small tumour areas consisting of 50-100 cells. This approach was used to investigate intratumour genetic heterogeneity in a case of metastatic prostatic adenocarcinoma and chromosomal changes in areas of prostatic intraepithelial neoplasia (PIN) adjacent to the invasive tumour. In four microdissected invasive tumour areas with different histological patterns (acinar, cribriform, papillary and solid) marked intratumour heterogeneity was found by CGH. Recurrent chromosomal imbalances detected in at least two microdissected tumour areas were gains on 1p32-->p36, 2p22, 3q21, 7, 8q21-->q24, 11q12-->q13, 16p12-->p13, 17, 19 and loss on 16q23. Additional chromosomal changes were found in only one of the microdissected areas (gains on 16q21-->q23, 20q22 and losses on 8p21-->p23, 12p11-->q12, 12q21-->q26, 13q21-->q34, 16q12, and 18q22). In PIN, gains on chromosomes 8q21-->q24 and 17 were found in both samples investigated (low and high grade PIN), while gains on chromosomes 7, 11q, 12q, 16p, and 20q and losses on 2p, 8p21-->p23, 12q were found only in one PIN area. Controls to ensure reliable CGH results consisted in CGH analyses of (i) approximately 80 microdissected normal epithelial cells, which showed no aberrations after DOP-PCR and (ii) larger cell numbers (approximately 10(5) or 10(7) cells) of the primary tumour investigated without DOP-PCR and partially displaying the chromosomal imbalances (gain on 16p12-->p13, losses on 2p25, 8p21-->p23, 12p11-->p12, 12q21-->q26, 18q22) found in the small microdissected areas. Microsatellite and FISH analyses further confirmed our CGH results from microdissected cells. The combined approach of laser-assisted microdissection, DOP-PCR and CGH is suitable to identify early genetic changes in PIN and chromosomal imbalances associated with the particular histological patterns of invasive prostatic adenocarcinoma.

Adenocarcinoma↗
PubMed1998

Typical and atypical carcinoid tumors of the lung are characterized by 11q deletions as detected by comparative genomic hybridization.

Neuroendocrine tumors of the lung represent a wide spectrum of phenotypically distinct entities with different biological characteristics such as typical carcinoid tumor (TC), atypical carcinoid tumor (AC), large-cell neuroendocrine carcinoma (LCNEC), and small-cell lung carcinoma (SCLC). The histogenetic relationships between TC, AC, LCNEC, and SCLC are still unclear. This study was carried out to provide cytogenetic data about pulmonary neuroendocrine tumors and to evaluate their characteristic alterations and histogenetic relations for an improved understanding of the mechanisms of tumor development. Twenty-nine paraffin-embedded tumor samples of TC (n = 17), AC (n = 6), LCNEC (n = 3), and SCLC (n = 3) were selected for isolation of tumor DNA and subsequent comparative genomic hybridization (CGH) analysis. To confirm the comparative genomic hybridization results for characteristic chromosomal imbalances, selected cases were additionally investigated by loss of heterozygosity analysis. For statistical evaluation, we also used comparative genomic hybridization data from 45 published SCLC cases. DNA underrepresentations of 11q were the most frequent findings in TC (8 of 17) and AC (4 of 6), whereas these aberrations were rare in LCNEC (1 of 3) and SCLC (0 of 3). Furthermore, AC showed DNA underrepresentation of 10q (3 of 6) and 13q (3 of 6). In contrast, SCLC and LCNEC were characterized by a different pattern of DNA losses (3p-, 4q-, 5q-, 13q-, and 15q-) and gains (5p+, 17p+, and +20). Statistical analysis revealed significantly different occurrences of 11q deletions in TC/AC versus SCLC (45 published cases of SCLC and our 3 cases; P = 0.002; Fisher's exact test). Thus, TC and AC display frequent loss of 11q material including the MEN1 gene locus, which represents a characteristic genetic alteration in these tumors. Losses of 10q and 13q sequences allow a further cytogenetic differentiation between TC and AC. These additional changes might be responsible for the more aggressive behavior of AC. Three cases of LCNEC, the first to be analyzed by comparative genomic hybridization, exhibited similar complex abnormal patterns (4q-, 5q-, 10q-, 13q-, 15q-) to those of SCLC. Although neuroendocrine tumors of the lung share common phenotypic features, suggesting a genotypic relationship, they differ remarkably in their cytogenetic characteristics, highlighting an early fundamental molecular divergence during the development of these tumors.

Adult↗
PubMed1998

Analysis of symmetrical translocations for retrospective biodosimetry in radiation workers of the Mayak nuclear-industrial complex (Southern Urals) using FISH-chromosome painting.

PURPOSE: Frequencies of symmetrical translocations were determined by fluorescence in situ hybridization (FISH) for retrospective biodosimetry in workers occupationally exposed to external gamma-rays and internal plutonium at the Mayak nuclear-industrial complex (Southern Urals, Russia). MATERIALS AND METHODS: Chromosome analyses were carried out on peripheral lymphocytes from 75 Mayak workers who had received their main exposures between 1948 and 1963. Cumulative external gamma-ray doses between 0.02 and 9.91 Sv and plutonium burdens ranging between 0.26 and 18.5 kBq are reported. As controls, 33 unexposed persons from non-contaminated areas of the Southern Urals were used. Whole-chromosome painting probes for chromosomes 1, 4 and 12 were used simultaneously with a pancentromeric probe. RESULTS: Compared with the control group, a significantly elevated translocation frequency was found for the total study group and for either of two subsets with (48 subjects) and without (27 subjects) plutonium incorporation. The dicentric frequency was not significantly different from the control level. In the pooled data set, translocation frequencies showed a significant dependence on cumulative external gamma-ray doses. Plutonium uptake had no substantial influence. Individual dose estimates for 21 cases exhibiting at least five translocations ranged between 0.5 and 1.8 Gy, which is substantially lower than the workers' registered personal doses. CONCLUSION: At 35-40 years after protracted exposure to low-dose rate external gamma-rays, the postulated lifetime stability of translocations cannot be confirmed. Apparently, the natural loss of translocation-bearing peripheral lymphocytes cannot be fully compensated so that a temporal decline even of transmissible aberrations takes place. As a consequence, individual retrospective biodosimetry estimates cannot be obtained reliably from the remaining fraction of translocations.

Aged↗
PubMed1998

DNA-proportional distribution of radiation-induced chromosome aberrations analysed by fluorescence in situ hybridization painting of all chromosomes of a human female karyotype.

PURPOSE: This is the extension of a previous study, showing deviations from a DNA-proportional involvement of 12 single chromosomes (1-4, 6-10, 12, 14 and X) in radiation-induced translocations and dicentrics measured by FISH-painting and classified by standard cytogenetic scoring criteria. By adding data on chromosomes 2, 4, 5, 9, 11-13, 15-22 and X the analysis now comprises all chromosomes of a human female karyotype evaluated with three nomenclature systems (PAINT, S & S and a conventional method). MATERIAL AND METHODS: Metaphase spreads were prepared from lymphocytes irradiated with 3 Gy 220 kV X-rays. FISH painting was performed with single chromosome-specific probes in combination with a pancentromeric probe. RESULTS: Deviations from a DNA-proportional distribution became apparent for all aberration parameters analysed with the three nomenclature systems. Chromosomes 2, 3 and 6 were less frequently involved and chromosomes 16, 17 and 20 were more frequently involved in exchange aberrations. Generally, smaller chromosomes (15-22, with the exception of chromosome 19) were more frequently involved in aberration formation than expected. CONCLUSION: The assumption that the probability of a chromosome being involved in an exchange aberration is proportional to its DNA content is not supported by the present data.

Cell Count↗
PubMed1998

LET dependence of yield ratios of radiation-induced intra- and interchromosomal aberrations in human lymphocytes.

PURPOSE: To investigate the LET dependence of various yield ratios of intra- and interchromosomal aberrations in order to quantify proximity effects for illegitimate rejoining of chromosome breaks. MATERIALS AND METHODS: Calculations are based on 13 human lymphocyte data sets obtained from in vitro irradiation experiments with various radiation qualities covering an LET range of 0.5 to 150keV/microm. A total of 93000 first-division metaphases was analysed for all categories of chromosome-type aberrations. RESULTS: No dose dependence was found for the various yield ratios of chromosomal aberrations. No LET dependence became apparent for the yield ratio of inter- to intrachromosomal exchange-type aberrations (dicentrics/centric rings = F value). However, a clear LET dependence was found for the yield ratio of intra-arm intrachanges to inter-arm intrachanges (interstitial deletions/centric rings = G ratio) and for the yield ratio of intra-arm intrachanges to interchromosomal exchanges (interstitial deletions/dicentrics = H ratio). CONCLUSION: Measurements of intrachanges are informative for interpretation and quantification of proximity effects in the formation of radiation-induced chromosome aberrations. In particular, yield ratios for intra-arm intrachanges are likely to have a high potential as an indicator of high-LET radiations.

Alpha Particles↗
PubMed1998

FISH-based analysis of stable translocations in a Techa River population.

PURPOSE: Measurements of symmetrical translocations by fluorescence in situ hybridization (FISH) were performed for retrospective biodosimetry in a Techa River population exposed to external (gamma-rays) and internal (90Sr, 137Cs) irradiation. MATERIALS AND METHODS: Chromosome analyses were carried out on peripheral lymphocytes from 73 radiation-exposed residents from settlements along the Techa River (Southern Urals, Russia) located 7-148 km downstream from the site of release of liquid radioactive waste from the plutonium production facility Mayak. Thirty-nine unexposed persons from non-contaminated areas were used as controls. Whole-chromosome painting probes for chromosomes 1, 4 and 12 were used simultaneously with a pancentromeric probe. RESULTS: A significantly elevated mean translocation frequency compared with controls was found for the total study group and for either of two subgroups of inhabitants residing in villages of the upper regions of the Techa River (7-60 km) during 1950 to 1951, or in villages of the lower regions (78-148 km) until the time of blood sampling. Within the first subgroup, subjects born between 1937 and 1949 showed higher translocation frequencies than those born between 1914 and 1936. Collective biodosimetry estimates for the various groups were between 0.24 and 0.54 Gy. Individual dose estimates for seven subjects with at least five translocations ranged between 0.77 and 1.80 Gy and compared well with doses reconstructed on the basis of 90Sr whole-body counts (WBC) and electronic paramagnetic resonance (EPR) measurements. Individual translocation frequencies from 40 subjects with existing WBC data and calculated cumulative red bone marrow doses below 0.6 Gy fall within the 95% prediction limits of the calibration curve. CONCLUSIONS: FISH-based translocation measurements can provide useful information for a retrospective biodosimetric interpretation. However, with the analysed number of cells, individual estimates required for a reliable evaluation of this highly variable exposure situation can be obtained only for a minority of subjects. In addition, influence of a temporal decline on the yield of stable translocations cannot be fully excluded.

Body Burden↗
PubMed1998

Chromosome aberration frequencies in human lymphocytes irradiated in a phantom by a mixed beam of fission neutrons and gamma-rays.

PURPOSE: To provide further information on the existence of a significant quadratic component of the dose response relationship for the production of dicentrics in human lymphocytes by mixed fission neutron and gamma-ray irradiation, which has been observed previously employing the same beam under free-in-air conditions. MATERIALS AND METHODS: Irradiation of blood samples and dosimetry was performed at 2 cm depth in a polymethyl methacrylate (PMMA) phantom of 16 cm side-length where the PMMA material fully surrounded the blood specimen. Chromosome analysis was carried out exclusively in complete first division metaphases. RESULTS: Dicentric yields induced by absorbed doses between 0.043 and 2.68 Gy fit a linear-quadratic model with a quadratic coefficient significantly different from zero. The distribution of dicentrics is overdispersed compared to Poisson at lower doses, but is poissonian at higher doses. CONCLUSION: The presence of a significant quadratic dose response coefficient for dicentrics, both for free-in-air and phantom irradiation, is caused by the various degraded fission spectra that produce neutrons or recoil protons over a broad energy range, rather than by the gamma-ray component of the beam.

Cells, Cultured↗
PubMed1998

FISH-based analysis of radiation-induced chromosomal aberrations using different nomenclature systems.

PURPOSE: Comparison of three nomenclature systems for the description of chromosomal aberrations involving painted chromosomes (PAINT, S&S and a conventional method) by parallel application to one data set. MATERIALS AND METHODS: Radiation-induced (3 Gy 220 kV X-rays) chromosome aberrations in human lymphocytes were analysed by FISH-painting of seven single chromosomes (1, 3, 6, 7, 8, 10 and 14) simultaneously with a pancentromeric probe. RESULTS: Each system is based on different prerequisites and uses different criteria for the classification and quantification of structural chromosome aberrations. Due to the frequent occurrence of complex exchanges (resulting from > or = 3 breaks on > or = 2 chromosomes), standard cytogenetic scoring criteria used for solid-stained preparations are inadequate for a precise and reproducible classification of aberrant painting patterns. S&S is particularly suitable if a mechanistic interpretation of aberration origins is required. The descriptive terminology of PAINT enables a rapid, reproducible description, even of most extensively rearranged chromosomes by classifying each abnormal painting pattern individually. CONCLUSION: A modification of PAINT criteria, allowing also for mechanistic aspects, would be most advantageous for practical application.

Chromosome Aberrations↗
PubMed1997

Cytogenetic evaluation of occupational exposure to external gamma-rays and internal 241Am contamination.

Chromosome analyses were performed on peripheral lymphocytes from seven radiation workers exposed to external gamma-radiation within maximum annual permissible dose limits during 11 to 22 years of employment. Five years prior to blood sampling, six workers had additionally incorporated the alpha-emitting radionuclide 241Am which contributed between five to 25% of the total accumulated whole body dose in five workers and about 70% in one worker. For the radiation workers as a group, both the mean frequencies of symmetrical translocations measured by fluorescence in situ hybridisation (FISH)/chromosome painting and dicentrics + ring chromosomes scored in first division metaphases of conventional preparations were significantly elevated compared with respective controls. The mean biodosimetry estimate for the group was 270 mSv when based on stable translocations which compares well with the mean dose of 247 mSv based on official dosimetry records. The lower mean dose estimate of 160 mSv based on unstable dicentrics is compatible with the well-known loss of dicentrics from the circulating blood with time after exposure.

Adult↗
PubMed1997

p53 mutations in childhood thyroid tumours from Belarus and in thyroid tumours without radiation history.

Mutations in the p53 tumour-suppressor gene (exons 5-8) were investigated in 31 Belarussian childhood thyroid tumours (24 cases of papillary thyroid carcinoma, 3 benign tumours and 2 cases each of thyroiditis and goiter); 33 thyroid tumours from juveniles and adults without radiation exposures (25 carcinomas of various histological types, including 11 papillary carcinomas and 8 adenomas) and 6 tumours from adults (4 papillary carcinomas, 1 adenoma, 1 goiter) served as controls. The mutational spectrum of p53 differed greatly between the childhood thyroid carcinomas from Belarus and the control groups. In the control groups of 29 malignant thyroid tumours, 7 different mutations were detected on exons 5-8, none of which occurred among the 15 papillary carcinomas in this group. Five mutations were found in tissue samples of the 24 childhood papillary carcinomas, and they were all the same p53 point mutation (CGA --> CGG) on codon 213 of exon 6. To determine whether this mutation is simply a polymorphism or whether it is specific to the tumour cells, laser-assisted microdissection was applied to collect various areas of tumorous and non-tumorous cells (10-20 cells per sample) from each paraffin-embedded tissue section of 8 of the papillary thyroid carcinomas. Using PCR-SSCP and sequence analysis on these cells, the very same p53 mutation on codon 213 was detected in various microdissected tumour samples of 2 cases, but it was not found in any microdissected non-tumorous sample. The exclusive occurrence of this p53 mutation in selective microdissected samples of tumour cells, even as homozygous mutation in 1 case, reflects a distinct tumour heterogeneity within papillary childhood thyroid carcinomas.

Adolescent↗
PubMed1997

Cytogenetic and molecular genetic characterization of a chromosome 2 rearrangement in a case of human papillary thyroid carcinoma with radiation history.

Karyotype analysis of a primary culture from a case of papillary thyroid cancer (PTC) showed an abnormal short arm of one homologue of chromosome 2 as sole abnormality in 4 of 16 metaphases. Based on G-banding analysis, two different aberration types on chromosome 2 could be assumed representing either a del(2)(p22-23) or a pericentric inversion. Further comparative genomic hybridization (CGH) analysis as well as fluorescence in situ hybridization (FISH) analysis were performed to confirm the assumed alterations. While CGH analysis showed no loss of chromosome 2 material, FISH with yeast artificial chromosome (YAC) probes homologous to the region 2p22-23 demonstrated two pericentric inversions of chromosome 2 involving different breakpoints on 2p in 6.8% and 4.2% of the metaphases, respectively. Polymerase chain reaction (PCR) analysis with degenerated oligonucleotide primers that bind within the conserved catalytic domain of tyrosine kinase (tk) genes resulted in amplification products with DNA of YAC 851D11 suggesting the presence of such genes at or near the translocation breakpoint.

Adult↗
PubMed1997

Chromosome aberration frequencies in human lymphocytes irradiated in a multi-layer array by protons with different LET.

PURPOSE: To provide data on the dose-and linear energy transfer (LET)-dependence of the production of dicentrics in human lymphocytes. MATERIALS AND METHODS: Track segment irradiation with 16.5 MeV protons was performed in a multi-layer array allowing the simultaneous exposure of human peripheral lymphocytes in three successive samples. Within these samples the dose-averaged linear energy transfer (LD) of protons increased with increasing depth, i.e. LD = 3.5, 5.3 and 19.0 keV/microm. Dicentrics were scored in first division solid-stained metaphases. RESULTS: Dicentric yields measured in the first and second samples fit the linear-quadratic model, those of the third sample fit a linear model of the dose response relationship. Relative to 137Cs gamma-rays, limiting RBE values of 2.9, 4.3 and 21.5 were determined from the ratios of the respective linear coefficients of the dose effect curves. The linear dose effect coefficient increases approximately proportionally with increasing LET. The quadratic coefficient is approximately constant for protons with LD of 3.5 and 5.3 keV/microm, and is not significantly different from zero at LD= 19.0 keV/microm. CONCLUSION: This result is well in line with theoretical predictions on the variations of dose-effect coefficients with radiation quality. However, such an evaluation can only be obtained from clearly defined track segment experiments.

Chromosome Aberrations↗
PubMed1997

Retrospective biodosimetry of Chernobyl clean-up workers using chromosome painting and conventional chromosome analysis.

Blood samples of 52 Chernobyl clean-up workers were analysed by fluorescence in situ hybridization (FISH) using whole-chromosome painting probes for chromosomes 1, 4 and 12, simultaneously with a pancentromeric probe and by conventional chromosome analysis, for radiation-induced symmetrical translocations and dicentrics in T-lymphocytes. Based on FISH measurements of translocations, individual biodosimetry estimates between 0.32 and 1.0 Gy were obtained from 18 cases. Pooled data for the total group of 52 workers provided a collective biodosimetry estimate of 0.23 Gy. For a group of 34 workers with documented doses, the mean dose estimate of 0.25 Gy compared well with the mean documented dose of 0.26 Gy. However, no correlation between individual translocation frequencies (FG) and documented doses could be found. A statistical analysis of the expected dose-response suggests exposures to higher doses than documented for a substantial fraction of workers with ascribed doses < 0.2 Sv. For subjects working repeatedly at the reactor site between 1986 and 1995 the mean translocation frequency was significantly higher than for those working only in 1986. A comparison of dicentric frequencies obtained by conventional scoring and by FISH measurements showed no significant difference, although only two of 52 cases revealed significantly higher yields than the mean control level. Based on conventionally scored dicentric frequencies, a collective biodosimetry estimate of 0.23 Gy could be derived only of the group of persons working at Chernobyl exclusively in 1986 for which a documented average dose of 0.19 Gy was reported.

Adult↗
PubMed1997

Is there reliable experimental evidence for a chromosomal "fingerprint" of exposure to densely ionizing radiation?

A selection of published data on the ratio, F, of interchromosomal to intrachromosomal stable (reciprocal translocations and pericentric inversions) and unstable (dicentric chromosomes and centric rings) exchange aberrations in human lymphocytes has recently been presented as evidence for F values of about 15 for X and gamma rays and about 6 for neutrons and alpha particles (D. J. Brenner and R. K. Sachs, Radiat. Res. 140, 134-142, 1994). On this basis it was proposed that low F values could serve as a chromosomal "fingerprint" of densely ionizing radiation. In the present commentary it is shown that some of the quoted data sets provide little support for this concept. It is further demonstrated that our own data, including a "head-to-head" experiment with gamma rays and alpha particles, reveal no LET dependence, even in the comparison of F values from low-LET radiation with those from low doses of high-LET radiation. In this context it is pointed out that a change in F values cannot be expected at doses of high-LET radiation where the linear component of the dose-effect relationship for exchange aberrations prevails. Additional data for the effects of high- and low-LET radiation which have not been considered in the discussion so far confirm that support of the concept of F-ratio "fingerprinting" by experimental data is insufficient.

Alpha Particles↗
PubMed1997

Health impacts of large releases of radionuclides. Cytogenetic effects as quantitative indicators of radiation exposure.

Scoring of dicentrics in metaphase preparations of human T lymphocytes is the method of choice for estimating individual whole-body doses of radiation exposure. A quantification of partial-body exposures or non-uniform distribution of the dose is more complicated but it can be achieved by using specific mathematical approaches. For retrospective biodosimetry, conventional scoring of dicentrics is less precise because these unstable aberrations are eliminated with time post-exposure. Symmetrical translocations are not selected against during mitotic division in the haematopoietic cell reproductive centres, so the frequencies of these stable aberrations are generally assumed to remain constant even for decades. They can now be analysed precisely by fluorescence in situ hybridization using whole chromosome-specific DNA probes (chromosome painting) with an alpha-satellite DNA probe for centromere detection. Based on in vitro calibration curves established with single or multicolour paints covering 4-22% of the total human genomic DNA content, scoring of translocations has been applied for dose reconstruction in smaller groups of atomic bomb survivors and victims of the Chernobyl and Goiania radiation accidents. However, prior to routine use, the method requires further validation. Such work includes the precise evaluation of the unexpectedly high frequency of complex exchanges (> or = 3 breaks in > or = 2 chromosomes) found both at > 2 Gy doses of low linear energy transfer (LET) radiation and generally for high LET alpha-particles. Data on the long-term stability of translocations and the appearance of clonal abberrations, as well as improved measurements of the linear coefficient of standard calibration curves, are also required.

Centromere↗