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  • 1995-1999  (9)
  • 1930-1934
  • Osteosarcoma  (4)
  • Spinacia  (3)
  • Fibronectin  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 110 (1997), S. 18-21 
    ISSN: 1437-1596
    Keywords: Key words Proteinase inhibitors ; Fibronectin ; Lysozyme ; Immunohistochemistry ; Autolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Notes: Abstract The influence of postmortem damage of tissues on the immunohistochemical diagnosis of wound age has not as yet been clarified. We utilized antibodies against the proteinase inhibitors α-1-antichymotrypsin and α-2-macroglobulin, fibronectin and lysozyme to study samples of skin which had been intact intravitally, but were damaged postmortem either by autolysis or compression with a surgical clamp at the time of dissection. Even in the absence of autolysis, antibodies against the proteinase inhibitors and fibronectin exhibited staining of tissue margins. Autolysis caused an increase in false positive results. In contrast, antibodies against lysozyme did not give false positive staining. There were no antigens sensitive to postmortem clamping and false positive results were not observed. Antibodies against proteinase inhibitors are not useful for the diagnosis of wound age because of a high number of false positive reactions in marginal areas. Fibronectin also showed false positive band-shaped staining patterns at the tissue margin. In addition, autolytic processes increase the number of false positives. The antibody against lysozyme is much less sensitive to autolysis and no false positive reactions were observed in our series of tests.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 110 (1997), S. 18-21 
    ISSN: 1437-1596
    Keywords: Proteinase inhibitors ; Fibronectin ; Lysozyme ; Immunohistochemistry ; Autolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Notes: Abstract The influence of postmortem damage of tissues on the immunohistochemical diagnosis of wound age has not as yet been clarified. We utilized antibodies against the proteinase inhibitors α-1-antichymotrypsin and α-2-macroglobulin, fibronectin and lysozyme to study samples of skin which had been intact intravitally, but were damaged postmortem either by autolysis or compression with a surgical clamp at the time of dissection. Even in the absence of autolysis, antibodies against the proteinase inhibitors and fibronectin exhibited staining of tissue margins. Autolysis caused an increase in false positive results. In contrast, antibodies against lysozyme did not give false positive staining. There were no antigens sensitive to postmortem clamping and false positive results were not observed. Antibodies against proteinase inhibitors are not useful for the diagnosis of wound age because of a high number of false positive reactions in marginal areas. Fibronectin also showed false positive band-shaped staining patterns at the tissue margin. In addition, autolytic processes increase the number of false positives. The antibody against lysozyme is much less sensitive to autolysis and no false positive reactions were observed in our series of tests.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Key words: Nitrate reductase ; Respiration inhibitors ; Spinacia ; Strobilurin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Spinach (Spinacia oleracea L.) leaf discs floating on buffer solution were treated with Kresoxim-methyl (KROM), an inhibitor of respiratory electron transport. In the leaf tissue, actual and maximal nitrate reductase (NR) activities, nitrite content and ATP levels were determined. In darkened leaf discs incubated without KROM (control) actual NR activity decreased to 20% after 6 h in the dark. Treatment with 10 μg ml−1 (corresponding to 32 μM) KROM totally prevented inactivation of NR in the dark and also diminished NR-protein degradation during prolonged darkness. Due to restricted nitrite reduction in darkened leaf tissues, nitrite accumulated in KROM-treated discs. Inhibition of respiration decreased ATP and increased AMP levels in KROM-treated discs. In illuminated leaf discs, NR was highly activated to 65%. Nevertheless, KROM-treatment caused an additional activation of NR (activation state 76%) in the light. Possible side-effects of KROM on nitrite reduction and photosynthesis were also checked in the leaf-disc system. Neither nitrite reduction nor photosynthesis were altered in KROM-treated discs. The extent of KROM-induced activation of NR was dependent on the applied concentration and on the pH of the external medium. The highest activation of NR was achieved at an external pH of 4.8, confirming previous results (Kaiser and Brendle-Behnisch, 1995, Planta 196: 1–6) that cytosolic acidification might play an important role in the modulation of NR activity.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Planta 196 (1995), S. 1-6 
    ISSN: 1432-2048
    Keywords: Acid-base loading ; Nitrate reductase ; pH regulation (intracellular) ; Protein phosphorylation ; Spinacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of acid or base-loading of spinach (Spinacia oleracea L.) leaf discs on the activation status of nitrate reductase (NR) in the dark and in the light was investigated. Activity of NR (NRA), measured in crude extracts of leaf discs with removed lower epidermis, which had been floating on Mes-buffer [2-(N-morpholino)ethane sulfonic acid] pH 5.2 in the dark, was at a similar low level as in whole, darkened leaves. By addition of acetate or propionic acid, butyric acid or benzoic acid, NR was activated to or beyond the light level. The pH of crude tissue extracts was decreased by 0.5–1 pH units. Tissue acidification caused an inhibition of photosynthesis and of dark CO2 fixation. The acid-induced activation of NR in vivo was largely prevented by okadaic acid, an inhibitor of Type 1 and Type 2A protein phosphatases. This indicates that acid-induced activation was mediated by protein dephosphorylation. When, on the other hand, leaf discs were illuminated on Ches-buffer (2-[ N-cyclohexylamino]ethane sulfonic acid) pH 9 in the presence of bicarbonate (80 mM), their NR was as active as in intact leaves. Addition of ammonium chloride (up to 6 mM) caused a pH increase of the tissue extract up to 0.9 pH units. At the same time NR was inactivated to the dark level. Methionine sulfoximine did not prevent the ammonium effect. Photosynthesis and dark CO2 fixation were stimulated at pH 9 by ammonium chloride (1–2· mol· m −3) and were only slightly inhibited by up to 6 mol· m−3. The modulation of NR by acid-base treatment in vivo was fully reversible. The response of the NR system to acid or base treatment is consistent with a proposed role of nitrate reduction in the cellular pH-stat. The observation also indicates that cytosolic pH changes may be involved the signal chain triggering the modulation of NR.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Knochentumoren ; Chondrosarkom ; Osteosarkom ; Zytogenetik ; DNA-Zytometrie ; Ploidie ; Key words Bone neoplasms ; Chondrosarcoma ; Osteosarcoma ; Cytogenetics ; DNA-cytometry ; Ploidy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary 10 chondrosarcomas and 10 osteosarcomas were examined using cytogenetics and DNA-image-cytometry. Cytogenetically 6 of 10 chondrosarcomas and 4 of 10 osteosarcomas showed hyperdiploid tumorcells. By DNA-cytometry in 8 of 10 chondrosarcomas and 9 of 10 osteosarcomas hyperdiploid tumorcells resp. hyperdiploid stemlines were detected. This discrepancy reflects an in-vitro-selection depending on the different entities. In 7 aneuploid clones of chondrosarcomas the chromosomal ploidy was calculated using the relative length of the chromosomes and compared with the DNA-ploidy of the native tumor. There was a close relation between both parameters of nuclear DNA-content. The interpretation of cytogenetic results is improved using a combination of karyotypic and DNA-cytometric examination. This is particularly important for the search for relations between numeric chromosomal aberrations and morphological parameters (grading).
    Notes: Zusammenfassung 10 Chondrosarkome und 10 Osteosarkome wurden tumorzytogenetisch und DNA-zytometrisch untersucht. Das Karyogramm erbrachte bei 6 von 10 Chondrosarkomen und bei 4 von 10 Osteosarkomen den Nachweis hyperdiploider Tumorzellklone. DNA-zytometrisch wurden am nativen Tumormaterial jedoch bei 8 von 10 Chondrosarkomen und bei 9 von 10 Osteosarkomen hyperdiploide Tumorzellen, häufig in Form eigenständiger hyperdiploider Stammlinien nachgewiesen. Diese Diskrepanz ist Ausdruck einer offenbar Entitäts-abhängigen In-vitro-Selektion. Bei insgesamt 7 aneuploiden Tumorzellklonen von Chondrosarkomen konnte die chromosomale Ploidie anhand der relativen Chromosomenlängen exakt errechnet und der zytometrisch bestimmten DNA-Ploidie gegenübergestellt werden, wobei sich eine sehr enge Abhängigkeit zwischen diesen beiden Parametern des nukleären DNA-Gehaltes ergab. Die Interpretation zytogenetischer Befunde bei Knochentumoren wird durch Kombination mit der DNA-Zytometrie verbessert. Dies ist besonders dann wichtig, wenn Zusammenhänge zwischen numerischen chromosomalen Aberrationen und morphologischen Parametern (z. B. Grading) dargestellt werden sollen.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Knochentumoren ; Osteosarkom ; Neoadjuvante Chemotherapie ; Regressionsgrad ; Histologische Typisierung ; Tumorgröße ; Key words Bone neoplasms ; Osteosarcoma ; Neoadjuvant chemotherapy ; Degree of regression ; Histological typing ; Tumor size
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary 207 osteosarcomas were examined morphologically after neoadjuvant chemotherapy according to the COSS-86 protocol using representative slides of one whole tumor plane. The rate of responders 63 %. In relapse-free patients both the whole tumors and the vital areas there of were smaller than in patients with relapse during a follow-up period of 5 years. Within the subgroup of osteoblastic osteosarcomas, metastases were observed following smaller tumors than in chondroblastic osteosarcomas. Therefore, in addition to degree of regression, histological subtype and tumor size should be considered in the prognostic evaluation of osteosarcomas.
    Notes: Zusammenfassung Im Rahmen der kooperativen Osteosarkomstudie (COSS 86) wurden 207 Resektionspräparate von Osteosarkomen anhand der Auswertung einer Gesamttumorebene zur Bestimmung des Regressionsgrads morphologisch untersucht. Innerhalb der protokollgerecht auswertbaren Fälle betrug das Verhältnis zwischen Respondern und Nonrespondern 63 % : 37 %. Relapse-freie Patienten wiesen zum Zeitpunkt der Resektion einen kleineren Gesamttumor und geringere vitale Tumoranteile auf als Patienten mit einem metastasierenden Verlauf. Bei Berücksichtigung des histologischen Subtyps traten metastasierende Verläufe innerhalb einer Nachbeobachtungszeit von mindestens 5 Jahren in der Gruppe der osteoblastischen Osteosarkome bereits bei einer niedrigeren Gesamttumorgröße auf als bei den chondroblastischen Osteosarkomen. Bei der prognostischen Einschätzung von Osteosarkomen sollten deshalb neben dem Regressionsgrad als Parameter für das Ansprechen auf die Chemotherapie auch der histologische Aufbau und die Gesamttumorgröße Beachtung finden.
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  • 7
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Chemotherapieresistenz ; P-Glykoprotein ; MDR ; Osteosarkom ; Key words Multidrug resistance ; P-glycoprotein ; Osteosarcoma ; Chemotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary One of the mechanisms by which multidrug resistance is mediated, is the mdr1 gene product, P-glycooprotein. Immunohistochemistry was performed for 63 osteosarcomas of 54 patients to investigate P-glycoprotein expression using the monoclonal antibody JSB-1. Most of the patients were children or adolescents who had received treatment under the framework of the Cooperative Osteosarcoma Study Group. In addition P-glycoprotein expression was assayed in five growth plates. Of all cases 68.5 % stained positive for P-glycoprotein. Cases that had received chemotherapy showed a higher incidence (80.9 %) of positive P-glycoprotein immunostaining than cases that had not received chemotherapy (66.6 %). No relation could be established between P-glycoprotein expression and the response to chemotherapy, since the majority of P-glycoprotein positive biopsies showed a good response in the surgical specimen after chemotherapy. Furthermore, 42.9 % of P-glycoprotein negative biopsies were classified as non-responders in the later surgical specimen. In addition to P-glycoprotein expression in osteosarcomas positive immunostaining was also detected in osteoblasts, osteocytes, osteoclasts as well as in some chondroblasts. The results indicate that P-glycoprotein expression in osteosarcomas also exists prior to chemotherapy and resembles the phenotype of normal bone tissue. However, the determination of P-glycoprotein by using immunohistochemistry in biopsies of osteosarcomas cannot predict the response to chemotherapy.
    Notes: Zusammenfassung Die Expression des mdr1-Genproduktes, das P-Glykoprotein, ist einer der Mechanismen der „multidrug resistance“. In 63 Osteosarkomen von 54 Patienten wurde die P-Glykoproteinexpression immunhistologisch mit Hilfe des monoklonalen Antikörpers JSB-1 untersucht. Bei den Patienten handelte es sich überwiegend um Kinder oder Jugendliche, die im Rahmen der kooperativen Osteosarkomstudie therapiert worden waren. Zusätzlich wurde die P-Glykoproteinexpression an 5 Wachstumsfugen untersucht. Für P-Glykoprotein positiv waren 68,5 % aller Osteosarkomfälle. Dabei waren Osteosarkome nach Chemotherapie mit 80,9 % häufiger positiv als Osteosarkome, bei denen keine Chemotherapie erfolgt war (66,6 %). Die untersuchten Metastasen zeigten alle eine positive Pgp-Expression. Die Pgp-Expression an der Biopsie ließ allerdings keine Rückschlüsse auf das spätere Ansprechen auf Chemotherapie zu, da die überwiegende Mehrheit P-Glykoprotein-positiver Osteosarkombiopsien am späteren Resektat ein gutes Ansprechen (Responder) auf die Chemotherapie zeigten. Darüber hinaus waren 42,9 % der P-Glykoprotein-negativen Biopsien später der Nonrespondergruppe zuzuordnen. Neben einer Pgp-Expression an Osteosarkomen konnte auch eine positive Reaktion mit dem JSB-1-Antikörper an Osteoblasten, Osteozyten, Osteoklasten sowie an einem Teil der Chondroblasten gefunden werden. Die Ergebnisse zeigen, daß eine P-Glykoproteinexpression beim Osteosarkom auch vor der Chemotherapie nachweisbar ist und den Phänotyp regelhaften Knochengewebes widerspiegelt. Das Ansprechen auf die Chemotherapie kann jedoch mit Hilfe einer immunhistologischen Bestimmung der P-Glykoproteinexpression an der Biopsie nicht vorausgesagt werden.
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  • 8
    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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  • 9
    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.
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