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  • 1
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Chondroblastom ; Knochentumoren ; Immunhistologie ; Proliferation ; Key words Chondroblastoma ; Bone tumors ; Immunohistochemistry ; Proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Representing only about 1 % of all primary bone tumors, chondroblastoma constitutes a very rare bone tumor entity. 56 cases of chondroblastoma, that had been collected by the Hamburg Bone Tumor Registry from 1972 to 1995, were examined histologically together with the radiological and clinical findings. In addition immunohistochemistry with antibodies against S 100, PGM1, LCA and the proliferationmarker MIB 1 was performed. The mean age was 20.4 years and male patients being the majority with a gender ratio of 2.7 : 1. Predominant localisation was the epiphyses of the long bones, although almost 40 % of the tumors were located at untypical sites. Usually a well-circumscribed lysis could be seen on plain X-Ray examination, however partial cortical destruction could be observed in one third of the cases. Histologically characteristic was a polygonal cell component with a weblike chonroid matrix, sometimes with a plane-like appearance. 5 cases showed a distinct nuclear polymorphism making a distinction from osteosarcoma difficult. Using immunohistochemistry all tumors except for one showed positive reaction for S 100 protein. Although the histogenesis of chondroblastoma is not completely understood, morphological findings as well as the observed reactivity with the S 100 protein indicate the chondroid origin. No reactivity for PGM 1 (CD 68) or LCA could be detected. All chondroblastoma showed a low rate of proliferation, thereby being distinguishable from high malignant bone tumors. In general chondroblastoma show a benign biological behavior. Different behavior was observed in 2 cases. One relapse located in the pelvis revealed local aggressive growth while in another case in the humerus a malignant transformation had taken place.
    Notes: Zusammenfassung 56 Chondroblastome, die im Hamburger Knochentumorregister im Zeitraum von 1972 bis 1995 archiviert wurden, wurden retrospektiv histologisch untersucht, unter Berücksichtigung des radiologischen Befunds sowie der klinischen Angaben. Zusätzlich wurden immunhistologische Färbungen für S 100, PGM 1, LCA und den Proliferationsmarker MIB 1 durchgeführt. Das Durchschnittsalter der Patienten betrug 20,4 Jahre unter Bevorzugung männlicher Patienten mit einem Geschlechtsverhältnis von 2,7 : 1. Bevorzugter Lokalisationsort waren die Epiphysen der langen Röhrenknochen. Radiologisch stellt sich typischerweise eine umschriebene Lyse mit umgebendem Randsaum dar. Charakteristisch ist histologisch eine polygonale Zellkomponente mit einer netzartigen chondroiden Matrix. In 5 Fällen lag eine deutliche Kernpolymorphie vor, die eine Abgrenzung zum Osteosarkom schwierig machte. Immunhistologisch waren mit Ausnahme eines Falle alle Tumoren positiv für S 100. Allen Chondroblastomen war eine niedrige Proliferationsrate gemeinsam, die diese deutlich von hochmalignen Knochentumoren unterschied. Chrondroblastome besitzen üblicherweise ein gutartiges biologisches Verhalten. Zwei Fälle dieser Studie zeigten einen davon abweichenden Verlauf. Bei einem Rezidivtumor im Becken zeigte sich ein lokal aggressives Wachstum, in einem anderen Fall im Humerus war es zu einer malignen Transformation gekommen.
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  • 2
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Chondrosarkom ; Knochentumoren ; Grading ; Morphometrie ; Histologie ; Key words Morphometry ; Bone neoplasma ; Chondrosarcoma ; Grading ; Histology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Chondrosarcomas are frequent malignant bone tumors. Aside from different subtypes, such as dedifferentiated, mesenchymal and clear-cell chondrosarcoma, chondrosarcomas (classical chondrosarcoma) show different grades of differentiation. The borderline between chondroma and classical chondrosarcoma is not clearly defined. The same chondrosarcoma can be graded differently at different institutes. Standardized therapy concepts are currently in preparation. As the Hamburg Bone Tumor Registry is often consulted for chondrogenic tumors, the histological criteria are based on a series of 74 chondrosarcomas recorded there. The emphasis has been laid on a classification which can be used in daily routine and which is reproducible and in agreement with the classifications of other international groups. Grade I chondrosarcomas (50 %) can be distinguished only by growth criteria. The nuclei are small and show high chromatin density. Grade II chondrosarcomas (42 %) have medium-sized, regular nuclei with loose chromatin structure. The chondrocytes of grade III cases (8 %) show polymorphic nuclei. Binucleas forms, the number of mitoses and cellularity all show considerable overlap for all three grades. So far there are no immunohistological and molecular biological methods for reliable differentiation. The therapeutic consequences of the classification into grades are thorough curettage, in the case of grade I tumors, or complete resection, for grade II and III cases. The long-term results, however, need to be confirmed by a larger number of cases. From 1991 to 1995 the method was applied and proved to be easily practicable in daily diagnostic routine. Some 104 cases of classical chondrosarcomas (grade I 53 %, grade II 39 %, grade III 8 %) were analyzed. Two pathologists both assigned the same grade in 90 % of cases.
    Notes: Zusammenfassung Das Chondrosarkom gehört zu den häufigen malignen Knochentumoren. Neben unterschiedlichen Subtypen treten unterschiedliche Differenzierungsgrade des klassischen Chondrosarkoms auf. Die Übergänge von einem Chondrom, der benignen Variante chondrogener Tumoren, und dem klassischen Chondrosarkom sind histologisch fließend. Über die Bewertung der einzelnen Tumorformen liegen unterschiedliche Befunde vor. Einheitliche Therapiekonzepte werden derzeit erarbeitet. Deshalb werden die histologischen Kriterien an einer Serie von 74 Chondrosarkomen zusammengestellt. Dabei wird Wert auf eine in der täglichen Routine brauchbare und reproduzierbare Klassifikation gelegt. Grad-I-Chondrosarkome (50 %) können nur aufgrund von Wachstumskriterien von Chondromen unterschieden werden. Die Zellkerne sind klein, chromatindicht. Grad-II-Chondrosarkome (42 %) besitzen mittelgroße, gleichmäßige Zellkerne mit einem lockeren Chromatingerüst. Bei den Grad-III-Fällen (8 %) tritt eine Kernpolymorphie der Tumorchondrozyten auf. Sog. Doppelkernformen, die Zahl an Mitosen sowie die Zelldichte sind Kriterien mit einer großen Überschneidung in den 3 Differenzierungsgraden. Die therapeutischen Konsequenzen aus der Gradeinteilung sind entweder eine sorgfältige Kürettage bei Grad-I-Tumoren oder eine weite Resektion bei Grad-II- und Grad-III-Fällen. Die Langzeitergebnisse bedürfen allerdings noch einer Bewertung an größeren Kollektiven. Das Verfahren hat sich in der täglichen diagnostischen Praxis bei 104 Fällen mit einem klassischen Chondrosarkom als sehr gut anwendbar und reproduzierbar erwiesen.
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  • 3
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Ki-67 ; MIB-1 ; Proliferation ; Osteosarkom ; Niedrig maligne Osteosarkome ; Knochentumoren ; Key words Ki-67 ; MIB-1 ; Proliferation ; Osteosarcoma ; Low grade osteosarcoma ; Bone tumors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Bone tumors represent a group of tumors of various dignity. In spite of this single tumor entities may display strong morphological resemblance to each other which can in turn result in profound difficulties in differential diagnosis. The biological behaviour of a tumor is mainly determined by its rate of proliferation. In this study the rate of proliferation of 64 bone tumors (30 high-grade central osteosarcomas, 6 low-grade osteosarcomas, 8 giant cell tumors, 8 aneurysmatic bone cysts, 5 osteoidosteomas/osteoblastomas, 7 fibrous dysplasias and 5 cases of a myositis ossificans) were analysed. Immunohistochemistry was performed on paraffin-embedded tissue sections using the MIB-1 monoclonal antibody. MIB-1 recognizes the proliferation-associated Ki-67 protein which is expressed during the active phases of the cell cycle but cannot be detected in senescent cells. Among high-grade central osteosarcomas a significantly higher rate of proliferation (average value 30 %) was found in comparison with low-grade osteosarcomas and other benign intraosseous bone tumors. This approach proved to be very useful in the distinction between high-grade and low-grade osteosarcomas as well as bone-forming intraosseous tumors. However distinguishing low-grade osteosarcomas from benign bone tumors by determining only the rate of proliferation was not possible, although interestingly, the proliferative rate of myositis ossificans, a purely reactive lesion, was in the range of the values determined for high-grade osteosarcoma.
    Notes: Zusammenfassung Knochentumoren stellen eine Gruppe von Tumoren mit unterschiedlicher Dignität dar. Trotzdem besteht zwischen den einzelnen Tumorentitäten z. T. eine starke morphologische Ähnlichkeit, die im Einzelfall zu erheblichen differentialdiagnostischen Schwierigkeiten führen kann. Da das biologische Verhalten eines Tumors wesentlich durch seine Wachstumsgeschwindigkeit bestimmt wird, wurde die Proliferation an 64 Knochentumoren (30 hochmaligne zentrale Osteosarkome, 6 niedrig maligne Osteosarkome, 8 Riesenzelltumoren, 8 aneurysmatische Knochenzysten, 5 Osteoid-Osteome/Osteoblastome, 7 fibröse Dysplasien) und an 5 Fällen einer Myositis ossificans immunhistologisch durch den monoklonalen Antikörper MIB-1 an Paraffinmaterial untersucht. MIB-1 erkennt das Ki-67-Protein, das in den aktiven Phasen des Zellzyklus exprimiert wird, in ruhenden Zellen aber nicht nachweisbar ist. Bei den hochmalignen zentralen Osteosarkomen fand sich eine hohe Proliferationsrate von durchschnittlich annähernd 30 %, die signifikant höher war als die der niedrigmalignen Osteosarkome und der übrigen benignen intraossären Knochentumoren. Eine Abgrenzung zwischen den niedrigmalignen Osteosarkomen und den benignen Knochentumoren war durch die Bestimmung der Proliferationsrate allerdings nicht möglich. Die Ermittlung der Proliferationsrate mit dem Antikörper MIB-1 ist hilfreich zur Abgrenzung hochmaligner Osteosarkome von niedrig malignen Osteosarkomen und benignen knochenbildenden intraossären Tumoren, innerhalb dieser Tumoren ist aber eine weitere Differenzierung mit Hilfe der Proliferationsrate nicht möglich. Eine Ausnahme bildet die Myositis ossificans, die zeigt, daß auch rein reaktive Läsionen eine Proliferationsrate besitzen können, die die Werte hochmaligner Tumore erreicht.
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  • 4
    ISSN: 1432-2048
    Keywords: Enzyme modulation ; Nitrate reductase ; Protein kinase ; Protein phosphorylation ; Protein purification ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using a three-step purification procedure, two protein fractions which catalyzed the ATP-dependent in-activation of nitrate reductase (NR) were obtained from spinach (Spinacia oleracea L.) leaf extracts. Purification involved ammonium-sulfate fractionation, anion-exchange chromatography and size-exclusion chromatography. The capacity of the fractions to inactivate NR by preincubation with ATP was examined by using as target either a crude NR-ammonium sulfate precipitate or partially purified NR (ppNR). The fractions were also examined for protein-kinase activity by measuring the phosphorylation of histone III S (or casein) with γ-[32P]ATP as substrate, and subsequent SDS-PAGE, autoradiography and liquid scintillation counting of cut-off histone bands. The two proteins had apparent molecular weights in the 67-kDa and 100-kDa region (termed P67 and P100, respectively). Neither P67 nor P100 alone was able to inactivate ppNR by preincubation with ATP. However, when P100 and P67 were added together to ppNR, ATP-dependent inactivation was observed, with a half-time of about 10 min. The P67, but not P100 had histone-kinase activity (casein was not phosphorylated). Using the partially purified system, various compounds were examined as possible effectors of NR inactivation. Sugar phosphates had little effect on the inactivation of NR. Addition of AMP at very high concentrations (5 mM), and removal of Mg2+ by excess EDTA also prevented the inactivation.
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  • 5
    ISSN: 1432-2048
    Keywords: Inhibitor protein ; Nitrate reductase ; Protein kinase ; Protein phosphatase ; Protein turnover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrate reductase activity and NR protein levels in various leaf tissues were drastically decreased (〈3.5% of normal activity) either by keeping detached leaves in continuous darkness for up to 6 d (spinach), or by growing plants (pea, squash) hydroponically on ammonium as the sole N-source, or by germinating and growing etiolated seedlings in complete darkness (squash). The presence of nitrate reductase protein kinase (NRPK), nitrate reductase protein phosphatase (NRPP) and inhibitor protein (IP) was examined by measuring the ability of NR-free desalted extracts to inactivate (ATP-dependent) and reactivate (5′-AMP/EDTA-dependent) added purified spinach NR in vitro. Extracts from low-NR plants (ammonium-grown pea and squash) were also prepared from leaves harvested at the end of a normal light or dark phase, or after treating leaves with anaerobiosis, uncouplers or mannose, conditions which usually activate NR in nitrategrown normal plants. Without exception, extracts from NR-deficient plant tissues were able to inactivate and reactivate purified spinach NR with normal velocity, irrespective of pretreatment or time of harvest. Considerable NRPK, NRPP and IP activities were also found in extracts from almost NR-free ripe fruits (cucumber and tomato). Activities were totally absent, however, in extracts from isolated spinach chloroplasts. The NRPK and IP fractions were partially purified with normal yields from NR-deficient squash or spinach leaves, following the purification protocol worked out for nitrate-grown spinach. The Ca2+/Mg2+-dependent kinase fraction from NR-deficient squash or spinach phosphorylated added purified spinach NR with γ-[32P]ATP and inactivated the enzyme after addition of IP. It is suggested (i) that the auxiliary proteins (NRPK, IP, NRPP) which modulate NR are rather species- or organ-unspecific, (ii) that they do not turn over as rapidly as does NR, (iii) that they are probably expressed independently of NR, and (iiii) that they are not covalently modulated, but under control of metabolic and/or physical signals which are removed by desalting.
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  • 6
    ISSN: 1432-2048
    Keywords: Enzyme modulation ; Nitrate reductase ; Protein kinase ; Protein phosphorylation ; Protein purification ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using a three-step purification procedure, two protein fractions which catalyzed the ATP-dependent in-activation of nitrate reductase (NR) were obtained from spinach (Spinacia oleracea L.) leaf extracts. Purification involved ammonium-sulfate fractionation, anion-exchange chromatography and size-exclusion chromatography. The capacity of the fractions to inactivate NR by preincubation with ATP was examined by using as target either a crude NR-ammonium sulfate precipitate or partially purified NR (ppNR). The fractions were also examined for protein-kinase activity by measuring the phosphorylation of histone III S (or casein) withγ-[32P]ATP as substrate, and subsequent SDS-PAGE, autoradiography and liquid scintillation counting of cut-off histone bands. The two proteins had apparent molecular weights in the 67-kDa and 100-kDa region (termed P67 and P100, respectively). Neither P67 nor P100 alone was able to inactivate ppNR by preincubation with ATP. However, when P100 and P67 were added together to ppNR, ATP-dependent inactivation was observed, with a half-time of about 10 min. The P67, but not P100 had histone-kinase activity (casein was not phosphorylated). Using the partially purified system, various compounds were examined as possible effectors of NR inactivation. Sugar phosphates had little effect on the inactivation of NR. Addition of AMP at very high concentrations (5 mM), and removal of Mg2+ by excess EDTA also prevented the inactivation.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2048
    Keywords: Inhibitor protein ; Nitrate reductase ; Protein phosphorylation ; Protein kinase ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The function of two proteins (P67 and P100) required for the MgATP-dependent inactivation of nitrate reductase (NR) from spinach leaves (Spinacia oleracea L.) was studied. When NR was incubated with γ-[32P]ATP and P67, NR-protein was phosphorylated, but without a change in NR activity. Protein P100 by itself was neither able to phosphorylate nor to inactivate NR, and when added together with P67 it did not change the extent of NR phosphorylation. However, when NR was first phosphorylated with MgATP and P67, subsequent addition of P100 after removal of unreacted ATP caused an immediate NR inactivation. In presence of both P67 and P100 the time-course of ATP-dependent NR phosphorylation paralleled the time course of inactivation. The extent of NR phosphorylation and of NR inactivation (in the presence of P67 plus P100) was similarly affected by metabolites or high salt concentrations. Magnesium (Mg2+) played a dual role in the inactivation process: the phosphorylation of NR by P67 was strictly Mg2+-dependent. Further, phospho-NR (+P100) was inactive only in the presence of Mg2+, but active in the presence of excess EDTA. Dephospho-NR appeared to be Mg2+-insensitive. The observations suggest that phosphorylation of NR by P67 is obligatory, but not sufficient for inactivation. In addition to protein phosphorylation, inactivation requires “binding” of an inhibitor protein (P100) to phospho-NR.
    Type of Medium: Electronic Resource
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