Zur Elektronenmikroskopie der Kerneinschlüsse im menschlichen Nebenhodenepithel

Zusammenfassung Es werden verschiedene Typen von Einschlußkörperchen im Karyoplasma der Epithelzellen des menschlichen Nebenhodenepithels beschrieben. Die verschieden strukturierten Kerninklusionen werden als eine Differenzierungsreihe aufgefaßt. Sie beginnt mit kleinen rundlichen Körperchen aus feinfädigem Material und führt über die Ausbildung von Anhäufungen dichter homogener Kugeln zu großen Vakuolen wechselnden Inhaltes.

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Summary Different types of inclusion bodies are described in the karyoplasm of the epithelial cells of the human epididymis. The structural differences between these inclusions are interpreted as being indicative of consecutive stages in the process of their formation. Thus small, spherical bodies consisting of a fine fibrous material are believed to be the initial stages in the formation of the inclusion bodies whereas the dense, homogenous globules are thought to represent a later stage. Large vacuoles containing different materials are regarded as the final stage in this process of differentiation.

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Herrn Professor Dr. W. Bargmann zum 60. Geburtstag gewidmet.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

The Electrophoretic Velocity of Human Red Cells, of Their Ghosts and Mechanically Produced Fragments, and of Certain Lipid Complexes

Ghosts prepared in CO₂-saturated water from unwashed human red cells can be fragmented mechanically, but ghosts from thrice washed cells cannot. If the ghosts are prepared by freezing and thawing, this difference is not observed. The electrophoretic velocity varies also with the way in which the ghosts are prepared. The pH-mobility dependence of washed red cells flatten off to a plateau at pH 9, and the electrophoretic velocity is zero at about pH 2. Ghosts prepared by freezing and thawing have almost the same pH-mobility dependence, but if the ghosts are prepared in CO₂-saturated hyptonic saline, the mobility at pH 9.4 is 0.75 times that of washed cells. Fragments of ghosts of unwashed red cells have a smaller mobility than that of the red cells. Trypsin reduces the mobility of washed red cells and of ghosts. Sols of lipid complexes (lecithin, cephalin, and lipositol), at varying pH's, have a mobility 1.2 times that of the washed red cell. The pH-mobility relation is otherwise similar. These complexes can be coated with dextran and trypsin.     

Rapid Differentiation Between Nocardia and Streptomyces by Paper Chromatography of Whole-Cell Hydrolysates

Whole-cell hydrolysates were prepared from 58 strains of nocardiae and streptomycetes. Strains morphologically intermediate between the two genera and morphological variants of the same strains were included. Paper chromatograms made from the whole-cell hydrolysates clearly demonstrated meso-diaminopimelic acid as a major constituent of cultures of Nocardia spp., and LL-diaminopimelic acid as a major constituent of cultures of Streptomyces spp. In cultures of ten strains of N. madurae and of three of N. pelletieri, meso-diaminopimelic acid predominated, thereby supporting the assignment of these species to the genus Nocardia.     

Genetic analysis of streptomycin-resistance and-dependence inChlamydomonas

Summary Three non-chromosomal and two chromosomal genes which influence resistance to streptomycin are described. Each of the non-chromosomal factors,sr-500,sr-1500, andsd, exhibits uniparental inheritance, with all progeny receiving the factor when it is carried by the parent of mating-typeplus, and none when it is carried by the mating-typeminus parent. The streptomycin-dependence factor,sd, shows zygotic dominance when contributed by the mating-typeplus parent, but not when coming from the mating-typeminus parent, indicating that the uniparental transmission results from events occurring within the zygote early in maturation and well before meiosis. The chromosomal geneA interacts both with chromosomal and non-chromosomal genes at the biochemical level, but does not alter their patterns of inheritance.With 1 Figure in the TextThis paper is dedicated to ProfessorL. C. Dunn in gratitude to him as teacher and advisor, on the occasion of his retirement.This work was supported by grants from the U.S. Public Health Service and the National Science Foundation. The generosity and interest of ProfessorFrancis J. Ryan in providing laboratory space is gratefully acknowledged, as is the technical assistance of MissFran Yablonsky.     

Effects of seven different mutations in the pho1 gene on enzymatic activity, glycosylation and secretion of acid phosphatase in Schizosaccharomyces pombe

Summary Structural gene mutants of the cell-surface glycoprotein acid phosphatase of Schizosaccharomyces pombe were analysed to define structural determinants that are responsible for enzymatic activity, N-glycosylation and secretion. All seven defined mutations cause a single amino acid substitution in the mature acid phosphatase protein and destroy the enzymatic activity. The mutational lesions are distributed throughout the pho1 gene. A ser to phe substitution at position 349 abolishes enzymatic activity only and does not affect glycosylation and secretion. Two mutations create a new N-glycosylation site by substitution of pro at position 56 by phe and ser, respectively. This new site is apparently used in the mutants. Their core-glycosylated acid phosphatase is slightly larger than that of the wild type. Overglycosylation seems not to affect secretion. Four different mutations (a gly to asp substitution at position 281 and ser to phe substitutions at positions 150, 271 and 277) cause intracellular accumulation of enzymatically inactive core-glycosylated acid phosphatase precursor. These mutational lesions apparently block transport of acid phosphatase from the endoplasmic reticulum to the Golgi apparatus.     

Mechanism of C-3 hydrogen exchange and the elimination of ammonia in the 3-methylaspartate ammonia-lyase reaction.

The enzyme 3-methylaspartate ammonia-lyase (EC 4.3.1.2) catalyzes the exchange of the C-3 hydrogen of the substrate, (2S,3S)-3-methylaspartic acid, with solvent hydrogen. The mechanism of the exchange reaction was probed using (2S,3S)-3-methylaspartic acid and its C-3-deuteriated isotopomer. Incubations conducted in tritiated water allowed the rate of protium or deuterium wash-out from the substrates to be measured as tritium wash-in. The primary deuterium isotope effects for the exchange under essentially Vmax conditions ( [S] much greater than Km) were 1.6, 1.5, and 1.5 at pH 9.0, 7.6, and 6.5. The deamination reaction, measured spectrophotometrically on the same incubations, showed isotope effects of 1.7, 1.6, and 1.4 at pH 9.0, 7.6, and 6.5, in agreement with the values of DV and D(V/K) reported previously [Botting, N.P., Akhtar, M., Cohen, M.A., & Gani, D. (1988) Biochemistry 27, 2956-2959]. The ratio of the rate of exchange to the rate of deamination, however, varied widely with pH. Together with the identical values of the primary isotope effects for the two reactions, this result indicates that the partition between reaction pathways occurs after the slowest steps in the common part of the reaction coordinate pathway, almost certainly after the cleavage of the C-N bond at the level of the enzyme-ammonia-mesaconic acid complex, and not at the putative carbanion level as was previously suggested. The enzyme requires both K+ and Mg2+ ions for activity, although ammonium ion is also able to bind in the K+ site and act as an activator. Variation of the metal ion concentration alters the magnitude of the primary deuterium isotope effects. The variation of potassium ion concentration causes the most marked changes: at 1.6 mM K+, DV and D(V/K) are 1.7, whereas at 50 mM K+, DV and D(V/K) are reduced to 1.0. The isotope effects are also reduced at low K+ concentration due to the emergence of a slow-acting high K+ affinity monopotassium form of the enzyme. The binding order and role of the metal ion cofactors and their influence in determining the formal mechanism of the reaction is discussed, and the failure of previous workers to observe primary deuterium isotope effects for the deamination process is explained. The product desorption order was tested by product inhibition, alternative product inhibition, and isotope exchange experiments. Ammonia and mesaconic acid debind in a random fashion.(ABSTRACT TRUNCATED AT 400 WORDS)     

Phospholipase C activation in rat pituitary adenoma (GH) cells

The presence of the pertussis toxin (PTX) insensitive GTP-binding proteins (G-proteins) G_q and/or G₁₁ has been demonstrated in three different prolactin (PRL) and growth hormone (GH) producing pituitary adenoma cell lines. Immunoblocking of their coupling to hormone receptors indicates that G_q and/or G₁₁ confer throliberin (TRH) responsive phospholipase C (PL-C) activity in these cells. The contention was substantiated by immunoprecipitation analyses snowing that anti G_q/11-sera coprecipitated PL-C activity. In essence, only G_q/11 (but neither G_i2, G_i3 nor G_o) seems to mediate the TRH-sensitive PL-C activity, while G_o may be coupled to a basal or constitutive PL-C activity. Immunoblocking studies imply that the B-complex also, to some extent, may stimulate GH₃ pituitary cell line PL-C activity. Finally, the steady state levels of G_q/11 mRNA and protein were downregulated upon long term exposure of the GH3 cells to TRH (but not to vasoactive intestinal peptide = VIP).