Electron Microscopic Observations on the Ribonucleic Acid of Murine Leukemia Virus

The ribonucleic acid (RNA) of murine leukemia virus (MLV) Rauscher strain was observed by the aid of electron microscopy with the use of the protein monolayer technique. RNA was observed directly after release from virus particles or after isolation by sedimentation in sucrose density gradients. Molecules were found in an extended linear form. Many of the RNA filaments released by detergent treatment contained curled regions, suggesting the linear filaments were originally coiled within the virus particle. The relationship of the curled areas to the containment of the RNA within the virus particle is discussed, and a mechanism for the inclusion of RNA in the budding virion is proposed. Treatment of the extended MLV-RNA with dimethyl sulfoxide resulted in the collapse of the molecule forming a tangled complex. Treatment with urea or heating at 50 C in 3 mm NaCl also produced this effect. Also under the conditions in which MLV-RNA was linear, RNA from Rous sarcoma virus also was linear, but Newcastle disease virus RNA and ribosomal RNA of rat liver had collapsed structures. The results indicated that the RNA of MLV, and perhaps other RNA-containing tumor viruses, has a specific unique conformation dependent upon hydrogen bonds.     

Zytologische Beobachtungen an der hyperplastischen Nebennierenrinde des Menschen

Summary Clinical observations and the light and electron microscopic appearance of the adrenal cortex are described in a case of Cushing's disease associated with adrenocortical hyperplasia and a basophilic adenoma of the pituitary gland. The electron microscopic investigation concentrates upon the occurrence of osmiophil material in the different cytoplasmic compartments of the cell type rich in organelles and relatively poor in paraplasmic inclusions. It seems that the observation of osmiophil material in the perinuclear space, the mitochondrial matrix, Golgi vacuoles, and cytoplasmic vesicles has been made possible by a rather long fixation in Wohlfarth-Bottermann's chromium-osmium fixative. Mitochondria and related particles (named X-Einschlüsse) may contain a light center with a few dense filaments and masses resembling those described by other authors as being of deoxyribonucleic acid nature. Relatively large Golgi complexes are found, sometimes in close topographic relation with a centriole. A large population of osmiophil bodies (lipid droplets, pigment granules, microbodies) is described. Apart from these findings the cytoplasmic organization in the tissue studied is similar to that of the adrenal cortex as described in other mammals. The observations are discussed with respect to hypotheses about lipid droplet formation and to the possibility of artifacts due to preparation procedures.

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Herrn Prof. Dr. med. W. Bargmann zum 60. Geburtstag gewidmet. — Herrn Prof. Dr. E. Uehlinger, Direktor des Pathologischen Instituts der Universität Zürich, danken wir für die Überlassung des Materials und für die freundlicherweise zur Verfügung gestellte pathologisch-anatomische Diagnose, dem Schweizerischen Nationalfonds für die Unterstützung und Frau Jäger, Mlle. Challamel und Herrn Krieger für die Ausführung der histologischen und photographischen Arbeiten.     

A Cytochemical Study of the Stem Cell Concept in Specimens of a Human Ovarian Tumor

The amounts of DNA in interphase nuclei were compared with the amounts of DNA in metaphase and anaphase figures in Feulgen-stained tissue sections of 5 specimens of the human ovarian papillary serous adenocarcinoma. The relative amounts of DNA per cell were determined by cytophotometric measurements of interphase nuclei at a single wavelength and of mitotic figures by the two wavelength method. The 5 specimens conformed to the stem cell concept of cell proliferation since anaphase distributions of amounts of DNA were restricted to a narrow range of DNA values indicating the successful mitosis of a single cell type (stem cell) out of several cell types whose presence were suggested by the wide spread of interphase and metaphase values. In addition, the data indicated that, in some instances, only the amounts of DNA in anaphase figures can reliably identify the stem cell. Changes in the frequency of dividing cells having doubled amounts of DNA, and/or the presence of cells resulting from endoreduplication can distort the interphase distribution of amounts of DNA and thus give rise to a modal DNA interphase value which is not the same as the DNA value of the stem cell (anaphase figures).     

Extensive changes in cytokeratin expression patterns in pathologically affected human gingiva

The stratified squamous epithelium of the oral gingiva and the hard palate is characterized by a tissue architecture and a cytoskeletal composition similar to, although not identical with, that of the epidermis and fundamentally different from that of the adjacent non-masticatory oral mucosa. Using immunocytochemistry with antibodies specific for individual cytokeratins, in situ hybridization and Northern blots of RNA with riboprobes specific for individual cytokeratin mRNAs, and gel electrophoresis of cytoskeletal proteins of microdissected biopsy tissue samples, we show changes in the pattern of expression of cytokeratins and their corresponding mRNAs in pathologically altered oral gingiva. Besides a frequently, although not consistently, observed increase in the number of cells producing cytokeratins 4 and 13 (which are normally found as abundant components in the sulcular epithelium and the alveolar mucosa but not in the oral gingiva) and a reduction in the number of cells producing cytokeratins 1, 10 and 11, the most extensive change was noted for cytokeratin 19, a frequent cytokeratin in diverse one-layered and complex epithelia. While in normal oral gingiva cytokeratin 19 is restricted to certain, sparsely scattered cells of --or near--the basal cell layer, probably neuroendocrine (Merkel) cells, in altered tissue of inflamed samples it can appear in larger regions of the basal cell layer(s) and, in apparently more advanced stages, also in a variable number of suprabasal cells. Specifically, our in situ hybridization experiments show that this altered suprabasal cytokeratin 19 expression is more extended at the mRNA than at the protein level, indicating that cytokeratin 19 mRNA synthesis may be a relatively early event during the alteration. These changes in cytokeratin expression under an external pathological influence are discussed in relation to other factors known to contribute to the expression of certain cytokeratins and with respect to changes occurring during dysplasia and malignant transformation of oral epithelia.     

Augmentation of host antitumor immunity by low doses of cyclophosphamide and mafosfamide in two animal tumor models

Summary By cloning in vitro we have obtained two sublines of the L5222 rat leukemia, one with high (L5222-S) and the other with low (L5222-R) in vivo sensitivities to non-toxic doses of mafosfamide, a stabilized derivative of 4-hydroxy-cyclophosphamide. This sensitivity in vivo was not related to the cytotoxic activity of the drug in vitro. Treatment of rats bearing the L5222-S and of mice transplanted with the MOPC-315 plasmocytoma with low doses of mafosfamide or cyclophosphamide resulted in a high percentage of surviving animals, which were resistant to a subsequent tumor challenge. Viable leukemic cells were needed to establish antitumor immunity, since it was not possible to induce resistance by injection of mitomycin-C-treated, non-viable L5222 cells. The adoptive transfer of spleen cells from animals immune against the L5222-S and the MOPC-315 resulted in resistance of the syngeneic recipients against a rechallenge with tumor cells, provided that the animals were treated with an immunosuppressive dose (100 mg/kg) of cyclophosphamide prior to the spleen cell implantation. In nude mice treatment of the L5222 with low doses of mafosfamide also resulted in surviving animals, however resistance to a second tumor challenge occurred only sporadically.The data presented confirm that therapy with cyclophosphamide or mafosfamide enhances host antitumor immunity but, contrary to previous reports, it could be demonstrated that successful tumor rejection was independent of T cells.Supported by the Federal Ministry of Research and Technology (BMFT), Bonn-Bad Godesberg, FRG