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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Der Pathologe 16 (1995), S. 128-138 
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Dreidimensionale Rekonstruktion ; Lichtmikroskopie ; Serienschnitte ; Megakaryozyten ; Primäre Osteomyelofibrose ; Knochenmark ; Key words Three-dimensional reconstruction ; Light microscopy ; Serial sections ; Megakaryocytes ; Primary osteomyelofibrosis ; Bone marrow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Computer-based three-dimensional reconstruction of serially cut light microscopic sections is being increasingly used in medical and biological research. Compared with conventional two-dimensional evaluation of histological sections, complex histotopographical relationships and structural details are easy to assess and could be imposingly visualized on the graphics screen. Because of the rapid progress in modern computer hardware, uncomplicated and fast reconstruction systems are available on standard personal computers. Therefore, 3D reconstruction is applicable for a wide range of investigations that warrant 3D exploration. Own results are presented and the principles of 3D reconstruction, as well as the problems and difficulties in this new technique, are discussed. The complex ameboid morphology of megakaryocytes in bone marrow of primary (idiopathic) osteomyelofibrosis is examined with this new method in combination with selective immunohistochemical staining procedures. 3D reconstruction can visualize the complexity of cytoplasmic and nuclear formation impressively. Moreover, the models generated permit exact quantitative measurements of cell morphology. The technique presented is a powerful tool for anatomical-morphological investigations and can furthermore increase our comprehension of complex histomorphological objects.
    Notes: Zusammenfassung Computergestützte dreidimensionale Rekonstruktionsverfahren von lichtmikroskopischen Serienschnitten haben sich in den letzten Jahren in allen Bereichen der medizinisch-biologischen Forschung etablie-ren können. Im Vergleich zur konventionellen zweidimensionalen Schnittbetrachtung lassen sich komplexe histotopographische Beziehungen sowie Strukturcharakteristika besser erfassen und auch eindrucksvoll darstellen. Aufgrund der schnell fortschreitenden Entwicklung im Bereich der Computertechnik sind heutzutage Rekonstruktionssysteme auch auf preisgünstigen Rechnersystemen verfügbar, so daß sich dieser neuen Methode ein zunehmendes Anwendungsgebiet erschließt. Anhand von eigenen Befunden werden die Grundlagen der 3 D-Rekonstruktion besprochen sowie Schwierigkeiten und Probleme dieser neuen Technik diskutiert. In Kombination mit kontrastreichen und selektiven immunhistochemischen Färbungen wird beispielhaft die komplexe amöbenartige Morphologie der Megakaryozyten bei der primären (idiopathischen) Osteomyelofibrose durch lichtmikroskopische Serienschnitte aus dem Knochenmark vorgestellt. Die erzeugten 3 D-Modelle vermitteln einen plastischen Eindruck von der Komplexität der Zell- und Kernstrukturen dieser Zellen und stellen die Basis für exakte quantitative Analysen der Zellmorphologie dar. Die vorgestellte Technik beinhaltet ein ergänzendes Werkzeug für anatomisch-morphologische Analysen und ist in der Lage, entscheidende Erkenntnisse bei der Untersuchung vielschichtiger histomorphologischer Objekte zu liefern.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0584
    Keywords: Key words Chronic myeloproliferative disorders ; Erythroid precursors ; Neutrophil granulopoiesis ; Megakaryocytes ; Macrophages ; Myelofibrosis ; Enzyme-immunohistochemistry ; Morphometry ; Bone marrow biopsy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of this review is to evaluate morphological characteristics of the different subtypes of chronic myeloproliferative disorders (MPDs) derived by applying immunohistochemical and morphometric techniques to bone marrow biopsies and to combine these results with relevant clinical parameters. In comparison to control specimens, a significant decrease in erythroid precursors is determinable in chronic myeloid leukemia (CML), while this cell lineage is most prominent in polycythemia vera (PV) and moderately to markedly reduced in idiopathic myelofibrosis (IMF). On the other hand, neutrophilic granulopoiesis shows a predominance in CML and a relevant increase in PV, but no conspicuous changes are detectable in essential thrombocythemia (ET). CML is characterized by a prevalent growth of dwarflike micromegakaryocytes, occurring in particular in the so-called megakaryocyte-rich subtypes (about 30%). This finding differs significantly from the pleomorphous aspect, i.e., clusters of small to giant-sized megakaryocytes in PV and the grossly abnormal (dysplastic) appearance of this cell lineage in patients with IMF. Similar cytological abnormalities of megakaryopoiesis consistent with maturation defects are never encountered in ET. The incidence of mature (resident) macrophages (phagocytic reticular cells) is significantly enhanced in IMF in comparison to the other MPDs and controls. Moreover, there is a striking difference in the density of reticulin-collagen fibers, ranging from normal (ET) to extreme values (IMF). In IMF more than 80% of the patients present with some degree of myelofibrosis-osteosclerosis at diagnosis, while the rest show an initial prefibrotic, hypercellular stage. This feature deserves special attention since, when accompanied by thrombocythemia, it may simulate ET. Sequential bone marrow biopsies in patients with IMF disclose that evolution of myelofibrosis is progressive, but occurs at a variable and unpredictable speed. A synoptical approach regarding clinical diagnosis and histological subtyping of MPDs is explicitly recommended and demonstrated by sets of diagnostic criteria. This rationale requires equal consideration of laboratory data and morphology by clinicians to include well-defined subtypes of MPDs into prospective management studies. Furthermore, it may even warrant follow-up studies and repeated bone marrow examinations in initially unclassifiable cases.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0584
    Keywords: CML ; Myelofibrosis ; Dynamics ; Megakaryocytes ; Morphometry ; Interferon ; Busulfan ; Sequential bone marrow biopsies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To evaluate treatment-related changes of the reticulin stain-measured fibrosis in Ph1+-CML, a clinicopathological study was performed on sequential trephine biopsies of the bone marrow following either interferon (IFN) or busulfan (BU) monotherapy. Using the monoclonal antibody CD61 for the identification of megakaryopoiesis and Gomori's silver impregnation method, number of megakaryocytes and density of argyrophilic (reticulin and collagen) fibers were determined by morphometry. We studied specimens from 26 patients with IFN-alpha 2b (including nine patients with additional IFN gamma) therapy and from 23 patients who had received BU. In both groups, repeated bone marrow biopsies (total 125) revealed a significant increase in the fiber content, as well as in the number of megakaryocytes during treatment. To assess the dynamics of myelofibrosis more precisely, computation of differences in the degree of fiber density between the first and last examination was carried out. Regarding the considerable variations in the biopsy intervals, a so-called myelofibrosis progression index (MPI) was calculated. Following this rationale, we were able to demonstrate that, in comparison to the BU-group, speed of progression of bone marrow fibrosis was significantly increased in CML patients treated with IFN. Preliminary statistical analysis indicated a relationship between myelofibrosis on admission, which was always associated with increased growth of megakaryocytes, and the MPI with survival. Even when these parameters were regarded, prognosis was significantly more favorable in the IFN-treated patients. The failure of IFN and BU to inhibit the evolution of myelofibrosis may be related to several conversely acting pathomechanisms. Among others, the inability of both therapeutic agents to reduce the number of megakaryocytes more effectively should be taken into consideration.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0584
    Keywords: Key words CML ; Myelofibrosis ; Dynamics ; Megakaryocytes ; Morphometry ; Interferon ; Busulfan ; Sequential bone marrow biopsies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  To evaluate treatment-related changes of the reticulin stain-measured fibrosis in Ph1+-CML, a clinicopathological study was performed on sequential trephine biopsies of the bone marrow following either interferon (IFN) or busulfan (BU) monotherapy. Using the monoclonal antibody CD61 for the identification of megakaryopoiesis and Gomori's silver impregnation method, number of megakaryocytes and density of argyrophilic (reticulin and collagen) fibers were determined by morphometry. We studied specimens from 26 patients with IFN-alpha 2b (including nine patients with additional IFN gamma) therapy and from 23 patients who had received BU. In both groups, repeated bone marrow biopsies (total 125) revealed a significant increase in the fiber content, as well as in the number of megakaryocytes during treatment. To assess the dynamics of myelofibrosis more precisely, computation of differences in the degree of fiber density between the first and last examination was carried out. Regarding the considerable variations in the biopsy intervals, a so-called myelofibrosis progression index (MPI) was calculated. Following this rationale, we were able to demonstrate that, in comparison to the BU-group, speed of progression of bone marrow fibrosis was significantly increased in CML patients treated with IFN. Preliminary statistical analysis indicated a relationship between myelofibrosis on admission, which was always associated with increased growth of megakaryocytes, and the MPI with survival. Even when these parameters were regarded, prognosis was significantly more favorable in the IFN-treated patients. The failure of IFN and BU to inhibit the evolution of myelofibrosis may be related to several conversely acting pathomechanisms. Among others, the inability of both therapeutic agents to reduce the number of megakaryocytes more effectively should be taken into consideration.
    Type of Medium: Electronic Resource
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