Immunochemical localization ofin Vivo bound immunoglobulins in pemphigus vulgaris epidermis

Summary A multi-step immunocytochemical method utilizing horseradish peroxidase (HRP) as an immunological marker was employed to demonstrate, at the ultrastructural level, IgG antiepithelial autoantibodies boundin vivo to pemphigus epidermis, and circulating IgG antibodies, using monkey oesophagus as substrate. To demonstrate IgG the following antisera were employed in sequential steps: Goat antihuman IgG; rabbit antigoat IgG; goatanti-HRP-serum. After incubating with the final antigen, HRP, the latter was visualized with a cytochemical method. Results paralleled those obtained by immunofluorescence, the antiepithelial antibody being demonstrated at the sites of the intercellular spaces of the epidermis. Electron microscopic cytochemistry showed IgG in the surface coat of epidermal cells and in the intercellular space, both in the desmosomal and the interdesmosomal areas. These results confirm the results of a previous study but the localization of the antibody is more exact than that obtained with HRP-conjugates. The present method is more versatile and specific than HRP methods that utilize HRP-conjugated antibodies.

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Zusammenfassung Zum elektronenmikroskopischen Nachweisin vivo an Pemphigusepidermis gebundener antiepithelialer IgG-Autoantikörper und zirkulierender IgG-Antikörper wurde eine immuncytochemische Mehrstufenmethode mit Meerrettichperoxidase (HRP) verwendet. Folgende Antisera wurden zur Demonstration von IgG in aufeinander folgenden Schritten eingesetzt: Ziegen-antihuman-IgG, Kaninchen-anti-Ziegen-IgG und Ziegen-anti-HRP-Serum. Nach der Inkubation mit dem letzten Antigen, HRP, wurde dieses mittels einer cytochemischen Methode sichtbar gemacht. Die Ergebnisse stimmten mit denen der Immunfluorescenz völlig überein: Antiepitheliale Antikörper konnten im Intercellularraum der Epidermis aufgefunden werden. Die elektronenmikroskopisch-cytochemische Reaktion stellte IgG an der Oberfläche von Epidermalzellen und im Intercellularraum dar, und zwar sowohl in Desmosomen als auch im interdesmosomalen Bereich. Diese Ergebnisse bestätigen die Befunde einer früheren Untersuchung mit HRP-Konjugaten, die Lokalisation des Antikörpers ist jedoch exakter; die Methode ist vielseitiger und spezifischer als jene Methoden, bei welchen HRP konjugierte Antikörper verwendet werden.

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Supported, in part, by Fonds zur Förderung der wissenschaftlichen Forschung, Wien, grant No. 1416, and a research grant from Schering AG, Berlin.

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The authors gratefully acknowledge the excellent technical assistance of Mrs. Susan Csegezi, Mrs. Hilde Martinjak and Mrs. Lotte Polasek.     

The preliminary study on HMBA-induced differentiation of the drug-pretreated murine erythroleukemia cells

Several drug-resistant variants have been developed by growing the parental MEL cells in presence of colchicine, adriamycin and vincristine respectively with stepwise increasing concentration. Both the colchicine-resistant Sc9(Co10) and vincristine-resistant Sc9(VCR5) cells displayed an accelerated HMBA-induced commitment to terminal cell differentiation, whereas the adriamycin-resistant SC9 (A120) showed no acceleration but rather a substantial delay in HMBA-induced differentiation. The studies provide more clues as well as experimental models for further study on the mechanism of induced differentiation of MEL cells.     

175a. Spätergebnisse nach transhepatischer-diaenteraler Ringdrainage bei benignen Gallengangsstenosen

Zusammenfassung Benigne Choledochusstenosen sind in der Regel einer einfachen Rekonstruktion durch Endzu-End-Naht nicht mehr sinnvoll zugänglich. Um die funktionstüchtige Papille als besten Schutz vor Cholangitis zu erhalten, entwickelten wir zunächst tierexperimentell eine Methode der Gallengangserweiterung durch ein gestieltes Jejunum-Transplantat.. Nach erfolgreichem Abschluß der Serie haben wir bisher drei Patienten mit iatrogenen kurzstreckigen Choledochusstenosen nach dieser Methode operiert, die Patienten sind nach einer Nachbeobachtungszeit von 8–30 Monaten Beschwerde- und symptomfrei.     

Cyclophosphamide therapy of severe systemic necrotizing vasculitis.

We studied 17 patients with severe systemic necrotizing vasculitis over an 11-year-period. Sixteen patients were treated daily with cyclophosphamide (2 mg per kilogram per day), and one was treated with azathioprine (2 mg per kilogram per day). Before entering the study, all patients had active and progressive disease, even though 16 patients had been receiving corticosteroids that had caused severe and often incapacitating toxic side effects. Three patients died during the study. Complete and often dramatic remissions occurred in the surviving 14 patients, who were then placed on alternate-day corticosteroid treatment with continuation of cyclophosphamide. Corticosteroids were later discontinued in seven patients. The mean duration of remission was 22 months (range, two to 61). No patient showed recurrence of disease during treatment with cytotoxic agents.     

Deficiency of carnitine palmitoyltransferase I

Defective activity of carnitine palmitoyltransferase I was demonstrated in fibroblasts derived from a patient with hypoketotic hypoglycemia. The level of activity observed was approximately 10% of the control mean. Oxidation of palmitate by intact fibroblasts was reduced to 5% of control values. The patient presented at age 14 months with seizures and was found to have marked hypoglycemia and no ketones in the urine. In response to fasting, she developed hypoglycemia, but the curves for acetoacetate and 3-hydroxybutyrate were flat. Administration of medium-chain triglycerides relieved the hypoglycemia and generated a brisk ketogenesis.