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  • 1
    ISSN: 1365-2559
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The Philadelphia translocation was demonstrated by two-colour fluorescence in situ hybridization (FISH) in decalcified paraffin sections of bone marrow from patients with chronic myelogenous leukaemia. FISH was combined with immunocytochemical detection of different membrane-bound or cytoplasmic antigens. With this new technique, the cells bearing the 9;22 translocation can be identified morphologically, as well as immunocyto-chemically, in tissue sections.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2307
    Keywords: Lymphoma ; Hodgkin's disease ; Polymerase chain reaction ; Immunohistochemistry ; Histological classification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Ninety-one Hodgkin's lymphomas (HD), 52 non-Hodgkin lymphomas (NHL) and 33 specimens of non-neoplastic lymphatic tissues were investigated by polymerase chain reaction (PCR) for the presence of the bcl-2/JH gene rearrangement. The majority of the HD cases were drawn from the files of the German Hodgkin trial where diagnoses are established by a panel of four independent histopathologists. Using the very sensitive PCR method which detected 1 positive among 10000 negative cells, the bcl-2/JH gene rearrangement was found in 7/52 NHL and 3/16 tonsils with follicular hyperplasia, but in none of the 91 HD. The bcl-2 protein, however, was expressed by malignant cells of B and T cell lymphomas and by the giant tumour cells in 2/13 HD lymphocyte predominant, 11/28 HD nodular sclerosing I, 14/17 HD nodular sclerosing II, 10/27 HD mixed cellularity and 3/3 HD lymphocyte depleted. The bcl-2/JH rearrangement is thus independent of protein over-expression, the latter being found in all types of lymphomas. Our results do not confirm the findings of others who have detected the bcl-2/JH rearrangement in HD. These discrepancies may be explained by differences in choice of material, the gene rearrangement actually occuring in bystander cells but not in Reed-Sternberg or Hodgkin cells, or by contamination.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2307
    Keywords: Chronic myelogenous leukaemia ; Philadelphia chromosome ; FISH ; Megakaryocyte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Histological examination of bone marrow biopsies shows that about one-third of chronic myeloid leukaemia (CML) patients exhibit an increase of megakaryocytes. The megakaryocytic predominance may be so striking that differentiation from other chronic myeloproliferative disorders (CMPD) may be difficult in some CML patients. Megakaryocytes in CML are clonal as demonstrated by loss of glucose-6-phosphate dehydrogenase isoenzymes. The Ph translocation, fusing the abl and bcr genes on chromosomes 9 and 22, however, obviously occurs as a second step in tumour development. So far, the Ph translocation has not been assigned explicitly to megakaryocytes. The question is whether the megakaryocytic cell lineage could harbour the bcr/abl fusion in those CML cases with striking proliferation of megakaryocytes but lack this genetic defect in cases with normal or decreased megakaryocyte counts. We therefore performed triple-colour fluorescence in situ hybridization (FISH) for portions of the bcr and abl genes flanking the breakpoint in CML in paraffin sections of CML cases with normal and with increased numbers of megakaryocytes. This method allows identification of the bcr/abl fusion in single, morphologically intact cells, whereas conventional cytogenetics requires lysis and thus destruction of the cell. Among the 21 CML patients examined by FISH, 10 were informative for bcr and abl genes and displayed distinct hybridization signals within nuclei of bone marrow cells. Besides the granulopoietic cells, megakaryocytes of all those patients (4 without and 6 with varying grades of megakaryocytic increase) displayed bcr/abl fusion signals indiciative of a Ph translocation. The lack of hybridization signals in the remaining 11 cases indicates that this technique is not of value diagnostically and should be reserved for scientific questions. Positive controls consisted of conventional chromosome preparations from bone marrow aspirates demonstrating the Ph chromosome in all patients examined, and negative controls of paraffin sections of bone marrow biopsies from non-CML patients. These showed no fusion signals in bone marrow cells, including megakaryocytes, using FISH. Our results demonstrate clearly that not only the transforming event but also the Ph translocation leading to the bcr/abl fusion happens prior to the differentiation of the pluripotent stem cell into different myeloid lineages. The megakaryocytic proliferation evident in some CML cases is probably a consequence of the disease progress.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2307
    Keywords: Hepatocellular carcinoma ; Cholangiocellular carcinoma ; Cytogenetics ; Chromosome 1 ; Fluorescence in situ hybridization (FISH)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Conventional cytogenetic studies revealed gains and structural aberrations of chromosome 1 to be the most consistent chromosomal aberrations in hepatocellular carcinoma (HCC). We investigated touch preparations of eight HCC, five cholangiocellular carcinomas (CCC), five liver cell adenomas (LCA), four focal nodular hyperplasias (FNH) as well as nine specimens of normal liver tissue using fluorescence in situ hybridization (FISH) with centromere specific probes for chromosomes 1 and 8. Polysomies of chromosome 1, especially trisomy 1, were found in five of eight HCC and four of five CCC but in no normal liver tissue or benign tumour. Only three of seven cases of HCC revealed trisomy 8 whereas the five benign liver tumours and all normal liver tissues examined had disomy 8. Our results confirm conventional cytogenetic findings in terms of chromosome 1 aberrations in HCC although they are not specific for these types of malignant liver tumours. Since α-satellite probes were used in our study, only gains or losses including the centromeric regions of the chromosomes 1 and 8 could be detected. Nevertheless, our findings suggest that FISH may help in the differential diagnosis of malignant versus benign neoplasms of the liver.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Chronische myeloproliferative Erkrankungen ; Philadelphia-Translokation ; Zytogenetik ; Molekulargenetik ; Fluoreszenz-in-situ-Hybridisierung ; Histopathologie ; Key words Chronic myeloproliferative disorders ; Philadelphia-translocation ; Cytogenetics ; Molecular genetics ; Fluorescence in situ hybridization ; Histopathology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary The histopathological classification of chronic myeloproliferative disorders can be supported by applying cytogenetics and molecular genetics to the analysis of bone marrow or blood cells, as demonstrated in 253 cases evaluated. The Philadelphia translocation (9;22) is the most important genetic parameter, being specific for chronic myeloid leukemia. Conventional methods for the detection of the t(9;22) are karyotyping and Southern blot analysis of the bcr gene. The newly established technique of fluorescence in situ hybridization (FISH) allows visualization of bcr-abl fusion even in non dividing cells. Molecular cytogenetics for t(9;22) yield results that are rapid and reliable as well as easily quantifiable.
    Notes: Zusammenfassung Zytogenetische und molekulargenetische Untersuchungen von Knochenmark- oder Blutzellen sind für die histopathologische Klassifikation der chronischen myeloproliferativen Erkrankungen hilfreich, was durch die simultane Auswertung von 253 Fällen gezeigt wird. Insbesondere die Analyse der Philadelphia-Translokation (9;22) ist dabei für die Bestätigung oder den Ausschluß einer chronischen myeloischen Leukämie wichtig. Für den Nachweis der t(9;22) stehen die konventionelle Karyotypisierung mit Bestimmung des Philadelphia-Chromosoms und das Southernblotverfahren zur Analyse einer Umlagerung des bcr-Gens zur Verfügung. Durch die neuere Methode der Fluoreszenz-in-situ-Hybridisierung (FISH) kann auch eine bcr-abl-Fusion an Interphasekernen dargestellt werden. Diese molekulare Zytogenetik ist ein rasches und zuverlässiges Verfahren zum Nachweis der Philadelphia-Translokation, das zudem leicht quantifizierbare Ergebnisse liefert.
    Type of Medium: Electronic Resource
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