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  • 1
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Bei 68 männlichen kastrierten Ratten, die teilweise zusätzlich adrenalektomiert waren, wurden nach unterschiedlicher Behandlung mit Östradiol die spezifischen Aktivitäten von alkalischer und saurer Nierenphosphatase in Rinde und Außenstreifen des Marks quantitativ bestimmt und die Resultate durch histochemische Enzymdarstellung veranschaulicht. Die Resultate stellen sich wie folgt dar: 1. Eine Aktivitätssteigerung der alkalischen Nierenphosphatase kann mit einer einmaligen Zufuhr von 2,5 μg Östradiol sc./100 g erzielt werden. Danach nimmt die Enzymaktivität maximal in der Rinde (18 MKH, Ausgangsniveau 13,7 MKH) um 31%, im Außenstreifen (29 MKH, Ausgangsniveau 12,0 MKH) um 142% zu. Bei täglich wiederholter Östradiolzufuhr von 0,5 μg sc./100 g beträgt nach 5–12 Tagen der Aktivitätszuwachs in der Rinde 27% (17,4 MKH) und im Außenstreifen 185% (34,2 MKH). Bei demselben Vorgehen mit 0,5 mg sc./100 g ist spätestens nach 5 Tagen die höchstmögliche Aktivierung der alkalischen Nierenphosphatase erreicht mit einer spezifischen Aktivität von 39,7 MKH in der Rinde und 44,9 MKH im Außenstreifen, das bedeutet einen Zuwachs von 180% bis 280% gegenüber den Kontrollen. Aus diesen Befunden geht hervor, daß die gestreckten Hauptstücke auf Östradiol wesentlich empfindlicher mit einer Steigerung der Enzymaktivität reagieren als die gewundenen Hauptstücke, daß aber bei hohen Dosen (mg!) auch in den gewundenen Hauptstücken eine massive Aktivitätssteigerung der alkalischen Phosphatase zu erzielen ist. Auf den Zeitpunkt der Aktivitätszunahme hat die Höhe der Östradioldosis jedoch keinen Einfluß. 2. Bei einer einmaligen Dosis von 2,5 μg sc./100 g ist die Aktivitätssteigerung der alkalischen Phosphatase im Außenstreifen frühestens nach etwa 6stündiger Östradioleinwirkung nachweisbar. Aus dem zeitlichen Verlauf geht jedoch hervor, daß der Prozeß der Aktivitätssteigerung spätestens etwa 3 Std früher einsetzt. Jenseits der 6. Std steigt die Aktivität monoton bis zu ihrem Höhepunkt zwischen der 40. und 50. Std an. Danach klingt die Enzymaktivität in Rinde und Außenstreifen wieder ab, hat aber ihren Ausgangswert nach 72 Std noch nicht wieder erreicht. 3. li]Eine starke Aktivitätszunahme der sauren Phosphatase in Rinde und Außenstreifen findet sich gleichzeitig nur bei den im mg-Maßstab mit Östradiol behandelten Ratten. Sie wird als Ausdruck eines sekundären zellulären Anpassungsvorganges gedeutet.
    Notes: Summary In 68 male castrated rats, some of them being additionally adrenalectomized, the specific activity of the alkaline and acid kidney phosphatase in cortex and outer stripe of the medulla was determined quantitatively following different treatment with estradiol. The results were visualized by the histochemical demonstration of the enzymes. The results are as follows: 1. An increase of the alkaline phosphatase activity in the kidney can be reliably reproduced by a single dose of 2,5 μg estradiol/100 g injected subcutaneously. Thereafter the enzyme activity shows a maximal increase of 31% in the cortex (18 MKH, basic level 13,7 MKH) and a maximal increase of 142% in the outer stripe (29 MKH, basic level 12 MKH). With repeated daily applications of 0,5 μg estradiol sc./100 g for a period of 5 to 12 days the increase of the activity amounts to 27% in the cortex (17,4 MKH) and 185% in the outer stripe (34,2 MKH). Using the same mode of application but a dose of 0,5 mg instead of 0,5 μg sc./100 g the highest possible activation of the alkaline phosphatase is reached on the 5th day at the latest with a specific activity of 39,7 MKH for the cortex and 44,9 MKH for the outer stripe. These values represent an increase of 180% and 280% respectively when compared with untreated animals. These findings demonstrate that there is a more sensible reaction of the enzyme activity to estradiol in the straight part of the proximal tubule than in the convoluted part. With high doses (mg-range!), however, a massive activation of the alkaline phosphatase can also be produced in the convoluted tubules. The amount of the estradiol dose has no influence on the onset of the enzyme activation. 2. Following a single dose of 2,5 μg estradiol sc./100 g the activation of the alkaline phosphatase in the outer stripe can at the earliest be detected 6 hours after injection. From the time course it results, however, that the process of activation starts 3 hours earlier. Beyond the 6th hour the activity rises steadily to its maximum value which is reached between the 40th and 50th hour. Thereafter the enzyme activity of the cortex and outer stripe decreases, but does not come down to its basic level even after 72 hours. 3. A marked increase of the acid phosphatase activity in cortex and outer stripe is simultaneously found only in rats treated with estradiol doses in the mg-range. This phenomemon is suggested to be the expression of a secondary cellular process of adaptation.
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  • 2
    ISSN: 1432-2307
    Keywords: Schönlein-Henoch's disease ; Glomerulonephritis ; Glomerular basement membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In glomerulonephritis accompanying the Schönlein-Henoch syndrome (SHS) a characteristic subepithelial basement membrane change is present in 85% of cases. The subepithelial change is a reaction to subepithelial deposits and consists of a garland or dome-like new formation of thin densa lamellae. This change is much more frequent in SHS than in IgA-nephritis or idiopathic glomerulonephritis or any other systemic disease. Furthermore, subepithelial deposits (50% of cases) are nearly as frequent as subendothelial deposits (65%) and more often present than formerly assumed.
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  • 3
    ISSN: 1432-2307
    Keywords: Monoclonal pan-epithelial antibody ; Immunohistochemistry ; Tumour diagnosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A mouse monoclonal antibody (mAB lu-5) was prepared using a lung cancer cell line as an antigen. The selected clone produces an IgG with a gamma-1 heavy chain and a kappa-light-chain. Immunohistochemical testing of mAB lu-5 on 117 normal tissue biopsies and 474 tumours revealed reactivity with an intracytoplasmic, formaldehyderesistant antigen present in most epithelial and mesothelial cells, but absent in mesenchymal cells. The antibody can therefore be used as a first order, pan-epithelial marker. It proved also useful for fast tumour diagnosis on frozen sections.
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  • 4
    ISSN: 1432-2307
    Keywords: Cytokeratin ; Intermediate Filament Proteins ; Conformation-dependent epitopes ; Immunocytochemistry ; Tumor diagnosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The epitope recognized by the murine monoclonal antibody (mAB lu-5) recently described as a formaldehyde-resistant, “pan-epithelial marker” of great value in tumour diagnosis is located on the surface of cytokeratin filaments. It has been preserved during vertebrate evolution from amphibia to man. As this epitope is not reactive after SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the epitope-bearing protein has been identified by a dot-blot antibody binding assay, using purified proteins in which the epitope is reconstituted. We show that the epitope is present in most cytokeratin polypeptides of both the acidic (type I) and basic (type II) subfamily but does not occur in other cytoskeletal proteins. The location of this widespread epitope is discussed with respect to homologies of amino acid sequences of cytokeratins and their conformations.
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  • 5
    ISSN: 1432-2307
    Keywords: Cardiac myxoma ; Immunohistochemistry ; Histogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immunohistochemical investigation of 11 cardiac myxomas (CMs) including one malignant metastasizing CM showed a co-expression of epithelial (lu-5 and CAM 5.2), mesenchymal (vimentin) and neuroendocrine antigens (neuron-specific enolase) in all tumour cells. Factor VIII was found in the endothelial cells of capillaries only. In the subendocardium of fetal heart tissue close to the foramen ovale myofibroblasts reacting with the panepithelial antibody lu-5 were detected. We conclude that CMs are neoplasms that may develop from embryonic cell remnants.
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  • 6
    ISSN: 1432-2307
    Keywords: Malignant mesothelioma ; Lung adenocarcinoma ; Ber-EP4 ; BMA-120 ; Blood-group-isoantigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Specimens of 27 histologically definite mesotheliomas and 34 proven adenocarcinomas were examined with a panel of 14 antibodies: pan-epithelial antibody Lu-5, anti-keratin-18, anti-keratin-7, Ber-EP4, anti-Leu-M1, HEA-125, anti-carcino-embryonic antigen (CEA), anti-blood group-related antigens (anti-BGR A, B, H), B 72.3, anti-placental alkaline phosphatase (PLAP), anti-vimentin and BMA-120 used to determine their value in the differentiation between pleural mesothelioma and lung adenocarcinoma. Lu-5, anti-cytokeratin-7 and -18, B 72.3 and PLAP reacted in a high percentage of cases with both mesothelioma and adenocarcinoma. Anti-CEA and anti-Leu-Ml did not react with any of the 27 mesotheliomas tested but showed a reaction in 75% (anti-CEA) and 66% (anti-Leu-M1) of the lung adenocarcinomas. Seventeen percent of the adenocarcinomas and 96% of the mesotheliomas showed a positive reaction with anti-vimentin. Ber-EP4 was demonstrated in all lung adenocarcinomas, but only in 2 mesotheliomas in a focal manner (7%). HEA-125 and anti-BGR A, B, H reacted with 83% (HEA-125) and 75% (anti-BGR A, B, H) of the lung adenocarcinomas. The statistical parameters, sensitivity and efficiency were estimated and a normogram for judging the diagnostic power of a single antibody for the differential diagnosis of mesothelioma versus adenocarcinoma was developed. According to this, Ber-EP4, HEA-125, anti-BGR A, B, H and anti-CEA were, in descending order, the most powerful discriminatory antibodies.
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  • 7
    ISSN: 1432-2307
    Keywords: Malignant melanoma ; MAGE-1 ; Immunohistology ; Tumour antigen ; Tumour markers ; Biological immunology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The MAGE-1 gene encodes a protein encompassing a HLA-A1-restricted target epitope for cytolytic T lymphocytes. Monoclonal antibodies directed against the MAGE-1 protein were tested for usage in immunohistology of routine pathology material. Seven formalinfixed, paraffin-embedded malignant melanomas were studied by the Avidin-Biotin complex (ABC) method with or without different antigen retrieval methods. Native, frozen tissues from the same tumours were used to validate the results by immunohistochemistry on frozen sections, by PCR for mRNA and by protein demonstration in tissue extracts using western blotting. Of 4 monoclonal antibodies tested, mAB 34B and mAB 77B were highly efficient in detecting MAGE-1 protein in deparaffinised sections with the regular ABC method after microwave pretreatment. In a series of an additional 28 patients 75% expressed MAGE-1, 50% in a substantial proportion. Follow-up studies in 6 patients indicate that the expression pattern remains stable but may change substantially within a short range. Immunohistology is thus a rapid and well-established method that might be used to select and monitor HLA-A1 positive patients with malignant melanoma and other candidate tumours for MAGE-1-directed immuno-therapy.
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  • 8
    ISSN: 1432-2307
    Keywords: Thromboxane ; Thromboxane synthase ; Immunohistochemistry ; Mononuclear phagocyte system ; Epithelia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using the monoclonal antibody Tü 300 we localized thromboxane synthase, a secondary enzyme of the arachidonic acid cascade, employing the alkaline phosphatase anti-alkaline phosphatase method and indirect double labelling immunofluorescence in frozen sections of human tissues. Aside from platelets, the source of the antigen, all cells of the mononuclear phagocytic system were positive, including epithelioid cells and associated giant cells, starry sky macrophages, dendritic cells of T-cell areas, Langerhans cells and Kupffer cells. In addition, some epithelial cells such as epithelia of tonsillar crypts, reticular epithelia of the thymic cortex and ductular epithelia in liver, pancreas, female breast and salivary glands showed occasional focal reactivity for thromboxane synthase. We suggest that the mAb Tü 300 is a key marker for the macrophage system and the thromboxane generating system in normal and pathological conditions. It may detect functional activities of as yet unknown significance in some specialized epithelial cells.
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  • 9
    ISSN: 1432-2307
    Keywords: Kidney ; Thromboxane ; Thromboxane synthase ; Tü 300 monoclonal antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Thromboxane, excreted in the urine in increased amounts in glomerular, vascular and tubulo-interstitial diseases, is considered to originate from the kidney. The localization of thromboxane synthase, a key enzyme of arachidonic acid metabolism, was studied in the human kidney by immunohistology using the monoclonal antibody Tü 300. In the interstitial tissue dendritic reticulum cells surrounding the tubules expressed high concentrations of the enzyme. In glomeruli the enzyme was weakly expressed in podocytes. This was confirmed by co-localization with an antiserum directed to podocalyxin, a marker of the visceral epithelial cells. In the study of various kidney diseases, massive accumulation of thromboxane synthase containing cells was observed in interstitial diseases, whereas in glomerular diseases there were no differences from normal kidney; in a case of thrombotic microangiopathy podocytes exhibited an increase in thromboxane-synthase. The thromboxane-synthase positive infiltrating interstitial cells were shown by conventional light microscopy to be mononuclear phagocytic cells. The physiological sources of renal thromboxane are dendritic reticular cells and podocytes. In interstitial renal disease infiltrating cells of the monocyte/macrophage system constitute the major site of thromboxane synthesis. In glomerular disease, a characteristic alteration of thromboxane-synthase was not found.
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  • 10
    ISSN: 1432-2307
    Keywords: Bladder neoplasm ; p53 ; erbB-2 ; Metastasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Overexpression of p53 anderbB-2 was studied by immunohistochemistry in formalin-fixed tissue samples of 179 patients with transitional cell carcinoma of the urinary bladder. p53 immunostaining was strongly correlated with tumour stage (P〈0.0001). This was driven by a marked difference in p53 expression between pTa (37% positive) and pT1 (71%) tumours, while there was no difference between pT1 and pT2-4 tumours. Similarly, a strong overall association between p53 expression and grade (P〈0.0001) was driven by a marked difference between grade 1 (28%) and grade 2 tumours (71%), and there was no significant difference between grade 2 and grade 3 tumours. Surprisingly, the frequency oferbB-2 overexpression was higher in pT1 tumours (74%) than in either pTa (49%;P=0.0265) or pT2-T4 (56%;P=0.0645) tumours. Both p53 anderbB-2 expression was also associated with metastasis. Metastases were found in 77% of patients with p53 positive primary tumours, but in only 50% of the patients with p53 negative primary tumours (P=0.022). Metastases were found in 66% of patients witherbB-2 positive primaries, but in only 37% of theerbB-2 negative primaries (P=0.020). Of 32 patients with positivity for both p53 anderbB-2, 84% developed metastases, as compared to 49% of patients with positivity for either one or neither positive (P=0.002). We conclude that both p53 anderbB-2 over-expression are associated with early invasion in bladder cancer. Furthermore, p53 anderbB-2 may be important predictors for metastasis.
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